WO2011109130A1 - Threads of hyaluronic acid and methods of use thereof - Google Patents

Abstract

This invention relates generally to threads of hyaluronic acid, methods of making such threads and uses thereof, for example, in aesthetic applications (e.g., facial contouring, dermal fillers), surgery (e.g., sutures), drug delivery, negative pressure wound therapy, moist wound dressing, and the like.

Description

THREADS OF HYALURONIC ACID AND METHODS OF USE THEREOF

Cross-Reference to Related Applications

[0001] This application claims the benefit under 35 U.S.C. § 119(e) of United States Provisional Patent Application 61/309,353, filed on March 1, 2010, United States Provisional Patent

Application 61/347,325, filed on May 21, 2010, and United States Provisional Patent Application 61/405,171, filed on October 20, 2010, each of which are hereby incorporated by reference in their entirety.

Field of the Invention

[0002] This invention relates generally to threads of hyaluronic acid, methods of making such threads and uses thereof, for example, in aesthetic applications (e.g., facial contouring, dermal fillers), surgery (e.g., sutures), drug delivery, negative pressure wound therapy, moist wound dressing, and the like.

State of the Art

[0003] Hyaluronic acid (HA) is a linear polysaccharide (i.e., non-sulfated glycosaminoglycan) consisting of a repeated disaccharide unit of alternately bonded β-D-N-acetylglucoamine and β-D- glucuronic acid which can be depicted by the formula:

where n is the number of repeating units. Hyaluronic acid is sometimes referred to by the nomenclature (-4GlcUA i-3GlcNAc i-)_n) and is a chief component of the extracellular matrix found, for example, in connective, epithelial and neural tissue. Natural hyaluronic acid is highly biocompatible because of its lack of species and organ specificity and is often used as a biomaterial in tissue engineering and as a common ingredient in dermal fillers.

[0004] Natural hyaluronic acid has poor in vivo stability due to rapid enzymatic degradation and hydrolysis and, accordingly, various chemically modified forms of hyaluronic acid (e.g., cross- linked forms, ionically modified forms, esterified forms, etc.) have been synthesized to address this problem. Currently, hyaluronic acid or cross-linked versions thereof are used in various gel forms, for example as dermal fillers, adhesion barriers, and the like.

[0005] However, issues exist with the use of gels of hyaluronic acid or its cross-linked versions. First, the force required to dispense gels of hyaluronic acid or its cross-linked versions is non- linear which can cause an initial ejection of a "glob" of gel that many physicians report when using hyaluronic acid gels. Second, precisely dispensing hyaluronic gels to specific locations can be difficult because such gels have little mechanical strength. Further, the gel will occupy the space of least resistance which makes its use in many applications (e.g., treatment of fine wrinkles) problematic as the gel will often migrate into unintended spatial areas rendering the cosmetic procedure difficult and possibly even dangerous. Many common dermal fillers which are injected into the treatment site as a liquid or a gel, such as Restylane® (hyaluronic acid), Juvederm® (hyaluronic acid) Radiesse® (calcium hydroxyl apatite), Sculptra® (poly-L-lactic acid) and Perlane® (hyaluronic acid), are capable of migration and/or causing unsightly "lumps" which are painful to treat. Furthermore, these dermal fillers are not recommended for use around the eyes as migration from the injection site can cause blindness, tissue necrosis, and in rare cases even stroke. Clinicians also find performing lip augmentations using these fillers time consuming, and patients find treatments in this area so painful that nerve blocks are routinely performed.

[0006] Accordingly, there is a need for new physical forms of hyaluronic acid which can be dispensed uniformly to specific locations regardless of tissue resistance, and without the risk of migration. Further, it would be beneficial to have non-crosslinked versions so as to avoid any toxicity from residual, unreacted cross-linking agents. Such new forms will have particular uses, for example, in aesthetic and surgical applications, rejuvenation procedures, drug delivery, wound therapy and wound dressing.

Summary [0007] Hyaluronic acid, like collagen, is known to form triple-helices through hydrogen bonding. It has now been surprisingly found that a secondary organization, referred to herein as "interlocked," can be made to occur with hyaluronic acid. As contemplated herein, these secondary structures of hyaluronic acid are "interlocked" when a matrix of hyaluronic acid is formed upon dehydration under non-denaturing conditions. Such a matrix can comprise one or multiple hyaluronic acid polymers wherein the polymers are substantially parallel to one another, and/or the helices are substantially parallel to each other and/or the polymers/helices are intertwined among each other. [0008] The exact nature of the interlocking is not critical. Rather, the criticality of the interlocked structures, when in the form of a thread, is manifested in one or more of the following: improved tensile strength, reduced biodegradation, improved ability to promote fibrogenesis, and the like. An improved ability to promote fibrogenesis and/or tissue repair in vivo is provided by forming a scaffold-like structure in the body for collagen deposition. This tissue repair could prolong the "filler" or "rejuvenation" effects of the thread when used to treat or fill a wrinkle or provide facial contouring in vivo far beyond the half-life of the hyaluronic acid-based thread.

[0009] In light of the above, the present invention is directed to a thread comprising hyaluronic acid wherein at least a portion of the hyaluronic acid is interlocked. It is contemplated that the interlocking of the hyaluronic acid can be confirmed by its ability to reflect polarized light, scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM) and/or x-ray diffraction (XRD). In certain aspects, the thread is substantially cylindrical, substantially D-shaped, or substantially ribbon shaped.

[0010] Hyaluronic acid forms a gel under aqueous conditions. This gel form can then be converted by the methods described herein to provide the novel threads of this invention. In one process of the invention, an aqueous gel composition comprising hyaluronic acid is dried under non-denaturing conditions, and preferably ambient conditions, to provide a dried thread.

Surprisingly, it has been found that other forms of drying, such as submersing in solvents, freezing, lyophilization, and heating, denature the hyaluronic acid such that the hyaluronic acid threads formed thereby have undesirable characteristics. These characteristics may include a low degree of interlocking and/or an insufficient tensile strength.

[0011] In one of its method embodiments, there is provided a method of treating a wrinkle in a subject in need thereof. In such an aspect, the thread is inserted into the dermis of a patient adjacent to or under the wrinkle. The thread is then applied under the wrinkle thereby treating the wrinkle. In one embodiment, upon exposure to body fluids or by manually hydrating, the thread expands upon hydration and such expansion is typically sufficient to fill-in the wrinkle. It is advantageous to have a thread expand upon hydration because the invasiveness of the insertion profile is minimized, however, threads designed to not expand can also be used to treat the wrinkle. [0012] In another embodiment, the invention is directed to providing facial contouring in a subject in need thereof. In this embodiment, the thread is inserted into the dermis at or adjacent to the desired treatment location, e.g., the lips, the nasolabial fold, the tear trough, etc. The thread is then applied thereby providing facial contouring. In one embodiment, a thread is applied to various planes of the dermal tissue. In one embodiment, several threads can be placed generally parallel to each other and additional threads places in a generally perpendicular direction with respect to the first set of parallel threads thereby forming a mesh structure whose aggregate effect is to contour a larger defect or more widespread defect such as the tear trough or the infraorbital region of the eye.

[0013] Also encompassed by this invention is a kit of parts comprising the thread. In some embodiments, the kit further comprises a means for delivering the thread. The means for delivery can either be a syringe or a needle.

[0014] In still other aspects, methods of using threads of hyaluronic acid as dermal fillers, facial contouring, adhesion barriers, wound dressings including negative pressure wound dressings, sutures, and the like is provided. Further provided are methods of using threads of hyaluronic acid for example, in surgery, ophthalmology, wound closure, drug delivery, and the like. These embodiments, as well as others, are discussed in more detail below.

Brief Description of the Drawings [0015] The invention is best understood from the following detailed description when read in conjunction with the accompanying drawings. It is emphasized that, according to common practice, the various features of the drawings are not to-scale. On the contrary, the dimensions of the various features are arbitrarily expanded or reduced for clarity. Included in the drawings are the following figures: [0016] Fig. 1 shows images of various HA compositions taken with a bench top polarization setup where the polarization angle was aligned. (A) non-cross-linked hyaluronic acid thread; (B) dried Restylane®; (C) wet Restylane®; (D) cross-linked hyaluronic acid thread (0.4% BDDE); (E) non-cross-linked hyaluronic acid (intramolecular cross-linking attempted by freezing and thawing). [0017] Fig. 2 shows scanning electron microscopy (SEM) images of a HA gel (Fig. 2A) compared to a HA thread (Figs. 2B, 2C, and 2D).

[0018] Fig. 3 illustrates a thread attached to the distal end of a needle, in its entirety (N = needle; T = thread).

[0019] Fig. 4 shows a needle attached to the thread (N = needle; T = thread). Fig. 4A illustrates a close-up view of a thread inserted into the inner-diameter of a needle; and Fig. 4B illustrates a close-up view of the distal end of a solid needle with the thread overlapping the needle. [0020] Fig. 5 shows treatment of a wrinkle. Fig. 5A illustrates a fine, facial wrinkle in the periorbital region of a human; Fig. 5B illustrates a needle and thread being inserted into the dermis of the wrinkle at the medial margin; Fig. 5C illustrates the needle being adjusted to traverse beneath the wrinkle; Fig. 5D illustrates the needle exiting at the lateral margin of the wrinkle; Fig. 5E illustrates the needle having pulled the thread into the location it previously occupied beneath the wrinkle; and Fig. 5F illustrates the thread implanted beneath the wrinkle, with excess thread having been cut off.

[0021] Fig. 6 shows treatment of baldness. Fig. 6A illustrates a top-down view of a male with typical male-pattern baldness; Fig. 6B illustrates where hair re-growth is desired, taking hair-lines into consideration; Fig. 6C illustrates a curved needle with attached thread being inserted into one imaginary line where hair re-growth is desired; Fig. 6D illustrates the needle traversing the imaginary line, and exiting the skin; Fig. 6E illustrates the needle pulled through distally, pulling along the thread into the desired location; and Fig. 6F illustrates scissors being used to cut excess thread. [0022] Fig. 7 shows treatment of a wrinkle. Fig. 7A illustrates a cross-sectional view of a fold or a wrinkle; Fig. 7B illustrates a thread implanted beneath a wrinkle that is not yet hydrated; and Fig. 7C illustrates a thread implanted beneath a wrinkle that is fully hydrated and has flattened the surface appearance of the wrinkle.

[0023] Fig. 8 shows treatment of a tumor. Fig. 8A illustrates a human pancreas with a tumor; Fig. 8B illustrates a curved needle with a thread attached thereto; Fig. 8C illustrates a curved needle traversing the tumor within the pancreas; and Fig. 8D illustrates the end-result of repeated implantations of thread.

[0024] Fig. 9 shows a nipple reconstruction. Fig. 9A illustrates multiple layers of concentric coils of thread, shaped to represent a human nipple; Fig. 9B illustrates the implant of Fig. 9A in cross-section; and Fig. 9C illustrates how an implant of coiled thread would be used for nipple reconstruction.

[0025] Fig. 10 illustrates how a needle and thread could be used to place a thread in a specific, linear location to promote nerve or vessel regrowth in a specific line.

[0026] Fig. 1 1 shows photographs of the cross sectional shape of various threads of the invention under a microscope. Fig. 11 A shows a substantially cylindrical thread; Fig. 1 IB shows a substantially D-shaped thread; and Fig. 11 C shows a substantially ribbon-shaped thread. The thread was taped onto an aluminum surface and cut to reveal the cross-sectional shape. [0027] Fig. 12 shows transmission electron microscopy (TEM) images of the gel (Figs. 12A and 12B) and a thread of the invention (Figs. 12C and 12D). Figs. 12A - 12D are discussed in Example 10.

[0028] Fig. 13A and 13B shows the front view a patient who has received lip augmentation with threads of the invention. Fig. 13A is before and Fig. 13B is after. In this patient, four threads were used in the patient's upper lip. On each side of the lip, one thread was placed at the white roll and one at the wet-dry junction. For the lower lip, one thread was placed in the white roll on both the left and right side of the lip. Also, there was one thread implanted in the middle at the wet-dry junction. [0029] Fig. 14A and 14B shows the same patient from the side view where Fig. 14A is before and Fig. 14B is after. Fig. 15A and 15B is the same patient from the opposite side where Fig. 15A is before and Fig. 15B is after.

[0030] Fig. 16A and 16B shows the front view of a patient who has had threads of the invention implanted into the tear trough. Fig. 16A is before and Fig. 16B is after. The patient in this photo had 3 threads implanted in the tear trough. The threads were placed substantially parallel at mid- depth (which is considered the deep dermis). The threads were placed about 1 millimeter apart in a substantially parallel fashion to the axis of the trough.

[0031] Fig. 17A and 17B shows the before and after of a patient who has had threads of the invention implanted into the nasolabial folds. In this patient, 4 threads were used on the right side (2 threads were implanted in approximately the subcutaneous space, 1 thread in the deep dermis, and 1 thread in the superficial dermis). On the left side, four threads were used (1 thread was implanted in approximately the subcutaneous space, 2 threads in the deep dermis, and 1 thread in the superficial dermis).

[0032] Fig. 18A and 18B shows the before and after of a patient who has had threads of the invention implanted into the forehead.

[0033] Fig. 19A shows placement of threads in a relatively parallel orientation for facial contouring in the tear trough (Thread 1 , 2, 3, 4, 5, and 6). This figure also shows placement of the thread for facial contouring of the nasolabial fold (Thread 7 and 8).

[0034] Fig. 19B shows an alternative placement of the threads for facial contouring in the tear trough (Thread 1, 2, 3, 4, 5, 6, 7, and 8). [0035] Figs. 20A and 20B show a schematic of the contemplated microanatomy of a thread implanted into a patient both in a cross-section of the skin and a three-dimensional cross-section.

Detailed Description

[0036] This invention is directed to threads of hyaluronic acid, methods for their preparation and uses thereof and to specific shapes formed there from. However, prior to describing this invention in greater detail, the following terms will first be defined.

[0037] It is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0038] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a thread" includes a plurality of threads.

1. Definitions [0039] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. As used herein the following terms have the following meanings.

[0040] As used herein, the term "comprising" or "comprises" is intended to mean that the compositions and methods include the recited elements, but not excluding others. "Consisting essentially of when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination for the stated purpose. Thus, a composition consisting essentially of the elements as defined herein would not exclude other materials or steps that do not materially affect the basic and novel characteristic(s) of the claimed invention. "Consisting of shall mean excluding more than trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0041] The term "about" when used before a numerical designation, e.g., temperature, time, amount, and concentration, including range, indicates approximations which may vary by ( + ) or ( - ) 10 %, 5 % or l %. [0042] As stated above, the invention is directed to a thread of hyaluronic acid wherein at least a portion is interlocked.

[0043] As used herein, the term "thread" refers to a long, thin, flexible form of a material. The thread of the invention can have a variety of shapes in the cross-section which are discussed below. [0044] The term "hyaluronic acid" or "HA" refers to the polymer having the formula:

where n is the number of repeating units. All sources of hyaluronic acid are useful in this invention, including bacterial and avian sources. Hyaluronic acids useful in this invention have a molecular weight of from about 0.5 MDa (mega Dalton) to about 3.0 MDa. In some

embodiments, the molecular weight is from about 0.6 MDa to about 2.6 MDa and in yet another embodiment, the molecular weight is from about 1.2 MDa to about 1.8 MDa, or from about 1.4 MDa to about 1.6 MDa.

[0045] The term "interlocked" refers to a matrix of hyaluronic acid that is formed upon dehydration under non-denaturing conditions. Such a matrix can comprise one or multiple hyaluronic acid polymers wherein the polymers are substantially parallel to one another, or the helices are substantially parallel to each other and/or the polymers/helices are intertwined among each other along an axis. In some embodiments, at least about 20% of the helices are substantially parallel to each other. In another embodiment, at least about 50%> of the helices are substantially parallel to each other. The interlocking can occur prior to, during, or after the hyaluronic acid's organization into triple helices. In one embodiment, at least about 10%> is interlocked. In another embodiment, at least about 30%> is interlocked.

[0046] The term "non-denaturing conditions" refers to conditions which preserve interlocking. In some embodiments, non-denaturing conditions include ambient conditions. In another embodiment, non-denaturing conditions include the use of a desiccant. [0047] The term "ambient conditions" is intended to refer to the typical environmental conditions and preferably, a pressure of about 1 atmosphere and/or temperature of 5 °C to about 40 °C, and preferably 20 °C to 30 °C. In some embodiments the ambient conditions comprise a relative humidity of from about 20% to about 80%>.

[0048] The term "percent hydration" is intended to refer to the total percent of water by weight. In one embodiment, the percent hydration of the thread is about 30%> or less, or alternatively, about 15%) or less, or alternatively, about 10%> or less. This can typically be measured by Karl Fisher titration.

[0049] The term "ultimate tensile strength" is intended to refer to the tensile strength of the thread which has been normalized with respect to cross-sectional area. The term "tensile strength" is intended to refer to the maximum stress a thread can withstand without failing when subjected to tension. In one embodiment, it is contemplated that the ultimate tensile strength is sufficient to pull the thread through the dermis and manipulate it once in the dermis such that the integrity of the thread is not substantially compromised by, for example, breaking or segmenting. It is contemplated that threads of the invention preferably have an ultimate tensile strength of about 3 kpsi ("kilopounds per square inch") or greater, or 5 kpsi or greater, or 10 kpsi or greater, or 15 kpsi or greater or 20 kpsi or greater or 50 kpsi or greater or 75 kpsi or greater. [0050] The threads of the invention can be made into a variety of shapes. The term

"substantially cylindrical" refers to a thread wherein the cross-section of the thread is round. The term "substantially" as used to refer to shapes of the threads means that at least 50%> of the thread has the shaped described. The term substantially is also used to encompass threads which have a variety shapes along the length of the thread. For example, a thread could be substantially cylindrical but the ends of the thread may be tapered. The substantially cylindrical threads can be provided when the contact angle of the gel composition and the substrate on which it is extruded have an equilibrium contact angle of greater than about 90 degrees.

[0051] The term "substantially D-shaped" refers to a thread wherein the cross-section is D- shaped or substantially semi-circular. The substantially D-shaped threads have one flat side and one substantially round side. The substantially D-shaped threads can be provided when the contact angle of the gel composition and the substrate on which it is extruded have an equilibrium contact angle of about 90 degrees.

[0052] The term "substantially ribbon-shaped" refers to a thread wherein the thickness of the thread is less than about 50%> of the width of the thread. In some embodiments, the cross-section is substantially rectangular. The ribbon-shaped threads can be provided when the contact angle of the gel composition and the substrate on which it is extruded have an equilibrium contact angle of less than about 90 degrees. Alternatively, the ribbon-shaped threads can be formed by cutting a wetted gel to achieve the desired cross-sectional shape. "Ribbon-shaped" may also include shapes that are substantially ellipsoidal. The term "substantially ellipsoidal" refers to a thread wherein the cross-section is substantially oblong or elliptical.

[0053] The term "therapeutic agent" can include one or more therapeutic agents. In still other of the above embodiments, the therapeutic agent is an anesthetic, including but not limited to, lidocaine, xylocaine, novocaine, benzocaine, prilocaine, ripivacaine, propofol or combinations thereof. In still other of the above embodiments, the therapeutic agent includes, but is not limited to, epinephrine, adrenaline, ephedrine, aminophylline, theophylline or combinations thereof. In still other of the above embodiments, the therapeutic agent is botulism toxin. In still other of the above embodiments, the therapeutic agent is laminin-51 1. In still other of the above

embodiments, the therapeutic agent is glucosamine, which can be used, for example, in the treatment of regenerative joint disease. In still other of the above embodiments, the therapeutic agent is an antioxidant, including but not limited to, vitamin E or all-trans retinoic acid such as retinol. In still other of the above embodiments, the therapeutic agent includes stem cells. In still other of the above embodiments, the therapeutic agent is insulin, a growth factor such as, for example, NGF (nerve growth factor),BDNF (brain-derived neurotrophic factor), PDGF (platelet- derived growth factor) or Purmorphamine Deferoxamine NGF (nerve growth factor),

dexamethasone, ascorbic acid, 5-azacytidine, 4,6-disubstituted pyrrolopyrimidine, cardiogenols, cDNA, DNA, RNAi, BMP-4 (bone morphogenetic protein-4), BMP -2 (bone morphogenetic protein-2), an antibiotic agent such as, for example, β lactams, quinolones including

fluoroquinolones, aminoglycosides or macrolides, an anti-fibrotic agent, including but not limited to, hepatocyte growth factor or Pirfenidone, an anti-scarring agent, such as, for example, anti- TGF-b2 monoclonal antibody (rhAnti-TGF-b2 mAb), a peptide such as, for example, GHK copper binding peptide, a tissue regeneration agent, a steroid, fibronectin, a cytokine, an analgesic such as, for example, Tapentadol HC1, opiates, (e.g., morphine, codone, oxycodone, etc.) an antiseptic, alpha- beta or gamma-interferon, EPO, glucagons, calcitonin, heparin, interleukin-1, interleukin-2, filgrastim, a protein, HGH, luteinizing hormone, atrial natriuretic factor, Factor VIII, Factor IX, or a follicle-stimulating hormone.

[0054] The term "diagnostic agent" refers to a therapeutic agent which is used as part of a diagnostic test (e.g., a fluorescent dye to be used for viewing the thread in vivo). In one embodiment, the diagnostic agent is soluble TB (tuberculosis) protein.

[0055] The term "lubricity-enhancing agent" is intended to refer to a substance or solution which when contacted with the dried thread, acts to lubricate the dried thread. A lubricity- enhancing agent can comprise, for example, water and/or an alcohol, an aqueous buffer, and may further comprise additional agents such as polyethylene glycol, hyaluronic acid, and/or collagen.

[0056] The term "biodegradation impeding agent" is intended to refer to a biocompatible substance that slows or prevents the in vivo degradation of the thread. For example, a

biodegradation impeding agent can include hydrophobic agents (e.g., lipids) or sacrificial biodegradation agents (e.g., sugars).

[0057] The term "failure stress" is intended to refer to the maximum weight which, when applied to the thread, causes the thread to fail. By "failing," it meant that the thread can break or segment or otherwise lose structural integrity. In some embodiments, the failure stress is about 0.1 pounds or 0.22 kilograms or greater.

[0058] The term "aqueous gel composition" or "gel composition" or "gel mixture" is intended to refer to an aqueous composition comprising water and hyaluronic acid. In some embodiments, the composition may further comprise a buffer such that that the pH of the solution changes very little with the addition of components of the composition. In these embodiments, the composition is referred to as an aqueous buffered gel composition. The pH of the buffered gel composition is typically from about 7 to about 10. In certain embodiments the pH is about 7. In certain embodiments, the pH is higher at about 9 or about 10. In some embodiments, the pH can be adjusted by adding an appropriate amount of a suitable base, such as Na₂C0₃ or NaOH. In some embodiments, the aqueous gel buffered composition comprises phosphate buffered saline. In some embodiments, the aqueous gel buffered composition comprises

tris(hydroxymethyl)aminomethane (Tris), which has the formula (HOCH₂)₃CNH₂. In some embodiments, additional solutes are added to adjust the osmolality and ion concentrations, such as sodium chloride, calcium chloride, and/or potassium chloride.

[0059] The term "buffer" is intended to refer to a solution comprising a mixture of a weak acid and its conjugate base or a weak base and its conjugate acid. Buffer solutions include, but are not limited to, 2-amino-2-methyl-l,3-propanediol, 2-amino-2-methyl-l-propanol, L-(+)-tartaric acid, D-(-)-tartaric acid, ACES, ADA, acetic acid, ammonium acetate, ammonium bicarbonate, ammonium citrate, ammonium formate, ammonium oxalate, ammonium phosphate, ammonium sodium phosphate, ammonium sulfate, ammonium tartrate, BES, BICINE, BIS-TRIS, bicarbonate, boric acid, CAPS, CHES, calcium acetate, calcium carbonate, calcium citrate, citrate, citric acid, diethanolamine, EPP, ethylenediaminetetraacetic acid disodium salt, formic acid solution, Gly-Gly-Gly, Gly-Gly, glycine, HEPES, imidazole, lithium acetate, lithium citrate, MES, MOPS, magnesium acetate, magnesium citrate, magnesium formate, magnesium phosphate, oxalic acid, PIPES, phosphate buffered saline, piperazine potassium D-tartrate, potassium acetate, potassium bicarbonate, potassium carbonate, potassium chloride, potassium citrate, potassium formate, potassium oxalate, potassium phosphate, potassium phthalate, potassium sodium tartrate, potassium tetraborate, potassium tetraoxalate dehydrate, propionic acid solution, STE buffer solution, sodium 5,5-diethylbarbiturate, sodium acetate, sodium bicarbonate, sodium bitartrate monohydrate, sodium carbonate, sodium citrate, sodium chloride, sodium formate, sodium oxalate, sodium phosphate, sodium pyrophosphate, sodium tartrate, sodium tetraborate, TAPS, TES, TNT, TRIS-glycine, TRIS-acetate, TRIS buffered saline, TRIS-HCl, TRIS phosphate- EDTA, tricine, triethanolamine, triethylamine, triethylammonium acetate, triethylammonium phosphate, trimethylammonium acetate, trimethylammonium phosphate, Trizma® acetate, Trizma® base, Trizma® carbonate, Trizma® hydrochloride or Trizma® maleate.

[0060] The term "aqueous solvent" is intended to refer to a non-toxic, non-immunogenic aqueous composition. The aqueous solvent can be water and/or an alcohol, and may further comprise buffers, salts and other such non-reactive solutes. [0061] The term "contact angle" or "equilibrium contact angle" refers to a measure of a liquid's affinity for a solid and quantifies the degree of a liquid drop's spread when placed on the solid. In the case of the invention, the liquid is the aqueous gel composition and the rigid or solid surface is the substrate on which the composition is extruded. The contact angle is a measure of the angle that the edge of an ideal drop makes with a flat surface. The lower that the contact angle is, the greater attraction between the surface and the liquid. For example, water spreads almost completely on glass and has a very low contact angle of nearly 0 degrees. Mercury, in contrast, beads up and spreads very little; its contact angle is very large.

2. Interlocked Hyaluronic Acid

[0062] The present invention is directed to a thread comprising hyaluronic acid wherein at least a portion of the hyaluronic acid is interlocked. The thread is formed by drying an aqueous gel composition which comprises hyaluronic acid under non-denaturing conditions and preferably ambient conditions so as to provide for the interlocking. Without being limited to any theory, it is contemplated that intramolecular or intermolecular cross-linking occurs after at least a portion of the polymer chains of the hyaluronic acid in the aqueous gel composition have interlocked. In some embodiments, at least a portion of the thread of the invention is intramolecularly and/or intermolecularly cross-linked. The term "intramolecularly or intermolecularly cross-linked" is intended to refer intermolecular or intramolecular dehydration which results in lactone or anhydride formation within a single polymer chain or between two or more chains. This is differentiated from two or more polymer chains of hyaluronic acid which have been covalently bonded via a cross-linking agent, such as butanediol diglycidyl ether (BDDE), divinyl sulfone (DVS), and l-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), or a combination thereof. [0063] Benefits exist in threads which are not cross-linked with a cross-linking agent, for example, toxicity associated with unreacted cross-linking agent is no longer an issue.

Accordingly, in one embodiment, the thread is substantially free of cross-linking agents.

[0064] It is further contemplated that that the portion that is interlocked is the outer surface or the outer surface and the inner surface of the thread. It is further contemplated that the thread is substantially interlocked uniformly along its length. In certain embodiments, it is contemplated that the threads of the invention are not viscoelastic. In one embodiment, the threads of the invention do not have an elasticity along their length of greater than 100%, or greater than 50%>.

[0065] The interlocking of the hyaluronic acid can be observed by the ability of the thread to reflect polarized light. This can be observed in Fig. 1. As can be seen in the figure, the thread of the invention reflects polarized light but the forms of HA which are not considered interlocked, such as the Restylane® gel, do not reflect polarized light.

[0066] It is also contemplated that the interlocking can be quantified by the use of one or more of the following: scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM) and/or x-ray diffraction (XRD). The physical properties of the thread of the invention can be tailored for a specific use by adjusting the components in the aqueous gel composition and adjusting the method of producing the thread as discussed below.

[0067] The half-life of the hyaluronic acid thread in vivo can be controlled by controlling the thickness of the thread, the density, the molecular weight of the hyaluronic acid and the degree of hydration, which can then be further controlled by adjusting the amounts of hyaluronic acid. [0068] The percent hydration of hyaluronic acid can range from about 1%> to greater than about 1000%) based on the total weight. The percent hydration of the thread of the present invention can be controlled, for example, by adjusting the percent hyaluronic acid in the gel. It is contemplated that a lower percent hydration thread would result in a thread with a higher tensile strength. In some embodiments, the thread has no more than about 30%> percent, or no more than 15%>, or no more than 10%> by weight hydration based on the total weight. The percent hydration will be determined by the environment to which the thread is subjected to during or after the drying process. [0069] In one embodiment, at least about 10% of the thread is interlocked. In another embodiment, at least about 30% is interlocked. It is further contemplated that a sufficient amount of the thread is interlocked so as to provide the improved mechanical properties of increased strength and/or an enhanced ability to promote fibrogenesis. In one embodiment, the thread is elastomeric. In one embodiment, the thread is not too rigid or too plastic-like so as it can be moved within the dermis during delivery when used as a dermal filler. The appropriate stiffness or elastic modulus is determined by the intended use of the thread.

3. Methods of Making the Threads of the Invention

[0070] The invention is also directed to a method of making the thread of the invention. The method comprises drying under non-denaturing and preferably ambient conditions an aqueous gel composition comprising hyaluronic acid to provide a dried thread.

[0071] Typically, the aqueous gel composition comprises water and can optionally comprise phosphate buffered saline (PBS) or tris(hydroxymethyl)aminomethane (Tris) buffer and optionally have a pH of about 7. The buffer can be selected based on the desired pH of the composition. In some embodiments, the pH is adjusted with the appropriate amount of a suitable base, such as Na₂C0₃ or NaOH to reach the desired pH.

[0072] Once the desired pH is reached, the desired amount of HA is added, which is from about 1%) to about 30%) by weight, and is preferably about 5 to about 10%> by weight. The relative amount of HA can be adjusted based on its molecular weight to provide a composition of desired viscosity. The molecular weight of the HA used in the threads of the invention is from about 0.5 MDa to about 3.0 MDa, or from about 1.2 MDa to about 1.8 MDa, or from about 1.4 MDa to about 1.6 MDa. After adding the HA, it is allowed to dissolve slowly to form a gel. The viscosity of the gel is typically from about 150 Pascal-seconds (Pa.s) to about 2,000 Pascal-seconds (Pa.s).

[0073] In some embodiments, the gel composition is degassed prior to extrusion to minimize air bubbles after extrusion. The degassing can be done by freeze-pump-thaw which procedure is known by one of skill in the art.

[0074] To form the thread, the gel composition is typically extruded onto a substrate which is more thoroughly discussed in Example 1 to form a wetted thread. The composition is extruded using a pressurized syringe affixed to a nozzle. The nozzle can have various geometries, such as various lengths, internal diameters and shapes. The nozzle may be circular or non-circular in shape, for example, a flattened shape or a "D" shape. The syringe nozzle may be anywhere from about a 15 gauge to a 25 gauge syringe nozzle. Typically, the pressure employed is from about 10 to about 2000 psi or from about 20 to about 240 psi. The pressure requirements are dictated by the nozzle geometry. The pressure can be applied hydraulically, for example using ambient air or nitrogen, or mechanically. The speed at which the gel is extruded is selected so as to minimize air bubbles in the length of the thread and maximize a consistent shape. Air bubbles can reduce the structural integrity of the thread by causing weak spots.

[0075] Various substrates are contemplated for use by methods of the invention. Substrates include by hydrophilic and hydrophobic substrates and may be selected from, but are not limited to, polytetrafluoroethylene (PTFE), expanded PTFE, nylon, polyethylene terephthalate (PET), polystyrene, silicon, polyurethane, and activated cellulose. [0076] The substrate employed, along with the viscosity of the gel composition, dictates the general shape of the thread. Various shapes are shown in Fig. 11. For example, if the gel and the substrate have an equilibrium contact angle of less than 90 degrees, it is contemplated that the thread formed will be substantially ribbon-shaped (Fig. 11C). Further, if the gel and the substrate have an equilibrium contact angle of about 90 degrees, the thread formed will be substantially D- shaped (Fig. 1 IB). Still further, if the gel and the substrate have an equilibrium contact angle of greater than 90 degrees, then the thread formed will be substantially round (Fig. 11C). For example, a 10% 1.5 MDa gel will have a substantially round cross-section (e.g., about 80% of a circle) when extruded on PTFE, while a 5% 1.5 MDa gel will form a substantially ribbon-shaped thread when extruded on PTFE. [0077] Alternatively to pressurized extrusion, the gel composition can be rolled out into an elongated cylinder and/or cut into elongated strips before drying.

[0078] The wetted thread is then dried to form a dried thread. The drying step is required to form threads with a sufficient tensile strength, as discussed below. As the thread may lose some of its interlocking properties when exposed to heat, in excess of water boiling temperature, it is preferred that the drying step be performed under ambient conditions. It is contemplated that by drying under ambient conditions, the hyaluronic acid is allowed to interlock. This drying procedure provides a thread with a higher tensile strength, such as, for example, an ultimate tensile strength of about 8 kpsi or greater or 20 kpsi or greater. In other words, the threads of the invention have a failure stress of about 0.1 pounds or 0.22 kilograms or greater. [0079] The thread is allowed to dry for anywhere from about 30 minutes to about 72 hours to form threads having a diameter of from 0.05 mm to about 1.0 mm and having no more than 30%> by weight hydration. In some embodiments, the thread can be dried for about 12 hours or about 24 hours. It is contemplated that the larger the molecular weight of HA employed or the more concentrated the HA in the composition, the longer the drying times that are required.

[0080] It is contemplated that the threads of the invention can be sterilized using typical sterilization methods known in the art, such as autoclave, ethylene oxide, electron beam (e-beam), supercritical CO₂ (with peroxide), freeze-drying, etc. For example, the threads of the invention can be sterilized using electron beam (e-beam) sterilization methods. In some embodiments, the threads are first washed in a buffer solution at high pH (i.e., pH 9 or pH 10) prior to sterilization. In some embodiments, the wash solutions further comprise ethanol, ascorbic acid, vitamin E and/or sodium phosphate. [0081] Optionally and as necessary, the thread is mechanically stretched prior to drying. The stretching or absence of stretching can provide a thread of the desired length and/or rehydration swelling volume. In some embodiments, the length of the thread can be from about 0.5 mm to about 15 mm. Upon drying, the thread, in some embodiments, heals to provide a more uniform surface of the thread. 4. Modification of Threads

[0082] In addition to washing the thread, it can also be further functionalized by adsorbing a sufficient amount of a member selected from the group consisting of a therapeutic agent, a diagnostic agent, a fibrogenesis-enhancing agent, a biodegradation impeding agent, a lubricity- enhancing agent and combinations thereof, optionally followed by re-drying the thread. Such therapeutic agents include antibacterials, anesthetics, dyes for viewing placement in vivo, and the like. In some embodiments, a dried or hydrated thread is coated to alter the properties with a bioabsorbable biopolymer, such as collagen, PEG, PLGA or a phase transfer Pluronic™ which can be introduced as a liquid and which solidifies in vivo.

[0083] In one embodiment, the thread can be coated to modulate the rate at which the thread is rehydrated. For example, the thread can be coated with a hydrophobic layer, such as a lipid. The thickness of the lipid layer can then be adjusted to achieve the desired rate of rehydration. In another embodiment, the thread can be coated with an aqueous composition of non-cross-linked hyaluronic acid. This can be performed just prior to implantation of the thread to act as a lubricant. It is also contemplated that this coating with non-cross-linked hyaluronic acid may slow the rate of hydration of the thread. In some embodiments, the thread is coated, either totally or in part, with the gel composition to form a layered material. For woven constructs, whether single layer or 3D, can be coated in their entirety to create weaves or meshes with altered physical properties from that of a free-woven mesh.

[0084] The threads as disclosed herein can be braided, coiled, layered or woven. In some embodiments, braids may be formed from the threads described above. A braid can be formed by intertwining three or more threads wherein each thread is functionally equivalent in zigzagging forward through the overlapping mass of the others. The braids can be a flat, three-strand structure, or more complex braids can be constructed from an arbitrary (but usually odd) number of threads to create a wider range of structures, such as wider ribbon-like bands, hollow or solid cylindrical cords, or broad mats which resemble a rudimentary perpendicular weave. [0085] In one embodiment, a plasticizer is added to adjust the stiffness of the thread.

Alternatively, or in addition to, threads of varying stiffness may be weaved together to produce a braided thread or material having the desired stiffness.

[0086] In some embodiments, a three-dimensional structure may be constructed by weaving or wrapping or coiling or layering the threads described above. In other embodiments, a three- dimensional structure may be constructed by weaving or wrapping or coiling or layering the braids described above. In still other embodiments, a three-dimensional structure may be constructed by weaving or wrapping or coiling or layering the cords described above. In still other embodiments, a three-dimensional structure may be constructed by weaving or wrapping or coiling or layering the meshes described above. [0087] In some embodiments, a three-dimensional, cylindrical implant is made of any of the threads is provided. An exemplary use for such an implant is for nipple reconstruction. In some embodiments, the threads used to make the cylindrical implant may include chrondrocyte adhesion compounds. In other embodiments, the cylindrical shape is provided by multiple, concentric coils of threads. 5. Methods of Using the Hyaluronic Acid Threads

[0088] The threads, braids, cords, woven meshes or three-dimensional structures described herein can be used, for example, to fill aneurysms, occlude blood flow to tumors, (i.e., tumor occlusion), in eye-lid surgery, in penile augmentation (e.g., for enlargement or for sensitivity reduction, i.e., pre-mature ejaculation treatment), inter-nasal (blood-brain barrier) delivery devices for diagnostic and/or therapeutic agents, corneal implants for drug delivery, nose augmentation or reconstruction, lip augmentation or reconstruction, facial augmentation or reconstruction, ear lobe augmentation or reconstruction, spinal implants (e.g., to support a bulging disc), root canal filler (medicated with therapeutic agent), glottal insufficiency, laser photo-refractive therapy (e.g., hyaluronic acid thread/weave used as a cushion), scaffolding for organ regrowth, spinal cord treatment (BDNF and NGF), in Parkinson's disease (stereotactic delivery), precise delivery of therapeutic or diagnostic molecules, in pulp implantation, replacement pulp root canal treatment, shaped root canal system, negative pressure wound therapy, adhesion barriers and wound dressings.

Methods of Treating a Wrinkle

[0089] Threads of the invention have an improved ability to promote fibrogenesis and/or tissue repair in vivo by forming a scaffold-like structure in the body for collagen deposition. This tissue repair could prolong the "filler" effects of the thread when used to treat or fill a wrinkle in vivo far beyond the half-life of the hyaluronic acid-based thread of the invention. This is described in Example 8.

[0090] In some embodiments, the present invention is directed to a method of treating a wrinkle in a patient in need thereof by 1) inserting the thread of the invention into dermis or subcutaneous space of the patient adjacent to or under the wrinkle; and 2) applying the thread adjacent to or under the wrinkle thereby treating the wrinkle. These steps can be performed at least once and up to 6 times to treat each wrinkle. In some embodiments, the thread is attached to the distal end of a syringe as shown in Figs. 3, 4A and 4B. The thread is inserted by a needle which needle is then removed. Optionally and as necessary, the thread is hydrated with water or saline, or by the fluids normally perfusing the surrounding tissue. Further, the remainder of the wrinkle can be filled with a biocompatible material such as a phase transfer Pluronic™ which can be introduced as a liquid and which solidifies in vivo. Alternatively, conventional hyaluronic acid gel can be introduced to fill the wrinkle. In either case, the formed web acts to maintain the biocompatible filler at the site of the wrinkle. [0091] In some embodiments, a method of treating a wrinkle in a subject is provided. In some embodiments, the attending clinician may numb the treatment area according to procedures known in the art using a variety of anesthetics, including, but not limited to, topical lidocaine, ice or a block with lidocaine injection. For example, the wrinkle may be in the peri-orbital region as illustrated in Fig. 5A. The thread may be attached to a needle as illustrated, for example, in Figs. 3, 4A and 4B. The distal end of the needle may be inserted through the skin surface of the subject into the dermis adjacent to or within the wrinkle as illustrated, for example, in Fig. 5B. In some embodiments, the thread is inserted into the subcutaneous space instead of the dermis. The needle then may traverse the dermis or subcutaneous space of the subject beneath the wrinkle as illustrated, for example, in Fig. 5C. The needle then may exit the skin of the subject at the opposite margin of the wrinkle, as illustrated, for example, in Fig. 5D. The needle may then be pulled distally until it is removed from the subject such that the thread is pulled into the location previously occupied by the needle beneath the wrinkle, as illustrated, for example, in Fig. 5E. Finally, excess thread is cut from the needle at the skin surface of the subject which leaves the thread implanted as illustrated, for example, in Fig. 5F.

[0092] While not wishing to be bound by theory, the method above may successfully treat wrinkles as shown in Figs. 7A, 7B and 7C. A typical wrinkle is illustrated in Fig. 7A. Fig. 7B illustrates a thread implanted beneath a wrinkle that is not yet hydrated. As the thread implanted beneath the wrinkle becomes fully hydrated the surface appearance of the wrinkle is concurrently flattened as illustrated in Fig. 7C.

[0093] In some embodiments, the thread is manipulated in such a fashion such that one end of the thread is sufficiently hard such that the thread is used to penetrate the skin. This may be accomplished by coating the thread with a hardening material, such as a sugar coating, In another embodiment, the thread is coated in its entirety, for example with a sugar coating, to provide the thread with increased columnar strength.

[0094] In some embodiments, the above method may be used to rejuvenate the skin of a subject in need thereof. In many of these embodiments, the thread includes substantial amounts of non-cross linked hyaluronic acid. In some of these embodiments, the thread includes

antioxidants, vitamin E or retinol or combinations thereof.

Facial Contouring

[0095] It is contemplated that the threads of the invention are useful in facial contouring. What is meant by facial contouring is that the threads can be applied to any area of the face or neck or chest that the patient desires to have augmented, including, by way of example only, the lips, the nasolabial fold (see, Fig. 17A and 17B), tear trough (see, Fig. 16A and 16B), and the forehead (see, Fig. 18A and l 8B).

[0096] Lip augmentation is a commonly desired aesthetic procedure. Typically, the aesthetic goal is fuller, plumper lips. Available treatment options for lip augmentation include temporary fillers such as Restylane® and Juvederm®, permanent fillers such as ArteFill®, Radiesse® and Goretex® implants, as well as surgical procedures. Areas of enhancement can include the vermillion border (or white roll) for lip effacement and contouring and the wet-dry mucosal junction for increasing fullness. Other techniques include more diffuse infiltration of the orbicularis oris muscle.

[0097] Lip contouring and augmentation by temporary dermal fillers is a popular, low risk option due to the minimal invasiveness and temporary nature of the procedure. The major shortcomings of dermal fillers currently used in lip procedures are that it is (a) painful, (b) difficult to consistently and homogenously inject the gel into the desired location, and (c) the gel can migrate over the lifetime of the implant causing the aesthetic results to change.

[0098] The present invention addresses the shortcomings described above. Beyond addressing the above-listed shortcomings for existing temporary dermal fillers described above, it has been found that the HA thread-based method of enhancing lip appearance is very quick. A typical patient may have 3 threads placed in their lip(s) in only 3 minutes. Current dermal filler lip procedures can take 15 to 20 minutes. See, Fig. 13A, 13B, 14A, 14B, 15A, and 15B.

[0099] In embodiments, directed to facial contouring, the attending clinician may numb the treatment area according to procedures known in the art using a variety of anesthetics, including, but not limited to, topical lidocaine, ice or a block with lidocaine injection. Threads made of HA (hyaluronic acid) can be attached to the proximal end of a needle and pulled into the lip. The needle can serve as a precise guide, and also be used to predict and correct the implant location prior to pulling the thread into the desired location. This precise delivery mechanism can be used to deliver threads along the vermillion border for contouring, superficially if desired, as well as at the wet-dry junction for plumping, deeper into the lip if desired.

[0100] It is contemplated that when the thread is used for facial contouring, any number of threads may be used depending on the desired effect and the size of the thread. For example, description of the procedure done for the lip augmentation and contouring is discussed below in Example 11. [0101] It is has been surprisingly and unexpectedly found that that threads may be implanted in various tissue planes of the patient to provide a more natural look when performing facial contouring. For example, the threads may be implanted in a manner that forms a hammock in the desired location. Given the unique properties of the threads of the invention, the attending clinician may deposit or implant the threads in the epidermis, the dermis, and/or the subcutaneous layer. This technique is referred to as stratifying.

[0102] This technique is enabled by the precision with which the threads can be placed, and their size relative to the dermis and underlying structures. Threads can impart different effects on facial features such as wrinkles, contours, folds and troughs depending on where they are implanted.

[0103] For example, recent clinical experience indicates that placing a thread (in this case one that was appx .008" in diameter) deeply, for example in the subcutaneous space, along the axis of a forehead wrinkle can help soften then appearance of the wrinkle that forms when the patient animates, by flexing their forehead, which would typically exacerbate the appearance of the wrinkle. These types of dynamic wrinkles are currently only well treated with Botox®, which has the undesirable effect of preventing the patient from expressing all facial expressions. Further, recent clinical experience shows that static wrinkles, ones that are visible in repose, can be effectively treated by placement of a thread (from 0.004" to 0.008" in diameter) superficially, for example within the dermis.

[0104] The technique of stratifying the thread implant tissue planes is also successfully used in improving the appearance of nasolabial folds (up to four 0.008" threads), glabellar lines, marionette lines, and lips. [0105] This is another technique that is enabled by the HA threads and their implantation method. To smooth the appearance of hollows or troughs such as the tear trough, or otherwise contour the face in areas such as the cheek bones, chin, for example, threads can be implanted in hatch (see, Fig. 19A) and/or cross-hatched patterns (see, Fig. 19B) to effect areas greater than the width of a single thread. As seen in Figs. 19A and 19B, two patients have their tear troughs effectively smoothed out by placing threads parallel in one case (Fig. 19A) and cross-hatched in another case (Fig. 19B). The cross-hatching could be done obliquely to the initial direction, as was the case in Fig. 19B, or perpendicularly. Further, the hatches can be in different tissue planes as well.

[0106] In another embodiment of this technique, the hatching can be done obliquely to the directionality of the area being treated. For example, in Fig. 19A the threads are placed aligned to the axis of the tear trough. Instead, the threads could be placed obliquely to the axis of the tear trough to support the tissue in the area differently.

[0107] It is contemplated that implanting the threads in various planes may also be done in the treatment of wrinkles as described above. Wound Therapy

[0108] In some embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are used in wound dressings including negative pressure wound dressings.

[0109] In some embodiments, wound dressing remains in contact with the wound for at least 72 hours. In other embodiments, the negative pressure wound dressing remains in contact with the wound for at least 1 week. In still other embodiments, the wound dressing remains in contact with the wound for at least 2 weeks. In still other embodiments, the wound dressing remains in contact with the wound for at least 3 weeks. In still other embodiments, the wound dressing remains in contact with the wound for at least 4 weeks. In the above embodiments, it should be understood that granulation tissue is not retaining the threads, braids, cords, woven meshes or three- dimensional structures described herein as these components are fully absorbable. In some of these embodiments, the wound dressing is between about 1 cm and about 5 cm thick.

Accordingly, in some of these embodiments, wound bed closure may be achieved without changing the dressing.

[0110] In some embodiments, the woven meshes described herein are used in wound dressings including negative pressure wound dressings. In other embodiments, the dressing include between 2 and about 10 layers of woven meshes.

[0111] In still other embodiments, the woven meshes comprise identical threads. In still other embodiments, the woven meshes comprise different threads.

[0112] In some embodiments, the woven meshes are between about 1 mm and about 2 mm thick when dry. In other embodiments, the woven meshes are between about 2 mm and about 4 mm thick when dry.

[0113] In some embodiments, the pore size of the woven mesh is between about 1 mm and about 10 mm in width. In other embodiments, the pore size of the woven mesh is between about 0.3 mm and about 0.6 mm in width. In still other embodiments, the pores of the woven mesh are aligned. In still other embodiments, the pores of the woven mesh are staggered. In still other embodiments, the woven meshes are collimated to create pores of desired size.

[0114] In some embodiments, the woven mesh is mechanically stable at a minimum vacuum level of about 75 mm Hg. In other embodiments, the woven mesh is mechanically stable at a vacuum up to about 150 mm Hg. [0115] In some embodiments, the woven mesh includes collagen. In other embodiments, the dressing is attached to a polyurethane foam. In still other embodiments, the polyurethane foam is open celled. In still other embodiments, the dressing is attached to a thin film. In still other embodiments, the thin film is silicone or polyurethane. In still other embodiments, the dressing is attached to the thin film with a water soluble adhesive.

[0116] In some embodiments, the thread used in the dressing includes a therapeutic agent or a diagnostic agent.

[0117] In some embodiments, a negative pressure wound dressing (Johnson et al., U.S. Patent No. 7,070,584, Kemp et al., U.S. Patent No. 5,256,418, Chatelier et al., U.S. Patent No. 5,449383, Bennet et al., U.S. Patent No. 5,578,662, Yasukawa et al., U.S. Patent Nos. 5,629,186, 5,780,281 and 7,611,500) is provided for use in vacuum induced healing of wounds, particularly open surface wounds (Zamierski U.S. Patent Nos. 4,969,880, 5,100,396, 5,261,893, 5,527,293 and 6,071,267 and Argenta et al., U.S. Patent Nos. 5,636,643 and 5,645,081). The dressing includes a pad which conforms to the wound location, an air-tight seal which is removably adhered to the pad, a negative pressure source in fluid communication with the pad and the threads, braids, cords, woven meshes or three-dimensional structures described herein attached to the wound contacting surface of the pad. The pad, seal and vacuum source are implemented as described in the prior art.

[0118] In other embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are mechanically stable at a minimum vacuum level of about 75 mm Hg. In still other embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are mechanically stable at a vacuum up to about 150 mm Hg. In still other embodiments, the dressing includes at least one layer of woven mesh. In still other embodiments, the dressing include between 2 and about 10 layers of woven mesh. [0119] In some embodiments a tube connects the pad to the negative pressure source. In still other embodiments, a removable canister is inserted between the pad and the negative pressure source and is in fluid communication with both the pad and the negative pressure source.

[0120] In some embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are not hydrated. Accordingly, in these embodiments, the dressing could absorb wound exudates when placed in contact with the wound. In other embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are hydrated. Accordingly, in these embodiments, the dressing could keep the wound moist when placed in contact with the wound.

[0121] In some embodiments, an input port attached to a fluid is connected with the pad.

Accordingly, in these embodiments, fluid could be dispensed in the wound. In some

embodiments, the fluid is saline. In other embodiments, the fluid contains diagnostic or therapeutic agents.

[0122] In some embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are used as adhesion barriers. In some embodiments, the woven meshes described herein are used in adhesion barriers. Hair Loss Treatment

[0123] In some embodiments, a method of treating hair loss in a subject is provided. A subject such as, for example, a male with typical male-pattern baldness is illustrated in Fig. 6A and the area where hair growth (with imaginary hairlines) is desired is shown in Fig. 6B. The thread may be attached to a needle as illustrated, for example, in Figs. 3, 4A, 4B and 6C. The distal end of the needle may be inserted into one of the hair lines as illustrated, for example, in Fig. 6C. The needle then may traverse the area beneath the hairline of the subject and then may exit the skin of the subject as illustrated, for example, in Fig. 6D. The needle may then be pulled distally until it is removed from the subject such that the thread is pulled into the location previously occupied by the needle as illustrated, for example, in Fig. 6E. Finally, excess thread is cut from the needle at the skin surface of the subject which leaves the thread implanted as illustrated, for example, in Fig. 6F.

Additional Medical and Surgical Treatments

[0124] In some embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are used as dermal fillers in various aesthetic applications as described above. In other embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are used as sutures in various medical and/or surgical applications. In still other embodiments, the threads, braids, cords, woven meshes or three-dimensional structures described herein are used in ophthalmologic surgery, drug delivery and intra-articular injection.

[0125] In some embodiments, a method for treating tumors in a subject in need thereof is provided. The thread may be attached to a needle as illustrated, for example, in Figs. 3, 4A and 4B. The distal end of the needle may be inserted into the tumor of the subject. The needle then may traverse the tumor and then may exit the tumor. The needle may then be pulled distally until it is removed from the tumor of the subject such that the thread is pulled into the location previously occupied by the needle. Finally, excess thread is cut from the needle which leaves the thread implanted in the tumor of the subject. In some of the above embodiments, the thread includes an anti-cancer agent. In some embodiments, the thread includes Bcl-2 inhibitors.

[0126] In an exemplary embodiment, methods of the current invention may be used to treat pancreatic tumors. Fig. 8A illustrates a human pancreas with a tumor while Fig. 8B illustrates a needle with a thread attached thereto. The pancreas may be accessed by surgery or minimally invasively methods such as by laparoscopy. The distal end of the needle may be inserted into the pancreatic tumor. The needle then may traverse the pancreatic tumor as illustrated in Fig. 8C and then may exit the tumor. The needle may then be pulled distally until it is removed from the pancreatic tumor such that the thread is pulled into the location previously occupied by the needle. Finally, excess thread is cut from the needle which leaves the thread implanted in the pancreatic tumor. The process may be repeated any number of times to provide, as illustrated in Fig. 8D, a pancreatic tumor which has been implanted with a number of threads. In some embodiments, the thread includes an anti-cancer agent.

[0127] In some embodiments, a method for treating a varicose vein in subject in need thereof is provided. The thread may be attached to a needle as illustrated, for example, in Figs. 3, 4A and 4B. The distal end of the needle may be inserted into the varicose vein of the subject. The needle then may traverse the varicose vein and then may exit the vein. The needle may then be pulled distally until it is removed from the varicose vein of the subject such that the thread is pulled into the location previously occupied by the needle. Finally, excess thread is cut from the needle which leaves the thread implanted in the varicose vein of the subject. In some embodiments, the needle is a flexible. In other embodiments, the thread coils when hydrated, more readily occluding the vessel.

[0128] In some embodiments, a method for nipple reconstruction is provided where a three- dimensional, cylindrical implant comprised of threads is implanted underneath the skin. The implant may include therapeutic agents, for example chrondrocyte adhesion compounds. Fig. 9A illustrates an implant of multiple layers of concentric coils of threads shaped to represent a nipple while Fig. 9B shows a cross-section of the implant of Fig. 9A. Fig. 9C illustrates how the implant of Fig. 9A could be used for nipple reconstruction. [0129] In some embodiments, methods for nerve or vessel regrowth are provided. As illustrated in Fig. 10, a needle can be used to place a thread in a specific line which could promote nerve or vessel regeneration.

6. Kits [0130] Also proved herein is a kit of parts comprising a thread of the invention. In some embodiments, the kit comprises a thread and a means for delivering or implanting the thread to a patient. In one embodiment, the means for delivery to a patient is a syringe or a needle. In another embodiment, the means for delivery to a patient is an air gun. The size (or diameter) of the needle may depend on the use of the thread, and therefore also be based on the cross-sectional area of the thread used. The outer diameter of the needle or syringe may be greater than or equal to the cross-sectional area of the thread used to lessen the tensile requirement of the thread as it is being applied to the dermis. It is further contemplated that the outer diameter of the thread may be larger than the outer diameter of the needle. Skin is quite pliable so by having a smaller diameter needle can allow the puncture size to be small even with the use of a larger diameter thread. Further, the thickness of the thread would be different in the case where the thread is a suture in comparison to the treatment of fine lines and wrinkles where it may be that a thinner thread is used. More than one thread may also be attached to a single needle.

[0131] Further, the size of the delivery device, a needle, will be dependent on its intended use and the size of the thread. It is contemplated that for use in facial contouring and or wrinkle filling a 0.006 to about 0.008" diameter thread or a 0.003 to about 0.004" diameter thread will be sufficient. In one embodiment, the needle is stainless steel. In other embodiments, the size of the thread is from about 0.01" to 0.02" in diameter.

[0132] The thread attachment to the needle can be either a mechanical attachment and/or with the use of an adhesive, such as cyanoacrylate. In one embodiment, the thread woven or looped through holes in the distal end of the needle, or alternatively, the thread wrapped around the distal end of the needle, or alternatively, the thread threaded thru an eyelet of the needle and either tied or bonded with an adhesive to form a loop, or alternatively, the thread secured (either mechanically or bonded with an adhesive) within a hole in the distal end of the needle. In another embodiment, the thread can be made to form a physical attachment to the needle during the drying process as the thread forms from the gel. For example, if a needle is used which has pores in the distal end, the pores can fill with the gel during the extrusion process and the thread would be thus be secured upon drying. The needle can be rigid or flexible to enable the user to track the needle under the wrinkle within the dermis. Further, the needle may be equipped with a ramp to guide the needle at a desired depth within the dermis, and after needle insertion, the guide may be unclasped as the needle is brought through the skin surface. In some embodiments, the thread is attached to a needle.

[0133] It is further contemplated that the kit comprises a needle and the thread attached thereto, is packaged sterile, and intended for single use. Alternatively, a kit can comprise several needles, each with an attached thread. In an additional embodiment, a kit includes threads of different sizes to enable treatment options for the physician while minimizing the number of required needle sticks. In yet another embodiment, the kit includes threads and needles of different length and curved shapes to simplify implantation in areas that are difficult to access or treat with a straight needle, for example near the nose, around the eyes and the middle portion of the upper lip.

Examples

[0134] The present invention is further defined by reference to the following examples. It will be apparent to those skilled in the art that many modifications, both to materials and methods, may be practiced without departing from the scope of the current invention. Hyaluronic acid is available from commercial sources.

Example 1 : Synthesis of a Thread

[0135] A hyaluronic acid thread of a diameter of up to 1 mm can be made by the following procedure. It is contemplated that a thread as prepared below can be stored under ambient conditions for greater than 9 months without a loss of its structural integrity or interlocking.

1. The desired amount of hyaluronic acid is weighed out into a suitable container and an aqueous solution, such as deionized water, is added to result in the desired % HA gel by weight.

2. The HA is allowed to slowly dissolve in the aqueous solution at a temperature of about 4-10 °C for 8 to 24 hours until the HA has completely swelled thus forming a gel. With higher molecular weight hyaluronic acid (e.g. >2MDa) and/or higher % gels (e.g. >10%), a longer swelling time may be required, or alternatively, the composition can me mechanically stirred. The viscosity of the gel composition is from about 150 Pascal- seconds (Pa.s) to about 2,000 Pascal-seconds (Pa.s). Optionally, the gel can be degassed by applying a vacuum or by freeze -pump-thaw cycles either prior to or after the addition of the cross-linking agent. 3. The gel composition is then transferred to a pressurized extruder (e.g., EFD Model XL 1500 pneumatic dispense machine). The nozzle of the extruder can have a tip ranging from a 15 gauge to about 25 gauge. The syringe pressure may be between about 10 psi and about 2,000 psi, depending on the viscosity of the gel composition. For very viscous gels, a pressure multiplier can be used.

4. The wetted thread is then be formed by extruding the gel composition onto a substrate by an extruder which is linearly translating at a speed commensurate with the speed of gel ejection from the syringe to achieve the desired wetted thread thickness.

5. The wetted thread is then dried under ambient conditions for about 12 hours to a percent hydration of less than about 30%, or less than about 15%, or less than about 10%, thus providing a dried thread. Optionally, the thread can be allowed to dry under a relative humidity of from about 20% to about 80%> at a temperature of from about 20 °C to about 37 °C.

6. Optionally, prior to step 7, the wetted thread can be stretched to a desired length and reduced diameter prior to dying. The stretching can be by either hanging the thread by one end and applying weight to the opposing end, or by horizontally stretching the wetted thread on a surface (either the same or different from the extrusion surface) and adhering the ends to the surface.

Example 2: Threads vs Restylane® Threads [0136] Restylane® was used as a gel composition in an attempt to form threads using the methods provided herein. Although a thread was produced, the thread did not show similar mechanical properties to the treads formed by the methods disclosed herein. A 0.007 inch x 0.020 inch Restylane® thread failed at 0.08 kg, which is far weaker than the threads of the present invention. In addition, the following observations were made: 1) Restylane® is of a lower viscosity than the gel composition of the present invention; 2) once extruded onto a surface and positioned vertically, the Restylane® formed beads which segmented the initially 6-12 inch wetted threads into smaller 1-2 inch segments; and 3) the Restylane® thread segments have an unsmooth surface and are translucent and milky-white in color, whereas the threads of the present invention are smooth and transparent. Example 3: Comparison of Tensile Strength of Different Hyaluronic Acid Threads

[0137] The tensile strength of an autocross-linked thread of hyaluronic acid (U.S. Patent 6,387,413) was compared to a thread prepared by the method of Example 1. For the autocross- linked thread, a thread of non-crosslinked hyaluronic acid was repeatedly frozen and thawed, replicating a method of autocross-linking hyaluronic acid (see, US 6,387,413). All such autocross-linked samples had less tensile force at failure than a thread prepared by the method of Example 1.

Example 4: Comparison of Ultimate Tensile Strength of Different Threads

[0138] Various threads prepared as described above were tested for tensile strength using a force gauge (e.g. Digital Force Gauge by Precision Instruments). The Restylane® threads were prepared from commercial Restylane® using the above methods. Monocryl® was used as purchased as a standard. Failure was determined by weight at which the thread broke. A zero measurement is the result of an inability to form a thread of testing quality.

Table 1

Failure stress

Example 5: Treatment of Wrinkles of a Cadaver with Hyaluronic Acid Threads

[0139] Hypodermic needles (22 to 25 Ga) were affixed with single or double strands of hyaluronic acid threads ranging from thicknesses of 0.004 in to 0.008 in with LocTite® 4014. The needles were able to traverse wrinkles in a cadaveric head of a 50 year old woman such as the naso-labial fold, peri-orals, peri-orbitals, frontalis (forehead), and glabellar. The needle was able to pull the thread through the skin such that the thread was located where the needle was previously inserted. More than one thread was used to treat the wrinkles in order to achieve the desired fill effect (two to four threads). Since cadaveric tissue does not have the same hydration characteristics as living tissue, the threads were then hydrated by applying a 0.9% saline solution to the treated area. The wrinkle was visibly lessened upon thread hydration.

Example 6: Placement of Hyaluronic Acid Threads in Dogs

[0140] Acute and chronic canine studies were performed. Hypodermic needles (22 to 25 Ga) were affixed with single or double strands of hyaluronic acid threads, ranging from thicknesses of 0.004 in to 0.008 in with LocTite® 4014. The samples were e-beam sterilized by NuTek Corp. at 29 kGy. In all cases, the needle was able to pull the attached thread or threads into the dermis. Within minutes most threads produced a visible impact on the skin surface of the animals in the form of a linear bump.

Example 7: Organization and Interlocking of the Threads via Scanning Electron

Microscopy (SEM) [0141] A 10% 1.5 MDa HA thread was prepared as described in Example 1 and was compared to a 10%) 1.5 MDa HA gel using scanning electron microscopy (SEM) (Fig. 2). The SEM (Scanning Electron Microscope) analysis was performed on the FEI Quanta 600 SEM. The thread was mounted on a double-sided carbon tape and coated with a thin layer of iridium (Ir) to avoid charging. A small portion of the HA gel was transferred to a piece of the clean silicon wafer. The gel (Fig. 2A) and the thread (Figs. 2B, 2C and 2D) have very different surface morphology. SEM images were acquired at multiple locations to ensure the morphology differences observed are representative sample differences.

[0142] As can be seen in Fig. 2, the striations in Figs. 2B, 2C and 2D (30 μιη, 10 μιη and 10 μιη, respectively) clearly show the organization and interlocking of the hyaluronic acid as compared to the gel composition before the thread is formed (Fig. 2A) (50 μιη).

Example 8: In Vitro or In Vivo Testing Regarding Increase in Fibrogenesis

[0143] The in vivo stimulation of collagen production caused by the threads of the invention can be accomplished using methods known in the art. For example, according to the methods of Wang et al. (Arch Dermatol. (2007) 143(2):155-163), the thread can be applied to a patient followed by a biopsy of the treatment area at one or more time intervals following treatment. The de novo synthesis of collagen can then be assessed using immunohistochemical analysis, quantitative polymerase chain reaction, and electron microscopy.

[0144] It is contemplated that the threads as disclosed herein will result in the synthesis of collagen at the treatment site, thus prolonging the wrinkle filling effects of the threads beyond the half-life the thread (Fig. 15 A and 15B) .

Example 9: Water Content of Dried Threads by Karl Fisher Titration

[0145] Hyaluronic acid (HA) is a water binding polymer that present in the mammalian tissues. The swelling and water intake within HA aggregates depend on propensity of water molecules to interact with the polar groups of this polymer. IR spectroscopy studies on HA films in the dried and hydrated states have demonstrated that the presence of intramolecular hydrogen-bonded organization in the dried state (Haxaire et al. (2003) Biopolymers, 72(3): 149-161). Upon interaction with water, this organization develops into hydrogen-bonded intermolecular structures where nano aggregates of water bridge the HA molecules. Intrachain hydrogen-bonded structure that exists in the dried states contain N-H...(-)0-C=0 pairs . At higher humidity, N-H and (-)O- C=0 groups are hydrated with nanodroplets containing 25 water molecules.

[0146] Threads made by the methods above were tested for the percent hydration via Karl Fisher titration. The threads were prepared with 1.5 MDa HA according to the methods of Example 1.

[0147] One water molecule per disaccharide unit will give 4.5% of water content in the HA preparation. Very low water content in the thread of Sample 2 (above) indicates that this thread it is highly dehydrated (less than one molecule of water per disaccharide unit). Percent water content (%> H₂0) data for Sample 1 and Sample 3 shows, in these threads one disaccharide unit contain 4- 5 water molecules. The reduced hydration (1-2 water molecules around the disaccharide units) in Sample 4 indicates a higher density packing of HA molecules. Example 10: Organization and Interlocking of the Threads via Transmission Electron Microscopy (TEM)

[0148] Samples of hyaluronic acid gel and thread as prepared in Example 1 were removed from refrigerator then capped with protective carbon, iridium metal, and local platinum. TEM-ready samples were then prepared by focused ion beam (FIB) milling. The fiber samples were cross sectioned in the longitudinal direction using the in situ FIB lift out method with a FEI 830 Dual Beam FIB fitted with an Omniprobe Autoprobe™ 2000. The gel sample was a random cut. TEM imaging was performed at room temperature in bright- field TEM mode using a FEI Tecnai TF-20 operated at 200kV. [0149] Some evidence of an internal microstructure was observed for the gel in Figs. 12A and 12B (dark bands). The thread, however, showed organization and interlocking of the hyaluronic acid helices. This can be seen in Figs. 12C and 12D. The hyaluronic acid helices are the dark horizontal bands observed in the direction of the thread axis. Interlocking of the helices can be observed, for example, in Fig. 12D as vertical helices can be seen in the image. Example 11: Lip Augmentation

[0150] A patient was implanted with HA threads for lip enhancement, for contouring and plumping. The patient received only topical anesthetic on the face, but it was not applied specifically to the lips. The following procedure was followed:

• Peal open the pouch and remove the sterile tray holding the HA (hyaluronic acid) threads. · Using sterile gloves or a sterile implement such as forceps, remove the desired HA thread from the tray.

• Insert the sharp end of the needle into one margin of the intended treatment area.

• Translate the needle within the dermis under or near the intended treatment area. If the needle is not in a desired location at any point, gently retract the needle and reinsert to correct the location.

• Exit the skin at the opposing margin of the intended treatment area using the sharp end of the needle. If the needle is not in the desired location, gently retract the needle and reinsert to correct the location.

• Upon confirming the desirable location of the needle, swiftly pull the needle distally, pulling the thread into place within the dermis. To avoid the thread separating from the needle before being pulled through fully, it is important to pull the thread through within approximately 2 seconds to avoid it hydrating, hence reducing its tensile strength. • Using sterile surgical scissors or scalpel, cut the excess thread protruding from the skin on both margins of the treatment area. This effectively separates the needle, which should be discarded appropriately.

• Alternatively, sterile primarily water solutions such as phosphate-buffered saline, or injection grade water can be applied directly or applied on a damp cloth to cause the excess thread to revert to a gel and the thread will separate from the needle.

[0151] Approximately 6 threads were injected into the patient's lips. Two in the lower lip, and two on each side of the upper lip. Results of the patient's augmentation is shown in Fig. 13 A, 13B, 14A, 14B, 15A, and 15B. [0152] Areas of enhancement include the vermillion border (or white roll) for lip effacement and contouring, the wet-dry mucosal junction for increasing fullness. Other techniques include more diffuse infiltration of the orbicularis oris muscle. The attending clinician is able to select the location of the thread placement, the number of threads and the size of the threads depending on desired effect. It is contemplated that each area is treated with 1 to 2 threads wherein each thread has a diameter of anywhere from 200 microns to about 500 microns when the thread is dry. After hydration, it is contemplated that the thread has a diameter of from 0.5 millimeters to about 5 millimeters.

[0153] It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all conditional language recited herein is principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art, and are to be construed as being without limitation to such specifically recited conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims.

Claims

What is claimed is:

1. A thread comprising hyaluronic acid wherein at least a portion of the hyaluronic acid is interlocked.

2. The thread of claim 1, wherein the thread reflects polarized light.

3. The thread of claim 1, wherein the thread has no more than about 30% percent by weight hydration.

4. The thread of claim 1, wherein the thread has an ultimate tensile strength about 8 kpsi or greater.

5. The thread of claim 4, wherein the thread has an ultimate tensile strength of about 20 kpsi or greater.

6. The thread of claim 1, wherein the thread has a failure of stress of about 0.1 pounds or greater.

7. The thread of claim 1, wherein the thread is braided, coiled, layered or woven to form a material.

8. The thread of claim 1, wherein the thread is substantially cylindrical.

9. The thread of claim 1, wherein the thread is substantially D-shaped.

10. The thread of claim 1, wherein the thread is substantially ribbon-shaped.

11. The thread of claim 1 , wherein the thread is substantially ellipsoidal.

12. The thread of claim 1, wherein the hyaluronic acid has a molecular weight of from about

0.5 MDa to about 2.6 MDa, from about 1.2 MDa to about 1.8 MDa, or from about 1.4 MDa to about 1.6 MDa.

13. The thread of claim 12, wherein the hyaluronic acid has a molecular weight of from about

1.4 MDa to about 1.6 MDa.

14. The thread of claim 1, wherein the thread further comprises a member selected from the group consisting of a therapeutic agent, a diagnostic agent, a fibrogenesis-enhancing agent, a lubricity-enhancing agent, a biodegradation impeding agent, and combinations thereof.

15. A method of making a thread comprising hyaluronic acid wherein at least a portion of the hyaluronic acid is interlocked, said method comprising drying under ambient conditions an aqueous gel composition comprising hyaluronic acid to provide a dried thread.

16. The method of claim 15, wherein the aqueous gel composition is buffered.

17. The method of claim 16, wherein the aqueous gel composition has pH of about 7.

18. The method of claim 15, wherein the composition is provided by adding the hyaluronic acid to an aqueous solution.

19. The method of claim 18, wherein aqueous solution comprises from about 1 % to about 30

% by weight hyaluronic acid.

20. The method of claim 19, wherein the hyaluronic acid has a molecular weight of from about 0.5 MDa to about 2.6 MDa, from about 1.2 MDa to about 1.8 MDa, or from about 1.4 MDa to about 1.6 MDa.

21. The method of claim 19, wherein the hyaluronic acid has a molecular weight of from about 1.4 MDa to about 1.6 MDa.

22. The method of claim 18, wherein the aqueous gel composition has a viscosity of from about 150 Pascal-seconds (Pa.s) to about 2,000 Pascal-seconds (Pa.s).

23. The method of claim 15, wherein prior to drying the gel is degassed.

24. The method of claim 15, wherein the composition is dried for from about 30 minutes to about 72 hours.

25. The method of claim 24, wherein the composition is dried for from about 12 hours to about 24 hours.

26. The method of claim 25, further comprising stretching the thread prior to drying.

27. The method of claim 26, wherein the drying is performed under non-denaturing conditions.

28. The method of claim 15 or 24-27, further comprising applying to the thread a sufficient amount of a member selected from the group consisting of a therapeutic agent, a diagnostic agent, a fibrogenesis-enhancing agent, a biodegradation impeding agent, a lubricity-enhancing agent and combinations thereof, optionally followed by re-drying the thread.

29. The method of any one of claims 15-28, wherein prior to the drying step, the composition is extruded from a syringe onto a substrate to provide a wetted thread.

30. The method of claim 29, wherein the composition is extruded from the syringe under pressure.

31. The method of claim 29, wherein the substrate is selected from the group consisting of polytetrafluoroethylene (PTFE), expanded PTFE, nylon, polyethylene terephthalate (PET), polystyrene, silicon, polyurethane, and activated cellulose.

32. The method of claim 31, wherein the composition and the substrate have an equilibrium contact angle of greater than about 90 degrees.

33. The method of claim 32, wherein the substrate is selected so as to provide a substantially cylindrical thread or a substantially ellipsoidal thread.

34. The method of claim 31, wherein the composition and the substrate have an equilibrium contact angle of about 90 degrees.

35. The method of claim 34, wherein the substrate is selected so as to provide a substantially

D-shaped thread.

36. The method of claim 31, wherein the composition and the substrate have an equilibrium contact angle of less than about 90 degrees.

37. The method of claim 36, wherein the substrate is selected so as to provide a substantially ribbon-shaped thread.

38. A method of treating a wrinkle in a patient in need thereof, said method comprising;

1) inserting the thread of any one of claims 1 to 14 into dermis or subcutaneous space of the patient adjacent to or under the wrinkle; and

2) applying the thread adjacent to or under the wrinkle thereby treating the wrinkle.

39. The method of claim 38, wherein steps 1) and 2) are performed 2 to 6 times.

40. The method of claim 38, wherein the thread is inserted by a needle.

41. The method of claim 40, further comprising removing the needle from the dermis.

42. The method of claim 38, further comprising hydrating the thread.

43. The method of claim 38, wherein prior to step 1), a lubricity enhancing agent is applied to the thread.

44. A kit of parts comprising the thread of claim 1.

45. The kit of claim 44, further comprising a means for delivery of the thread to a patient.

46. The kit of claim 45, where the means for delivery to a patient is a syringe, a needle or an air gun.

47. A kit of parts for use in treating a wrinkle in a patent, said kit comprising the thread of claim 1.

48. The kit of claim 47, further comprising a means for delivery of the thread to a patient.

49. A wound dressing comprising the thread of any one of claims 1 -14.

50. The wound dressing of claim 49 wherein the thread further comprises collagen.

51. A wound dressing comprising at least one woven mesh of the thread of any one of claims

1-14.

52. The wound dressing of claim 51, wherein the dressing include between 2 and about 10 layers of woven meshes.

53. The wound dressing of claim 51, wherein the woven meshes comprise identical threads.

54. The wound dressing of claim 51, wherein the woven meshes comprise different threads.

55. An adhesion barrier comprising the thread of any one of claims 1-14.

56. An adhesion barrier comprising at least one woven mesh of the thread of any one of claim

1-14.

57. A wound dressing comprising a pad which conforms to a wound location, an air-tight seal removably adhered to the pad, a negative pressure source in fluid communication with the pad and the thread of any one of claims 1 -14 attached to the wound contacting surface of the pad.

58. The wound dressing of claim 57, wherein the dressing includes at least one layer of

woven mesh comprised of the thread.

59. A method of treating a wound in a subject comprising attaching the wound dressing of claim 58 to the wound of the subject in need thereof.

60. A method of treating a wound in a subject comprising attaching the wound dressing of claim 58 to the wound of the subject in need thereof.

61. A suture comprised of any one of the threads of claims 1-14.

62. A method of using the suture of claim 61 in surgical applications.

63. A method of using any one of the threads of claims 1 -14 in surgery applications,

ophthalmologic surgery, wound closure, drug delivery and intra-articular injection.

64. A method of providing facial contouring in a patient in need thereof, said method

comprising;

1) inserting the thread of claim 1 into dermis or subcutaneous space of the patient adjacent to or under a treatment location; and

2) applying the thread adjacent to or under the treatment location thereby providing facial contouring.

65. The method of claim 64, wherein the treatment location is selected from the lips, the nasolabial fold, and the tear trough.

66. The method of claim 64, wherein steps 1) and 2) are performed 2 to 6 times.

67. The method of claim 64, wherein the thread is inserted by a needle.

68. The method of claim 67, further comprising removing the needle from the dermis.

69. The method of claim 64, further comprising hydrating the thread.

70. The method of claim 66, wherein each thread may be implanted into the epidermis, the dermis, or subcutaneous layer.

71. The method of claim 66, wherein threads are implanted relatively parallel to one another.

72. The method of claim 66, wherein the threads are implanted relatively perpendicular to one another.

73. The method of claim 66, wherein the threads are placed in a cross-hatch pattern.

74. The method of claim 66, wherein the threads are placed in a hatch pattern.