WO2023084005A1

WO2023084005A1 - Resorbable collagen scaffold for dural repair

Info

Publication number: WO2023084005A1
Application number: PCT/EP2022/081570
Authority: WO
Inventors: Louis Pascarella; Gwendolyn NIEBLER; Alexandra DIETRICH
Original assignee: Innocoll Pharmaceuticals Limited
Priority date: 2021-11-12
Filing date: 2022-11-11
Publication date: 2023-05-19

Abstract

A drug delivery device comprises a collagen matrix and at least one drug substance.

Description

RESORBABLE COLLAGEN SCAFFOLD FOR DURAL REPAIR

BACKGROUND

Field

[0001] The present disclosure relates generally to the resorbable collagen scaffolds, and in particular to scaffolds that can be used for repairing dura mater or stopping leakage of cerebrospinal fluid.

Background

[0002] Traumatic brain injury is a leading cause of death and disability in the US, can be associated with long-term disability (2,870,000 TBI-related emergency department (ED) visits in 2014 in the US). One aspect of treating traumatic brain injuries is repairing any tears or damage to the dura mater so prevent leakage of cerebrospinal fluid (CSF).

[0003] Certain chemotherapies can also cause damage to dura mater, which can also cause leakage of CSF. Repair of the dura mater may be needed as a follow-up after the chemotherapy.

[0004] Dura reconstruction is often performed with synthetic materials associated with transmission of viral infections and hydrodynamic complications, some cross-linked collagen- based products are also available. Despite product availability, there remains unmet need for improved products that can help reduce complications (e.g. CSF leakage) and consequently improve length of hospital stay and other administrative metrics.

DETAILED DESCRIPTION

[0005] The embodiments disclosed herein stem from the realization that collagen based resorbable scaffolds can be constituted to form a suitable replacement for dura mater. Such resorbable scaffolds may be surgically implanted, e.g., by suturing the scaffold to a torn portion of dura mater or adhering the matrix over a damaged portion of dura mater, to stop or prevent leakage of CSF during or after neurosurgery. Accordingly, disclosed herein is a resorbable scaffold comprising a non-crosslinked collagen gel. The scaffold may be constituted in the form of a patch that can be sutured or adhered to dura mater at a position where dura mater is torn or damaged. Accordingly, the resorbable scaffolds disclosed herein have a mechanical strength and flexibility similar to that of native dura mater. Additionally, the resorbable scaffolds disclosed herein are impermeable to water, and more specifically to CSF.

Definitions

[0006] 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 the invention pertains. Although any methods and materials similar or equivalent to those described herein may be used in the practice for testing of the present invention, the preferred materials and methods are described herein. In describing and claiming the present invention, the following terminology will be used.

[0007] 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.

[0008] A reference to an element in the singular is not intended to mean “one and only one” unless specifically stated, but rather “one or more.” Pronouns in the masculine (e.g., his) include the feminine and neuter gender (e.g., her and its) and vice versa. The term “some” refers to one or more. Underlined and/or italicized headings and subheadings are used for convenience only, do not limit the subject technology, and are not referred to in connection with the interpretation of the description of the subject technology. All structural and functional equivalents to the elements of the various configurations described throughout this disclosure that are known or later come to be known to those of ordinary skill in the art are expressly incorporated herein by reference and intended to be encompassed by the subject technology. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the above description.

[0009] As used herein, the articles “a” and “an” are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0010] As used herein, the term “about” preceding a quantity indicates a variance from the quantity. The variance may be caused by manufacturing tolerances or may be based on differences in measurement techniques. The variance may be up to 10% from the listed value in some instances. Those of ordinary skill in the art would appreciate that the variance in a particular quantity may be context dependent and thus, for example, the variance in a dimension at a micro or a nano scale may be different than variance at a meter scale. For instance, when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and still more preferably ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

[0011] As used herein, the phrase “at least one of’ preceding a series of items, with the term “and” or “or” to separate any of the items, modifies the list as a whole, rather than each member of the list (i.e., each item). The phrase “at least one of’ does not require selection of at least one of each item listed; rather, the phrase allows a meaning that includes at least one of any one of the items, and/or at least one of any combination of the items, and/or at least one of each of the items. By way of example, the phrases “at least one of A, B, and C” or “at least one of A, B, or C” each refer to only A, only B, or only C; any combination of A, B, and C; and/or at least one of each of A, B, and C.

[0012] Terms such as “top,” “bottom,” “front,” “rear” and the like as used in this disclosure should be understood as referring to an arbitrary frame of reference, rather than to the ordinary gravitational frame of reference. Thus, a top surface, a bottom surface, a front surface, and a rear surface may extend upwardly, downwardly, diagonally, or horizontally in a gravitational frame of reference.

[0013] The word “exemplary” is used herein to mean “serving as an example, instance, or illustration.” Any embodiment described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0014] Furthermore, to the extent that the term “include,” “have,” or the like is used in the description or the claims, such term is intended to be inclusive in a manner similar to the term “comprise” as “comprise” is interpreted when employed as a transitional word in a claim.

[0015] As used herein, the terms “comprising,” “including,” “containing” and “characterized by” are exchangeable, inclusive, open-ended and do not exclude additional, unrecited elements or method steps. Any recitation herein of the term “comprising,” particularly in a description of components of a composition or in a description of elements of a device, is understood to encompass those compositions and methods consisting essentially of and consisting of the recited components or elements.

[0016] As used herein, the term “consisting of’ excludes any element, step, or ingredient not specified in the claim element. [0017] The term “maturing” as used herein refers to processing the dehydrated collagen under conditions suitable to allow ageing of the dehydrated collagen without substantial degradation or contamination.

[0018] A “subject” or “patient,” as used therein, may be a human or non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably, the subject is a human.

[0019] It is to be appreciated that certain features of the invention which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination. Further, reference to values stated in ranges include each and every value within that range.

[0020] Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

Description

[0021] The present invention may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures and examples, which form a part of this disclosure. It is to be understood that this invention is not limited to the specific methods, applications, conditions or parameters described and/or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed invention.

[0022] The present disclosure provides a resorbable scaffold having a non-crosslinked collagen gel. The scaffold may be constituted as a suturable or adherable patch that can be surgically implanted at a site where dura mater is to be repaired. In some embodiments, the resorbable scaffolds disclosed herein is impermeable to water, and specifically to CSF. In some embodiments, the resorbable scaffold disclosed herein have mechanical properties similar to that of native dura mater.

[0023] The present disclosure further provides a method of manufacturing the resorbable scaffolds disclosed herein. In some embodiments, the resorbable scaffolds are prepared by lyophilizing an aqueous dispersion of matured collagen to obtain lyophilized non-crosslinked collagen and reconstituting the lyophilized non-crosslinked collagen into a gel.

[0024] The present disclosure further provides a method of using the resorbable scaffolds disclosed herein. The resorbable scaffolds disclosed herein may be used for repairing dura mater in some embodiments. The repairing may be performed by surgically implanting the resorbable scaffold at a site where dura mater requires repair and/or reinforcement.

[0025] The present disclosure also provides modified resorbable scaffolds that can assist healing of dura mater. In some embodiments, the resorbable scaffolds disclosed herein may be modified by incorporating an active substance such as a drug (e.g., dexamethasone, progesterone or a combination thereof), an antibiotic, an anti-inflammatory agent, an anti-fungal agent, a cytokine (e.g., a pro-angiogenic cytokine), a fibroblast growth factor (FGF) receptor agonist, FGF, an FGF analogue, or other suitable peptides, small molecules, or pharmaceutically acceptable salts, solvates, hydrates, cocrystals, or prodrugs thereof. In some embodiments, the resorbable scaffolds may be seeded with fibroblasts that can proliferate upon implantation of the scaffold at a site where dura mater requires repair and/or reinforcement.

[0026] Thus, the following disclosure describes the methods, systems, devices and kits associated with a resorbable scaffold usable for repairing and/or reinforcing dura mater. However, those of skill in the art, upon understanding of the present disclosure, will be able to suitably modify the methods, systems, devices and kits disclosed herein for implanting other types of implants designed for repairing and/or reinforcing other collagenous membranes of the body.

[0027] Additional features and advantages of the subject technology will be set forth in the description below, and in part will be apparent from the description, or may be learned by practice of the subject technology. The advantages of the subject technology will be realized and attained by the structure particularly pointed out in the written description and embodiments hereof. [0028] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the subject technology.

[0029] In the following detailed description, numerous specific details are set forth to provide a full understanding of the subject technology. It should be understood that the subject technology may be practiced without some of these specific details. In other instances, well-known structures and techniques have not been shown in detail so as not to obscure the subject technology.

[0030] Further, while the present description sets forth specific details of various embodiments, it will be appreciated that the description is illustrative only and should not be construed in any way as limiting. Additionally, it is contemplated that although particular embodiments of the present disclosure may be disclosed or shown in the context of delivery of anticancer drugs, such embodiments can be used with various devices and implants. Furthermore, various applications of such embodiments and modifications thereto, which may occur to those who are skilled in the art, are also encompassed by the general concepts described herein.

[0031] The disclosed methods, compositions, kits and implantable devices include a resorbable scaffold comprising a non-crosslinked collagen matrix. In some embodiments, the noncrosslinked collagen matrix comprises a collagen gel. The resorbable scaffold may further include at least one drug substance that can aid in repair and/or reinforcement of dura mater by, e.g., recruiting fibroblasts to produce collagen or inducing angiogenesis to provide blood supply to the damaged or torn portion of dura mater. In some embodiments, the at least one drug substance is substantially homogenously dispersed in the non-crosslinked collagen matrix.

[0032] In some embodiments, the at least one drug substance may include dexamethasone and progesterone, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof. In some embodiments, the at least one drug substance may include a fibroblast growth factor (FGF) agonist.

[0033] In some embodiments, the at least one drug substance may include an antibiotic, an antifungal, and/or an antimicrobial. The antibiotic, the antifungal, and/or the antimicrobial is selected from at least one of amoxicillin, amoxicillin/clavulanate, cephalexin, ciprofloxacin, clindamycin, metronidazole, azithromycin, levofloxacin, sulfamethoxazole/trimethoprim, tetracycline(s), minocycline, tigecycline, doxycycline, rifampin, triclosan, chlorhexidine, penicillin(s), aminoglycides, quinolones, fluoroquinolones, vancomycin, gentamycin, cephalosporin(s), carbapenems, imipenem, ertapenem, antimicrobial peptides, cecropin-mellitin, magainin, dermaseptin, cathelicidin, a-defensins, and a-protegrins, ketoconazole, clortrimazole, miconazole, econazole, intraconazole, fluconazole, bifoconazole, terconazole, butaconazole, tioconazole, oxiconazole, sulconazole, saperconazole, voriconazole, terbinafine, amorolfine, naftifine, griseofulvin, haloprogin, butenafine, tolnaftate, nystatin, cyclohexamide, ciclopirox, flucytosine, terbinafine, and amphotericin B.

[0034] In some embodiments, the at least one drug substance may include an antiinflammatory agent selected from at least one of steroids, prednisone, betamethasone, cortisone, dexamethasone, hydrocortisone and methylprednisolone, non-steroidal anti-inflammatory drugs (NSAIDs), aspirin, Ibuprofen, naproxen sodium, diclofenac, diclofenac-misoprostol, celecoxib, piroxicam, indomethacin, meloxicam, ketoprofen, sulindac, diflunisal, nabumetone, oxaprozin, tolmetin, salsalate, etodolac, fenoprofen, flurbiprofen, ketorolac, meclofenamate, mefenamic acid, and COX-2 inhibitors.

[0035] In some embodiments, the resorbable scaffold may include at least one of one or more peptides. Examples of one or more peptides include cytokines such as, for example, proangiogenesis cytokines (e.g., VEGF, FGF-2, PDGF, PD-ECGF/TP, angiopoietins, and the like), FGF or an FGF analogue. Other suitable cytokines are contemplated.

[0036] In some embodiments, the resorbable scaffold may include a bioresorbable polymer for providing structural strength or integrity to the resorbable scaffold. In some embodiments, the polymer is a hydrophilic polymer designed to retain a certain amount of moisture. In some embodiments, the polymer is included in the resorbable scaffold in an amount ranging from about 1 wt% to about 10 wt%. For example, the amount of the polymer in the resorbable scaffold may be about 1 wt%, about 1.5 wt%, about 2 wt%, about 2.5 wt%, about 3 wt%, about 3.5 wt%, about 4 wt%, about 4.5 wt%, about 5 wt%, about 5.5 wt%, about 6 wt%, about 6.5 wt%, about 7 wt%, about 7.5 wt%, about 8 wt%, about 8.5 wt%, about 9 wt%, about 9.5 wt%, about 10 wt%, or any amount between any two of these values.

[0037] In some embodiments, the resorbable scaffold may have a water content comparable to that of native dura mater. For example, the resorbable scaffold may have a water content in a range from about 60 wt% to about 85 wt%. In some embodiments, the resorbable scaffold has a water content of about 60 wt%, about 62 wt%, about 64 wt%, about 66 wt%, about 68 wt%, about 70 wt%, about 72 wt%, about 74 wt%, about 76 wt%, about 78 wt%, about 80 wt%, about 82 wt%, about 84 wt%, about 85 wt%, or any other value between any two of these values.

[0038] Ins some embodiments, the polymer is a polyethylene glycol (PEG). In some embodiments, the content of PEG may be in a range from about 4 wt% to about 6 wt% in the resorbable scaffold. For example, the resorbable scaffold may include PEG in a range from about 4 wt%, 4.1 wt%, about 4.2 wt%, about 4.3 wt%, about 4.4 wt%, about 4.5 wt%, about 4.6 wt%, about 4.7 wt%, about 4.8 wt%, about 4.9 wt%, about 5.0 wt%, about 5.1 wt%, about 5.2 wt%, about 5.3 wt%, about 5.4 wt%, about 5.5 wt%, about 5.6 wt%, about 5.7 wt%, about 5.8 wt%, about 5.9 wt%, about 6 wt% or any other value between any two of these values.

[0039] In some embodiments, the polymer is a copolymer. In some embodiments, the polymer is a terpolymer.

[0040] In some embodiments, the polymer includes at least one of polyglycolide (PGA), polycaprolactone (PCL), poly(DL-lactic acid) (PLA), poly(alpha-hydroxy acids), poly(lactide-co-glycolide)(PLGA or DLG), poly(DL-lactide-co-caprolactone) (DL-PLCL), poly(trimethylene carbonate) (PTMC), polydioxanone (PDO), poly(4-hydroxy butyrate) (PHB), polyhydroxyalkanoates (PHA), poly(phosphazene), polyphosphate ester), poly(amino acid), pol- ydepsipeptides, poly (butylene succinate) (PBS), polyethylene oxide, polypropylene fumarate, polyiminocarbonates, poly(lactide-co-caprolactone) (PLCL), poly(glycolide-co-caprolactone) (PGCL) copolymer, poly(D,L-lactic acid), polyglycolic acid, poly(L-lactide-co-D,L-lactide), poly(L-lactide-co-glycolide), poly(D,L-lactide-co-glycolide), poly(gycolide-trimethylene carbonate), poly(ethyl glutamate-co-glutamic acid), poly(tert-butyloxy-carbonylmethyl glutamate), poly(glycerol sebacate), tyrosine-derived polycarbonate, poly l,3-bis-(p- carboxyphenoxy) hexane-co-sebacic acid, polyphosphazene, ethyl glycinate polyphosphazene, polycaprolactone co-butylacrylate, a copolymer of polyhydroxybutyrate, a copolymer of maleic anhydride, a copolymer of poly(trimethylene carbonate), polyethylene glycol (PEG), hydroxypropylmethylcellulose and cellulose derivatives, polysaccharides (such as hyaluronic acid, chitosan and starch), proteins (such as gelatin and collagen) or PEG derivatives, polyaspirins, polyphosphagenes, collagen, starch, pre-gelatinized starch, hyaluronic acid, chitosans, gelatin, alginates, albumin, fibrin, vitamin E analogs, such as alpha tocopheryl acetate, d-alpha tocopheryl succinate, D-lactide, D,L-lactide, L-lactide, D,L-lactide-caprolactone (DL-CL), D,L-lactide- glycolide-caprolactone (DL-G-CL), dextrans, vinylpyrrolidone, polyvinyl alcohol (PVA), PVA- g-PLGA, PEGT-PBT copolymer (polyactive), methacrylates, poly(N-isopropylacrylamide), PEO- PPO-PEO (pluromcs), PEO-PPO-PAA copolymers, PLGA-PEO-PLGA, PEG-PLG, PLA-PLGA, poloxamer 407, PEG-PLGA-PEG triblock copolymers, SAIB (sucrose acetate isobutyrate)hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxyethyl methylcellulose, carboxymethylcellulose or salts thereof, Carbo-pol®, poly(hydroxyethylmethacrylate), poly(methoxyethylmethacrylate), poly(methoxyethoxy- ethylmethacrylate), polymethylmethacrylate (PMMA), methylmethacrylate (MMA), gelatin, polyvi-nyl alcohols, propylene glycol, and poly(DL-lactide-co-glycolide-co-caprolactone).

[0041] In some embodiments, the polymer is ester-terminated. In some embodiments, the polymer is a terpolymer that includes three polymers selected from the following: polyglycolide (PGA), polycaprolactone (PCL), poly(L-lactic acid) (PLA), poly(DL-lactic acid) (PLA), poly(trimethylene carbonate) (PTMC), poly-dioxanone (PDO), poly(4-hydroxy butyrate) (PHB), polyhydroxyalkanoates (PHA), poly(phosphazene), and polyethylene glycol.

[0042] In some embodiments, the resorbable scaffold is constituted such that the scaffold is resorbed over a period ranging from about 6 weeks to about 1 year. As will be appreciated, the amount of time needed for resorption of the resorbable scaffold may be dependent on the size and constitution of the scaffold.

[0043] The resorbable scaffold may be generally constituted as a membrane or a film. In some embodiments, the resorbable scaffold may have a thickness in a range from about 1 pm to about 100 pm. For example, the resorbable scaffold may have a thickness of about 1 pm, about 5 pm, about 10 pm, about 15 pm, about 20 pm, about 25 pm, about 30 pm, about 35 pm, about 40 pm, about 45 pm, about 50 pm, about 55 pm, about 60 pm, about 70 pm, about 75 pm, about 80 pm, about 85 pm, about 90 pm, about 95 pm, about 100 pm or any thickness between any two of these thicknesses. While, embodiments with resorbable scaffold having higher thickness than 100 pm are contemplated within the scope of the disclosure, such applications may be rare in practice.

[0044] In some embodiments, the resorbable scaffold may be constituted into a patch having an area ranging from about 5 cm² to about 250 cm². For example, the resorbable scaffold may have an area of about 5 cm², about 10 cm², about 15 cm², about 20 cm², about 25 cm², about 30 cm², about 35 cm², about 40 cm², about 50 cm², about 55 cm², about 60 cm², about 70 cm², about 80 cm², about 90 cm², about 100 cm², about 110 cm², about 120 cm², about 130 cm², about 140 cm², about 150 cm², about 160 cm², about 170 cm², about 180 cm², about 190 cm², about 200 cm², about 210 cm², about 220 cm², about 230 cm², about 240 cm², about 250 cm², or any area between any two of these values.

[0045] Without wishing to be bound by theory, where the damage to dura mater is confined to an area smaller than 5 cm², dura mater may be repaired by suturing the damaged area together during a surgery.

[0046] In some embodiments, the resorbable scaffold may have any suitable shape and/or may be cut into any suitable shape prior to surgery. Examples of the shape of the resorbable scaffold include, but are not limited to, triangle, quadrilateral (e.g., rectangle, square, etc.), pentagon, hexagon, octagon, or any other regular or irregular polygon, and circle.

[0047] In some embodiments, the edges of the resorbable scaffold may have a slightly higher thickness than the rest of the resorbable scaffold so as to provide additional strength and/or integrity to the scaffold to allow the scaffold to be sutured at the implant site. In some embodiments, the edges of the resorbable scaffold may include a layer of a suitable resorbable adhesive that can seal the edges of the resorbable scaffold to the adjoining dura mater.

[0048] In some embodiments, the resorbable scaffold has mechanical properties similar to that of native dura mater. For example, the resorbable scaffold may have an elastic modulus in a range from about 30 MPa to about 120 MPa. In some embodiments, the resorbable scaffold may have an elastic modulus of about 30 MPa, about 35 MPa, about 40 MPa, about 45 MPa, about 50 MPa, about 55 MPa, about 60 MPa, about 65 MPa, about 70 MPa, about 75 MPa, about 80 MPa, about 85 MPa, about 90 MPa, about 95 MPa, about 100 MPa, about 105 MPa, about 110 MPa, about 115 MPa, about 120 MPa, or any elastic modulus between any two of these values.

[0049] In some embodiments, the resorbable scaffold may have a tensile strength in a range from about 2 MPa to about 12 MPa. For example, the resorbable scaffold has a tensile strength of about 2 MPa, about 2.5 MPa, about 3 MPa, about 3.5 MPa, about 4 MPa, about 4.5 MPa, about 5 MPa, about 5.5 MPa, about 6 MPa, about 6.5 MPa, about 7 MPa, about 7.5 MPa, about 8 MPa, about 8.5 MPa, about 9 MPa, about 9.5 MPa, about 10 MPa, about 10.5 MPa, about 11 MPa, about 11.5 MPa, about 12 MPa, about or any other value between any two of these values.

[0050] In some embodiments, the collagen in the resorbable scaffold comprises a modified collagen obtained by providing isolated collagen, optionally an isolated collagen dispersion; freezing the isolated collagen; and dehydrating the frozen collagen. In some embodiments, the modified collagen is obtained by providing isolated collagen, optionally an isolated collagen dispersion; freezing the isolated collagen; dehydrating the frozen collagen; and maturing the dehydrated collagen.

[0051] The term “dispersion” as used herein refers to a mixture in which collagen particles are dispersed in a fluid, optionally a liquid, further optionally an aqueous, medium. The collagen particles may comprise collagen molecules, or aggregates thereof; which are dispersed in a fluid, optionally a liquid, further optionally an aqueous, medium. Optionally, the collagen particles, which are dispersed in a fluid, optionally a liquid, further optionally an aqueous, medium; have a length (or maximum dimension) of at least one micrometer.

[0052] In some embodiments, the modified collagen is obtained by: (a) providing isolated collagen, optionally an isolated collagen dispersion; (b) freezing the isolated collagen; and (c) dehydrating the frozen collagen.

[0053] In some embodiments, the modified collagen is obtained by: (a) providing isolated collagen, optionally an isolated collagen dispersion; (b) freezing the isolated collagen; (c) dehydrating the frozen collagen; and (d) maturing the dehydrated collagen.

[0054] In some embodiments, the providing step comprises the step of removing the fluid prior to the providing step. In some embodiments, the providing step comprises the step of removing at least some of the fluid prior to the providing step. In some embodiments, the providing step comprises the step of removing at least some of the fluid prior to the providing step; to provide an isolated collagen dispersion. In some embodiments, the fluid is a liquid. In some embodiments, the liquid is an aqueous medium.

[0055] In some embodiments, the providing step comprises the step of removing the fluid prior to the providing step to provide a dispersion having a concentration of about 3-30%, optionally 3-4%, (w/w) collagen particles. In some embodiments, the fluid is a liquid. In some embodiments, the liquid is an aqueous medium. In some embodiments, the collagen particles may have a size (e.g., average diameter) in a range from about 50 nm to about 50 pm. For example, the collagen particles may have an average diameter of about 50 nm, about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 950 nm, about 1 pm, about 1.05 pm, about 1.1 pm, about 1.25 pm, about 1.5 pm, about 2 pm, about 2.5 pm, about 3 pm, about 3.5 pm, about 4 pm, about 4.5 pm, about 5 pm, about 5.5 pm, about 6 pm, about 6.5 pm, about 7 pm, about 7.5 pm, about 8 pm, about 8.5 pm, about 9 pm, about 9.5 pm, about 10 pm, about, 10.5 pm, about 11 pm, about 12 pm, about 13 pm, about, 14 pm, about 15 pm, about 16 pm, about 17 pm, about 18 pm, about 19 pm, about 20 pm, about 21 pm, about 22 pm, about 23 pm, about 24 pm, about 25 pm, about 26 pm, about 27 pm, about 28 pm, about 29 pm, about 30 pm, about 32 pm, about 34 pm, about 36 pm, about 38 pm, about 40 pm, about 42 pm, about 44 pm, about 46 pm, about 48 pm, about 50 pm, about or any other value between any two of these values. In some embodiments, the size dispersion of the collagen particles is in a range from about 5 nm to about 50 nm, about 50 nm to about 100 nm, about 100 nm to about 200 nm, about 200 nm to about 500 nm, about 500 nm to about 1 pm, or any other range between or within any two of these ranges.

[0056] In some embodiments, the freezing step comprises freezing to a temperature of about -33 °C to about -42 °C In some embodiments, the freezing step comprises freezing to a temperature of about -38 °C In some embodiments, the freezing step comprises freezing at a rate of about 0.3 °C to about 1.5 °C per minute, optionally a rate of about 0.5 °C per minute.

[0057] In some embodiments, the dehydrating step comprises removing the aqueous phase. In some embodiments, the dehydrating step comprises removing the aqueous phase by reducing the pressure. In some embodiments, the dehydrating step comprises removing the aqueous phase by reducing the pressure to about 0.05 to about 0.5 mbar. In some embodiments, the dehydrating step comprises removing the aqueous phase by applying a vacuum.

[0058] Optionally or additionally, the dehydrating step comprises increasing the temperature of the frozen collagen. Further optionally or additionally, the dehydrating step comprises increasing the temperature of the frozen collagen under vacuum. Still further optionally or additionally, the dehydrating step comprises increasing the temperature of the collagen to about +30 °C Still further optionally or additionally, the dehydrating step comprises increasing the temperature of the collagen to about +30 °C under vacuum.

[0059] Optionally or additionally, the dehydrating step comprises increasing the temperature of the collagen to about +30 °C at a rate of about 0.3 °C to about 1.5 °C per minute, further optionally at a rate of about 0.5 °C per minute. Further optionally or additionally, the dehydrating step comprises increasing the temperature of the collagen to about +30 °C at a rate of about 0.3 °C to about 1.5 °C per minute, further optionally at a rate of about 0.5 °C per minute, under vacuum.

[0060] In some embodiments, the dehydrating step comprises at least one equilibrating step.

[0061] In some embodiments, the at least one equilibrating step comprises maintaining the temperature at a constant temperature, sufficient to allow the frozen collagen to reach a desired temperature. Optionally, the at least one equilibrating step comprises maintaining the temperature at a constant temperature for a sufficient period of time to allow the frozen collagen to reach a desired temperature. Still further optionally, the at least one equilibrating step comprises maintaining the temperature at a constant temperature for at least 10 mins, optionally at least 20 mins, further optionally at least 30 mins, still further optionally at least 45 mins, still further optionally at least 60 mins; to allow the frozen collagen to reach a desired temperature.

[0062] In some embodiments, the at least one equilibrating step is conducted when the temperature is increased to at least -20 °C. Optionally or additionally, the at least one equilibrating step is conducted when the temperature is increased to at least -10 °C. Optionally or additionally, the at least one equilibrating step is conducted when the temperature is increased to at least 0 °C. Optionally or additionally, the at least one equilibrating step is conducted when the temperature is increased to at least +10 °C. Optionally or additionally, the at least one equilibrating step is conducted when the temperature is increased to at least +20 °C Optionally or additionally, the at least one equilibrating step is conducted when the temperature is increased to at least +30 °C.

[0063] In some embodiments, the dehydrating step comprises six equilibrating steps, each equilibrating step being conducted when the temperature is increased by about 10 °C Optionally, the dehydrating step comprises six equilibrating steps, each equilibrating step being conducted when the temperature is increased to about -20 °C, about -10 °C, about 0 °C, about +10 °C, about +20 °C, and about +30 °C.

[0064] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 2 °C. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 10 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 20 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 30 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 40 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 50 °C. In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 60 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 70 °C. In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 80 °C.

[0065] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 30 °C. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 40 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of at least 65 °C.

[0066] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of 30 °C. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 40 °C. Still further optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 65 °C.

[0067] In some embodiments, the maturing step is conducted for a period of at least one week, optionally at least two weeks, further optionally at least three weeks, still further optionally at least four weeks, still further optionally at least five weeks, still further optionally at least six weeks.

[0068] In some embodiments, the maturing step is conducted for a period of at least two months, optionally at least four months, further optionally at least six months, still further optionally at least twelve months.

[0069] In some embodiments, the maturing step is conducted for a period of one week, optionally two weeks, further optionally three weeks, still further optionally four weeks.

[0070] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 2 °C for a period of at least six months. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 2 °C for a period of six months.

[0071] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 30 °C for a period of at least two months. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 30 °C for a period of two months. [0072] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 40 °C for a period of at least six weeks. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 40 °C for a period of six weeks.

[0073] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 65 °C for a period of at least one week. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 65 °C for a period of one week.

[0074] In some embodiments, the maturing step is conducted at a relative humidity of less than 100%, optionally less than 90%, further optionally less than 80%, still further optionally less than 70%, still further optionally less than 60%, still further optionally less than 50%, still further optionally less than 40%, still further optionally less than 30%.

[0075] The term “relative humidity” as used herein refers to a measure of the maximum amount of water in a mixture of gas and water vapor, optionally at a given gas temperature and atmospheric pressure, optionally at constant atmospheric pressure, optionally expressed as a percentage of the maximum amount of water vapor within the gas at the given gas temperature and atmospheric pressure. For the purposes of this specification, the term “relative humidity” is intended to mean a measure of the amount of water vapor in a mixture of environmental air and water vapor, in which the maturing step is conducted, at a constant atmospheric pressure, and expressed as a percentage. For the purposes of this specification, atmospheric pressure understood to be about 980 to about 1040 millibars.

[0076] It is understood that, in conducting the maturing step, the parameters of temperature, time, pressure, and relative humidity are not necessarily mutually exclusive, and the skilled person would recognize that as one parameter is varied, one or both of the other parameters may also be varied accordingly.

[0077] In some embodiments, the maturing step comprises storing the dehydrated collagen at a temperature of at least 40 °C for a period of at least six weeks, and at a relative humidity of less than 80%. Optionally, the maturing step comprises storing the dehydrated collagen at a temperature of 40 °C for a period of 6 weeks, and at a relative humidity of 75%.

[0078] In some embodiments, the isolated collagen is fibrillar collagen. Fibrillar collagen from different sources may be used including commercially available fibrillar collagen, for example, biomedical collagen from Devro Biomedical Collagen, Australia. Currently there are five known types of fibrillar collagen; Type I, II, III, V and XI. Alternatively, collagen can be extracted from tendons or hides of different mammals, including human, horse, cattle, sheep and pigs. Collagen can also be extracted from a non-mammal such as fish. Details on the various types of collagen are described by Gelse et al., (Advanced Drug Delivery Reviews 55 (2003), 1531-1546), the whole contents of which are incorporated herein by reference. The present inventors have used a bovine-derived collagen Type I for the manufacture of bupivacaine- collagen sponges. Equine-derived collagen Type I is also suitable for use in the present invention, as are fibrillar collagen such as type I collagen from pigs and sheep. Type I collagen is a connective tissue extracted from animal tendons and other sources; in this case, the collagen is derived from bovine tendons. The Type I collagen consists of three approximately 1,050 amino- acid-long polypeptide chains, two alpha- 1 chains, and one alpha-2 chain. These are coiled to form a right-hand helix (known as a triple helix) around a common axis. The rod-shaped molecule has a length of 2900 Angstrom, a diameter of 14 Angstrom and a molecular weight of approx. 300,000 Daltons. Type I collagen can be typified by its reaction with the protein core of another connective tissue component known as a proteoglycan. Type I collagen contains signaling regions that facilitate cell migration.

[0079] Optionally, the isolated collagen is selected from Type I collagen, Type II collagen, Type III collagen, and a mixture thereof. Still further optionally, the isolated collagen is Type I collagen.

[0080] In some embodiments, the matured collagen is powderized to form matured collagen particles. In some embodiments, the matured collagen particles may have a size (e.g., average diameter) in a range from about 50 nm to about 50 pm. For example, the collagen particles may have an average diameter of about 50 nm, about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 950 nm, about 1 pm, about 1.05 pm, about 1.1 pm, about 1.25 pm, about 1.5 pm, about 2 pm, about 2.5 pm, about 3 pm, about 3.5 pm, about 4 pm, about 4.5 pm, about 5 pm, about 5.5 pm, about 6 pm, about 6.5 pm, about 7 pm, about 7.5 pm, about 8 pm, about 8.5 pm, about 9 pm, about 9.5 pm, about 10 pm, about, 10.5 pm, about 11 pm, about 12 pm, about 13 pm, about, 14 pm, about 15 pm, about 16 pm, about 17 pm, about 18 pm, about 19 pm, about 20 pm, about 21 pm, about 22 pm, about 23 pm, about 24 pm, about 25 pm, about 26 pm, about 27 pm, about 28 pm, about 29 pm, about 30 pm, about 32 pm, about 34 pm, about 36 pm, about 38 pm, about 40 pm, about 42 pm, about 44 pm, about 46 pm, about 48 pm, about 50 pm, about or any other value between any two of these values. In some embodiments, the size dispersion of the matured collagen particles is in a range from about 5 nm to about 50 nm, about 50 nm to about 100 nm, about 100 nm to about 200 nm, about 200 nm to about 500 nm, about 500 nm to about 1 pm, or any other range between or within any two of these ranges.

[0081] In some embodiments, the powderization process is performed by grinding or pulverizing the matured collagen.

[0082] Method of Manufacture

[0083] The following general method of manufacture refers to manufacturing of the resorbable scaffold disclosed herein, without departing from the scope of the teaching of this disclosure.

[0084] Collagen

[0085] Production of Type I Collagen from Bovine Tendons: The collagen is extracted from bovine Achilles tendon. During the manufacturing process, bovine tendons are first treated with IN sodium hydroxide (NaOH) to clean and purify the material and to deplete the fat content followed by neutralization with IN HC1. This step is followed by treatment with 0.9% sodium chloride (NaCl) solution to remove low molecular weight soluble components of the collagen. A treatment with hydrogen peroxide solution ensures bleaching of the tendons. Reduction of the particle size of the collagen material is followed by fermentative breakdown using pepsin. Treatment with pepsin is used to degrade contaminating serum protein components, primarily bovine serum albumin and causes the detachment of non-helical portions of the collagen molecule (telopeptides). After filtration, precipitation of the collagen is accomplished by means of manipulation of the pH (from acidic pH to neutral pH). The fibrillar Type I collagen material is finally precipitated out of solution, washed again with distilled water to remove residual pepsin and then concentrated by means of centrifugation.

[0086] Compounding Process and Equipment

[0087] The fibrillar Type I collagen material is prepared and added to preheated water (below 42 °C) in a stainless steel (SS) vessel. Collagen swelling and subsequent dispersion formation is performed using, e.g., a high-shear homogenizer. For example, the homogenizer may have a rotor-stator head that is designed to create high shear forces by pulling the collagen material through the rotating homogenizer head and forcing it against the proximal stationary stator head. It is this design that facilitates the high shear forces required to separate the fibrous collagen mass at the beginning of dispersion preparation.

[0088] Following completion of collagen dispersion formation, the dispersion is transferred to a closed heated jacketed vessel for final compounding. The jacket temperature is maintained at a range of about 36 °C to about 38 °C. Optionally, a drug substance or one or more active ingredient raw material is first dissolved in a portion of water at room temperature and is then introduced into the heat-jacketed SS vessel under low shear mixing to achieve homogeneity in the “loaded” collagen dispersion. The collagen (or the loaded collagen) dispersion is a free- flowing opaque white to off-white liquid.

[0089] The dispersion is subsequently freeze-dried yielding a membrane containing collagen and optionally a drug substance or other active ingredient. Examples of drug substances and/or active ingredients that can be included in the dispersion are provided elsewhere herein.

[0090] Filling/Lyophilization Process and Equipment

[0091] The collagen (or the loaded collagen) dispersion is filled into appropriately sized lyophilization molds or blister trays for freeze-drying. The filling process is performed using a positive displacement pump. The pump may be valve-less, and may have ceramic pistons.

[0092] Upon completion of tray filling, the filled molds or blister trays are placed into the lyophilizer. Thermocouples are placed both in product and on shelves and a conductivity probe is also employed to provide in-process feedback on process temperatures and conductivity. The lyophilization process cycle used for the collagen membrane involves freezing down to a temperature of about -38 °C over a period of about 3.5 hours, followed by drying to a temperature of about 30 °C over a period of about 14.5 hours.

[0093] Ethylene Oxide (EtO) Sterilization Process

[0094] The lyophilized membrane is packed into suitable packaging material, which may comprise of a sealed polyethylene blister or low density polyethylene (LDPE) sachet in an outer pouch consisting of polyethylene/LDPE laminate or aluminum foil. The product is then subjected to terminal sterilization, which can be gas-mediated ethylene oxide sterilization or radiation (gamma or electron beam). In the preferred embodiment, sterilization by ethylene oxide gas has been selected. [0095] Ethylene oxide (C2H4O) is a gas at operating temperature and sterilizes via its action as a powerful alkylating agent. Under the appropriate conditions, cellular constituents of organisms such as nucleic acid complexes, functional proteins and enzymes react with ethylene oxide, causing the addition of alkyl groups. As a result of the alkylation, cell reproduction is prevented and cell death ensues. Specific processing conditions and parameters must be met to achieve this effect within the target product; including but not limited to, acceptable concentration of ethylene oxide in the chamber and a minimum water activity level within the organism. The process is essentially a chemical reaction and is therefore temperature dependent; the rate of reaction increases with temperature.

[0096] The optimum temperature is within the range of about 30 °C to about 40 °C. These properties define the key characteristics of the ethylene oxide sterilization process.

[0097] The process is dependent on the water content existing in the membranes and a consistent range of moisture content is achieved by equilibration of the product with atmospheric humidity prior to sterilization. An optimum water content is not less than 9%. The product is loaded into stainless steel wire mesh baskets and placed into the stainless-steel sterilizer chamber using a defined loading pattern. The sterilization chamber is then evacuated to remove air and ethylene oxide is introduced until the required concentration is achieved.

[0098] Product is held under these conditions for a defined period and, on completion of the pre-determined dwell period, ethylene oxide from the chamber is exhausted to the atmosphere via catalytic converters. These units ensure catalytic conversion of ethylene oxide to carbon dioxide and water with high efficiency. The sterilization chamber and its contents arc then repeatedly flushed with air to remove the remaining ethylene oxide from the chamber. After completion of post sterilization flushing, the product is transferred to a holding area for longer term aeration. This phase of the process serves to further scavenge low level residual ethylene oxide from the product and packaging. The product is held at room temperature until the limits for ethylene oxide derivative residues have been reached.

[0099] Alternative Sterilization Process and Equipment

[0100] Radiation sterilization including gamma and electron beam may be used instead of the EtO sterilization process mentioned above.

[0101] In some embodiments, the method of manufacturing the modified collagen further comprises the step of mechanically degrading the modified collagen prior to the maturing step. Optionally, the mechanical degrading step comprises milling. Further optionally, the mechanical degrading step is selected from milling, cutting, grinding, and a mixture thereof.

[0102] In some embodiments, a method for isolating collagen may include: (a) providing a collagen source; and (b) increasing the pH of the collagen source to about 6.5 to about 7.5. In some embodiments, the collagen source is a collagen dispersion.

[0103] In some embodiments, the providing step comprises the step of removing the fluid prior to the providing step. In some embodiments, the providing step comprises the step of removing at least some of the fluid prior to the providing step. Optionally, the providing step comprises the step of removing at least some of the fluid prior to the providing step; to provide an isolated collagen dispersion. In some embodiments, the fluid is a liquid. In some embodiments, the liquid is an aqueous medium.

[0104] In some embodiments, the pH of the collagen source, optionally the collagen dispersion, is increased to about 7.5.

[0105] In some embodiments, the collagen source is a fibrous tissue, optionally connective tissue. Further optionally, the collagen source is tendon, optionally animal tendon, further optionally equine or bovine tendon, preferably equine tendon.

[0106] In some embodiments, the method further comprises the step of degrading the collagen source prior to the pH-increasing step. Optionally, the degrading step comprises mechanically degrading the collagen source prior to the pH-increasing step. Optionally or additionally, the degrading step comprises chemically degrading the collagen source prior to the pH-increasing step.

[0107] In some embodiments, the mechanical degrading step comprises milling. Further optionally, the mechanical degrading step is selected from milling, cutting, grinding, granulating, and a mixture thereof. Optionally or additionally, the chemical degrading step comprises contacting the collagen source with an enzyme, optionally a proteolytic enzyme. Optionally, the proteolytic enzyme is selected from chymosin, cathepsin E, and pepsin; preferably pepsin.

[0108] In some embodiments, the chemical degrading step is conducted at a pH of about 2.5.

[0109] In some embodiments, the method further comprises the step of removing contamination from the collagen source. Optionally, the removing step comprises contacting the collagen source with a base, optionally a strong base, further optionally sodium hydroxide, still further optionally an aqueous solution of sodium hydroxide.

[0110] In some embodiments, the method comprises the step of filtering the degraded collagen source, optionally the degraded collagen dispersion, prior to the pH-increasing step.

[0111] In some embodiments, the method comprises the step of concentrating the collagen. In some embodiments, the concentrating step comprises isolating the collagen. Further optionally, the concentrating step comprises isolating the collagen by centrifugation.

[0112] In some embodiments, the concentrating step comprises the step of removing the fluid to provide a dispersion having a concentration of about 3-30%, optionally 3-4%, (w/w) collagen particles. In some embodiments, the fluid is a liquid. In some embodiments, the liquid is an aqueous medium.

[0113] In some embodiments, the isolated collagen is frozen. Optionally, the isolated collagen is frozen at less than -20 °C. Optionally, the frozen isolated collagen is thawed prior to preparing the modified collagen.

[0114] In some embodiments, use of the resorbable scaffolds disclosed herein comprises the implanting the resorbable scaffold at a site where dura mater requires repair and/or reinforcement. Implanting may include placing the resorbable scaffold at the site and suturing (or otherwise fastening) the scaffold to the dura mater adjoining the site such that the portion of dura mater that requires repair and/or reinforcement is covered by the scaffold. Such implantation of the resorbable scaffold may, e.g., plug a hole in dura mater, thereby preventing leakage of cerebrospinal fluid (CSF).

[0115] In some instances, dura mater may not be torn, but may have thinned at certain sites because of trauma, lifestyle choices (e.g., smoking), or age. In such instances, the thinned dura mater may not effectively prevent leakage of CSF, and thus, may require reinforcement. The resorbable scaffolds disclosed herein may be implanted at sites where dura mater has thinned so as to provide reinforce and thereby prevent leakage of CSF.

[0116] According to at least some embodiments of the present disclosure, a method for manufacturing a resorbable scaffold comprising a modified collagen may include: (a) providing a modified collagen; (b) preparing an aqueous dispersion of the modified collagen; (c) degrading the aqueous dispersion; and (d) dehydrating the aqueous dispersion. The modified collagen may be obtained using any of the methods disclosed herein. [0117] In some embodiments, the preparing step comprises adding heated water, optionally heated purified water, to the modified collagen. Optionally, the water, optionally the purified water is heated to about 35 to about 42 °C prior to adding to the modified collagen.

[0118] In some embodiments, the preparing step is conducted at a pH of about 4.0.

[0119] In some embodiments, the degrading step comprises mechanically degrading the aqueous dispersion.

[0120] In some embodiments, the mechanical degrading step comprises shear mixing.

[0121] In some embodiments, the composition comprises modified collagen in an amount of about 0.4% to 1.5% (w/w).

[0122] In some embodiments, the composition has a pH of about 4.0.

[0123] In some embodiments, the dehydrating step comprises removing liquid from the aqueous dispersion such that the composition comprises liquid in an amount of less than 30%, optionally less than 20%, further optionally less than 15% (w/w) of the composition. Further optionally, the dehydrating step comprises removing liquid from the aqueous dispersion such that the composition comprises liquid in an amount of less than 13%, preferably less than 12%, (w/w) of the composition.

[0124] In some embodiments, the dehydrating step comprises removing liquid from the aqueous dispersion using a convective drying cabinet.

[0125] In accordance with at least some embodiments of the present disclosure, a resorbable scaffold is obtainable by providing isolated collagen, optionally an isolated collagen dispersion; freezing the isolated collagen; and dehydrating the frozen collagen.

[0126] In accordance with at least some embodiments of the present disclosure, a drug delivery composition is obtainable by providing isolated collagen, optionally an isolated collagen dispersion; freezing the isolated collagen; dehydrating the frozen collagen; and maturing the dehydrated collagen.

[0127] In accordance with at least some embodiments of the present disclosure, a method of preparing a resorbable scaffold for sustained drug release may include: (a) providing isolated collagen, optionally an isolated collagen dispersion; (b) freezing the isolated collagen; and (c) dehydrating the frozen collagen.

[0128] In accordance with at least some embodiments of the present disclosure, a method of preparing a resorbable scaffold for sustained drug release may include: (a) providing isolated collagen, optionally an isolated collagen dispersion; (b) freezing the isolated collagen; (c) dehydrating the frozen collagen; and (d) maturing the dehydrated collagen.

[0129] In some embodiments, the method further comprises the step of lyophilizing and/or dehydrating, the drug-, optionally drug solution-, containing drug delivery composition.

[0130] Specific embodiments of the invention will now be demonstrated by reference to the following general methods of manufacture and examples. It should be understood that these examples are disclosed solely by way of illustrating the invention and should not be taken in any way to limit the scope of the present invention.

EXAMPLES

Example 1: Collagen isolation

[0131] Collagen can be isolated from a number of sources, for example, animal hides and animal tendons. In a preferred embodiment, the collagen is isolated from animal tendon, for example equine or bovine tendon; although any known source of collagen, including fibrous tissue, optionally connective tissue, may be used and selected by one skilled in the art. Preferably, the collagen is isolated from equine tendon. In the method of isolation, equine tendons were milled to degrade the collagen source. The milled equine tendons were treated with a number of reagents, including IN sodium hydroxide (NaOH) to remove microbiological contamination such as prions at the beginning of the process. Treatment steps with hydrogen peroxide and washing steps at different pH values were conducted, followed by a milling step, which was used to increase the surface for the next treatment step. The molecular weight of the collagen source was additionally reduced by treatment with the proteolytic enzyme pepsin at an approximate pH of 2.5. The pH was adjusted using an aqueous solution of IN HC1. The pepsin was used to degrade contaminating serum components such as equine serum albumin (ESA) and resulted in the detachment of nonhelical portions of the collagen molecule (telopeptides). During this process, the collagen material was also partially solubilized in the acidic medium. After filtration, the pH level was increased from 2.5 to 7.5 by addition of IN sodium hydroxide (NaOH). This pH adjustment resulted in precipitation of the fibrillar collagen out of solution, which was then concentrated by means of centrifugation to provide a collagen dispersion having a concentration of about 3-30% (w/w). The resulting material was designated fresh collagen. The fresh collagen can be processed in several ways. [0132] The fresh collagen can be packaged in suitable portions and frozen to -20 °C. to be stored in a freezer until required for use. The resulting material was designated frozen collagen (FWC). The frozen collagen is thawed prior to use in the same manner as fresh collagen.

[0133] Alternatively, frozen collagen can be freeze-dried (lyophilized), and optionally subsequently milled. For this purpose, frozen collagen was manually distributed onto a flat surface, for example a polystyrene mold, the frozen collagen having a layer thickness of between about 5 mm and about 10 mm. The collagen-filled molds were transferred onto the shelves of a commercially available freeze dryer (Christ Epsilon) and frozen to a temperature of about -38 °C with a ramp rate between 0.3 °C and 1.5 °C. After an equilibration period of approximately 30 minutes vacuum was initiated and the shelf temperature was sequentially increased from about -38 °C to about +30 °C at a rate of about 0.5 °C per minute. The combination of vacuum and sequentially increasing the shelf temperature from about -38 °C to about +30 °C facilitated sublimation of the ice from the frozen collagen up until the collagen reached a temperature of 0 °C. To ensure that the temperature of the collagen increased uniformly, at least one equilibrating step was conducted, in which the shelf temperature was maintained at a constant desired temperature for approximately 30 mins, or until the collagen reached the desired temperature. For example, an equilibrating step was conducted every 10 °C between the temperatures of -20 °C and +30 °C to ensure that the temperature of the collagen increased uniformly. The equilibrating step, for example at -20 °C comprised maintaining the shelf temperature at a constant temperature of -20 °C for about 30 mins. Once the ice had been removed by sublimation, and the collagen reached a temperature of 0 °C, the residual water content was further reduced by continuing to sequentially increase the shelf temperature to about +30 °C at a rate of about 0.5 °C per minute. The lyophilized collagen was then milled using a commercially available cutting mill (Rotoplex, Hosokawa Alpine). The resulting material was designated non-matured lyophilized milled collagen (non-matured LMC).

[0134] Optionally, the non-matured lyophilized milled collagen was matured by storing in polyethylene containers (bags) under ambient conditions of about 2-8 °C at atmospheric pressure for periods of about 1-3 years until required for use. The resulting material was designated old lyophilized milled collagen (old LMC).

[0135] Alternatively, the non-matured lyophilized milled collagen (non-matured LMC) was matured by storing in polyethylene containers (bags) as described herein until required for use, for example stored at 40 °C for 2-6 weeks. The resulting material was designated matured lyophilized milled collagen (matured LMC).

Example 2: Compounding process and equipment

[0136] An aqueous modified collagen dispersion was prepared in a stainless steel vessel using pre-heated (35-42 °C.) purified water, which was adjusted to pH 4.0±0.2. High shear mixing was required to break up the modified collagen mass and expose the collagen fibers to the acidic medium. The high shear mixer (homogenizer) comprised a rotor/stator head that is designed to create high shear forces by pulling the modified collagen through the rotating homogenizer head and forcing the modified collagen against the proximal stationary stator head. It is this design that provided the high shear forces required to separate the fibrous collagen mass at the beginning of the aqueous dispersion preparation. However, other comparable mixing equipment may also be used; and can be selected by one skilled in the art. For example, an IKA Ultra-Turrax mixer may be used at a high speed for about 2 to about 5 minutes.

[0137] If required, although not essential, the resulting aqueous dispersion can be filtered and degassed, for example by using 250 micron filters and a suitable means of degassing, for example ultrasonication.

[0138] The collagen concentration in the final aqueous dispersion can be in the range of 0.4% to 1.5% and the pH can be in the range of 4.0±0.2. The final aqueous dispersion can be subsequently transferred to a closed jacketed stainless steel vessel, optionally where the jacket temperature is maintained at 37 °C and the aqueous dispersion is slowly agitated using a low shear setting.

[0139] The dispersion was filled into, for example 10x10 cm, blister trays or lyophilization molds using, for example, a positive displacement pump. The pump can be a valveless pump, optionally having ceramic pistons. Alternatively, a peristaltic pump could also be used. The fill weight was adjusted based on the collagen content of the aqueous dispersion to achieve the target collagen content per area, for example about 0.1 to about 10.0 mg/cm², optionally about 4 mg/cm². Upon completion of the filling process, the filled blisters or molds were placed into a convective drying cabinet. A commercially available drying cabinet (LabAir; Bleymehl) at 31 °C was utilized for this drying process. The drying step can typically require between 1 and 3 days to remove the excess water, which results in the finished composition, for example membrane, being retained in the blisters or molds. [0140] Following completion of the drying process, the blisters or molds were removed from the drying cabinet. The resulting composition, for example membrane, was cut to the desired size, for example using a pneumatic dye. The packaging process was a two-step process comprising introduction to an inner and outer pouch packaging (ethylene oxide; EO type; PMS MEDICAL LTD) followed by pneumatic heat sealing. One side of the outer pouch comprised a transparent polyester or low-density polyethylene (LDPE) foil laminate with a high-density polyethylene (HDPE) strip seal. The other side was an opaque polyester or LDPE laminate. Other outer pouch packaging material can be used, including aluminum oxide coated polyethylene materials or, if E-beam radiation is used for sterilization, an aluminum outer pouch can be used. The pneumatic heat sealer facilitated the formation of a continuous seal at the open end of the pouch. The top part of the pouch included two holes or strips lined with a high-density polyethylene (HDPE) strip seal. These openings/windows were specifically designed for the EO gas sterilization process and were gas permeable only. The permeability of the window facilitated permeation of theEO gas during the terminal EO sterilization process. Following sterilization and ventilation, the outer pouch was resealed below the gas permeable openings/windows, and this gas permeable (top) portion was then removed from the pouch. This resulted in a fully sealed outer pouch containing a terminally sterilized finished composition, for example membrane.

[0141] Ethylene Oxide (EO; C2H4O) is a gas that, at appropriate operating temperatures, sterilizes via the action as a powerful alkylating agent. Under the correct conditions, cellular constituents of organisms such as nucleic acid complexes, functional proteins, and enzymes will react with ethylene oxide, causing the addition of alkyl groups. As a result of the alkylation, cell reproduction is prevented and cell death ensues. The sterilizer used in the present Examples was a DMB 15009 VD (DMB Apparatebau GmbH, Germany). A mixture of EO/CXhat a ratio of 15:85 was used as the sterilization gas over a period of 6 hours at 4 bar pressure. For successful completion of this process, the product needs to contain a moisture level of not less than 9%, which can be achieved by holding it in an area under controlled environmental conditions. Following the EO sterilization process, the product was ventilated for a minimum of 3 to 4 weeks to reduce the level of remaining ethylene oxide gas and any residues from the composition, for example membrane, and packaging materials.

Example 3: Characterization [0142] All compositions (membranes) were prepared from a 0.6% dispersion using the method described herein above. All tests on the collagen dispersion were conducted within 1 day after compounding; and all characterization experiments with the membranes were performed within 1 month after membrane manufacture using unsterilized membranes.

[0143] Dispersion Viscosity

[0144] The viscosity values of 0.9% collagen dispersions prepared from each of the fresh collagen, frozen collagen, non-matured lyophilized milled collagen, and matured lyophilized milled collagen according to Example 2 were measured using a Brookfield viscometer (Digital Rheometer DV-III+ with associated TC-501 Circulating Bath). The viscosity values were measured at a constant shear rate (15 s^-1) and over a temperature range from 25 to 40 °C. at 5 °C. increments. 60 measurements per temperature were averaged to obtain reliable results.

[0145] The dispersion viscosity depends on the temperature and decreases when heating up the dispersion. The viscosity profiles of fresh and frozen collagen are comparable over the temperature range tested. The lyophilized milled collagen, which was stored at a temperature of 2-8 °C. for 3 years before compounding (old LMC), showed significantly lower viscosity at all investigated temperatures compared to the fresh collagen and the frozen collagen. Lyophilized milled collagen, which was matured (stored at a temperature of 40 °C. before compounding; matured LMC), showed lower viscosity compared to non-matured LMC and comparable with old lyophilized milled collagen.

[0146] Maturing the lyophilized milled collagen as described herein resulted in improved viscosity at all investigated temperatures compared to non-matured lyophilized milled collagen, which is not subjected to the maturing step described herein.

[0147] Without wishing to be bound by theory, the difference in viscosity is an advantage for processing of the membranes. Collagen with lower viscosity can be more easily degassed, and filled or casted; and the drying time is also reduced as collagens having higher concentrations can be processed. The modified collagen disclosed herein provides improved viscosity characteristics compared to fresh collagen and frozen collagen; and the maturing step provides comparable viscosity characteristics compared to lyophilized milled collagen, which was stored at a temperature of 2-8 °C for 3 years before compounding (old LMC), thereby providing the improved viscosity characteristics of aged collagen (old LMC) without the extended ageing period. [0148] Water Uptake and Swelling

[0149] Three rectangular samples (1.5x4 cm in size) were cut from 5 membranes prepared from each of the fresh collagen, frozen collagen, old lyophilized milled collagen, and matured lyophilized milled collagen. Each of these samples was soaked in WFI (water for injection) for 10 minutes, and analyzed regarding water uptake (wet weight — dry weight) and swelling (wet thickness — dry thickness). The sample thickness was measured using a Mitutoyo Micrometer IP54.

[0150] Membranes prepared from lyophilized milled collagen, which was stored at a temperature of 2-8 °C. for 3 years before compounding (old LMC), showed lower water uptake and swelling than membranes prepared from fresh collagen and from frozen collagen. The variability of results was substantially lower for membranes prepared from lyophilized milled collagen, which was stored at a temperature of 2-8 °C for 3 years before compounding (old LMC) than for the membranes prepared from fresh collagen and from frozen collagen.

[0151] The thickness change for each collagen membrane tested demonstrated that the improved water uptake and swelling characteristics of membranes prepared from matured lyophilized milled collagen over membranes prepared from lyophilized milled collagen, which was stored at a temperature of 2-8 °C for 3 years before compounding (old LMC), are comparable to the improved water uptake and swelling characteristics of membranes prepared from matured lyophilized milled collagen over membranes prepared from frozen collagen.

[0152] The reduced swelling characteristics of membranes prepared from matured lyophilized milled collagen may be advantageous as the membranes may be implanted into restricted anatomical spaces with a lower risk of pressurizing and potentially damaging vital organs. Thus, for use in treating or preventing surgical adhesions, membranes prepared from the modified collagen may be used in a greater variety of anatomical geometries and surgical procedures.

[0153] Degradation with Collagenase

[0154] Degradation studies were conducted using 4 to 5 membranes per batch of each of the fresh collagen, frozen collagen, old lyophilized milled collagen, and matured lyophilized milled collagen. One membrane (4.5x4.5 cm in size) was placed into a beaker and covered with 15 mL of 0.2 N Phosphate buffer (pH 7.4 with CaCb). Collagenase (Collagenase Type IA-S, sterile, 50 mg, SIGMA, REP C5894) was reconstituted with 5 mL of WFI, and 0.5 mL of the resulting solution was added to the mixture. The solution in the beaker was agitated using a shaking water bath (Julabo SW 22) at 37 °C (120 rpm) for 60 minutes. The degradation was documented by taking photographs of the samples every 5 minutes. Membranes prepared from lyophilized milled collagen degraded the fastest with no residue, while membranes prepared from fresh collagen and frozen collagen degraded considerably slower, and left behind small fiber agglomerates.

[0155] In a further study, 3.1 *3.1 cm membrane samples were immersed in 15 mL of the buffer described above; to which 100 pL of reconstituted collagenase solution was added. 1 mL samples were removed after 5, 10, 15, 25, 40, 60, and 90 minutes; samples were filtered through a 0.45 pm syringe filter, and a 100 pL aliquot was diluted 1 :30. UV absorption spectra between 210 and 230 nm (2 nm increments) were measured against a blank solution using a UV- VIS Photometer Specord 205 (Analytic Jena). The degraded fraction at each time point was calculated from the maximum absorption relative to the 90- minute time point (defined as 100%).

[0156] A composition for use in treating or preventing surgical adhesions, for example a membrane for use as an adhesion barrier, needs to stay intact for a certain time in order to effectively inhibit adhesion. Prolonged presence of the membrane could lead to increased risk of infections, given that collagen is known to be a medium for bacterial growth. These in vitro experiments demonstrate that the membranes prepared from matured lyophilized milled collagen degrade faster than membranes prepared from old lyophilized milled collagen, and yet faster than membranes prepared from fresh collagen and frozen collagen, suggesting that this effect will also be true for the in vivo behavior. Accordingly, a composition comprising a modified collagen in accordance with any of the embodiments of the present disclosure, for use in treating surgical adhesions, can reduce the probability of infections as an adverse effect of the use of the adhesion barrier.

[0157] Example 5: Storage

[0158] Non-matured lyophilized milled collagen (non-matured LMC) was prepared as described in Example 1; and matured by storing in polyethylene containers (bags) as described herein for up to 4 weeks. The resulting material was designated matured lyophilized milled collagen (matured LMC).

[0159] The viscosity values were measured at each of the time periods noted (1, 2, 3, and 4 weeks storage) as described in Example 3. In short, the viscosity values were measured using a Brookfield viscometer (Digital Rheometer DV-III+ with associated TC-501 Circulating Bath) at a constant shear rate (15 s-1) and over a temperature range from 30 to 65°C. The viscosity values of matured lyophilized milled collagen having a low moisture content of 1-2% and a high moisture content of 13-15% were measured.

[0160] Generally, the viscosity of the matured lyophilized milled collagen is unaffected by the moisture content of the matured lyophilized milled collagen. Moreover, increasing the storage temperature accelerates the viscosity reduction of the matured lyophilized milled collagen. Certainly, maturing the lyophilized milled collagen as described herein results in improved viscosity at all investigated storage times. At lower storage temperature, the time required to reach the target viscosity is extended.

[0161] Taken together, the examples provided herein demonstrate that a composition comprising a modified collagen according to a first aspect of the present invention, or a modified collagen prepared according to a second aspect of the present invention — for example, membranes prepared from matured lyophilized milled collagen — exhibit significantly altered properties compared to membranes made from fresh collagen, frozen collagen, or non-matured lyophilized milled collagen. The maturing step providing the altered properties of aged collagen without the extended ageing period; and so can be particularly useful in the manufacture of compositions that require a non-crosslinked gel-like collagen matrix such as, for example, resorbable scaffolds for repairing and/or reinforcing dura mater.

Further Considerations

[0162] In some embodiments, any of the clauses herein may depend from any one of the independent clauses or any one of the dependent clauses. In one aspect, any of the clauses (e.g., dependent or independent clauses) may be combined with any other one or more clauses (e.g., dependent or independent clauses). In one aspect, a claim may include some or all of the words (e.g., steps, operations, means or components) recited in a clause, a sentence, a phrase or a paragraph. In one aspect, a claim may include some or all of the words recited in one or more clauses, sentences, phrases or paragraphs. In one aspect, some of the words in each of the clauses, sentences, phrases or paragraphs may be removed. In one aspect, additional words or elements may be added to a clause, a sentence, a phrase or a paragraph. In one aspect, the subject technology may be implemented without utilizing some of the components, elements, functions or operations described herein. In one aspect, the subject technology may be implemented utilizing additional components, elements, functions or operations.

[0163] The foregoing description is provided to enable a person skilled in the art to practice the various configurations described herein. While the subject technology has been particularly described with reference to the various figures and configurations, it should be understood that these are for illustration purposes only and should not be taken as limiting the scope of the subject technology.

[0164] There may be many other ways to implement the subject technology. Various functions and elements described herein may be partitioned differently from those shown without departing from the scope of the subject technology. Various modifications to these configurations will be readily apparent to those skilled in the art, and generic principles defined herein may be applied to other configurations. Thus, many changes and modifications may be made to the subject technology, by one having ordinary skill in the art, without departing from the scope of the subject technology.

[0165] It is understood that the specific order or hierarchy of steps in the processes disclosed is an illustration of exemplary approaches. Based upon design preferences, it is understood that the specific order or hierarchy of steps in the processes may be rearranged. Some of the steps may be performed simultaneously. The accompanying method claims present elements of the various steps in a sample order, and are not meant to be limited to the specific order or hierarchy presented.

Claims

32 CLAIMS

1. A resorbable scaffold comprising a non-crosslinked collagen matrix.

2. The resorbable scaffold of claim 1 , further comprising at least one drug substance.

3. The resorbable scaffold of claim 2, wherein the drug substance is selected from dexamethasone and progesterone, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof.

4. The resorbable scaffold of claim 1, for use in dural repair surgery.

5. The resorbable scaffold of any one of claims 1-4, for use in a method of repairing dura mater, comprising surgically implanting the resorbable scaffold at a location where dura is damaged.

6. The resorbable scaffold of any one of claims 2-4, wherein the drug substance is substantially homogeneously dispersed in the non-crosslinked collagen matrix.

7. The resorbable scaffold of claim 6, wherein the non-crosslinked collagen matrix comprises Type I collagen and/or Type III collagen.

8. The resorbable scaffold of any one of claims 1-7, wherein the non-crosslinked collagen matrix of the resorbable scaffold comprises a collagen gel containing from about 3.6 to about 8.0 mg/cm³ type I collagen.

9. The resorbable scaffold of any one of claims 1-8, wherein the non-crosslinked collagen matrix comprises a composition including matured collagen. 33

10. The resorbable scaffold of claim 9, wherein the matured collagen comprises collagen stored at a temperature of at least about 40 °C for a period of at least one week or at a temperature of about 2-8 °C for about 1-3 years.

11. The resorbable scaffold of claim 9, wherein the matured collagen comprises the collagen stored at a temperature of about 40 °C to about 65 °C for a period of at least one week.

12. The resorbable scaffold of claim 9, wherein the matured collagen comprises the collagen stored at a temperature of at least 65 °C for a period of at least one week.

13. The resorbable scaffold of claim 9, wherein the matured collagen comprises the collagen stored at a temperature of about 2-8 °C for about 1-3 years.

14. The resorbable scaffold of claim 9, wherein a 0.9% dispersion of the composition has a viscosity of less than 500 cP at a temperature of 40 °C, wherein viscosity is measured using a Brookfield Digital Rheometer DV-III at a constant shear rate of 15 s^-1 at a temperature of 40 °C.

15. The resorbable scaffold of claim 9, having a water uptake of about 60 mg of water when tested for water uptake by weighing a 1.5 cm x 4 cm sample of the composition in a dry state to obtain a dry weight, inserting the sample in water for injection for about 10 minutes, weighing the sample after 10 minutes of insertion in water to obtain a wet weight, and wherein the difference between the wet weight and dry weight is the amount of water absorbed.

16. The resorbable scaffold of claim 9, swelling to a thickness of less than 60 pm when tested for swelling by measuring the thickness of a 1.5 cm x 4 cm sample of the composition in dry state, inserting the sample in water for injection for about 10 minutes, measuring the thickness of the sample after 10 minutes of insertion in water to obtain thickness in a wet state, and wherein swelling is the difference between the thickness in the wet state and dry state.

17. The resorbable scaffold of claim 9, degrading in about 25 minutes when tested for degradation by a method comprising: (a) placing a 4.5 cm x 4.5 cm sample of the composition in 15 mL of 0.2 N phosphate buffer (pH 7.4 with CaCL); (b) adding 0.5 mb of 10 mg/mL Collagenase Type IA-S to the solution in step (a); (c) agitating the solution of step (b) using a shaking water bath at 37 °C at 120 rpm for 60 minutes; and (d) measuring the degradation of the sample every 5 minutes throughout step (c).

18. The resorbable scaffold of claim 9, wherein about 50% of the composition is degraded in about 15 to 20 minutes upon when tested for degradation by a method comprising: (a) placing a 3.1 cm x 3.1 cm sample of the composition in 15 mL of 0.2 N phosphate buffer (pH 7.4 with CaCb); (b) adding 100 pL of 10 mg/mL Collagenase Type IA-S to the solution in step (a); and (c) measuring the degradation of the sample after 5, 10, 15, 25, 40, 60, and 90 minutes.

19. The resorbable scaffold of claim 9, wherein the composition comprising matured collagen is prepared by a method comprising the steps of: (a) providing isolated collagen; (b) freezing the isolated collagen; (c) dehydrating the frozen collagen; and (d) maturing the dehydrated collagen, wherein the maturing step comprises storing the dehydrated collagen at a temperature of at least 40 °C for at least one week or at a temperature of about 2-8 °C for about 1-3 years.

20. The resorbable scaffold of claim 19, wherein step (a) comprises providing an isolated collagen dispersion having a concentration of about 3-30% (w/w) collagen particles.

21. The resorbable scaffold of claim 19, wherein step (d) comprises storing the dehydrated collagen at a temperature of about 40 °C to about 65 °C for a period of at least one week.

22. The resorbable scaffold of claim 19, wherein the maturing step comprises storing the dehydrated collagen at a temperature of at least 65 °C for a period of at least one week.

23. The resorbable scaffold of claim 19, wherein the matured collagen comprises the collagen stored at a temperature of about 2-8 °C for about 1-3 years.

24. The resorbable scaffold of claim 19, wherein the method further comprises: (e) preparing an aqueous dispersion of the matured collagen of step (d); (f) degrading the aqueous dispersion; and (g) dehydrating the aqueous dispersion.

25. The resorbable scaffold of claim 19, wherein the dehydrated collagen of step (c) is milled before step (d).

26. The resorbable scaffold of claim 19, wherein step (d) comprises storing the dehydrated collagen at a temperature of at least 40 °C for a period of at least four weeks; or comprises storing the dehydrated collagen at a temperature of at least 65 °C for a period of at least one week.

27. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix has an average pore size in a range from 5 pm to 50 pm.

28. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix has adhesive edges enabling adhesion to dura mater of a subject.

29. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix is formed into a film having a thickness in a range from about 1 pm to about 100 pm.

30. The resorbable scaffold of claim 1, wherein the scaffold is constituted in the form of a suturable graft.

31. The resorbable scaffold of claim 1, wherein the scaffold is constituted in the form of an adhesive graft.

32. The resorbable scaffold of claim 1, further comprising one or more active ingredients.

33. The resorbable scaffold of claim 32, wherein the active ingredient is a cytokine. 36

34. The resorbable scaffold of claim 33, wherein the cytokine is selected from the group consisting of pro-angiogenic cytokines.

35. The resorbable scaffold of claim 32, wherein the active ingredient is selected from an anti-fungal agent, an antibacterial agent and an anti-inflammatory agent.

36. The resorbable scaffold of claim 32, wherein the one or more active ingredients comprises an fibroblast growth factor (FGF) receptor agonist.

37. The resorbable scaffold of claim 1, further comprising one or more peptides.

38. The resorbable scaffold of claim 37, wherein the one or more peptide is fibroblast growth factor (FGF), or an analogue thereof.

39. The resorbable scaffold of claim 1, wherein the scaffold has a size in a range from about 5 cm² to about 250 cm².

40. The resorbable scaffold of claim 1, wherein the scaffold is resorbed over a period ranging from about 6 weeks to about 1 year.

41. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix is impermeable to water.

42. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix has an elastic modulus in a range from about 30 MPa to about 120 MPa.

43. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix has a tensile strength in a range from about 2 MPa to about 12 MPa.

44. The resorbable scaffold of claim 1, wherein the non-crosslinked collagen matrix has maximum strain in a range from about 6% to about 15%. 37

45. The resorbable scaffold of claim 1, wherein the scaffold has a water content in a range from about 60 wt% to about 85 wt%.

46. The resorbable scaffold of claim 1, further comprising a polyethylene glycol (PEG).

47. The resorbable scaffold of claim 46, wherein content of PEG is in a range from about 4 wt% to about 6 wt%.