# US 11,439,663 B2 · Methods of generating cross-linked protein foams in situ > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,439,663 B2 - Document ID: US11439663B2 - Status: Granted - Kind: B2 - Application number: 16/706,300 - Filed: 2019-12-06 - Published: 2020-07-16 - Granted: 2022-09-13 - Earliest claimed: 2015-04-03 - Assignees: Biochange Ltd. - Applicants: Biochange Ltd. - Inventors: Ishay Attar - Web page: https://patentgrep.com/patent/US11439663B2 ## CPC - A61K 35/12: Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells (vaccines or medicinal preparations containing antigens or antibodies A61K39/00) - A61B 17/064: Surgical staples, {i.e. penetrating the tissue} - A61K 9/122: {Foams; Dry foams (edible foams A61K9/0056)} - A61K 47/42: Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein (oligopeptides having up to five amino acids {A61K47/183}; polyamino acids A61K47/34) - A61L 27/3604: {characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel} - A61L 27/3843: {Connective tissue} - A61P 1/12: Antidiarrhoeals - A61P 11/00: Drugs for disorders of the respiratory system - A61P 13/10: of the bladder - A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like - A61P 19/00: Drugs for skeletal disorders - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D) - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract In one embodiment, the present invention provides a composition, wherein the composition is a porous scaffold, wherein the pores of the scaffold are from 2 to 500 microns, the composition comprising: a) a cross-linkable protein selected from the group consisting of collagen and gelatin; b) a cross-linker which induces cross-linking of the cross-linkable protein; and c) a liquid. ## Claims ### Claim 1 A method, comprising: obtaining a composition, the composition comprising: a hygroscopic cross-linkable gelatin, wherein the hygroscopic cross-linkable gelatin is present in the composition in a range of 0.5 wt % to 25 wt % based on a total weight of the composition; at least one cross-linker, wherein the at least one cross-linker is present in the composition in a range of 0.0001 wt % to 25 wt % based on a total weight of the composition; a liquid; introducing the hygroscopic cross-linkable gelatin into the composition as a hygroscopic cross-linkable gelatin powder having a particle size between 5 to 200 microns, introducing the composition in a patient at a site where the patient is in need of treatment of a tissue defect, crosslinking the composition at the site where the patient is in need of treatment of the tissue defect, so as to form a porous scaffold in situ, wherein the porous scaffold is a closed cell foam, wherein the closed cell foam has a pore size of from 2 microns in diameter to 500 microns in diameter. ### Claim 2 (depends on 1) The method of claim 1, further comprising milling the hygroscopic cross-linkable gelatin with at least one milling apparatus so as to form the hygroscopic cross-linkable gelatin powder. ### Claim 3 (depends on 1) The method of claim 1, wherein the liquid is a physiological buffer. ### Claim 4 (depends on 1) The method of claim 1, wherein the at least one cross-linker comprises at least one transglutaminase enzyme. ### Claim 5 (depends on 4) The method of claim 4, wherein the at least one transglutaminase enzyme comprises at least one of: microbial transglutaminase (mTG), tissue transglutaminase (tTG), keratinocyte transglutaminase, epidermal transglutaminase prostate transglutaminase, neuronal transglutaminase, human transglutaminase, Factor XII, or any combination thereof. ### Claim 6 (depends on 5) The method of claim 5, further comprising stabilizing the at least one transglutaminase enzyme with at least one stabilizer, wherein the at least one stabilizer comprises at least one of maltodextrin, mannitol, sucrose, trehalose, or any combination thereof. ### Claim 7 (depends on 2) The method of claim 2, wherein the hygroscopic cross-linkable-gelatin powder solubilizes at a temperature range of 5° C.-37° C. ### Claim 8 (depends on 2) The method of claim 2, wherein the hygroscopic cross-linkable gelatin powder dissolves in liquid in 0.01 second-60 seconds. ### Claim 9 A method, comprising: obtaining a composition, the composition comprising: a hygroscopic cross-linkable gelatin, wherein the hygroscopic cross-linkable gelatin is present in the composition in a range of 0.5 wt % to 25 wt % based on a total weight of the composition; at least one cross-linker, wherein the at least one cross-linker is present in the composition in a range of 0.0001 wt % to 25 wt % based on a total weight of the composition; a liquid; introducing the hygroscopic cross-linkable gelatin into the composition as a hygroscopic cross-linkable gelatin powder having a particle size between 5 to 200 microns, introducing the composition to cells in vitro, wherein the composition is cross-linked so as to form a porous scaffold, wherein the porous scaffold is a foam, wherein the foam has a pore size of from 2 microns in diameter to 500 microns in diameter. ### Claim 10 (depends on 9) The method of claim 9, wherein the introducing comprises mixing the composition with cells.