# US 10,369,284 B2 · Kit, use thereof and method for filling connective tissue of the skin > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,369,284 B2 - Document ID: US10369284B2 - Status: Granted - Kind: B2 - Application number: 14/417,402 - Filed: 2013-07-15 - Published: 2015-07-23 - Granted: 2019-08-06 - Earliest claimed: 2012-07-31 - Assignees: Meidrix Biomedicals GmbH - Applicants: Amedrix GmbH - Inventors: Thomas Graeve - Web page: https://patentgrep.com/patent/US10369284B2 ## CPC - A61M 5/19: having more than one chamber - A61K 9/0021: {Intradermal administration, e.g. through microneedle arrays or needleless injectors} - A61K 9/0024: {Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue (compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L)} - A61K 38/39: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG] - A61L 27/24: Collagen - A61L 27/60: Materials for use in artificial skin - A61M: DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY (introducing media into or onto the bodies of animals A61D7/00; means for inserting tampons A61F13/26; devices for administering food or medicines orally A61J; containers for collecting, storing or administering blood or medical fluids A61J1/05); DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY (surgery A61B; chemical aspects of surgical articles A61L); DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR {(Electrotherapy, e.g. producing anaesthesia by the use of alternating or intermittent currents A61N1/36021)} - 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) - 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) ## Abstract The invention concerns a kit and a syringe for use in a therapeutic method for intracutaneous filling of the collagen structure in connective tissue of the skin, and the use thereof, particularly as part of the treatment of skin defects, particularly skin diseases and signs of skin aging. ## Claims ### Claim 1 A method for intracutaneous filling of a collagen structure in connective tissue of a patient's skin with a collagen gel, the method comprising the steps: a) separately supplying a collagen solution and a neutralizing buffer solution, and separately tempering the collagen solution and the neutralizing buffer solution by bringing the collagen solution and the neutralizing buffer solution each to a temperature of 20° C. to 37° C., wherein the collagen solution comprises native collagen at a concentration of from 9 mg/ml to 16 mg/ml; b) mixing the tempered collagen solution and the tempered neutralizing buffer solution by merging a stream of the tempered collagen solution and a stream of the tempered neutralizing buffer solution together in a mixing device coupled to a syringe to obtain a liquid gellable collagen composition having a native collagen concentration of from 6 mg/ml to 12 mg/ml and a viscosity of 2 to 9 Pa·s; c) intracutaneously injecting the liquid gellable collagen composition through a needle coupled to the mixing device and into an intracutaneous target site before gelling of the liquid gellable collagen composition, the needle having an outer diameter of 0.3 to 0.4 mm; and d) shaping the injected liquid gellable collagen composition at the intracutaneous target site, wherein the shaped collagen gel cures at the target site. ### Claim 2 (depends on 1) The method according to claim 1, wherein the mixing includes mixing for a maximum period of 5 seconds. ### Claim 3 (depends on 1) The method according to claim 1, further comprising gelling of the prepared collagen composition starting within 10 seconds after the mixing. ### Claim 4 (depends on 1) The method according to claim 1, wherein the intracutaneous injection and the shaping of the collagen composition collectively last at most 4 minutes. ### Claim 5 (depends on 1) The method according to claim 1, wherein the intracutaneous injection and the shaping of the collagen composition collectively last at most 2 minutes. ### Claim 6 (depends on 1) The method according to claim 1, wherein the mixing of the collagen solution and the neutralizing buffer solution is achieved by expulsion of the solutions from chambers of the syringe and merging of solution streams in a mixing device allocated to the syringe, wherein the prepared collagen composition exits the mixing device. ### Claim 7 (depends on 1) The method according to claim 1, wherein the pH of the collagen solution at a temperature of 21° C. is 6 or lower. ### Claim 8 (depends on 1) The method according to claim 1, wherein the collagen solution is produced from collagen-containing tissue by acid extraction without treating with enzyme. ### Claim 9 (depends on 1) The method according to claim 1, wherein at least one of the collagen solution and the liquid gellable collagen composition is cell-free. ### Claim 10 (depends on 1) The method according to claim 1, wherein step a) includes providing at least a first container including the collagen solution and supplying at least a second container including the neutralizing buffer solution. ### Claim 11 (depends on 1) The method according to claim 1, wherein step c) includes instantaneous injection with a syringe having at least two separate containers configured as chambers, the at least two separate containers in fluid connection with a mixing device arranged in the syringe and with the needle of the syringe, wherein at least a first chamber is filled with the collagen solution and at least a second chamber separate therefrom is filled with the neutralizing buffer solution. ### Claim 12 (depends on 1) The method according to claim 1, wherein the shaping of the injected liquid gellable collagen composition at the intracutaneous target site includes molding the injected liquid gellable collagen composition at the intracutaneous target site into a three-dimensional structure with a desired shape before the liquid gellable collagen composition completely cures. ### Claim 13 (depends on 1) The method according to claim 1, wherein the neutralizing buffer solution is a buffer salt solution having a pH of 8.3. ### Claim 14 (depends on 13) The method according to claim 13, wherein the buffer salt solution is HEPES buffered saline. ### Claim 15 (depends on 1) The method according to claim 1, wherein the mixing of the collagen solution and the neutralizing buffer solution comprising mixing the collagen solution and the neutralizing buffer solution at a collagen solution:buffer solution ratio of 1:1 to 9:1, such that the liquid gellable collagen composition obtained from the mixing has a pH of 7.0 to 7.5. ### Claim 16 A method for intracutaneous filling of the collagen structure in connective tissue of a patient's skin with a collagen gel, the method comprising: a) disposing a collagen solution comprising 9 to 16 mg/mL native collagen and having a pH of 6 or lower, and a neutralizing buffer solution comprising a buffer, a salt, and having a pH of 8.3 in separate chambers of a syringe comprising a needle having an outlet channel in fluid communication with the separate chambers by way of a mixing device, and coupled syringe plungers for emptying the separate chambers; b) tempering the syringe such that the collagen solution and the neutralizing buffer solution are heated to a temperature of from 20° C. to 37° C. to obtain a tempered collagen solution and a tempered neutralizing buffer solution; c) piercing the patient's skin with the needle and positioning the outlet channel at a target site; d) depressing the coupled syringe plungers, wherein the depressing causes: mixing of the tempered collagen solution with the tempered neutralizing buffer solution in the mixing device by expelling the tempered collagen solution and the tempered neutralizing buffer solution from their respective chambers and merging the tempered collagen solution and the tempered neutralizing buffer solution into the mixing device to obtain a liquid gellable collagen composition comprising 6 to 12 mg/mL native collagen, a pH of 7.0 to 7.5, and a viscosity of 2 to 9 Pa·s, and injecting the liquid gellable collagen composition intracutaneously at the target site of the patient's skin through the outlet channel of the needle, wherein the needle has an outer diameter of 0.3 to 0.4 mm; and e) molding the liquid gellable collagen composition within the patient's skin into a three-dimensional structure with a desired shape, wherein the liquid gellable collagen composition cures in situ within the patient's skin and retains the desired shape. ### Claim 17 (depends on 16) The method according to claim 16, wherein the needle has an outer diameter of 0.32 to 0.38 mm. ### Claim 18 (depends on 16) The method according to claim 16, wherein: the neutralizing buffer solution is 5× concentrated, the syringe comprises two separate chambers with a chamber volume ratio of 1:4 for receiving the 5× neutralizing buffer solution and the collagen solution, respectively, the tempering comprises bringing the collagen solution and the neutralizing buffer solution each to a temperature of about 30° C., and the liquid gellable collagen composition has a collagen content of about 8 mg/m L.