# US 9,518,106 B2 · Collagen fibrillar construction > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,518,106 B2 - Document ID: US9518106B2 - Status: Granted - Kind: B2 - Application number: 14/664,137 - Filed: 2015-03-20 - Published: 2015-10-22 - Granted: 2016-12-13 - Earliest claimed: 2008-04-11 - Assignees: Northeastern University - Applicants: Northeastern University - Inventors: Nima Saeidi; Jeffrey W. Ruberti - Web page: https://patentgrep.com/patent/US9518106B2 ## CPC - C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] - C08H 1/06: derived from horn, hoofs, hair, skin or leather - C08L 89/06: derived from leather or skin {, e.g. gelatin} - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C08H: DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS (polysaccharides C08B; natural rubber C08C; natural resins or their derivatives C09F; bituminous materials C10) - C08L: COMPOSITIONS OF MACROMOLECULAR COMPOUNDS (compositions based on polymerisable monomers C08F, C08G; artificial filaments or fibres D01F; textile treating compositions D06) ## Abstract Methods and compositions are described for organizing collagen into fibrillar networks, e.g, short and long-range organization. Collagen produced by the disclosed methods can be used for tissue engineering. ## Claims ### Claim 1 A method of producing an organized array of collagen fibrils, the method comprising: applying a tension to a plurality of collagen fibrils in a solution, wherein the tension is controlled to generate a strain of about 1% to about 10% such that a controlled load on collagen monomers in the solution is produced; and adding a collagen lytic protease to the solution, thereby producing an organized array of collagen fibrils. ### Claim 2 (depends on 1) The method of claim 1, further comprising neutralizing the pH of the solution. ### Claim 3 (depends on 2) The method of claim 2, further comprising neutralizing the pH of the solution at about 10° C. to about 39° C. ### Claim 4 (depends on 1) The method of claim 1, wherein the tension is applied to both ends of the plurality of collagen fibrils. ### Claim 5 (depends on 4) The method of claim 4, further comprising adding supplemental collagen monomers to the solution continuously when applying the tension. ### Claim 6 (depends on 5) The method of claim 5, wherein the collagen lytic protease and the supplemental collagen monomers are added simultaneously to the solution. ### Claim 7 (depends on 5) The method of claim 5, wherein the collagen lytic protease and the supplemental collagen monomers are added sequentially to the solution. ### Claim 8 (depends on 5) The method of claim 5, wherein the collagen lytic protease and the supplemental collagen are added more than once to the solution. ### Claim 9 (depends on 1) The method of claim 1, further comprising organizing the array of collagen fibrils into a tissue. ### Claim 10 (depends on 5) The method of claim 5, further comprising continuously extending a collagen fibrillar structure at a rate of from about 0.1 μm/min to about 100 μm/min. ### Claim 11 (depends on 10) The method of claim 10, further comprising adding collagen lytic protease to the solution while extending the collagen fibrillar structure. ### Claim 12 (depends on 10, 11) The method of claim 10 or 11, further comprising extending the collagen fibrillar structure in the presence of a co-nonsolvency agent. ### Claim 13 (depends on 12) The method of claim 12, wherein the co-nonsolvency agent is polyethylene glycol, hyaluronic acid, a glycosaminoglycan, a proteoglycan, or a combination thereof. ### Claim 14 (depends on 13) The method of claim 13, wherein the proteoglycan is lumican, decorin, biglycan, perlecan, versican, fibromodulin, aggrecan, sydecan or a combination thereof. ### Claim 15 (depends on 1, 11) The method of claim 1 or claim 11, wherein the collagen lytic protease is a bacterial collagenase, a matrix metalloproteinase, or cathepsin. ### Claim 16 (depends on 1) The method of claim 1, wherein the solution further comprises a collagen binding agent. ### Claim 17 (depends on 16) The method of claim 16, wherein the collagen binding agent is a proteoglycan, a glycoprotein, a collagen-binding portion thereof, or a combination thereof. ### Claim 18 (depends on 1) The method of claim 1, wherein the controlled load on each collagen monomer is from 1.0 piconewtons to 10.0 piconewtons. ### Claim 19 (depends on 1) The method of claim 1, wherein the controlled load on each collagen monomer is greater than 10.0 piconewtons. ### Claim 20 (depends on 1) The method of claim 1, further comprising adding a cross-linking agent to the solution. ### Claim 21 (depends on 20) The method of claim 20, wherein the cross-linking agent is added before applying the tension. ### Claim 22 (depends on 20) The method of claim 20, wherein the cross-linking agent is added after applying the tension. ### Claim 23 (depends on 20) The method of claim 20, wherein the cross-linking agent is formaldehyde, hexamethylene diisocyanate, glutaraldehyde, polyepoxy compounds, gamma irradiation, or ultraviolet irradiation with riboflavin.