# US 11,298,443 B2 · Innervated artificial skin > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,298,443 B2 - Document ID: US11298443B2 - Status: Granted - Kind: B2 - Application number: 16/314,169 - Filed: 2017-06-30 - Published: 2020-10-01 - Granted: 2022-04-12 - Earliest claimed: 2016-07-01 - Assignees: Trustees of Tufts College - Applicants: Trustees of Tufts College - Inventors: David L. Kaplan; Sarah Lightfoot Vidal; Rosalyn Abbott; Siwei Zhao; Dana Cairns; Fiorenzo G. Omenetto - Web page: https://patentgrep.com/patent/US11298443B2 ## CPC - A61L 27/60: Materials for use in artificial skin - A61L 27/227: {Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24} - A61L 27/3637: {characterised by the origin of the biological material other than human or animal, e.g. plant extracts, algae} - A61L 27/3641: {characterised by the site of application in the body (materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60)} - A61L 27/3813: {Epithelial cells, e.g. keratinocytes, urothelial cells} - A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells} - A61L 27/3886: {comprising two or more cell types} - A61L 27/3891: {as distinct cell layers} - C12N 5/0698: {Skin equivalents} - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention provides, in some embodiments, multi-layer silk compositions including a first layer comprising silk fibroin and keratinocytes, a second layer comprising silk fibroin and fibroblasts, a third layer comprising silk fibroin and adipocytes, and a plurality of nervous system cells, wherein at least some of the plurality of nervous system cells span at least two layers, and methods of making and using the same. In some embodiments, provided methods and compositions further include immune cells and/or endothelial cells. ## Claims ### Claim 1 A multi-layer silk composition comprising a first layer comprising silk fibroin and keratinocytes; a second layer comprising silk fibroin and fibroblasts; a third layer comprising silk fibroin and adipocytes; and a plurality of nervous system cells, wherein at least some of the plurality of nervous system cells span at least two layers. ### Claim 2 (depends on 1) The composition of claim 1, wherein all of the cells in each of the first, second and third layers are human cells. ### Claim 3 (depends on 2) The composition of claim 2, wherein the human cells are primary cells or cell lines. ### Claim 4 (depends on 1) The composition of claim 1, wherein the nervous systems cells comprise at least one of neurons, glia, and neural stem cells. ### Claim 5 (depends on 1) The composition of claim 1, wherein at least a plurality of the nervous system cells are functional. ### Claim 6 (depends on 1) The composition of claim 1, further comprising a plurality of at least one type of immune cells. ### Claim 7 (depends on 6) The composition of claim 6, wherein the at least one type of immune cells includes macrophages. ### Claim 8 (depends on 1) The composition of claim 1, further comprising a plurality of at least one type of endothelial cell. ### Claim 9 (depends on 8) The composition of claim 8, wherein the endothelial cells comprise a monolayer. ### Claim 10 (depends on 1) The composition of claim 1, further comprising an electrical device that is functionally connected to at least some of the plurality nervous system cells. ### Claim 11 (depends on 10) The composition of claim 10, wherein activation of the electrical device results in the firing of one or more neurons. ### Claim 12 (depends on 10) The composition of claim 10, wherein the electrical device comprises at least one electrode. ### Claim 13 (depends on 10) The composition of claim 10, wherein the electrical device comprises silk fibroin. ### Claim 14 (depends on 1) The composition of claim 1, further comprising at least one of collagen, laminin, fibronectin, hyaluronic acid, fibrinogen, sulfated glycosaminoglycans, and/or one or more growth factors, in at least one layer. ### Claim 15 (depends on 14) The composition of claim 14, wherein the one or more growth factors comprise epidermal growth factor, fibroblast growth factor, nerve growth factor, platelet-derived growth actor, insulin-like growth factor, or tumor necrosis factor-β, and combinations thereof. ### Claim 16 (depends on 1) The composition of claim 1, wherein at least one layer is exposed to an air-liquid interface. ### Claim 17 (depends on 1) The composition of claim 1, wherein the third layer comprises lipoaspirate. ### Claim 18 A method of making a multi-layer silk composition comprising providing a first layer comprising silk fibroin and keratinocytes; providing a second layer comprising silk fibroin and fibroblasts; providing a third layer comprising silk fibroin and adipocytes; providing a plurality of nervous system cells; and associating the first layer, second layer, third layer, and plurality of nervous system cells to form a multi-layer silk composition, wherein at least some of the plurality of nervous system cells span at least two layers. ### Claim 19 A method comprising a) providing a multi-layer silk composition comprising a first layer, a second layer, a third layer, and a plurality of nervous system cells, the first layer comprising silk fibroin and keratinocytes, the second layer comprising silk fibroin and fibroblasts, the third layer comprising silk fibroin and adipocytes, wherein at least some of the plurality of nervous system cells span at least two layers, wherein at least some of the cells in the composition are cancer cells; b) exposing the composition to one or more therapeutic agents and/or inflammatory agents; and c) characterizing the response of the cells in the composition to the one or more therapeutic agents. ### Claim 20 (depends on 19) The method of claim 19, wherein the one or more therapeutics agents comprise at least one cancer therapeutic agent.