# US 10,864,300 B2 · Method for preparing bilayer scaffold through single process and method for regenerating tissue using bilayer scaffold obtained by preparing method

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- Number: US 10,864,300 B2
- Document ID: US10864300B2
- Status: Granted
- Kind: B2
- Application number: 14/905,827
- Filed: 2013-07-16
- Published: 2016-10-06
- Granted: 2020-12-15
- Earliest claimed: 2013-07-16
- Assignees: Research Cooperation Foundation of Yeungnam University
- Applicants: Research Cooperation Foundation of Yeungnam University
- Inventors: Sung Soo Han; Soon Mo Choi; Deep Ti Singh
- Web page: https://patentgrep.com/patent/US10864300B2

## CPC

- A61L 27/3687: {characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents}
- A61F 2/105: {Skin implants, e.g. artificial skin}
- A61F 2/28: Bones
- A61F 2/30756: {Cartilage endoprostheses (A61F2/3603 takes precedence)}
- A61L 27/26: Mixtures of macromolecular compounds
- A61L 27/3691: {characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation}
- A61L 27/3813: {Epithelial cells, e.g. keratinocytes, urothelial cells}
- A61L 27/3817: {Cartilage-forming cells, e.g. pre-chondrocytes}
- A61L 27/3821: {Bone-forming cells, e.g. osteoblasts, osteocytes, osteoprogenitor cells}
- A61L 27/56: Porous materials, {e.g. foams or sponges}
- A61L 27/60: Materials for use in artificial skin
- B32B 5/32: {at least two} layers being foamed {and next to each other}
- C08J 3/24: Crosslinking, e.g. vulcanising, of macromolecules (mechanical aspects B29C35/00; crosslinking agents C08K)
- C08J 9/0061: {characterized by the use of several polymeric components}
- C08J 9/28: by elimination of a liquid phase from a macromolecular composition or article, e.g. drying of coagulum
- C08J 9/30: by mixing gases into liquid compositions or plastisols, e.g. frothing with air
- C08J 9/36: After-treatment (C08J9/22 takes precedence)
- A61F 2002/30011: {differing in porosity}
- A61F 2210/0004: bioabsorbable
- A61F 2210/0076: multilayered, e.g. laminated structures
- A61F 2240/001: Designing or manufacturing processes
- A61L 2300/412: Tissue-regenerating or healing or proliferative agents
- A61L 2430/34: for soft tissue reconstruction
- B32B 2250/22: All layers being foamed
- C08J 2201/026: Crosslinking before of after foaming
- C08J 2201/0484: the liquid phase being aqueous
- C08J 2207/10: Medical applications, e.g. biocompatible scaffolds
- C08J 2301/00: Characterised by the use of cellulose, modified cellulose or cellulose derivatives
- C08J 2305/00: Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J 2323/08: Copolymers of ethene (C08J2323/16 takes precedence)
- C08J 2325/06: Polystyrene
- C08J 2329/02: Homopolymers or copolymers of unsaturated alcohols (C08J2329/14 takes precedence)
- C08J 2329/04: Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids
- C08J 2333/02: Homopolymers or copolymers of acids; Metal or ammonium salts thereof
- C08J 2339/06: Homopolymers or copolymers of N-vinyl-pyrrolidones
- C08J 2367/04: Polyesters derived from hydroxy carboxylic acids, e.g. lactones (C08J2367/06 takes precedence)
- C08J 2371/02: Polyalkylene oxides
- C08J 2375/06: from polyesters
- C08J 2379/02: Polyamines
- C08J 2389/00: Characterised by the use of proteins; Derivatives thereof
- C08J 2489/00: Characterised by the use of proteins; Derivatives thereof
- 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)
- A61F: FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS (dental prosthetics A61C)
- B32B: LAYERED PRODUCTS, i.e. PRODUCTS BUILT-UP OF STRATA OF FLAT OR NON-FLAT, e.g. CELLULAR OR HONEYCOMB, FORM
- C08J: WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H (working, e.g. shaping, of plastics B29)

## Abstract

Disclosed is a method for preparing a bilayer scaffold through single process comprising: preparing a first polymer aqueous solution; adding a second polymer into the first polymer aqueous solution and stirring a reactant; adding a surfactant into the stirred reactant and stirring the reactant at high temperature and high speed; freeze-drying the stirred reactant thereby obtaining a sponge; dipping the sponge in a cross-linking agent thereby rendering be cross-linked; and freeze-drying the cross-linked reactant.

## Claims

### Claim 1

A method for preparing a bilayer scaffold through single process, comprising:
preparing a gelatin aqueous solution by stirring gelatin in water at a temperature of 60 to 70° C. so as to dissolve completely;
preparing a polymer blend solution by adding polyvinyl alcohol into the gelatin aqueous solution and stirring to create a polymer blend solution;
adding sodium dodecyl sulfate into the polymer blend solution and stirring at a temperature of 80 to 120° C.;
freeze-drying the stirred solution thereby obtaining a sponge;
dipping the sponge in a cross-linking agent selected from the group consisting of glutaraldehyde or ethyldiethylaminopropylcarbodimide thereby rendering the sponge cross-linked; and
freeze-drying the cross-linked sponge,
wherein the gelatin aqueous solution includes 2.5% by weight of gelatin,
wherein the polymer blend solution includes 10% by weight of polyvinylalcohol.

### Claim 2 (depends on 1)

A method for regenerating tissue by culturing cells into the bilayer scaffold prepared by the method according to claim 1.

### Claim 3 (depends on 2)

The method for regenerating tissue according to claim 2, wherein the type of cell is any one of or two or more cell types selected from the group consisting of cartilage cells, bone cells, and skin cells.

### Claim 4 (depends on 2)

The method for regenerating tissue according to claim 2, comprising culturing a first cell type in a top layer and culturing a second cell type in a bottom layer of the bilayer scaffold.

### Claim 5 (depends on 1)

A composition for tissue regeneration and wound treatment comprising a bilayer scaffold prepared by the method of claim 1.