Skin substitutes, preparation methods and uses thereof

• US 8,097,274 B2
• Filed: 10/27/2006
• Issued: 01/17/2012
• Est. Priority Date: 10/27/2006
• Status: Active Grant

• Current Assignee: National Defense Medical Center (Government of The Republic of China)
• Original Assignee: National Defense Medical Center (Government of The Republic of China)
• Inventors: Adams, Eric Frank, Dai, Niann-Tzyy, Yeh, Ming-Kung, Coombes, Allan Gerald Arthur, Liu, Tsung-Hsun
• Primary Examiner(s): Arnold, Ernst
• Assistant Examiner(s): Levin, Miriam A
• Application Number:
  US12/447,434
• Publication Number:
  US 20100062041A1
• Time in Patent Office: 1,908 Days
• Field of Search: 424/443, 424/93.7, 514/772.7, 435/396, 435/366
• US Class Current: 424/443
• CPC Class Codes: 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
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3813 Epithelial cells, e.g. keratinocytes, urothelial cells
• CPC Class Codes: A61L 27/386 Ligaments, tendons
• CPC Class Codes: A61L 27/3865 Dental/periodontal tissues
• CPC Class Codes: A61L 27/3878 Nerve tissue, brain, spinal cord, nerves, dura mater
• CPC Class Codes: A61L 27/3891 as distinct cell layers
• CPC Class Codes: A61L 27/48 with macromolecular fillers
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence
• CPC Class Codes: C12N 2502/094 keratinocytes
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691
• CPC Class Codes: C12N 5/0698 Skin equivalents
• CPC Class Codes: Y10S 602/90 Method of making bandage structure

Abstract

Provided herein are skin substitutes suitable for use in a living subject for purpose of repairing damaged tissues, methods of producing the skin substitutes and their uses. A biocomposite membrane comprising poly(ε-caprolactone) (PCL) and at least one material selected from collagen and gelatin is provided. In one embodiment, the biocomposite is a 2-component membrane of PCL and gelatin. In another embodiment, the biocomposite is a 3-component membrane of PCL, collagen and gelatin. The bio-composite membrane may be used directly in vivo as a wound dressing, or as a support for cell growth on each side of the membrane to produce an in vitro cultivated artificial skin for future in vivo and/or in vitro applications.

Claims

1. A porous biocomposite membrane for promoting at least one of wound healing and tissue regeneration, comprising poly(E-caprolactone) (PCL) and gelatin in a weight ratio between 4:

1 to 20;

1, with the proviso that collagen is not present in the biocomposite membrane, wherein the biocomposite membrane is obtained by a method comprising;

providing a freeze-dried gelatin mat; and

pouring a poly (e-caprolactone) solution onto the freeze-dried gelatin mat and subsequently evaporating the solvent therein to form the porous biocomposite membrane.

2. An artificial skin for grafting onto a subject in need of at least one of skin treatment and wound coverage, comprising:

the biocomposite membrane of claim 1;

a layer of keratinocytes on one side of the biocomposite membrane; and

a layer of fibroblasts on the other side of the biocomposite membrane.

3. The artificial skin of claim 2, wherein the cells are of mammalian origin, and the cells being normal, genetically modified or malignant.

4. The artificial skin of claim 3, wherein the mammalian origin is human.

5. The artificial skin of claim 2, wherein the subject in need of at least one of skin treatment and wound coverage is the one with ear drum defect, disruption of tendon/nerve and/or skin damage that includes burn injuries, ulcers, impaired pigmentation, excisions or other dermatological conditions.

6. A method for cultivating an artificial skin for grafting onto a subject in need of at least one of skin treatment and wound coverage, comprising:

growing a layer of keratinocytes on one side of the biocomposite membrane of claim 2; and

growing a layer of fibroblasts on the other side of the biocomposite membrane of claim 2.

7. The method of claim 6, wherein the step of growing a layer of keratinocytes is performed before or after the step of growing a layer of fibroblasts.

8. The method of claim 6, wherein the cells are of mammalian origin.

9. The method of claim 8, wherein the cells are from human.

10. The method of claim 6, wherein the subject in need of at least one of skin treatment and wound coverage is the one with ear drum defect, disruption of tendon/nerve and/or skin damage that includes burn injuries, ulcers, impaired pigmentation, excisions or other dermatological conditions.

11. An artificial skin prepared in accordance with the method of claim 6.

12. A biocomposite product, comprising:

the biocomposite membrane of claim 1; and

at least one layer of cells grown on at least one side of the biocomposite membrane;

wherein the cells are selected from the group consisting of fibroblasts, keratinocytes, melanocytes, cells originated from hair follicle and/or sweat gland, endothelial cells originated from the blood, blood cells, chondrocytes, osteocytes, osteoblasts and stem cells originated from the cord blood and/or bone marrow, and the cells being normal, genetically modified or malignant.

13. The biocomposite product of claim 12, wherein the biocomposite membrane comprises two layers of cells, with each layer being grown on one side of the membrane.

14. The biocomposite product of claim 13, wherein one layer of cells is keratinocytes, and the other layer of cells is fibroblasts.

15. The biocomposite product of claim 12, wherein the biocomposite product is useful for grafting onto a subject in need of skin treatment and/or wound coverage.

16. The biocomposite product of claim 15, wherein the subject in need of skin treatment and/or wound coverage is the one with ear drum defect, disruption of tendon/nerve and/or skin damage that includes burn injuries, ulcers, impaired pigmentation, excisions or other dermatological conditions.