Biocompatible perforated membranes, processes for their preparation, their use as a support in the in vitro growth of epithelial cells, the artificial skin obtained in this manner, and its use in skin grafts

• US 5,326,356 A
• Filed: 02/16/1993
• Issued: 07/05/1994
• Est. Priority Date: 06/01/1990
• Status: Expired due to Term

• Current Assignee: Fidia S.P.A.
• Original Assignee: Fidia S.P.A.
• Inventors: Della Valle, Francesco, Romeo, Aurelio, Calderini, Gabriella, Rastrelli, Alessandro
• Primary Examiner(s): Green, Randall L.
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US08/019,818
• Time in Patent Office: 504 Days
• Field of Search: 623/15, 623/66, 623/1, 623/11, 128/155, 128/156, 604/304, 604/307, 424/447, 424/448, 424/422, 424/423, 424/424, 424/425
• US Class Current: 623/15.12
• CPC Class Codes: A61F 2/105 Skin implants, e.g. artificial skin
• 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 27/60 Materials for use in artificial skin
• CPC Class Codes: C12N 2533/70 Polysaccharides
• CPC Class Codes: C12N 2533/80 Hyaluronan
• CPC Class Codes: C12N 2539/10 Coating allowing for selective detachment of cells, e.g. thermoreactive coating
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0629 Keratinocytes; Whole skin
• CPC Class Codes: Y10T 428/24273 including aperture

Abstract

It is provided biocompatible membranes constructed of materials of natural, synthetic or semisynthetic origin, and having a thickness of between 10 and 500μ, characterised by containing an ordered series of holes of a constant size between 10 and 1000μ, separated from each other by a constant distance of between 50 and 1000μ, and obtained by perforation by mechanical, thermal laser or ultraviolet radiation means, they being suitable for use as a support for the in vitro growth of epithelial cells. Artificial skins obtained thereby and its use in grafts are also provided.

Claims

1. Biocompatible membranes consisting of materials of natural, synthetic or semisynthetic original, and having a thickness of between 10 and 500μ

, which comprise an ordered series of holes of a defined and constant size between 10 and 1000μ

, separated from each other by a constant distance of between 50 and 1000μ

, said membrane enabling in vitro culture of keratinocytes within the holes and on both sides of said membrane.

2. Biocompatible membranes as claimed in claim 1, in which the hole size is between 40 and 70μ

.

3. Biocompatible membranes as claimed in claim 1, in which the distance between holes is 80μ

.

4. Biocompatible membranes as claimed in claim 1, in which the biocompatible material of natural origin is chosen from the group consisting of collagen or coprecipitates of collagen and glycosaminoglycans, cellulose, gelled polysaccharides and natural rubbers, either alone or in mixture with each other or with polymers of synthetic or semisynthetic origin, in the presence of suitable precipitating or gelling agents.

5. The biocompatible membranes according to claim 4 in which the gelled polysaccharides are selected from the group consisting of chitin, chitosan, pectins or pectic acids, agar, agarose, xanthan gum, gellano, alginic acid or alginates, polymannans or polyglucans and starches.

6. Biocompatible membranes as claimed in claim 1, in which the biocomcompatible material of synthetic origin is chosen from the group consisting of polylactic acid, polyglycolic acid or copolymers thereof or their derivatives, polydioxanones, polyphosphazenes, polysulphones and polyurethanes.

7. Biocompatible membranes as claimed in claim 1, in which the biocompatible material of semisynthetic origin is chosen from the group consisting of semisynthetic derivatives of natural polymers crosslinked with crosslinking agents and derivatives of cellulose, of alginic acid, of starch, of hyaluronic acid, of chitin or chitinosan, or gellan, of xanthan, of pectins or pectin acids, of polyglucans, of polymannans, of agar, of agarose, of natural rubbers or of glycosaminoglycans.

8. Biocompatible membranes as claimed in claim 7, in which the biocompatible membrane consists of hyaluronic acid benzyl ester with 100% esterification.

9. The biocompatible membranes according to claim 7 in which the semi-synthetic derivative of a natural polymer is collagen.

10. The biocompatible membranes according to claim 7 in which the crosslinking agent is selected from the group consisting of dialdehydes or their precursors, bicarboxylic acids or derivatives thereof and diamines.

11. The biocompatible membranes according to claim 1 which have a thickness of 20-40μ

.

12. Artificial skin composed of a biocompatible support membrane consisting of materials of natural, synthetic or semisynthetic origin, and having a thickness of between 10 and 500μ

, which contains an ordered series of holes of a defined and constant size between 10 and 1000μ

, separated from each other by a constant distance of between 50 and 1000μ

, together with autologous or heterologous keratinocyte microcolonies in the active proliferation stage present within said holes and on both sides of said membrane.

13. Artificial skin as claimed in claim 12, in which the hole size is between 40 and 70μ

.

14. Artificial skin as claimed in claim 12, in which the distance between holes is 80μ

.

15. Artificial skin as claimed in claim 12, in which the biocompatible material of natural origin is chosen from the group consisting of collagen or coprecipitates of collagen and glycosaminoglycans, cellulose, gelled polysaccharides and natural rubbers, either alone or in mixture with each other or with polymers of synthetic or semisynthetic origin, in the presence of suitable precipitating or gelling agents.

16. Artificial skin as claimed in claim 15 in which the gelled polysaccharides are selected from the group consisting of chitin, chitosan, pectins or petic acids, agar, agarose, xanthan gum, gellano, alginic acid or alginates, polymannans or polyglucans and starches.

17. Artificial skin as claimed in claim 12, in which the biocompatible material of synthetic origin is chosen from the group consisting of polylactic acid, polyglycolic acid or copolymers thereof or their derivatives, polydioxanones, polyphosphazenes, polysulphones and polyurethanes.

18. Artificial skin as claimed in claim 12, in which the biocompatible material of semisynthetic origin is chosen from the group consisting of semisynthetic derivatives of natural polymers crosslinked with crosslinking agents and derivatives of cellulose, of alginic acid, of starch, of hyaluronic acid, of chitin or chitosan, of gellano, of xanthan, of pectins or pectic acids, of polyglucans, of polymannans, of agar, of agarose, of natural rubbers or of glycosaminoglycans.

19. Artificial skin as claimed in claim 18, in which the biocompatible material consists of hyaluronic acid benzyl ester with 100% esterification.

20. Artificial skin according to claim 19 which has a thickness of 20-40μ

and the holes are separated from each other by a constant distance of 80μ

.