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Claims for Patent: 6,803,037

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Summary for Patent: 6,803,037
Title: Hyaluronic acid derivative based cell culture and biodegradable three-dimensional matrix
Abstract:The present invention relates to a biological material having a matrix which contains at least one derivative of hyaluronic acid on which endothelial cells, glandular cells such as islets of Langerhans and liver cells, skin adnexa, germinative cells of hair bulbs, and kerinatocytes are grown, optionally in presence of a medium treated with fibroblasts or in a co-culture with fibroblasts. A process for the production of said biologic materials and the use of such materials for human and veterinary applications such as cardiovascular and oncological surgery, in connection with transplants, for enhancing the biological process of tissue vascularization and for aesthetic use, and also for the screening of medicaments or toxic substances and as a support in the process of gene transfection. The biological material is based on an efficacious cell culture and a biocompatible and biodegradable three-dimensional matrix containing a hyaluronic acid derivative.
Inventor(s): Abatangelo; Giovanni (Saccolongo, IT), Callegaro; Lanfranco (Theine, IT)
Assignee: Fidia Advanced Biopolymers S.r.l. (Brindisi, IT)
Application Number:09/445,604
Patent Claims:1. A biological material comprising: a) at least one autologous or homologous cellular line selected from the group consisting of endothelial cells, glandular cells, skin adnexa and germinative cells of hair bulbs, b) a biocompatible and biodegradable three-dimensional matrix, on which said cellular line is seeded and grown, said matrix comprising a hyaluronic acid derivative selected from the group consisting of: A) an ester of hyaluronic acid wherein part or all of the carboxylic groups of said hyaluronic acid are esterified with an alcohol of aliphatic, aromatic, arylaliphatic, cycloaliphatic series; B) an autocrosslinked ester of hyaluronic acid wherein part or all of the carboxylic moieties of said hyaluronic acid are esterified with the alcoholic groups of the same or a different hyaluronic acid chain; C) a hemiester of succinic acid or a heavy metal salt of the hemiester of succinic acid with hyaluronic acid or with a hyaluronic acid ester having part or all of the carboxy groups of hyaluronic acid esterified with an alcohol of aliphatic, aromatic, arylaliphatic, cycloaliphatic series; and D) an O-, an N-sulphated hyaluronic acid and a derivative thereof, c) optionally collagen and/or fibrin d) optionally autologous or homologous keratinocytes said autologous or homologous cellular line is cultivated in presence of a medium treated with autologous or homologous human fibroblasts or in a coculture with autologous or homologous human fibroblasts.

2. The biological material according to claim 1 further containing keratinocytes, fibrin and collagen.

3. The biological material according to claim 1, wherein said endothelial cells are isolated from the umbilical vein from dermis or other tissue wherein blood vessels are present.

4. The biological material according to claim 1 wherein the glandular cells are liver or Langerhans' islet cells.

5. The biological material according to claim 1 wherein the skin adnexa are sebaceous glands, sweat glands or hair bulbs and the germinative cells are isolated from autologous or homologous hair bulbs.

6. The biological material according to claim 1, wherein the hyaluronic acid ester is a benzyl ester with a degree of esterification of between 25 and 100%.

7. The biological material according to claim 1, wherein component (b) is in the form of a non woven fabric, sponges, granules, gauzes, microspheres guide-channels or combination with one another.

8. The biological material according to claim 7 wherein component (b) is in the form of a non woven fabric.

9. A process for preparing a biological material according to claim 1 comprising the following steps: i) isolating at least one autologous or homologous cellular line selected from the group consisting of endothelial cells, glandular cells, skin adnexa, and germinative cells of hair bulbs, ii) preparing a biocompatible and biodegradable three-dimensional matrix, comprising at least one hyaluronic acid derivative and optionally fibrin and/or collagen, and iii) seeding said cellular line on said matrix in the presence of a medium treated with autologous or homologous human fibroblasts or in a co-culture with autologous or homolgous human fibroblasts.

10. The process according to claim 9 wherein, when in step (i) the cellular line is selected from the group consisting of autologous or homologous skin adnexa and germinative cells of hair bulbs, the cellular line is optionally seeded in association with autologous or homologous keratinocytes.

11. A process for the preparation of the biological material according to claim 3 comprising the following steps: i) isolating endothelial cells from human umbilical vein by enzymatic digestion with collagenase; ii) amplifying said cells on collagen treated dishes, iii) preparing a biocompatible and biodegradable three-dimensional matrix, comprising at least one hyaluronic acid derivative and optionally fibrin and/or collagen, and iv) seeding said endothelial cells in association with a cellular line selected from the group consisting of glandular cells, skin adnexa and germinative cells of hair bulb in the presence of a medium treated with autologous or homologous human fibroblasts in primary culture or in a coculture with autologous or homologous human fibroblasts.

12. The biological material according to claim 1, for use in human and veterinary surgery.

13. The biological material according to claim 1 wherein component (a) comprises skin adnexa optionally in association with keratinocytes for use in skin transplants.

14. The biological material according to claim 13 wherein component (a) further comprises autologous or homologous endothelial cells facilitating the mechanism of neovascularization of the transplanted skin.

15. The biological material according to claim 1 wherein component (a) comprises germinative cells of hair bulbs for use in scalp transplants.

16. The biological material according to claim 1, wherein component (a) comprises islets of Langerhans for use in case of insufficient insulin production.

17. The biological material according to claim 1, wherein component (a) comprises endothelial cells for use in surgery.

18. The biological material according to claim 17, for use in cardiovascular, aesthetic and oncological surgery.

19. The biological material according to claim 18 for use in surgery to enhance the biological process of tissue vascularisation.

20. The biological material according to claim 1, wherein component (a) is an autologous cellular line as support for gene transfection.

21. The biological material according to claim 1, wherein component (a) is an autologous cellular line for use in gene transfection.

22. An in vitro biological material comprising: a) at least one cellular line selected from the group consisting of endothelial cells, glandular cells, skin adnexa and germinative cells of hair bulbs, b) a biocompatible and biodegradable three-dimensional matrix, on which said cellular line is seeded and grown, said matrix comprising a hyaluronic acid derivative selected from the group consisting of: A) an ester of hyaluronic acid wherein part or all of the carboxylic groups of said hyaluronic acid are esterified with alcohols of the aliphatic, aromatic, arylaliphatic, cycloaliphatic series, B) an autocrosslinked ester of hyaluronic acid wherein part or all of the carboxylic moieties of said hyaluronic acid are esterified with the alcoholic groups of the same or a different hyaluronic acid chain, C) a hemiester of succinic acid or a heavy metal salt of the hemiester of succinic acid with hyaluronic acid or with a hyaluronic acid ester having part or all of the carboxy groups esterified with an alcohol of aliphatic, aromatic, arylaliphatic, cycloaliphatic series and D) an O-, an N-sulphated hyaluronic acid and a derivative thereof, c) optionally collagen and/or fibrin d) autologous and/or homologous keratinocytes, said cellular line being cultivated in presence of a medium treated with autologous or homologous human fibroblasts or in a coculture with autologous or homologous human fibroblasts.

23. The biological material according to claim 22, containing autologous or homologous keratinocytes, collagen and/or fibrin.

24. The biological material according to claim 22, wherein said endothelial cells are isolated from the umbilical vein from dermis or other tissue wherein blood vessels are present.

25. The biological material according to claim 22, wherein the glandular cells are liver or Langerhans' islet cells.

26. The biological material according to claim 22, wherein the skin adnexa are sebaceous glands, sweat glands or hair bulbs and germinative cells are isolated from autologous, homologous and heterologous hair bulbs.

27. The biological material according to claim 22, wherein the hyaluronic acid ester is a benzyl ester with a degree of esterification of between 25 and 100%.

28. The biological material according to claim 22, wherein component (b) is in the form of a non woven fabric, sponges, granules, gauzes, microspheres, guide channels or combination with one another.

29. The biological material according to claim 22 wherein component (b) is in the form of a non woven fabric.

30. A process for preparing a biological material according to claim 22 comprising the following steps: i) isolating at least one cellular line selected from the group consisting of endothelial cells, glandular cells, skin adnexa, and germinative cells of hair bulbs, ii) preparing a biocompatible and biodegradable three-dimensional matrix, comprising at least one hyaluronic acid derivative and optionally fibrin and/or collagen, and iii) seeding said cellular line on said matrix in the presence of a medium treated with autologous or homologous human fibroblasts or in a co-culture with autologous or homolgous human fibroblasts.

31. The process according to claim 30 wherein, when in step (i) the cellular line is selected from the group consisting of skin adnexa and germinative cells of hair bulbs, the cellular line is optionally seeded in association with keratinocytes.

32. A process for preparing the biological material according to claim 24 comprising: i) isolating said endothelial cells from human umbilical vein by enzymatic digestion with collagenase; ii) amplifying said cells on collagen treated dishes, iii) preparing a biocompatible and biodegradable three-dimensional matrix, comprising at least one hyaluronic acid derivative and optionally fibrin and/or collagen, and iv) seeding said endothelial cells optionally in association with a cellular line selected from the group consisting of glandular cells, skin adnexa and germinative cells of hair bulb and in the presence of a medium treated with autologous or homologous human fibroblasts in primary culture or in a coculture with autologous or homologous human fibroblasts.

33. The biological material according to claim 22 for the screening of medicaments or toxic substances.

34. A biological material comprising: a) at least one autologous or homologous cellular line selected from the group consisting of endothelial cells, glandular cells, skin adnexa and germinative cells of hair bulbs, b) a biocompatible and biodegradable three-dimensional matrix, on which said cellular line is seeded and grown, said matrix comprising a hyaluronic acid derivative selected from the group consisting of: A) an ester of hyaluronic acid wherein part or all of the carboxylic groups of said hyaluronic acid are esterified with alcohols of the aliphatic, aromatic, arylaliphatic, cycloaliphatic series; B) an autocrosslinked ester of hyaluronic acid wherein part or all of the carboxylic moieties of said hyaluronic acid are esterified with the alcoholic groups of the same or a different hyaluronic acid chain; and C) a hemiester of succinic acid or a heavy metal salt of the hemiester of succinic acid with hyaluronic acid or with a hyaluronic acid ester having part or all of the carboxy groups of hyaluronic acid esterified with an alcohol of aliphatic, aromatic, arylaliphatic, cycloaliphatic series, c) optionally collagen and/or fibrin d) optionally autologous or homologous keratinocytes said autologous or homologous cellular line is cultivated in presence of a medium treated with autologous or homologous fibroblasts human or in a coculture with autologous or autologous human fibroblasts.

35. A biological material comprising: a) at least one autologous or homologous cellular line selected from the group consisting of endothelial cells, glandular cells, skin adnexa and germinative cells of hair bulbs, b) a biocompatible three-dimensional matrix, on which said cellular line is seeded and grown, said matrix comprising an ester of hyaluronic acid wherein part or all of the carboxylic groups of said hyaluronic acid are esterified with alcohols of the aliphatic, aromatic, arylaliphatic, cycloaliphatic series, c) optionally collagen and/or fibrin d) optionally autologous or homologous keratinocytes said cellular line being cultivated in presence of a medium treated with autologous or homologous human fibroblasts or in a coculture with autologous or homologous human fibroblasts.

Summary for Patent:   Start Trial

Foriegn Application Priority Data
Foreign Country Foreign Patent Number Foreign Patent Date
Italy97A000122Jun 11, 1997
PCT Information
PCT FiledJune 10, 1998PCT Application Number:PCT/EP98/03510
PCT Publication Date:December 17, 1998PCT Publication Number:WO98/56897

Details for Patent 6,803,037

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Advance Biofactures SANTYL collagenase VIAL 101995 001 1965-06-04   Start Trial Fidia Advanced Biopolymers S.r.l. (Brindisi, IT) 2017-06-11 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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