Support with crosslinked marine collagen for tissue engineering and manufacture of biomaterials

a technology of crosslinked marine collagen and biomaterials, which is applied in general culture methods, epidermal cells/skin cells, skeletal/connective tissue cells, etc., can solve the problems of not providing collagen supports which satisfactorily comply with all research results

Inactive Publication Date: 2005-09-22
COLETICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0029] The present invention makes it possible to solve all these technical problems in a particularly simple, inexpensive manner applicable to the industrial scale, particularly in cosmetics, dermopharmaceutics or pharmaceutics. DETAILLED DESCRIPTION OF THE INVENTION
[0036] In yet another advantageous embodiment of the invention, it can be of particular value to prepare either “young” reconstructed skin using cells taken substantially exclusively from young subjects, or “aged” reconstructed skin obtained from cells taken substantially exclusively from elderly subjects. These models will make it possible to improve our knowledge of the skin ageing process and study the influence of active agents on this process.
[0063] To obtain the strongest collagen materials, the inventors carried out more particularly the process described in U.S. Pat. No. 5,333,092 granted on 19 Jul. 1994. This technique affords a mixture of soluble and insoluble type I and type III native collagens which are very strong from the mechanical point of view and very resistant to enzymatic digestion. These last two characteristics may optionally be improved by any crosslinking technique or by the addition of substances which interact strongly with collagen and do not exhibit toxicity towards the cells. Furthermore, this collagen production process makes it possible virtually to eliminate the risk of biological contamination due to bacteria, viruses or prions.

Problems solved by technology

The researches carried out hitherto have not provided collagen supports which satisfactorily comply with all the constraints listed above.

Method used

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  • Support with crosslinked marine collagen for tissue engineering and manufacture of biomaterials
  • Support with crosslinked marine collagen for tissue engineering and manufacture of biomaterials
  • Support with crosslinked marine collagen for tissue engineering and manufacture of biomaterials

Examples

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example 1

OF THE INVENTION

Preparation of a Porous Collagen Layer of Native Collagen by the Technique of U.S. Pat. No. 5,331,092

A—Preparation of the Native Collagen

[0078] A gel is prepared from calf skins which have previously been washed (2 hours) and then depilated with a lime / sulfide mixture (lime: 3.5%, sodium sulfide: 2.5%) at a rate of 400 g of skin (solids content: about 30%) to 250 ml of water. This bath lasts for 30 minutes with rotation at 4 rpm.

[0079] The total depilation time is 36 hours.

[0080] The skins are then unlimed in a bath containing ammonium chloride (3%) and sodium metabisulfite (0.5%) at a rate of 400 g of skin to 50 ml of bath.

[0081] The total duration of this bath is 2 hours thirty minutes.

[0082] The salts are removed by two successive washes with water (15 minutes per wash) at a rate of 200 ml of water to 100 g of tissue.

[0083] The skins are subsequently ground and then washed by agitation for 1 hour with phosphate buffer of pH 7.8 (0.78 g / l of potassium dihyd...

example 2

OF THE INVENTION

Preparation of a Porous Collagen Layer Crosslinked with Diphenylphosphorylazide (DPPA) by the Technique Described in European Patent No. 466 829 of 24 Jul. 1996

[0089] The collagen lyophilizate is incubated for 24 h in a solution containing 5 to 250 μl DPPA / g collagen in 100 ml of dimethylformamide (DMF). The collagen is then rinsed in 100 ml of DMF to remove the DPPA. The DMF is then removed by rinsing in 100 ml of a borate buffer solution of pH 8.9 (0.04 M sodium tetraborate, 0.04 M boric acid).

[0090] The collagen is finally incubated overnight in the same borate buffer, the borate buffer then being removed by continuous rinsing with softened water for 6 h.

example 3

OF THE INVENTION

Preparation of a Porous Collagen Layer Crosslinked with Carbodiimide and N-hydroxysuccinimide

[0091] The collagen is crosslinked with EDC (ethyldlmethylaminopropylcarbodilmide) at a concentration of 0.23 to 0.69 g / g collagen and with NHS (N-hydroxysuccinimide) at a concentration of 0 to 0.42 g / g collagen.

[0092] After rinsing with softened water, the collagen is lyophilized again.

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Abstract

The invention relates to a method of in vitro testing of the efficacy of a potentially active substance comprising monitoring the effect of said potentially active substance on an artificial skin, comprising a composite product forming a collagen support comprising at least one porous collagen layer covered on at least one side with a collagen membrane component selected from the group consisting of a collagen membrane prepared by compression of a collagen sponge at a pressure of at least about 50 bar and of a collagen membrane comprising a collagen film prepared by drying a collagen gel separately from the porous collagen layer, thereby providing a reliable method for finding new potentially active substances.

Description

[0001] This application is a Divisional and Continuation in Part of U.S. patent application Ser. No. 09 / 616,282 filed on Jul. 14, 2002, and of U.S. patent application Ser. No. 09 / 616,526 filed on Jul. 14, 2002. [0002] The invention relates to a support with collagen base for tissue engineering and manufacture of biomaterials and to different methods including a method of in vitro testing of the efficacy of a potentially active substance using such a support.DISCUSSION OF THE PRIOR ART [0003] For many years collagen has proved to be an irreplaceable substrate for the production of artificial tissues containing living cells. [0004] The biomaterials obtained are increasingly used in the field of pharmaceutics and they appear to have a very promising future for the preparation of injured connective tissues or for gene therapy by allowing the introduction and survival of modified cells in a living organism. [0005] Furthermore, for “in vitro” tests, the cosmetic and dermopharmaceutical in...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61L27/24A61L27/34A61L27/36A61L27/38A61L27/40A61L27/48A61L27/56A61L27/60C12N5/00C12N5/071C12Q1/00G01N33/50
CPCA61L27/24A61L27/34G01N2333/78A61L27/36A61L27/3839A61L27/40A61L27/48A61L27/56A61L27/60C12N5/0068C12N5/0698C12N2502/094C12N2502/1323C12N2503/06C12N2533/54C12N2533/72G01N33/5085C08L89/06
Inventor ANDRE, VALERIEABDUL MALAK, NABILHUC, ALAIN
Owner COLETICA
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