TWI235670B - Method for manufacturing a composite collagen film - Google Patents

Abstract

A method for manufacturing a composite collagen film is provided. The method includes providing a collagen film and dipping the collagen film in a solution of collagen and glycosaminoglycans (GAGs). The method further includes sequentially performing a freezing process, a vacuum freezing drying process, a condensing process, and a drying process. Finally, the method comprises using a cross-linking agent for making the collagen film cross link with the collagen and GAGs in the solution so as to produce a composite collagen film.

Description

1235670 Case No. 92109800 Day M. Amendment V. Description of Invention U) Technical Field to which the Invention belongs The present invention provides a method for guiding tissue regeneration (Guided

Tissue Regeneration, a method for manufacturing a composite collagen film of GTR ’, especially a method for manufacturing a biocompatible slow-resorbing composite collagen film that has suture ability (suturab 1 e) and can be slowly absorbed.

Guidance Tissue Regeneration (GTR) is a surgical procedure that is currently widely used, the purpose of which is to modify the type of tissue or organ damaged by disease or trauma (m 〇rph ο 1 〇gy) and The function is to restore or regenerate.

In a successful tissue regeneration process, the regenerated tissue must be repopulated in the location and space where the tissue was originally healthy. In particular, in order for the type and function relationships between different regenerative tissues in the region to be properly repaired, cell division and replication of the region and subsequent differentiation of the regenerative cells must be performed. It is an orderly and consistent (c ο ncerted) program. However, when a void is generated in a living tissue due to partial tissue damage, the void will be filled by the adjacent and fastest replicating cell line (c e 1 1 1 i n e).

1235670 Amendment No. 92109800 a. V. Description of the invention (2) Take periodontal disease as an example. In a periodontal lesion, there are four main sources of wound healing cells, which are from the teeth. Silver epithelial cells (epithel iiim) and connective tissue, as well as cells from bone, and weaving and periodontal τ (periradicu 1 ar 1 igament), of which only cells from periodontal ligament have the ability to tissue regeneration. However, because the regeneration rate of epithelial cells is much faster than that of periodontal ligament cells, it makes repair of damaged periodontal ligament tissue very difficult. The aforementioned technology of guided tissue regeneration (GTR) can enable appropriate tissue cells to perform a sequential and consistent division and replication to achieve the purpose of damaging tissue regeneration. In order to achieve this, a barrier is currently placed between the regenerating tissue and the tissue that may interfere with the regeneration process of the regenerating tissue until the damaged part of the tissue is completely: regeneration and the regeneration Until the organization reaches maturity. Taking the periodontal lesion mentioned above as an example, a biomedical material (Biomateria 1) membrane is placed to guide tissue regeneration to prevent epidermal cells and connective tissue from the gums from coming into contact with the root of the tooth. Only periodontal blades are allowed. Growth with cells. I _. I 'At present, there are two main biomedical materials used to guide tissue regeneration technology. The first is e X panded polytetrafluoroethylene (EPTFE) membrane and high-density porous polyethylene. (MEDPOR) membranes and other non-absorbable polymer membranes. Since the EPTFE membranes are not absorbed, they can maintain their integrity during implantation (imp 1 anted) in the body, so they have excellent barriers. effect. However, 3 1235670 Case No. 92109800 5. Description of the invention (3) The EPTFE membrane cannot be left in the body indefinitely, so six to eight weeks after the first operation of the EPTFE membrane It needs to be removed for a second operation.

I The second type is an absorbable (resorbab 1 e) membrane composed of collagen or other polymers, such as po 1 yg 1 y co 1 at es. The membrane is eventually absorbed and disappears from the implant, so no additional surgery is required to remove it. In contrast, the absorbable membrane may not remain intact in the body for a sufficient period of time to guide the completion of a successful tissue regeneration procedure. SUMMARY OF THE INVENTION Therefore, the object of the present invention is to provide: · A biocompatible composite collagen film with suture ability (su t urab 1 e) and slow absorption (s 1 ow-r esorb i ng) In order to apply to the technology of guided tissue regeneration (GTR) and solve the above problems. In a preferred embodiment of the present invention, the method of the present invention first provides a collagen film, and immerses the collagen film in a collagen material and GAG s (g 1 yc 〇sami η 〇g 1 ycans) In the mixed solution, a freeze treatment process, a vacuum freeze-drying process, a compression process cleaning, and a drying process are sequentially performed, and then a cross-l inking agent is used to mix the collagen film with the The collagen raw materials and GAGs in the solution undergo a cross-linking reaction to form a collagen composite film. 1235670 Case No. 92 i Amended on 09/09, V. Explanation of the invention (4) Since the collagen film provided by the present invention has a composite structure, it can slow down the rate of absorption by the implant and prolong its retention in the implant. Time, thereby increasing its barrier effect. On the other hand, the composite collagen film of the present invention further contains at least one antibiotic (antimicrobial agent), anti-inflammatory agent (anti-inflammatory agent) or growth factor (growth factor), so as to be released over time. Specific agents required for different treatments to improve wound healing and tissue regeneration.

Embodiments In a preferred embodiment of the present invention, a collagen film is first provided. The manufacturing method of the collagen film is firstly subjecting collagen-rich bovine skin (tendon or placenta) to a constant temperature treatment with gastrin (pepsi η), and then purifying the gastrin The obtained collagen molecules were dissolved in a 0.05 M acetic acid solution at a concentration of (1-10 mg / m 1), and placed in an environment of 4 ° C for reaction.

After a suitable period of time, 0.1 M sodium hydroxide solution was added for acid-base neutralization, and poured into a mold to continue the action for 24 hours at a temperature range of 20 to 37 ° C. Then, a piston is used to squeeze out the water in the solution until a collagen film with a predetermined thickness is obtained. Finally, the collagen membrane was immersed in a dimethylphosphonate containing 0.5% diphenylphosphoryl azide (DPPA).

Page 9 1235670 Case No. 92109800 Amended on March 5, V. Description of the Invention (5) (dimethylformamide, DMF) solution, so that the fibrils in the collagen membrane i are cross-linked with each other, enhancing its effect on enzyme decomposition ( enzyme degradation) to obtain the collagen film.

After the collagen film is obtained, a mixed solution composed of glycosaminoglycans (GAGs) and type I collagen is then prepared, in which the proportion of GAGs to collagen is about 10%. The mixed solution is prepared by first preparing GAGs and collagen in an aqueous solution having a weight concentration of about 0.5 to 2%. The collagen aqueous solution is a solution in which collagen membrane fibers are dissolved in a weakly acidic aqueous solution (for example, 0.1 Μ). Acetic acid in water). Finally, the two aqueous solutions were mixed, and the acidity was neutralized with an aqueous sodium hydroxide solution to adjust the pH of the mixed solution to about 6.5 to 8. : · Next, the collagen membrane is immersed in the mixed solution, and liquid nitrogen is used to perform a freezing treatment process at a temperature of about -180 ° C. The purpose of this freezing process is to make the gelatin film shape.

I

| Porous (micro-porous), so when the collagen film covers the tissue damage area, water and nutrients can be passed through the micropores to supply nutrients needed during the tissue regeneration process, while blocking May interfere with regenerative cells dividing and replicating in this area. Then, a vacuum freeze-drying process and a compression process, which take about 18 hours, are sequentially performed in order to remove water from the solution and obtain a pre-treatment.

Page 10 1235670 Case No. 92109800 Early Month Day Amendment V. Description of Invention (6) Film thickness determined. Then use the natural cross-linking agent-genipi η or glutaric acid (g 1 utara 1 dehyde) as the cross-linking agent to make the collagen film and the collagen in the mixed solution and g 1 yc 〇sami η 〇 g 1 ycans undergo a cross-1 inking reaction, so that the collagen membrane absorbs the collagen-GAGs mixed solution to obtain a collagen membrane with a composite structure. Finally, the composite collagen film is washed and dried, and the above steps can be repeated as needed to make a composite collagen protein film with multiple composite structures. In addition to the advantages that the composite collagen film produced by the present invention can be slowly absorbed to enhance its barrier effect, it can be incorporated into the cross-linking agent during the cross-linking reaction between the composite collagen films. One or more kinds of antibiotics (anti-microbial agent), anti-inflammatory agent (anti-inflammatory agent) or growth factor (growth factor) are induced, and then immobilized in the composite collagen membrane. Taking a composite collagen film with multiple composite structures as an example, we can use a cross-linking agent mixed with different agents to make the collagen film undergo at least one compounding process to form a compound containing different agents in different structural regions. Multiple composite collagen membranes. The mixed agent may be an antibiotic, an anti-inflammatory agent, or an inducer of tissue growth factor. Therefore, during the gradual absorption and decomposition of the multiple composite collagen film, these specific agents with different functions can be released over time. Improve wound healing and tissue regeneration. 1235670 Case No. 92109800 Fifth, invention description (7) | Compared with the conventional collagen film, the composite collagen | protein film provided by the present invention can slow the absorption rate and prolong the time it stays in the body. It can improve the barrier effect, and can release specific agents required for different treatments over time, thereby improving the effects of wound healing and tissue regeneration. The above description is only a preferred embodiment of the present invention. Any equivalent changes and modifications made in accordance with the scope of the patent application of the present invention shall belong to the present invention patent.

Page 12 1235670 Case No. Θ2Ι09800 ___________ Year J Day j Positive Schematic Description Page 13

Claims (1)

1235670 Case No. 92109800 Amendment VI. Patent application scope; (i) Inject the above-dissolved collagen into a mold and perform a second isothermal treatment to make the above-dissolved collagen Forming a film-like shape; (j) compressing the collagen-like film of the film shape until a predetermined film thickness is obtained; and (k) reacting the compressed collagen white film with a cross-linking agent to crosslink the fibers (fi br i 1 s) in the compressed collagen film with each other to form a matrix that is not easily decomposed ). 4. The method according to item 3 of the patent application, wherein the collagen is a collagen of type I. 5 .; For the method of claim 3 in the scope of patent application, wherein the solvent is acetic acid ° I! 6. For the method of claim 1 in the scope of patent application, wherein the freezing process i uses liquid nitrogen to reduce the temperature To -1.8 0 ° C. | * 7. The method according to item 6 of the patent application, wherein the freezing process is used to form a plurality of micropores (micr 0-porous) in the first collagen film. 8 Method wherein the vacuum freeze-drying Page 15 1235670 Case No. 92 丨 3/09800 Rash is positive 6. Scope of patent application The manufacturing process takes 18 hours. 9. The method according to item 1 of the patent application, wherein the cross-linking agent is glutaraldehyde. 10. The method according to item 1 of the scope of patent application, wherein the cross-linking agent further comprises at least one antibiotic (anti-microbial agent), anti-inflammatory agent (anti-inflammatory agent), or tissue growth factor (factor) And can be immobilized in the composite collagen membrane by the cross-linking agent. 1 1. The method according to item 10 of the scope of patent application, wherein the antibiotic, the anti-inflammatory agent or the growth-inducing factor can utilize each of the cross-linking reactions to embed the multiple composite structures in the composite collagen membrane, respectively. In different areas 1 2. The method according to item 11 of the scope of patent application, wherein the antibiotic, the anti-inflammatory agent or the induced tissue growth factor can be decomposed in the composite collagen membrane, according to the The decomposition time of different areas of the multiple composite structure is released into the implanted body environment. 1 3. The method according to item 1 of the patent application scope, wherein the collagen composite membrane is used as a guided tissue regeneration 1235670 Case No. 92109800 a. Amendment VI. Application for patent (regeneration, GTR) film. 14. The method according to item 1 of the patent application scope, further comprising a cleaning and drying process after completing step (e).