WOUND COVERING AND PREPARATION METHOD THEREOF

Abstract

The present invention relates to a wound covering with prominent biocompatible and accelerated wound-healing and its preparation method thereof. The wound covering comprises a film prepared from collagen and Dopa-containing protein—mussel adhesive protein, which is immobilized on collagen by chemical cross-linking, thus enhances the stability of protein structure and maintains the activity of collagen and mussel adhesive protein. The wound covering has excellent mechanical strength and can be trimmed into any shape; it accelerates tissue epithelisation and promotes wound healing with good biocompatibility, non-adhesive to the wound and no further wound damages.

Claims

1. A wound covering comprising a film, wherein the film is prepared from a biocompatible and high-mechanical-strength carrier and a Dopa-containing protein, wherein the carrier is collagen, and the Dopa-containing protein is mussel adhesive protein.

2. The wound covering according to claim 1, wherein the mass ratio of collagen to mussel adhesive protein is 3:1˜20:1.

3. The wound covering according to claim 2, wherein the film is made of collagen and mussel adhesive protein through chemical cross-linking.

4. The wound covering according to claim 3, wherein the dry weight of the unit area of the film is 1-5 mg/cm.sup.2.

5. The wound covering according to claim 4, wherein the wound covering further comprises a preservation solution, the preservation solution is a mixed solution containing glycerol and sodium hydrogen phosphate.

6. The preparation method of the wound covering according to claim 1, comprising the following steps: (a) preparing for a film containing collagen and mussel adhesive protein: (i) dissolving mussel adhesive protein in a glacial acetic acid solution, configuring a mussel adhesive protein preservation solution with a concentration of 0.4%; (ii) taking 1/20-¼ of the mussel adhesive protein preservation solution of the step (i), and adding collagen to dissolve to obtain a mixed solution; (iii) transferring the mixed solution of step (ii) into a stainless steel plate with a solution height of 2.4-12.5 mm; and lyophilizing for 7-15 hours at a temperature of −10° C. and under a vacuum degree of 10-50 Pa; then quickly increasing the temperature of the plate to 10° C. and the vacuum degree to 300-500 Pa and drying for 3 hours; and finally adjusting the temperature to 20 V and the vacuum degree 10-50 Pa and drying for 8-14 hours to for a dried film; (iv) transferring the dried film of the step (iii) into a crosslink agent, cross-linking for 2 hours, taking out and rinsing several times for later use; (b) preparing for a preservation solution: dissolving glycerol and sodium dihydrogen phosphate in purified water, adjusting the pH to 4-7 at a constant volume of 1 L; (c) finishing producing: placing the film containing collagen and mussel adhesive protein of the step (a) in a packet, adding the preservation solution of the step (b), sealing and stored in an aluminum foil bag.

7. The preparation method of the wound covering according to claim 6, wherein the mixed solution in the step (ii) comprises collagen at a concentration of 0.3-0.4% and mussel adhesive protein at a concentration of 0.02-0.1%.

8. The preparation method of the wound covering according to claim 6, wherein in the step (b), dissolving parts by weight of glycerol and 10 parts by weight of sodium dihydrogen phosphate in 900 mL purified water.

9. The preparation method of the wound covering according to claim 6, wherein in the step (c), the adding amount of the preservation solution is 2-5 mL/cm.sup.2.

10. The preparation method of the wound covering according to claim 6, wherein the crosslink agent used in the step (iv) is selected from the group consisting of formaldehyde, glutaraldehyde, kynepin and carbodiimide.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0031] The invention is further illustrated in detail below by means of embodiments, but it is understood by those skilled in the art that the embodiments of the invention are not a limitation on the scope of protection of the invention and that any improvements and variations made on the basis of the invention are within the scope of protection of the invention. All materials in the present invention can be acquired from the market.

Embodiment 1

[0032] This embodiment provides a wound covering and its preparation method comprising the following steps: [0033] (a) preparing for a film containing collagen and mussel adhesive protein [0034] (i) dissolving mussel adhesive protein in 0.0001 mol/L glacial acetic acid solution, configuring a mussel adhesive protein preservation solution with a concentration of 0.4%; [0035] (ii) taking 5 parts of the mussel adhesive protein preservation solution of the step (i) and diluting to 20 parts with water; and adding collagen to dissolve to obtain a mixed solution with sufficient stirring to give the collagen concentration of 0.5% and the mussel adhesive protein concentration of 0.1%; [0036] (iii) transferring the mixed solution of step (ii) into a 30*30 cm stainless steel plate with a solution height of 5 mm; and lyophilizing for 10 hours at a temperature of −10° C. and under a vacuum degree of 10-50 Pa, where approximately 40% of the water still contained in the sample is in the form of ice crystals due to the temperature and pressure of the system. Then quickly increasing the temperature of the plate to 10° C. and the vacuum degree to 300-500 Pa and drying for 3 hours so that the ice crystals become liquid and the carrier collapses at low temperatures to form a uniform gel-like film. Then finally adjusting the temperature to 20° C. and the vacuum degree 10-50 Pa and drying for 10 hours, thus further removes the residual moisture from the sample to less than 10% to form a dry and strong film. Then slitting of the collagen-MAP film into the required size and shape by passing it through a slitting die; [0037] (iv) transferring the dried collagen-MAP film into a 0.1% formaldehyde solution with adding amount 5 mL/cm.sup.2 of the film and cross-linking for 2 hours, taking out and rinsing 8 times for later use; [0038] (b) preparing for a preservation solution is dissolving 10 g glycerol and 31.2 g sodium dihydrogen phosphate in purified water of 900 mL, adjusting the pH to 5.0 by using sodium hydroxide solution, and adding water at a constant volume to 1 L; [0039] (c) finishing producing placing the rinsed collagen-MAP film in a packet, adding the preservation solution at a ratio of 2 mL/cm.sup.2, sealing and stored in an aluminum foil bag.

Embodiment 2

[0040] This embodiment provides a wound covering and its preparation method comprising the following steps: [0041] (a) preparing for a film containing collagen and mussel adhesive protein [0042] (i) dissolving mussel adhesive protein in 0.0001 mol/L glacial acetic acid solution, configuring a mussel adhesive protein preservation solution with a concentration of 0.4%; [0043] (ii) taking 5 parts of the mussel adhesive protein preservation solution of the step (i) and diluting to 20 parts with water; and adding collagen to dissolve to obtain a mixed solution with sufficient stirring to give the collagen concentration of 0.3% and the mussel adhesive protein concentration of 0.1%; [0044] (iii) transferring the mixed solution into a 30*30 cm stainless steel plate with a solution height of 12.5 mm; and lyophilizing for 15 hours at a temperature of −10° C. and under a vacuum degree of 10-50 Pa; then quickly increasing the temperature of the plate to 10° C. and the vacuum degree to 300-500 Pa and drying for 3 hours; adjusting the temperature to 20° C. and the vacuum degree 10-50 Pa and drying for 14 hours, Then slitting of the collagen-MAP film into the required size and shape by passing it through a slitting die. [0045] (iv) transferring the dried collagen-MAP film into a 0.1% formaldehyde solution with adding amount 5 mL/cm.sup.2 of the film and cross-linking for 2 hours, taking out and rinsing 8 times; [0046] (b) preparing for a preservation solution dissolving 10 g glycerol and 31.2 g sodium dihydrogen phosphate in purified water of 900 mL, adjusting the pH to 5.0 by using sodium hydroxide solution, and adding water at a constant volume to 1 L; [0047] (c) finishing producing placing the rinsed collagen-MAP film in a packet, adding the preservation solution at a ratio of 2 mL/cm.sup.2, sealing and stored in an aluminum foil bag.

Embodiment 3

[0048] This embodiment provides a wound covering and its preparation method comprising the following steps: [0049] (a) preparing for a film containing collagen and mussel adhesive protein [0050] (i) dissolving mussel adhesive protein in 0.0001 mol/L glacial acetic acid solution, configuring a mussel adhesive protein preservation solution with a concentration of 0.4%; [0051] (ii) taking 1 parts of the mussel adhesive protein preservation solution of the step (i) and diluting to 20 parts with water; and adding collagen to dissolve to obtain a mixed solution with sufficient stirring to give the collagen concentration of 0.4% and the mussel adhesive protein is concentration of 0.02%; [0052] (iii) transferring the mixed solution into a 30*30 cm stainless steel plate with a solution height of 2.4 mm; and lyophilizing for 7 hours at a temperature of −10° C. and under a vacuum degree of 10-50 Pa; then quickly increasing the temperature of the plate to 10° C. and the vacuum degree to 300-500 Pa and drying for 3 hours; adjusting the temperature to 20° C. and the vacuum degree 10-50 Pa and drying for 8 hours, Then slitting of the collagen-MAP film into the required size and shape by passing it through a slitting die. [0053] (iv) transferring the dried collagen-MAP film into a 0.1% formaldehyde solution with adding amount 5 mL/cm.sup.2 of the film and cross-linking for 2 hours, taking out and rinsing 8 times; [0054] (b) preparing for a preservation solution dissolving 10 g glycerol and 31.2 g sodium dihydrogen phosphate in purified water of 900 mL, adjusting the pH to 5.0 by using sodium hydroxide solution, and adding water at a constant volume to 1 L; [0055] (c) finishing producing

[0056] placing the rinsed collagen-MAP film in a packet, adding the preservation solution at a ratio of 2 mL/cm.sup.2, sealing and stored in an aluminum foil bag.

[0057] The results of the tests carried out on the wound covering prepared in Embodiments 1-3 are shown in Table 1.

TABLE-US-00001 TABLE 1 Test Result Embodiment 1 Embodiment 2 Embodiment 3 Sensory Flexible and Flexible and Flexible and Evaluation transparent film transparent film transparent film Dry 3 mg/cm.sup.2 5 mg/cm.sup.2 1 mg/cm.sup.2 Weights Tensile No breakage of No breakage of No breakage of Strength the sample (1 cm the sample (1 cm the sample (1 cm wide) hung with wide) hung with wide) hung with a 100 g weight a 150 g weight a 50 g weight

[0058] As shown in Table 1, the present invention mixes collagen and mussel adhesive protein, makes a dry film by half-lyophilisation at low temperature, then immobilize mussel mucin on collagen by chemical cross-linking, rinses to remove residual crosslink agent and stores in preservation solution, which enhances the stability of protein structure and maintains the activity of collagen and mussel adhesive protein, the collagen substrate has excellent mechanical strength and can be trimmed into any shape; it accelerates tissue epithelisation and promotes wound healing with good biocompatibility, non-adhesive to the wound and no further wound damages.

[0059] The above schematically describes the present invention and its embodiments, the description is not limiting, and what is shown in the attached drawings is only one of the embodiments of the present invention, and the actual structure is not limited to it. Therefore, if a person of ordinary skill in the art is inspired by it and designs, without departing from the creative purpose of the invention, structural ways and embodiments similar to this technical solution without inventiveness, they shall fall within the scope of protection of the present invention.