Disclosure of Invention
The invention aims to provide a collagen composition for repairing skin injury and a production method thereof, wherein modified antibacterial cellulose is prepared, zinc chloride is used as a cross-linking agent, and the modified antibacterial cellulose is cross-linked with active substances such as sodium alginate, collagen and the like to prepare a collagen composition gel dressing which has excellent antibacterial property, can promote wound healing and can improve scars at the wound.
The aim of the invention can be achieved by the following technical scheme:
A collagen composition for repairing skin injury comprises, by weight, 50-60 parts of sodium alginate, 30-40 parts of polyvinyl alcohol, 10-20 parts of glycerol, 5-10 parts of collagen, 4-8 parts of zinc chloride, and 2-6 parts of modified antibacterial cellulose;
the modified antibacterial cellulose is prepared by cellulose esterification and then amino acid antibacterial agent connection through ammonolysis reaction.
Further, the production method of the modified antibacterial cellulose specifically comprises the following steps:
S1, dispersing hydroxyethyl cellulose in tetrahydrofuran, stirring uniformly, adding dimethyl succinate, regulating the pH of a system to 8-9 by using a sodium hydroxide solution, stirring for 2-6 hours at the temperature of 60-70 ℃, cooling materials, regulating the pH of the system to be neutral by using dilute hydrochloric acid, filtering, washing and vacuum drying to prepare esterified cellulose;
s2, mixing esterified cellulose with N, N-dimethylformamide, adding L-arginine, regulating the pH value of the system by using a hydrochloric acid solution, stirring for 4-12 hours at the temperature of 50-60 ℃, after the reaction is finished, settling, washing the materials by using an ethanol solution with the volume fraction of 70%, and carrying out vacuum drying to prepare the modified antibacterial cellulose.
According to the technical scheme, in an alkaline environment, hydroxyl in a hydroxyethyl cellulose structure can perform ester exchange reaction with ester groups in a dimethyl succinate structure, and esterified cellulose containing the ester groups in the structure can be prepared by controlling the consumption of the hydroxyethyl cellulose and the dimethyl succinate, and the ester groups in the structure can perform ammonolysis reaction with amino in an L-arginine structure, so that carboxyl and guanidine groups in the L-arginine structure are introduced into the cellulose structure, and the modified antibacterial cellulose is prepared.
Further, in the step S1, the mass ratio of the hydroxyethyl cellulose to the dimethyl succinate is 1:0.1-0.25.
Further, in step S1, the volume fraction of the sodium hydroxide solution is 2-4%.
Further, in the step S1, the concentration of the dilute hydrochloric acid is 0.4-0.5 mol/L.
Further, in the step S2, the concentration of the hydrochloric acid solution is 1-1.5mol/L.
Further, in step S2, the pH is adjusted to 5-6.
A method of producing a collagen composition for skin lesion repair, comprising the steps of:
SS1, mixing sodium alginate, collagen, polyvinyl alcohol and glycerol with deionized water, stirring, standing and defoaming to obtain gel;
SS2, pouring the modified antibacterial cellulose and anhydrous zinc chloride into deionized water, adding glacial acetic acid to promote dissolution, uniformly stirring, standing and defoaming to obtain a cross-linking agent solution;
And SS3, pouring the gel solution into a mould, refrigerating for 1-3 hours at the temperature of 0-5 ℃, taking out, adding the cross-linking agent solution into the mould, stirring for 12-24 hours at room temperature, pouring out the material, and cleaning with deionized water to obtain the gel collagen composition.
Through the technical scheme, the sodium alginate is a natural polymer material with good non-toxicity, biodegradability and biocompatibility, is widely applied to hemostatic dressings, and the polyvinyl alcohol is a non-toxic and non-irritant hydrophilic polymer, contains a large number of hydroxyl functional groups in the structure, can absorb a large amount of water to form firm hydrogen bonds, has good film forming property, and is favorable for forming gel. The sodium alginate and the modified antibacterial cellulose both contain carboxyl groups, ionization can occur in aqueous solution to generate carboxyl anions, and the carboxyl anions can react with polyvalent metal ions to generate a three-dimensional crosslinked network of sodium alginate-Zn2+ -modified antibacterial cellulose, and collagen is wrapped in gel in the crosslinking reaction process to form the gel-like collagen composition wrapping the collagen.
The invention has the beneficial effects that:
1) According to the invention, L-arginine is introduced into a cellulose structure, so that guanidine groups in the arginine structure are introduced into the cellulose structure, wherein guanidine groups can show extremely strong electropositivity, a protonation structure can be adsorbed on the surface of electronegative bacterial cell walls to form a polymer film, so that nutrient substances are prevented from entering and metabolites are prevented from being discharged, the metabolism of bacterial microorganisms is disturbed, bacterial death is finally caused, good antibacterial performance is provided for cellulose, and the composition gel dressing prepared by crosslinking the cellulose with sodium alginate has good antibacterial performance, and bacterial infection at wounds can be effectively prevented, thereby promoting wound healing.
2) According to the invention, after L-arginine is introduced into a cellulose structure, carboxyl is generated in the cellulose structure, the principle that carboxyl anions and high-valence metal ions generate crosslinking reaction is utilized, zinc chloride is used as a crosslinking agent, so that the prepared composition gel dressing contains a large amount of zinc ions, and the zinc ions are combined with active protease in bacteria after contacting with bacteria to deactivate the active protease, so that bacteria death is caused, a synergistic antibacterial effect can be generated with guanidine groups in the cellulose structure, and the antibacterial effect of the gel dressing is further enhanced.
3) According to the invention, collagen is added in the process of preparing the gel dressing, can be combined with skin epithelial cells through a stratum corneum, participates in skin cell metabolism, enhances collagen activity in skin, assists cells in producing collagen, promotes skin cells to grow rapidly, is beneficial to leaving no scar at a wound, and is coated in a three-dimensional crosslinked network of sodium alginate-Zn2+ -modified antibacterial cellulose, so that the effect of slow release can be achieved, wound healing is facilitated, and additional pain brought to a patient by frequent dressing change is avoided.
Of course, it is not necessary for any one product to practice the invention to achieve all of the advantages set forth above at the same time.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the embodiments, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
A collagen composition for repairing skin injury comprises the following raw materials, by weight, 50 parts of sodium alginate, 30 parts of polyvinyl alcohol, 10 parts of glycerol, 5 parts of collagen, 4 parts of zinc chloride and 2 parts of modified antibacterial cellulose;
wherein the production method of the collagen composition comprises the following steps:
SS1, mixing sodium alginate, collagen, polyvinyl alcohol and glycerol with deionized water, stirring, standing and defoaming to obtain gel;
SS2, pouring the modified antibacterial cellulose and anhydrous zinc chloride into deionized water, adding glacial acetic acid to promote dissolution, uniformly stirring, standing and defoaming to obtain a cross-linking agent solution;
And SS3, pouring the gel solution into a mould, refrigerating at 0 ℃ for 1h, taking out, adding the cross-linking agent solution into the mould, stirring at room temperature for 12h, pouring out the material, and cleaning with deionized water to obtain the gel collagen composition.
Wherein the production method of the modified antibacterial cellulose comprises the following steps:
S1, dispersing 10g of hydroxyethyl cellulose in tetrahydrofuran, stirring uniformly, adding 2g of dimethyl succinate, regulating the pH of a system to 8.5 by using a sodium hydroxide solution with the volume fraction of 3%, stirring for 4 hours at the temperature of 70 ℃, cooling materials, regulating the pH of the system to be neutral by using dilute hydrochloric acid with the concentration of 0.5 mol/L, filtering, washing and drying in vacuum to prepare esterified cellulose;
S2, mixing 5g of esterified cellulose with N, N-dimethylformamide, adding 1.5g of L-arginine, regulating the pH of the system to 5 by using a hydrochloric acid solution with the concentration of 1 mol/L, stirring for 8 hours at the temperature of 50 ℃, after the reaction is finished, settling, washing the materials by using an ethanol solution with the volume fraction of 70%, and vacuum drying to prepare the modified antibacterial cellulose.
Example 2
A collagen composition for repairing skin injury comprises the following raw materials, by weight, 55 parts of sodium alginate, 35 parts of polyvinyl alcohol, 15 parts of glycerol, 8 parts of collagen, 6 parts of zinc chloride and 4 parts of modified antibacterial cellulose;
wherein the production method of the collagen composition comprises the following steps:
SS1, mixing sodium alginate, collagen, polyvinyl alcohol and glycerol with deionized water, stirring, standing and defoaming to obtain gel;
SS2, pouring the modified antibacterial cellulose and anhydrous zinc chloride into deionized water, adding glacial acetic acid to promote dissolution, uniformly stirring, standing and defoaming to obtain a cross-linking agent solution;
And SS3, pouring the gel solution into a mould, refrigerating at 5 ℃ for 2 hours, taking out, adding the cross-linking agent solution into the mould, stirring at room temperature for 16 hours, pouring out the material, and cleaning with deionized water to obtain the gel collagen composition.
Wherein the modified antibacterial cellulose was produced in the same manner as in example 1.
Example 3
A collagen composition for repairing skin injury comprises 60 parts by weight of sodium alginate, 40 parts by weight of polyvinyl alcohol, 20 parts by weight of glycerol, 10 parts by weight of collagen, 8 parts by weight of zinc chloride and 6 parts by weight of modified antibacterial cellulose;
wherein the production method of the collagen composition comprises the following steps:
SS1, mixing sodium alginate, collagen, polyvinyl alcohol and glycerol with deionized water, stirring, standing and defoaming to obtain gel;
SS2, pouring the modified antibacterial cellulose and anhydrous zinc chloride into deionized water, adding glacial acetic acid to promote dissolution, uniformly stirring, standing and defoaming to obtain a cross-linking agent solution;
and SS3, pouring the gel solution into a mould, refrigerating at 5 ℃ for 3 hours, taking out, adding the cross-linking agent solution into the mould, stirring at room temperature for 24 hours, pouring out the material, and cleaning with deionized water to obtain the gel collagen composition.
Wherein the modified antibacterial cellulose was produced in the same manner as in example 1.
Comparative example 1
A collagen composition for repairing skin injury comprises the following raw materials, by weight, 55 parts of sodium alginate, 35 parts of polyvinyl alcohol, 15 parts of glycerol, 8 parts of collagen, 6 parts of zinc chloride and 4 parts of carboxymethyl cellulose;
wherein the production method of the collagen composition comprises the following steps:
SS1, mixing sodium alginate, collagen, polyvinyl alcohol and glycerol with deionized water, stirring, standing and defoaming to obtain gel;
SS2, pouring carboxymethyl cellulose and anhydrous zinc chloride into deionized water, adding glacial acetic acid to promote dissolution, uniformly stirring, standing and defoaming to obtain a cross-linking agent solution;
And SS3, pouring the gel solution into a mould, refrigerating at 5 ℃ for 2 hours, taking out, adding the cross-linking agent solution into the mould, stirring at room temperature for 16 hours, pouring out the material, and cleaning with deionized water to obtain the gel collagen composition.
Performance detection
A. Cutting the collagen compositions prepared in the examples 1-3 and comparative example 1 into round gel samples with the diameter of 10mm and the thickness of 2mm, placing 0.3mL of staphylococcus aureus into beef broth, culturing at the constant temperature of 37 ℃ for 24 hours, diluting 10-6 times to prepare bacterial suspension to be tested, transferring 0.3mL of bacterial suspension to drop on the surface of the gel sample, culturing for 12 hours, washing the gel sample with sterilized water, sucking 0.3mL of washing liquid, continuously culturing on a culture dish for 24 hours, recording the colony number on the culture dish, and performing a blank test, wherein the formula is used
The antibacterial rate of the samples is calculated, wherein S0 is the colony number of a blank control experiment group, S1 is the colony number of each sample, P is the antibacterial rate, and the test results are as follows:
As is clear from the above table, the gel-like collagen compositions prepared in examples 1 to 3 of the present invention are excellent in antibacterial property, the average antibacterial rate is 99% or more, while the gel-like collagen composition prepared in comparative example 1 is slightly inferior in antibacterial property, presumably, after crosslinking with zinc chloride, the gel-like collagen composition contains a large amount of zinc ions, which are bound with active protease in bacteria to deactivate the protease after contacting with the bacteria, thereby causing bacterial death, but the components do not contain guanidine group synergistic antibacterial groups, resulting in slightly inferior antibacterial property.
B. Randomly selecting 50 rats with the weight of about 200g, dividing the average into 5 groups, 10 rats in each group are respectively in an example 1-an example 3 group, a comparison example 1 group and a blank control experiment group, removing hair on the back of the mice, scanning the damaged wound surface of 1cm multiplied by 1cm at the same position on the back of the mice by laser, respectively covering the gel-like collagen compositions prepared in the invention example 1-example 3 and the comparison example 1, wherein the wound surface of the mice in each group is exposed to air for 1 time per day, 30min each time, and observing the healing condition of the wound surface on the back of the mice, and the test results are as follows:
as can be seen from the above table, the gel-like collagen compositions prepared in examples 1-3 and comparative example 1 of the present invention can heal the wound surface of the mice in about 7d, and no obvious scar is generated after the wound surface is completely healed, but the gel-like collagen composition prepared in comparative example 1 has slightly weak antibacterial performance, has slight red swelling phenomenon in 3d, and the wound surface of mice in blank control group is obviously infected in 3d, and obvious scar is left after the wound surface heals, presumably, the addition of collagen can be combined with skin epithelial cells through stratum corneum, participate in skin cell metabolism, enhance collagen activity in skin, assist cells to make collagen, promote rapid growth of skin cells, and be favorable for leaving no scar at the wound.
The foregoing is merely illustrative and explanatory of the principles of the invention, as various modifications and additions may be made to the specific embodiments described, or similar thereto, by those skilled in the art, without departing from the principles of the invention or beyond the scope of the appended claims.