CN-122005954-A - Antioxidant and antibacterial sexual collagen repair film

CN122005954ACN 122005954 ACN122005954 ACN 122005954ACN-122005954-A

Abstract

The invention discloses an antioxidant and antibacterial collagen repair film and a preparation method thereof. The repair membrane is prepared by sequentially carrying out polylysine grafting modification and synchronous covalent grafting on catechol and aminoglycoside antibiotics on the purified collagen membrane. By controlling parameters such as polylysine molecular weight and concentration, functional molecular proportion and load, reaction conditions and the like, the precise regulation and control of film thickness, functional release rate, antioxidation and antibacterial performance can be realized. The invention also provides a double-layer composite membrane structure, which is used for compositing the rapid antibacterial layer and the long-acting antioxidant layer to realize functional decoupling and time sequence coordination. The repairing film has excellent oxidation resistance and broad-spectrum antibacterial property, good biocompatibility and adjustable degradation rate, can be customized according to different clinical requirements, and is suitable for repairing oral wounds, regenerating periodontal guiding tissues, inducing bone tissues to regenerate, preventing periimplantitis and treating chronic refractory wounds.

Inventors

GAO HONGYU
WANG YULU
Cheng Shuimiao
ZHANG YUFU
SHAN YANGANG
ZHANG SHUO
DING JIYUAN

Assignees

山东博达医疗用品股份有限公司

Dates

Publication Date: 20260512
Application Date: 20260228

Claims (10)

1. An antioxidant and antibacterial collagen repair membrane, which is characterized in that the membrane preparation method comprises the following steps: (1) Sequentially washing animal head skin with water, deashing, softening, oxidizing, nuclease treating, freeze drying, and shaving to obtain purified collagen film with thickness of 0.2-0.5 mm; (2) Immersing the purified collagen membrane obtained in the step (1) into a polylysine solution with the concentration of 0.1% -5%, regulating the pH to 5.5-6.0, then adding 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide to ensure that the concentration of the purified collagen membrane in the solution is 2-20 mg/mL, reacting at 4-8 ℃ for 30-240 min, adding glycine to the final concentration of 0.1% -2.5%, continuously reacting for 20-60 min, washing, and freeze-drying to obtain the polylysine modified collagen membrane; (3) Immersing the polylysine modified collagen membrane obtained in the step (2) into a mixed solution containing catechol and aminoglycoside antibiotics, wherein the mass ratio of the catechol to the aminoglycoside antibiotics in the mixed solution is 1:2-2:1, the total concentration is 5-50 mg/mL, then adding 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide to make the concentrations of the 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and the N-hydroxysuccinimide be 2-20 mg/mL and 1-10 mg/mL respectively, reacting at 4-25 ℃ for 4-24 h ℃, taking out the membrane after the reaction is finished, dialyzing for 24-72 h, freeze-drying, and forming to obtain the collagen repair membrane with antioxidant and antibacterial properties.
2. The antioxidant and antibacterial collagen repair film according to claim 1, wherein the animal's scalp in step (1) is selected from the group consisting of bovine, porcine, ovine and deer's scalp.
3. The antioxidant and antibacterial collagen repair film according to claim 1, wherein the oxidation treatment in the step (1) uses a hydrogen peroxide solution with a concentration of 1.5% -3%, and the soaking time is 2-10 min.
4. The antioxidant and antibacterial collagen repair membrane according to claim 1, wherein the nuclease treatment in step (1) uses dnase I and rnase a, the final concentration of dnase I is 5-100U/mL, the final concentration of rnase a is 1-20U/mL, and the treatment time is 1-2 h.
5. The antioxidant and antibacterial collagen repair film according to claim 1, wherein the molecular weight of the polylysine in step (2) is 1000-70000 Da.
6. The antioxidant and antibacterial collagen repair membrane of claim 1, wherein the mass ratio of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride to N-hydroxysuccinimide in step (2) is 5:1-1:5.
7. The antioxidant and antibacterial collagen repair membrane according to claim 1, wherein the aminoglycoside antibiotic in step (3) is selected from one or more of tobramycin, gentamicin sulfate, neomycin sulfate, netilmicin sulfate, and arbekacin sulfate.
8. The antioxidant and antibacterial collagen repair film according to claim 1, wherein the mixed solution in step (3) further contains citric acid or carnosine.
9. The antioxidant and antibacterial collagen repair membrane according to claim 1, wherein the dialysis in step (3) uses a dialysis bag having a molecular weight cut-off of 3500-14000 Da, and a dialysis time of 24-72 h.
10. The antioxidant and antibacterial collagen repair film according to claim 1, wherein the freeze-drying time in step (3) is 20-48 h.

Description

Antioxidant and antibacterial sexual collagen repair film Technical Field The invention relates to the technical field of tissue repair, in particular to an antioxidant and antibacterial collagen repair film. Background Collagen is widely used in clinical scenes such as oral cavity repair films, guided tissue regeneration and the like due to excellent biocompatibility and degradability, however, the existing collagen films generally lack inherent antibacterial and antioxidant activities, and wound infection, premature degradation of films and delayed healing are easy to occur in complex microenvironments of the oral cavity. Aiming at the defects, the prior art is mainly divided into two types, namely, the method is simple and convenient to operate and has the inherent defects of drug burst, short effective concentration maintaining time, easy induction of drug resistance and the like by physical adsorption or blending of antibiotics, and has poor function durability, and the method can improve the function stability by covalent fixation of an antibacterial agent or an antioxidant on a collagen network through chemical grafting, but the prior reports are concentrated on single function modification and lack the synchronous intervention capability on dual pathological processes of infection and oxidative stress. Few researches try to load antibacterial and antioxidant simultaneously, but the antibacterial and antioxidant are mostly physically mixed or independently grafted, the antibacterial and antioxidant activities are simply added at molecular level space separation and no interaction, all functional molecules in a single-layer membrane share the same cross-linked network, release dynamics are governed by a single system, and the quick release requirement of the antibacterial agent and the long-acting slow release requirement of the antioxidant cannot be independently regulated, so that the functions are seriously mismatched with a wound healing time axis. In addition, the inherent amino density of collagen is limited, the surface available ratio is less than 30%, the theoretical ceiling exists in the maximum grafting amount of functional molecules by directly taking the collagen as a reaction substrate, the bottleneck cannot be broken through by simply increasing the feeding amount or prolonging the reaction time, and high load is inevitably accompanied with high crosslinking, high rigidity and low biocompatibility. The same formula is difficult to adapt to the differentiated requirements of different patient subgroups (such as ultra-thin and soft children, high load and long-acting severe infection, strong antioxidation for diabetes, low renal toxicity for renal insufficiency and the like), so that the technical scheme and clinical accurate medical rupture are caused. In summary, the core bottleneck faced by the prior art can be attributed to the technical defects of three layers, namely, the lack of a multifunctional synergistic chemical platform on the molecular design layer, incapability of realizing synchronous, controllable-proportion and molecular-level adjacent covalent grafting of catechol and aminoglycoside antibiotics and triggering a conformational interaction mechanism, the lack of an active regulation and control means of reactive site density on the interface engineering layer, the inherent amino density of collagen becomes an insurmountable physical barrier for high-load grafting, the lack of a physical framework for functional decoupling on the structural engineering layer, and the single-layer membrane cannot simultaneously satisfy the time sequence contradiction of 'quick release' and 'slow release' in principle. Therefore, how to realize the precise regulation and control of the antioxidation/antibiosis function of the collagen membrane, and cover the differentiated requirements of multiple clinical scenes in the oral cavity and wound repair with a single technical system, becomes a key technical problem to be broken through in the field. Disclosure of Invention The embodiment of the application solves the problems in the prior art by providing the collagen repair film with oxidation resistance and antibacterial property The embodiment of the application provides a preparation method of an antioxidant and antibacterial collagen repair film, which comprises the following steps: (1) Sequentially washing animal head skin with water, deashing, softening, oxidizing, nuclease treating, freeze drying, and shaving to obtain purified collagen film with thickness of 0.2-0.5 mm; (2) Immersing the purified collagen membrane obtained in the step (1) into a polylysine solution with the concentration of 0.1% -5%, regulating the pH to 5.5-6.0, then adding 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide to ensure that the concentration of the purified collagen membrane in the solution is 2-20 mg/mL, reacting at 4-8 ℃ for 30-240 min, adding glycine to the final co