| 宋沐泽,王园园,宋文山,等.低免疫原性鳕鱼皮胶原蛋白的提取、表征及生物相容性研究[J].中国海洋药物,2025,(2):-. |
| 低免疫原性鳕鱼皮胶原蛋白的提取、表征及生物相容性研究 |
| Extraction, characterization and biocompatibility study of low immunogenic collagen from cod skin |
| 投稿时间：2023-12-08 修订日期：2024-01-31 |
| DOI： |
| 中文关键词: 胶原蛋白 端肽 提取 低免疫原性 生物相容性 |
| English Keywords:collagen telopeptide extraction low immunogenicity biocompatibility |
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| 中文摘要: |
| 目的 制备具有三螺旋结构的低免疫原性鳕鱼皮去端肽胶原蛋白(CAC),讨论海洋源胶原替代哺乳动物源胶原用于生物材料的可行性。方法 采用低温碱洗、氧化处理及限制性酶解制备得到CAC,并对其进行理化性能表征和生物相容性评价。结果 CAC的得率为45.3±1.30%；氨基酸分析和凝胶电泳分析均表明制备得到的CAC符合I型胶原蛋白特征；进一步通过傅里叶变换红外光谱检测证明CAC为保有三螺旋结构胶原；对CAC进行性能表征,其干燥失重为13.5±0.67%,酸碱度为4.73±0.16,吸湿性为10.61±0.87%；安全性方面,其DNA残留量为29.31±0.76 ng/mg,内毒素0.35±0.04 EU/mL,细胞增殖率为128.9±0.11%、无急/亚慢性全身毒性、无皮内刺激性且无免疫原性；以CAC为材料制备海绵,扫描电子显微镜(SEM)结果证实其具有疏松多孔结构,证明其作为医用材料的可行性。结论 从鳕鱼皮成功制备了CAC,为典型的I型胶原蛋白,去端肽工艺未影响其三螺旋结构,端肽去除后的胶原具有低免疫原性和良好的生物相容性,有望成为新的组织工程级胶原蛋白材料资源。 |
| English Summary: |
| Objective To extract low immunogenicity atelocollagen with triple helix structure from cod skin, identify it, and discuss the feasibility of marine collagen replacing mammalian collagen for biomaterials. Methods Atelocollagen of cod skin was extracted by alkaline washing, oxidation treatment, and restrictive enzymatic hydrolysis in low temperature, and its physicochemical properties were characterized and biocompatibility was evaluated. Results The extraction rate of CAC is 45.3±1.30%. Fourier transform infrared spectroscopy detection results indicate that the collagen has a triple helix structure. Amino acid analysis and SDS-PAGE analysis showed that CAC conforms to the characteristics of type I collagen. The loss on drying was 13.5±0.67%, pH was 4.73±0.16, hygroscopicity was 10.61±0.87%. In terms of biosafety, the DNA residue of CAC was 29.31±0.76 ng/mg, endotoxin level was 0.35±0.04 EU/mL, cell proliferation rate was 128.9±0.11%, and there was no acute/subchronic systemic toxicity, intradermal irritation, or immunogenicity. Scanning electron microscopy (SEM) results confirmed CAC has a porous structure and the feasibility of using CAC sponge as a medical material. Conclusion Successfully extracted CAC from cod skin, and the process of removing terminal peptides did not affect its triple helix structure. The collagen, after the removal of terminal peptides, exhibits low immunogenicity and good biocompatibility, making it a promising tissue engineering grade collagen material resource. |
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