基于分子自聚集的鱼皮胶原水凝胶的生物学性能研究

Collagen can self-assemble into hydrogel, which plays a vital role in its application in biological materials. Up to now, studies were scarce on the hydrogel formed by the self-assembly of marine collagen. How are the biological properties of marine collagen gel? Whether can it be used in the exploitation of collagen-based biomaterials? Especially, marine collagen shows poor thermal stability, then which modification method can cover the shortage and improve the gel performance? Aiming at these problems, studies are made centering on the following three aspects..(1) Fish skins of tilapia, tuna and squid, which are rich and concentrated in China, are taken as the research objects. The process of collagen self-assembling into hydrogel and the structure and biological properties of gels are investigated to determine which collagen can be used to further study..(2) The effects of cross-linking technology, blending technology and heat treatment on the structure and biological properties of hydrogels formed by fish skin collagen, especially the effects of γ-poly glutamic acid blending technology and heat treatment, are discussed, so as to obtain the new collagen hydrogel..(3) The hydrogel formed by fish skin collagen is used as the matrix to culture fibroblasts by two-dimensional and three-dimensional pattern, and its potentional is analysed as the skin wound healing material..This project will provide the foundation for the study, development and utilization of marine collagen–based biomaterials, and expand the application fields of marine collagen.

胶原能够自聚集形成水凝胶，这对其在生物材料领域的应用具有重要意义。目前水产胶原自聚集水凝胶的相关研究比较少。水产胶原水凝胶的生物学性能如何？是否可用于胶原基生物材料的研究和开发？特别是水产胶原的热稳定性较差，通过什么改性方法能够弥补不足，提高凝胶性能？针对上述问题，本项目主要围绕三个方面进行研究。.（1）以我国资源丰富并集中的罗非鱼、金枪鱼和鱿鱼的鱼皮为对象，研究其中胶原自聚集形成水凝胶的过程及凝胶的结构和生物学性能，确定后续研究所用胶原的来源。.（2）研究交联、共混和热处理技术对鱼皮胶原水凝胶结构和生物学性能的影响，特别探讨γ-聚谷氨酸共混技术和热处理技术的影响，以期能够获得新型胶原水凝胶。.（3）以鱼皮胶原水凝胶为支架通过二维和三维模式构建皮肤成纤维细胞培养系统，分析其作为皮肤创伤修复材料的潜力。.本项目将为水产胶原基生物材料的研究、开发和应用提供基础和依据，拓展水产胶原的应用领域。

鱼皮胶原通过自聚集能够形成水凝胶，以应用于生物材料等领域。但该水凝胶机械学性能和耐酶性较差，需通过改性手段来提高其生物学性能。针对这一问题，本项目研究了多糖和交联剂对自聚集鱼皮胶原纤维水凝胶性能的影响，所得结果如下：.（1）比较研究了罗非鱼皮和金枪鱼皮胶原的结构和性能，发现罗非鱼皮胶原自聚集水凝胶的性能优于金枪鱼皮胶原水凝胶。在此基础上，研究NaOH溶液前处理对罗非鱼皮胶原的脱色效果，确定了1.0 M的NaOH溶液在30 ℃下处理罗非鱼皮150 min后，鱼皮色素完全被脱除，并且胶原得率和自聚集比率未受到显著影响。.（2）罗非鱼皮胶原在无NaCl的中性磷酸缓冲液中于28 ℃下易发生自聚集。当海藻酸钠、硫酸软骨素和透明质酸被引入自聚集体系后，胶原水凝胶的结构和性能发生了显著变化。海藻酸钠可使胶原纤维网络结构更加致密，而硫酸软骨素和透明质酸分别促使大纤维束和同向排列纤维的出现，并且三种多糖均使胶原纤维的周期性增加。海藻酸钠和透明质酸使自聚集胶原水凝胶的最大凝胶强度分别增大到39.64和34.39 KN/m2，显著高于纯胶原水凝胶（14.53 KN/m2），而硫酸软骨素仅能使最大凝胶强度增大到17.20 KN/m2。然而，三种多糖对自聚集胶原水凝胶的耐酶性没有显著提高，透明质酸对其无显著影响，而海藻酸钠和硫酸软骨素却使其显著降低。综上，透明质酸改性的自聚集罗非鱼皮胶原水凝胶性能优于其它二者。.（3）EDC/NHS和京尼平对自聚集罗非鱼皮胶原纤维水凝胶表现较好的交联作用。但由于二者交联机理的差异，京尼平的交联效果远优于EDC/NHS。二者交联后，自聚集海藻酸钠/胶原纤维水凝胶和透明质酸/胶原纤维水凝胶的吸水性、保水性和耐酶性显著提高，并且该水凝胶对L929小鼠成纤维细胞没有毒性，且细胞在其上表现良好的增殖性。.上述研究结果可为罗非鱼皮胶原在生物材料领域的应用提供基础和参考。