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Title:: Preparation of silk fibron/chitosan/artemisia subsinensis seed hydrogel with surface composite micro/nano topologies and its effect on Schwann cells

作者:: 韩琦1; 2#; 梁如宇3#; 张李玲1; 2; 张鲁中1; 2; 吴红1; 2; 杨宇民1; 2△; 李贵才1; 2△; 1.南通大学神经再生江苏省和教育部重点实验室，南通 226001;2.南通大学神经再生协同创新中心，南通 226001;3.南通大学生命科学学院，南通 226001

Author(s):: HAN Qi1; 2; LIANG Ruyu3; ZHANG Liling1; 2; ZHANG Luzhong1; 2; WU Hong1; 2; YANG Yumin1; 2; LI Guicai1; 2; 1. Key Laboratory of Neural Regeneration, Jiangsu Province and Ministry of Education, Nantong University,Nantong 226001,China；2. Co-innovation Center of Neuroregeneration,Nantong University, Nantong 226001;3. School of Life Science, Nantong University,Nantong 226001

Keywords:: Peripheral nerve injury; Repair; Hydrogel; Micro/nano patterning; Schwann cells

摘要:: 本研究构建了表面具有复合微/纳拓扑结构的丝素/壳聚糖/沙蒿籽胶（SF/CS/AS）水凝胶，并初步探讨该复合水凝胶对雪旺细胞生长行为的调控作用。配制沙蒿籽胶溶液，筛选最优条件，利用微模塑技术，将具复合微/纳拓扑结构的PDMS印章压印到水凝胶上，制成具有微图形结构的SF/CS/AS水凝胶膜。脱酸处理后，培养雪旺细胞，评价微/纳拓扑结构对雪旺细胞的粘附、铺展等行为的调控作用。结果表明，通过联合采用溶胶凝胶技术及微模塑技术可以构建表面具有复合微/纳拓扑结构的SF/CS/AS水凝胶膜，光学显微镜可以观察到不同宽度的微/纳米凹槽，通过接种成纤维细胞证明该水凝胶膜无细胞毒性，通过接种雪旺细胞，可以发现雪旺细胞能够在具有微图形结构的复合水凝胶膜上很好地生长，并且对雪旺细胞的生长取向具有明显引导作用。该研究的开展有望为微/纳拓扑化人工神经移植物修复周围神经损伤提供实验依据。

Abstract:: We constructed a silk fibroin/chitosan/artemisia subsinensis seed (SF/CS/AS) hydrogel with a composite micro/nano topology on the surface, and preliminary explored the regulation effect of the composite hydrogel on the growth behavior of Schwann cells. The artemisia subsinensis seed gum solution was firstly prepared，the optimal conditions were obtained. After that, the micro-molding technology was used to imprint the PDMS stamps with a composite micro/nano topology on the hydrogel to fabricate SF/CS/AS film with micro-graphic structure. After deacidification, Schwann cells were cultured to evaluate the regulatory effect of micro/nano topology on the adhesion and spread of Schwann cells. The results showed that SF/CS/AS hydrogel film with composite micro/nano topology could be constructed by combining sol-gel technology and micro-molding technology, and micro/nano grooves with different widths could be observed through optical microscope. Fibroblasts culture showed that the hydrogel membrane was non-cytotoxic, while the results of Schwann cells exhibited that Schwann cells could grow well on the composite hydrogel membrane with micro-pattern structure, and the growth orientation had a certain guiding effect. The development of our research is expected to provide an experimental basis for the application of micro-nano topographic implants in peripheral nerve regeneration.