中药白芨/聚乙烯醇纳米静电纺丝膜的制备和生物相容性评价

Preparation of electrospinning nanofibers membrane of Bletilla striata extraction/polyvinyl alcohol and its biocompatibility evaluation

目的采用静电纺丝法制备白芨/聚乙烯醇复合纳米静电纺丝膜,研究其微观形貌,评价其生物活性和生物相容性。方法采用无针静电纺丝技术,以100目细筛白芨粉溶液和聚乙烯醇为主要原料,制备中药白芨和聚乙烯醇纳米静电纺丝膜。用扫描电子显微镜表征纳米纤维,细胞活死荧光染色和计数实验评价其活性,流式检测细胞周期实验评价生物相容性。结果通过优化工艺参数,最终制备了直径为800 nm的均匀纳米纤维膜。最优参数为转速3 r/min、采集距离20 cm、外加电压40 kV。扫描电子显微镜下该纳米纤维表面光滑且形貌均一;细胞活性荧光染色和细胞计数实验表明细胞存活率提高至93%。采用流式细胞术测量了HEK293细胞在有无中药白芨和聚乙烯醇纳米静电纺丝膜干预情况下的细胞周期,结果显示在此两种情况下HEK293细胞的细胞周期相同,生物相容性良好。结论疏水性中药白芨通过优化工艺参数,和聚乙烯醇混合,可以大规模制备成纳米静电纺丝膜,可提高附着细胞的活性,生物相容性好。此法拓宽了传统中药白芨在外科生物医用领域的应用,其促进创面愈合、组织修复等功能。

Objective To study the microscopic morphology of Bletilla striata/polyvinyl alcohol composite electrospinning nanofibers membrane made by electrospinning technology,and to evaluate its biological activity and biocompatibility.Methods Using needle-free electrospinning technology,with 100 mesh fine sieve Bletilla striata powder solution and polyvinyl alcohol as the main raw materials,the traditional Chinese medicine Bletilla striata and polyvinyl alcohol nano-electrospun sheet were prepared.The nanofibers were characterized by scanning electron microscopy,the viability was evaluated by fluorescence staining and counting,and the biocompatibility was evaluated by flow cytometry.Results By optimizing parameters,we successfully fabricated surface coating sheet with homogenous nanofibers at diameter of 800 nm.The optimal working condition was at a rotating rate of 3 r/min,a collection distance of 20 cm and an applied voltage of 40 kV.The surface of the nanofibers was smooth and uniform under scanning electron microscope.The results of fluorescence staining and cell counting showed that the survival rate of cells increased to 93%.Flow cytometry was used to measure the cell cycle of HEK293 cells with or without the intervention of traditional Chinese medicine Bletilla striata and polyvinyl alcohol nanofibers and the results showed that the cell cycle of HEK293 cells was the same in the two cases,both with good biocompatibility.Conclusion The hydrophobic traditional Chinese medicine Bletilla striata,mixed with polyvinyl alcohol,can be made into nano electrospinning membrane on a large scale by optimizing parameters,which can enhance the viability and biocompatibility of the attached cells.The application of traditional Chinese medicine Bletilla striata in the field of surgical biology and medicine has been widened,and its functions of promoting wound healing and tissue repair are the research directions in the future.