抗菌多糖水凝胶负载细胞治疗皮肤创伤的实验研究

Experimental study on the treatment of skin wounds with antibacterial polysaccharide hydrogel loaded cells

目的:构建负载成纤维细胞的抗菌多糖类水凝胶,探讨其对皮肤创伤的促进作用。方法:将L929成纤维细胞载入水凝胶中进行共培养1、2、3 d,采用活死细胞染色,通过共聚焦荧光显微镜观察成纤维细胞在水凝胶内的生长情况,计算细胞存活率,评价其作为细胞3D培养支架的效果。利用大肠埃希菌和金黄色葡萄球菌与水凝胶共培养检测其抗菌特性。构建大鼠背部直径2 cm全层皮肤缺损创面模型,分别注射生理盐水(A组)、海藻酸钠水凝胶(B组)、负载L929细胞的SCM水凝胶(C组)。第1、4、7天观察创面愈合情况,计算创面愈合率,采用HE染色观察组织修复效果。结果:L929细胞在水凝胶中培养3 d,细胞均可保持良好生长,存活率均超过80%。抑菌实验显示水凝胶具有良好的抑菌性能,可杀死超过75%的大肠埃希菌和金黄色葡萄球菌。动物实验显示,C组小鼠背部创面愈合在1、4 d显著优于A组(P<0.01)和B组(P<0.05)。与B组相比,C组中细胞能更快地从伤口周围组织迁移到水凝胶中,在第7天时细胞完全浸润水凝胶。结论:负载细胞的抗菌多糖水凝胶够有效促进皮肤创伤修复。

Objective:To construct an antibacterial polysaccharide hydrogel loaded with fibroblasts and explore its promoting effect on skin wounds.Methods:L929 fibroblasts were loaded into the hydrogel for co-cultivation for 1,2,and 3 days,stained with live dead cells,and the growth of fibroblasts in the hydrogel was observed through a confocal fluorescence microscope to calculate the cell survival rate for evaluating its effectiveness as a 3D cell culture scaffold.Using E.coli and S.aureus aureus co-cultivation with hydrogel to detect its antibacterial properties.A full-thickness skin defect model with a diameter of 2 cm in the back of the rat was constructed,then respectively treated with normal saline(group A),sodium alginate hydrogel(group B),and SCM hydrogel loaded with L929 cells(group C).On days 1,4,7,observe the wound healing,calculate the wound healing rate and observe the tissue repair effect by HE staining.Results:L929 cells were cultured in hydrogel for 3 days,and the cells could maintain good growth with survival rate exceeding 80%.Antibacterial experiments showed that hydrogels have good antibacterial properties and can kill more than 75%of E.coli and S.aureus.Animal experiments demonstrated that the wound healing of the back of mice in group C is significantly better than that in group A(P<0.01)and group B(P<0.05)on day 1 and day 4.Compared with group B,the cells migrated faster from the tissues around the wound into the hydrogel in group C.On day 7,the cells completely infiltrated the hydrogel.Conclusion:Cell-loaded antibacterial polysaccharide hydrogel is effective enough to promote skin wound repair.