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目的应用表面修饰技术处理的明胶海绵与颌下腺种子细胞复合培养,研究表面修饰技术对颌下腺种子细胞在支架材料上附着、生长和分泌功能的影响。方法选用明胶海绵36块,随机分A、B、C 3组,每组12块。分别用单纯明胶海绵、层粘连蛋白表面修饰的明胶海绵材料、转化生长因子β3表面修饰的明胶海绵材料与鼠颌下腺细胞在体外复合培养。培养后第3、7、15天行组织学观察、扫描电镜观察,第15天行免疫组织化学鉴定细胞来源,测定上清淀粉酶含量。结果组织学和扫描电镜观察可见培养后第3天各组细胞分散于材料表面,无细胞突起形成;第7天B组细胞数量明显多于A组和C组,细胞突起形成并锚定于胶原海绵表面;第15天B组细胞数量仍多于A组和C组,并可见细胞之间有触突联系和腺管样结构形成。免疫组织化学染色观察复合培养的颌下腺细胞B rdU呈强阳性。随着接种后培养时间的延长,各组颌下腺细胞分泌的淀粉酶含量均有不同程度的增加。第15天C组淀粉酶含量明显高于A组和B组(P<0.05)。结论应用层粘连蛋白和转化生长因子β3对明胶海绵进行表面修饰,可促进颌下腺种子细胞在材料上的附着、增殖和分泌功能。
Objective To study the effect of surface modification on the attachment, growth and secretion of submandibular gland seed cells on scaffolds by means of composite culture of gelatin sponge and submandibular gland seed cells treated with surface modification technique. Methods 36 gelatin sponges were randomly divided into groups A, B and C 3, with 12 in each group. Gelatin sponge material modified by simple gelatin sponge and laminin surface, gelatin sponge material modified by transforming growth factor β3 and rat submandibular gland cells were cultured in vitro. On the 3rd, 7th and 15th day after culturing, the histological observation, the scanning electron microscope observation and the immunohistochemistry were used to identify the source of the cells on the 15th day. The content of amylase in the supernatant was measured. Results On the 3rd day after culture, the cells in each group were dispersed on the surface of the material without cell protrusion. On the 7th day, the number of cells in group B was more than that in group A and C, and the cells formed and anchored to collagen Sponge surface; the number of cells in group B on day 15 was still more than that of group A and group C, and there was contact between the cells and the formation of duct-like structures. B rdU was strongly positive in submandibular gland cells by immunohistochemical staining. With the extension of incubation time after inoculation, the amylase content of the submandibular gland cells in each group increased in varying degrees. On the 15th day, the content of amylase in group C was significantly higher than that in group A and B (P <0.05). Conclusion The surface modification of gelatin sponge by laminin and transforming growth factor β3 can promote the attachment, proliferation and secretion of the submandibular gland seed cells.