Ji Qiu-xia, Yu Xin-bo, Xu Quan-chen, Wu Hong
Department of Stomatology, the Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, Shandong Province, China.
Shanghai Kou Qiang Yi Xue. 2012 Dec;21(6):632-6.
The aim of this investigation was to evaluate the cytocompatibility of an in situ chitosan-quaternized chitosan/α, β-glycerophosphate (CS-HTCC/GP) thermosensitive hydrogel in vitro.
The primary cells were isolated from human periodontal ligament and cultured. The role of different concentrations of CS-HTCC/GP extract to HPDLCs was evaluated by MTT assay and alkaline phosphatase (ALP) activity. Also, the ultra-architecture of HPDLCs was determined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) respectively. SPSS13.0 software package was used for statistical analysis.
By immunocytochemical method, the cells were stained positively to antibodies against vimentin, and negatively to antibodies against cytokeratin, which indicated that they were external embryo mesenchymal cell without epithelial cell mixure. CS-HTCC/GP thermosensitive hydrogel promoted proliferation of HPDLCs,especially at 3d and 5d, the results was significantly different (P<0.001). ALP activity was significantly greater in group 2 and 3 than in group 4 after 5d (P<0.001). Also, no negative influence to ultrastructure of HPDLCs was found through SEM and TEM.
The results indicate that CS-HTCC/GP thermosensitive hydrogel exhibits excellent cytocompatibility and has potential to be used as an in situ injectable local periodontal drug delivery vehicle and a tissue-engineering scaffold for periodontal disease therapy.
本研究旨在体外评价原位壳聚糖-季铵化壳聚糖/α,β-甘油磷酸酯(CS-HTCC/GP)热敏水凝胶的细胞相容性。
从人牙周膜分离原代细胞并进行培养。通过MTT法和碱性磷酸酶(ALP)活性评估不同浓度的CS-HTCC/GP提取物对人牙周膜细胞(HPDLCs)的作用。此外,分别通过扫描电子显微镜(SEM)和透射电子显微镜(TEM)观察HPDLCs的超微结构。使用SPSS13.0软件包进行统计分析。
免疫细胞化学方法显示,细胞波形蛋白抗体染色呈阳性,细胞角蛋白抗体染色呈阴性,表明它们是外胚层间充质细胞,无上皮细胞混合。CS-HTCC/GP热敏水凝胶促进HPDLCs增殖,尤其是在第3天和第5天,结果有显著差异(P<0.001)。5天后,第2组和第3组的ALP活性明显高于第4组(P<0.001)。此外,通过SEM和TEM未发现对HPDLCs超微结构有负面影响。
结果表明,CS-HTCC/GP热敏水凝胶具有优异的细胞相容性,有潜力用作原位注射局部牙周药物递送载体和用于牙周疾病治疗的组织工程支架。