人牙周膜细胞复合壳聚糖-胶原温敏水凝胶异种移植构建可注射骨组织的研究

Transplantation of Human Periodontal Ligament Cells Embedded Thermosensitive Chitosan-collagen Hydrogel in an Immunocompetent Xenogeneic Animal Model

目的观察人牙周膜细胞复合壳聚糖-胶原温敏水凝胶异种移植小鼠皮下的组织再生,评价其作为可注射骨组织的可行性。方法将壳聚糖、胶原、β-磷酸甘油钠按比例混合构建温敏性水凝胶。通过活/死细胞染色法检测人牙周膜细胞(periodontal ligament cells,PDLCs)在壳聚糖-胶原水凝胶三维培养中的存活。PDLCs成骨预诱导后复合水凝胶注入小鼠背部皮下,6周后取出植入物,通过HE、Masson染色法观察组织的再生以及胶原纤维的形成。结果壳聚糖-胶原水凝胶可支持PDLCs的三维培养,体内实验表明无刺激性。复合细胞后的凝胶支架在动物皮下可形成血管化包块,结缔组织和胶原纤维形成明显高于无细胞的对照组。结论壳聚糖-胶原温敏水凝胶是一种理想的可注射支架,复合PDLCs有望构建可注射骨组织,修复骨缺损。

Objective To access the potential application of human periodontal ligament cells （PDLCs） embedded thermosensitive chitosan - collagen hydrogel for injectable bone tissue engineering. Methods Live/dead staining was used to access the survival of the PDLCs cultured in the hydrogel scaffold. Osteo - induced PDLCs embedded hydrogel was injected in the subcutaneous dorsum of ICR mice. After 6 weeks the implants were separated for HE and Masson staining. Results Most PDL cells can survive in the hydrogel scaf- fold after 3D culture. Cell - embeded hydrogel formed vascularized implants under the skin. Histological staining showed that increasing connective tissue and collagen fiber formation in the implants. Conclusion The thermosensitive chitosan - collagen hydrogel might be an ideal injectable scaffold. Combining it with PDLCs is promising to injectable tissue engineering for bone regeneration.