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目的观察矿化诱导培养能否促进冻存骨髓基质细胞(BMSCs)在裸鼠体内形成Ⅰ型胶原,为牙周组织工程中选择适宜的种子细胞的保存和培养条件提供参考。方法体外分离培养Beagle犬BMSCs,经冻存12个月后,在矿化诱导培养液(矿化诱导组)或基础培养液(非矿化诱导组)中形成BMSCs-胶原膜支架复合物,将矿化诱导培养液处理的单纯胶原膜支架(空白对照组)、2种BMSCs-胶原膜复合物分别植入裸鼠背部皮下组织中,术后第4周行组织病理学和免疫组织化学观察,分析各组标本中的Ⅰ型胶原形成量。结果空白对照组植入胶原膜后,新生胶原分布很少;非矿化诱导组胶原分布明显增多;矿化诱导组中见大量新生的胶原形成。免疫组织化学光密度测量显示,矿化诱导组(0.1963±0.0126)>非矿化诱导组(0.1489±0.0037)>空白对照组(0.0775±0.0058)(P<0.05)。结论矿化诱导培养能够促进冻存的BMSCs在裸鼠体内形成Ⅰ型胶原。
Objective To investigate whether mineralized induction can promote the formation of type Ⅰ collagen in nude mice by cryopreserved bone marrow stromal cells (BMSCs), and provide references for the selection of appropriate seed cells for periodontal tissue engineering. Methods Beagle dog BMSCs were isolated and cultured in vitro. After 12 months of storage, BMSCs-collagen membrane scaffold complex was formed in mineralized induction medium (mineralization induction group) or basal medium (non-mineralization induction group) The bone marrow-derived collagen membrane scaffolds (blank control group) treated with mineralization-induced culture medium were implanted into the subcutaneous tissue of the back of nude mice respectively. The histopathological and immunohistochemical observation of the two BMSCs- The amount of type I collagen in each group was analyzed. RESULTS: After the collagen membrane was inserted into the blank control group, the distribution of new collagen was very little. The distribution of collagen in non-mineralized group was significantly increased. A large number of nascent collagen was formed in the mineralized group. Immunohistochemical densitometry showed that there was no significant difference between mineralization induction group (0.1963 ± 0.0126) and non-mineralization induction group (0.1489 ± 0.0037)> blank control group (0.0775 ± 0.0058) (P <0.05). Conclusion Mineralized induction culture can promote the formation of type Ⅰ collagen in nude mice by cryopreserved BMSCs.