冻存对组织工程骨膜生物活性的影响
Effect of cryopreservation on bioactivity the tissue engineered periosteum
目的 观察冻存复苏过程对组织工程骨膜生物学特性的影响.方法 体外培养新西兰大白兔骨髓间充质干细胞(BMSCs),与猪小肠黏膜下层(SIS)构建组织工程骨膜.冻存后复苏培养,扫描电镜(SEM)观查复苏后种子细胞生长状况、噻唑蓝(MTT)比色法绘生长曲线、钙-钴法检测碱性磷酸酶(ALP)、生物化学法定量分析ALP的表达.结果 冻存复苏后BMSCs仍能在SIS上保持良好的生长状况;组织工程骨膜复苏后成骨诱导培养5d时ALP含量达峰值,10~15d时稳定在较低的量,与对照组比较差异有统计学意义(P<0.05). 结论组织工程骨膜冻存复苏后仍保持较稳定的生物活性.
更多Objective To analyze the changes in bioactivity of cryopreserved tissue-engineered periosteum.Methods The bone marrow mesenchymal stem cells (BMSCs) from New Zealand rabbits were cultivated as seeding cells in vitro,then co-cultured with porcine small intestinal submucosa (SIS) matrix,which served as scaffold,to construct the tissue-engineered periosteum.They were cryopreserved.The bioactivity change in cryopreserved tissue-engineered periosteum was analyzed by scan electronic microscopy (SEM),MTT and ALP detection after resuscitation.Results After resuscitation BMSCs still kept good growth status in SIS.The content of alkaline phosphatase (ALP) in the tissue engineered periosteum reached peaks when they were cultivated in medium containing osteogenic inducers at 5th day,then the ALP content maintained at the relatively high level.Conclusion The bioactivity of tissue engineered periosteum kept stable osteogenetic activity.
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