Effects of a Novel Inoculation Method on Cell Distribution, Mineralization, and Vascularization of Tissue-Engineered Constructs
Effects of a Novel Inoculation Method on Cell Distribution, Mineralization, and Vascularization of Tissue-Engineered Constructs
复制标题
DOI：
10.1089/wound.2014.0613
复制
发表时间：
2016-03-01
影响因子：
4.9
通讯作者：
Li, Zubing
中科院分区：
文献类型：
--
作者：
Cheng, Gu;Li, Zhi;Li, Zubing
目的：尽管有一些潜在的优势，但由于种子细胞的不均匀分布，组织工程化构建体在移植到骨缺损部位后具有低的治疗功效，这在临床环境中呈现出主要障碍。本研究的目的是改善细胞在支架内的分布，提高接种效率，促进组织长入和血管化，最终提高移植的成功率。方法：设计了一种辅助接种的方法，通过反复正抽吸将骨髓基质细胞（BMSCs）锚定在壳聚糖支架的中心孔上。采用WST-8、CD 31免疫荧光染色和von Kossa染色，分别观察组织工程化构建体在常规双侧接种法和常规双侧接种法处理后的中心区域的细胞分布、细胞数量、血管化和矿化情况。经琼脂糖辅助接种法处理的组织工程化构建体中BMSCs数量较多且分布均匀，同时，在体内和体外实验中，与双侧接种方法相比，中央区域的血管化和矿化更加稳健。创新：这是首次评估了双接种方法对植入移植物血管化和矿化的影响。结论：本研究中使用的细胞辅助接种方法可能提供一种方法，通过该方法可以改善细胞在支架内的分布，促进随后的组织长入支架，并促进组织工程移植物的血管化和矿化。
Objective: Despite some potential advantages, tissue-engineered constructs have low therapeutic efficacy after transplantation into bone defect sites due to uneven distribution of seed cells, which presents a major obstacle in a clinical setting. The aim of this study was to improve cell distribution within the scaffold, to increase seeding efficiency, to facilitate tissue ingrowth and vascularization of the resultant grafts, and finally to boost the successful rate of transplantation.Approach: A syringe-aided inoculation method was designed to anchor bone marrow stromal cells (BMSCs) onto the central pores of the chitosan scaffold by repeat positive aspiration. The cell distribution, cell number, vascularization and mineralization at the central region of the tissue-engineered constructs treated by syringe-aided and regular two-side inoculation methods were evaluated side by side using the WST-8, immunofluorescence staining of CD31 and von Kossa staining.Results: The tissue-engineered constructs treated by the syringe-aided inoculation methods exhibited a larger number of BMSCs with uniform distribution, as well as more robust vascularization and mineralization in the central regions when compared with those treated by the two-side inoculation method both in vitro and in vivo.Innovation: This is the first time that the influence of a syringe-aided inoculation method has been evaluated with respect to the vascularization and mineralization of implanted grafts.Conclusion: The syringe-aided inoculation methods used in this study might provide an approach through which to improve cell distribution within the scaffold, facilitate the subsequent tissue ingrowth into the scaffold, and promote the vascularization and mineralization of tissue-engineered grafts.