Synergistic enhancement of ectopic bone formation by supplementation of freshly isolated marrow cells with purified MSC in collagen-chitosan hydrogel microbeads.
Synergistic enhancement of ectopic bone formation by supplementation of freshly isolated marrow cells with purified MSC in collagen-chitosan hydrogel microbeads.
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DOI：
10.3109/03008207.2015.1072519
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发表时间：
2016-11
影响因子：
2.9
通讯作者：
Stegemann JP
中科院分区：
文献类型：
--
作者：
Wise JK;Alford AI;Goldstein SA;Stegemann JP
骨髓间充质干细胞（MSC）具有向成骨细胞分化的能力，但其组织再生能力不一致。成人骨髓的骨髓单个核细胞（BMMC）部分含有多种祖细胞，可增强组织再生。本研究考察了BMMC单独使用和与纯化MSC联合使用作为骨再生细胞来源的效用。新鲜的BMMC，培养扩增的MSC，以及BMMC和MSC的组合被封装在胶原-壳聚糖水凝胶微珠中用于预培养和微创递送。将微珠在生长培养基中培养3天，然后在生长或成骨培养基中培养17天，然后皮下注射到大鼠背部。MSC在预培养中在微珠中保持存活超过17天，而一些BMMC部分是不可存活的。植入5周后，microCT和组织学显示，与单独的细胞来源相比，BMMC与MSC的补充对异位骨形成的体积产生了强烈的协同作用。仅含新鲜BMMC或仅含培养MSC的微珠在成骨培养基中培养比在生长培养基中培养的微珠导致更多的骨形成。组织学染色显示未分化样品中残留微珠基质的证据和分化样品中更高级组织重塑的指示。这些数据表明，BMMC组分可以与预分化的MSC协同作用，以增强异位骨形成。微珠系统可用于在骨再生应用中递送所需的细胞群。
Bone marrow-derived mesenchymal stem cells (MSC) can differentiate osteogenic lineages, but their tissue regeneration ability is inconsistent. The bone marrow mononuclear cell (BMMC) fraction of adult bone marrow contains a variety of progenitor cells that may potentiate tissue regeneration. This study examined the utility of BMMC, both alone and in combination with purified MSC, as a cell source for bone regeneration. Fresh BMMC, culture-expanded MSC, and a combination of BMMC and MSC were encapsulated in collagen–chitosan hydrogel microbeads for pre-culture and minimally invasive delivery. Microbeads were cultured in growth medium for 3 days, and then in either growth or osteogenic medium for 17 days prior to subcutaneous injection in the rat dorsum. MSC remained viable in microbeads over 17 days in pre-culture, while some of the BMMC fraction were nonviable. After 5 weeks of implantation, microCT and histology showed that supplementation of BMMC with MSC produced a strong synergistic effect on the volume of ectopic bone formation, compared to either cell source alone. Microbeads containing only fresh BMMC or only cultured MSC maintained in osteogenic medium resulted in more bone formation than their counterparts cultured in growth medium. Histological staining showed evidence of residual microbead matrix in undifferentiated samples and indications of more advanced tissue remodeling in differentiated samples. These data suggest that components of the BMMC fraction can act synergistically with predifferentiated MSC to potentiate ectopic bone formation. The microbead system may have utility in delivering desired cell populations in bone regeneration applications.