Differences between in vitro viability and differentiation and in vivo bone-forming efficacy of human mesenchymal stem cells cultured on PCL-TCP scaffolds
Differences between in vitro viability and differentiation and in vivo bone-forming efficacy of human mesenchymal stem cells cultured on PCL-TCP scaffolds
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DOI：
10.1016/j.biomaterials.2010.07.001
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发表时间：
2010-11-01
期刊：
影响因子：
14
通讯作者：
Cool, Simon M.
中科院分区：
文献类型：
--
作者：
Rai, Bina;Lin, Jane L.;Cool, Simon M.
人骨髓间充质干细胞（humanmesenchymalstemcells，hMSCs）在治疗骨疾病和骨折不愈合方面具有巨大的潜力。然而，太多时候，hMSC和所选生物材料之间相互作用的体外证据单独被用作继续开发材料进入临床的理由。显然，为了使基于hMSC的再生医学成功治疗骨科创伤，至关重要的是用合适的载体移植hMSC，以促进其在体外和体内的存活、最佳增殖和成骨分化。这促使我们评估使用的聚己内酯-20%磷酸三钙（PCL-TCP）的融合沉积建模生产的支架的hMSCs的交付。当hMSCs培养的PCL-TCP支架和成像相衬，扫描电子显微镜和激光共聚焦显微镜的组合，我们观察到五个不同的阶段的殖民超过21天的时间内，其特征在于细胞附着，扩展，细胞桥接，形成致密的细胞质量和矿化的细胞外基质的积累时，诱导成骨刺激剂。已经确定PCL-TCP支架能够支持hMSC增殖和成骨分化，我们接下来测试了负载hMSC的PCL-TCP支架在裸大鼠临界尺寸股骨缺损中的体内功效。我们发现荧光标记的hMSCs在移植后的缺损部位存活长达3周。然而，只有50%的hMSCs治疗的股骨缺损的反应良好，确定新的骨体积。因此，我们表明，hMSC的活力和分化在体外的验证是不足以预测移植的干细胞，以一贯促进骨形成原位缺损在体内的功效。(c)2010爱思唯尔有限公司版权所有。
Human mesenchymal stem cells (hMSCs) possess great therapeutic potential for the treatment of bone disease and fracture non-union. Too often however, in vitro evidence alone of the interaction between hMSCs and the biomaterial of choice is used as justification for continued development of the material into the clinic. Clearly for hMSC-based regenerative medicine to be successful for the treatment of orthopaedic trauma, it is crucial to transplant hMSCs with a suitable carrier that facilitates their survival, optimal proliferation and osteogenic differentiation in vitro and in vivo. This motivated us to evaluate the use of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds produced by fused deposition modeling for the delivery of hMSCs. When hMSCs were cultured on the PCL-TCP scaffolds and imaged by a combination of phase contrast, scanning electron and confocal laser microscopy, we observed five distinct stages of colonization over a 21-day period that were characterized by cell attachment, spreading, cellular bridging, the formation of a dense cellular mass and the accumulation of a mineralized extracellular matrix when induced with osteogenic stimulants. Having established that PCL-TCP scaffolds are able to support hMSC proliferation and osteogenic differentiation, we next tested the in vivo efficacy of hMSC-loaded PCL-TCP scaffolds in nude rat critical-sized femoral defects. We found that fluorescently labeled hMSCs survived in the defect site for up to 3 weeks post-transplantation. However, only 50% of the femoral defects treated with hMSCs responded favorably as determined by new bone volume. As such, we show that verification of hMSC viability and differentiation in vitro is not sufficient to predict the efficacy of transplanted stem cells to consistently promote bone formation in orthotopic defects in vivo. (c) 2010 Elsevier Ltd. All rights reserved.