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Effects of Fiber Alignment and Coculture with Endothelial Cells on Osteogenic Differentiation of Mesenchymal Stromal Cells.
Tissue Engineering, Part C: Methods ( IF 3 ) Pub Date : 2019-12-27 , DOI: 10.1089/ten.tec.2019.0232
Tianyu Yao 1 , Honglin Chen 2 , Matthew B Baker 1 , Lorenzo Moroni 1
Affiliation  

Vascularization is a critical process during bone regeneration. The lack of vascular networks leads to insufficient oxygen and nutrients supply, which compromises the survival of regenerated bone. One strategy for improving the survival and osteogenesis of tissue-engineered bone grafts involves the coculture of endothelial cells (ECs) with mesenchymal stromal cells (MSCs). Moreover, bone regeneration is especially challenging due to its unique structural properties with aligned topographical cues, with which stem cells can interact. Inspired by the aligned fibrillar nanostructures in human cancellous bone, we fabricated polycaprolactone (PCL) electrospun fibers with aligned and random morphology, cocultured human MSCs with human umbilical vein ECs (HUVECs), and finally investigated how these two factors modulate osteogenic differentiation of human MSCs (hMSCs). After optimizing cell ratio, a hMSCs/HUVECs ratio (90:10) was considered to be the best combination for osteogenic differentiation. Coculture results showed that hMSCs and HUVECs adhered to and proliferated well on both scaffolds. The aligned structure of PCL fibers strongly influenced the morphology and orientation of hMSCs and HUVECs; however, fiber alignment was observed to not affect alkaline phosphate (ALP) activity or mineralization of hMSCs compared with random scaffolds. More importantly, cocultured cells on both random and aligned scaffolds had significantly higher ALP activities than monoculture groups, which indicated that coculture with HUVECs provided a larger relative contribution to the osteogenesis of hMSCs compared with fiber alignment. Taken together, we conclude that coculture of hMSCs with ECs is an effective strategy to promote osteogenesis on electrospun scaffolds, and aligned fibers could be introduced to regenerate bone tissues with oriented topography without significant deleterious effects on hMSCs differentiation. This study shows the ability to grow oriented tissue-engineered cocultures with significant increases in osteogenesis over monoculture conditions. Impact statement This work demonstrates an effective method of enhancing osteogenesis of mesenchymal stromal cells on electrospun scaffolds through coculturing with endothelial cells. Furthermore, we provide the optimized conditions for cocultures on electrospun fibrous scaffolds and engineered bone tissues with oriented topography on aligned fibers. This study demonstrates promising findings for growing oriented tissue-engineered cocultures with significant increase in osteogenesis over monoculture conditions.

中文翻译:

纤维排列和与内皮细胞共培养对间质基质细胞成骨分化的影响。

血管化是骨骼再生过程中的关键过程。缺乏血管网络会导致氧气和营养供应不足,从而损害再生骨骼的生存。改善组织工程骨移植物存活和成骨的一种策略涉及内皮细胞(EC)与间质基质细胞(MSC)的共培养。此外,由于骨再生具有独特的结构特性以及对齐的地形线索,干细胞可以与之相互作用,因此骨再生尤其具有挑战性。受人松质骨中对齐的原纤维纳米结构的启发,我们制造了具有对齐和随机形态的聚己内酯(PCL)电纺纤维,将人MSC与人脐静脉EC(HUVEC)共培养,最后研究了这两个因素如何调节人MSC(hMSC)的成骨分化。优化细胞比例后,hMSCs / HUVECs比例(90:10)被认为是成骨分化的最佳组合。共培养结果表明,hMSCs和HUVECs在两个支架上均粘附并良好增殖。PCL纤维的排列结构强烈影响hMSC和HUVEC的形态和方向。然而,与随机支架相比,观察到纤维排列不会影响碱性磷酸酶(ALP)活性或hMSC的矿化。更重要的是,在随机支架和对齐支架上共培养的细胞均具有比单培养组更高的ALP活性,这表明与纤维对齐相比,与HUVEC的共培养对hMSC的成骨作用提供了更大的相对贡献。综上所述,我们得出的结论是,hMSC与EC的共培养是促进电纺支架上成骨的有效策略,并且可以引入对齐的纤维以定向的拓扑结构再生骨骼组织,而对hMSC的分化没有明显的有害影响。这项研究表明,在单一培养条件下,具有显着提高成骨能力的定向组织工程共培养物的生长能力。影响陈述这项工作证明了通过与内皮细胞共培养来增强电纺支架上的间充质基质细胞成骨的有效方法。此外,我们为在静电纺丝纤维支架和工程骨组织上进行共培养提供了优化的条件,并在对齐的纤维上定向了地形。这项研究表明,对于定向生长的组织工程共培养物,与单一培养条件相比,成骨作用显着增加,这些结果令人鼓舞。
更新日期:2019-11-01
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