Department of Prosthodontics, Guanghua School of Stomatology, Affiliated Stomatological Hospital, Guangdong Province Key Laboratory of Stomatology, Sun Yat-Sen University, Guangzhou, China.
J Cell Physiol. 2018 Dec;233(12):9458-9472. doi: 10.1002/jcp.26838. Epub 2018 Jul 11.
Craniofacial defects can cause morbidness. Adipose-derived stem cells (ADSCs) have shown great promise for osteogeneration and vascularization; therefore cocultures of differentiated ADSCs are explored to increase bone and vessel formation. In this study, ADSCs were induced into osteogenic ADSCs (os-ADSCs) and endothelial ADSCs (endo-ADSCs) cells, which were then cocultured in variable proportions (os-ADSCs/endo-ADSCs = 2:1, 1:1, 1:2). The os-ADSCs in a ratio of 1:1 expressed more ALP, RUNX2 and COL-I, whereas VEGF, vWF and CD31 were upregulated in the endo-ADSCs of this group. Next generation RNA sequencing (RNA-seq) was performed to evaluate the molecular mechanisms of cocultured ADSCs. The os-ADSCs and endo-ADSCs interacted with each other during osteogenic and angiogenic differentiation, especially at the ratio of 1:1, and were regulated by vascular-related genes, cell-mediated genes, bone-related genes and the transforming growth factor β signaling pathway (TGF-β), mitogen-activated protein kinase signaling pathway (MAPK) and wnt signaling pathway (Wnt). Angptl4, apoe, mmp3, bmp6, mmp13 and fgf18 were detected to be up-regulated, and cxcl12 and wnt5a were down-regulated. The results showed that the gene expression levels were consistent with that in RNA-seq. The cells were then seeded into self-assembling peptide RADA16-I scaffolds as cocultures (1:1) and monocultures (ADSCs, os-ADSCs, endo-ADSCs). The results showed that the cells of all groups grew and proliferated well on the scaffolds, and the cocultured group exhibited better osteogeneration and vascularization. In conclusion, cocultured os-ADSCs and endo-ADSCs at the ratio of 1:1 showed strong osteogenic and angiogenic differentiation. There is a great potential for osteogenesis and vascularization by 3D culturing cells in a 1:1 ratio in self-assembling peptide RADA16-I scaffolds, which requires evaluation for bone regeneration in vivo.
颅面畸形可导致病态。脂肪来源干细胞（ADSCs）在成骨和血管生成方面显示出巨大的潜力；因此，探索分化的 ADSC 共培养以增加骨和血管形成。在这项研究中，ADSCs 被诱导为成骨 ADSC（os-ADSCs）和内皮 ADSC（endo-ADSCs）细胞，然后以不同比例共培养（os-ADSCs/endo-ADSCs=2:1、1:1、1:2）。在比例为 1:1 的 os-ADSCs 中，ALP、RUNX2 和 COL-I 的表达更高，而该组中内皮 ADSC 的 VEGF、vWF 和 CD31 上调。接下来进行下一代 RNA 测序（RNA-seq）以评估共培养 ADSC 的分子机制。在成骨和血管生成分化过程中，os-ADSCs 和 endo-ADSCs 相互作用，尤其是在 1:1 的比例下，受血管相关基因、细胞介导基因、骨相关基因和转化生长因子β信号通路（TGF-β）、丝裂原激活蛋白激酶信号通路（MAPK）和 Wnt 信号通路（Wnt）调节。检测到 Angptl4、apoe、mmp3、bmp6、mmp13 和 fgf18 上调，cxcl12 和 wnt5a 下调。结果表明基因表达水平与 RNA-seq 一致。然后将细胞接种到自组装肽 RADA16-I 支架中作为共培养（1:1）和单培养（ADSCs、os-ADSCs、endo-ADSCs）。结果表明，所有组的细胞在支架上生长和增殖良好，共培养组表现出更好的成骨和血管生成。总之，比例为 1:1 的共培养 os-ADSCs 和 endo-ADSCs 表现出强烈的成骨和血管生成分化。通过在自组装肽 RADA16-I 支架中以 1:1 的比例对细胞进行 3D 培养，具有很强的成骨和血管生成潜力，需要在体内评估其骨再生能力。
