Exosomes from conditioned media of bone marrow-derived mesenchymal stem cells promote bone regeneration by enhancing angiogenesis
Exosomes from conditioned media of bone marrow-derived mesenchymal stem cells promote bone regeneration by enhancing angiogenesis
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DOI：
10.1371/journal.pone.0225472
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发表时间：
2019-11-21
期刊：
影响因子：
3.7
通讯作者：
Kobayashi, Tadaharu
中科院分区：
文献类型：
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作者：
Takeuchi, Ryoko;Katagiri, Wataru;Kobayashi, Tadaharu
来自人骨髓间充质干细胞（MSC-CM）的无血清条件培养基中的生长因子已知在骨再生中有效。然而，MSC-CM中作为骨再生活性成分的分泌体及其机制仍不清楚。外泌体作为MSC-CM的组成部分，为受体细胞提供遗传信息，并增强受体细胞旁分泌刺激，从而有助于组织再生。我们假设MSC-CM衍生的外泌体（MSC-Exo）促进骨再生，血管生成是关键步骤。在此，我们制备了MSC-Exo组、MSC-CM组和Exo-抗VEGF组（MSC-Exo+血管生成抑制剂），并检测了MSC的成骨和血管生成潜力。此外，我们使用了大鼠颅骨骨缺损模型，并植入每个样品，每周评估骨形成，直到治疗后第4周。结果显示，与其他组相比，MSC-Exo增强了MSCs的细胞迁移和成骨及血管生成基因的表达。在体内，也证实了MSC-Exo的早期骨形成。植入两周后，MSC-Exo组的新形成骨面积为31.5 +/- 6.5%，而对照组和Exo-抗VEGF组的新形成骨面积分别为15.4 +/- 4.4%和8.7 +/-1.1%。植入4周后，MSC-Exo组与Exo-antiVEGF组或对照组之间的面积差异进一步扩大。组织学观察，MSC-Exo组成骨样细胞和血管内皮细胞明显增多，而Exo-antiVEGF组和对照组成骨样细胞和血管内皮细胞明显减少。考虑到移植细胞及其分泌物的组织再生，本研究表明，外泌体可能发挥重要作用，特别是在血管生成中。
Growth factors in serum-free conditioned media from human bone marrow-derived mesenchymal stem cells (MSC-CM) are known to be effective in bone regeneration. However, the secretomes in MSC-CM that act as active ingredients for bone regeneration, as well as their mechanisms, remains unclear. Exosomes, components of MSC-CM, provide the recipient cells with genetic information and enhance the recipient cellular paracrine stimulation, which contributes to tissue regeneration. We hypothesized that MSC-CM-derived exosomes (MSC-Exo) promoted bone regeneration, and that angiogenesis was a key step. Here, we prepared an MSC-Exo group, MSC-CM group, and Exo-antiVEGF group (MSC-Exo with angiogenesis inhibitor), and examined the osteogenic and angiogenic potential in MSCs. Furthermore, we used a rat model of calvaria bone defect and implanted each sample to evaluate bone formation weekly, until week 4 after treatment. Results showed that MSC-Exo enhanced cellular migration and osteogenic and angiogenic gene expression in MSCs compared to that in other groups. In vivo, early bone formation by MSC-Exo was also confirmed. Two weeks after implantation, the newly formed bone area was 31.5 +/- 6.5% in the MSC-Exo group while those in the control and Exo-antiVEGF groups were 15.4 +/- 4.4% and 8.7 +/- 1.1%, respectively. Four weeks after implantation, differences in the area between the MSC-Exo group and the Exo-antiVEGF or control groups were further broadened. Histologically, notable accumulation of osteoblast-like cells and vascular endothelial cells was observed in the MSC-Exo group; however, fewer cells were found in the Exo-antiVEGF and control groups.In conclusion, MSC-Exo promoted bone regeneration during early stages, as well as enhanced angiogenesis. Considering the tissue regeneration with transplanted cells and their secretomes, this study suggests that exosomes might play an important role, especially in angiogenesis.