Department of Trauma/Joint Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Shandong, China.
PeerJ. 2022 Jul 11;10:e13722. doi: 10.7717/peerj.13722. eCollection 2022.
Osteochondral decellularization can promote local vascular regeneration, but the exact mechanism is unknown. The aim of this study is to study osteogenic microvascular regeneration in single cells.
The scRNA-seq dataset of human periosteal-derived cells (hPDCs) were analyzed by pySCENIC. To examine the role of TBX3 in osteogenesis and vascularization, cell transfection, qRT-PCR, western blot, and CCK-8 cell proliferation assays were performed.
TCF7L2, TBX3, FLI1, NFKB2, and EZH2 were found to be transcription factors (TFs) most closely associated with corresponding cells. The regulatory network of these TFs was then visualized. Our study knocked down the expression of TBX3 in human osteoblast cell lines. In the TBX3 knockdown group, we observed decreased expression of VEGFA, VEGFB, and VEGFC. Moreover, Western blot analysis showed that downregulating TBX3 resulted in a reduction of VEGFA expression. And TBX3 stimulated osteoblast proliferation in CCK-8 assays.
TBX3 regulates VEGFA expression and promotes osteoblast proliferation in skeletal microvasculature formation. The findings provide a theoretical basis for investigating the role of TBX3 in promoting local vascular regeneration.
脱细胞软骨可以促进局部血管再生，但确切机制尚不清楚。本研究旨在研究成骨微血管的单细胞再生。
通过 pySCENIC 分析人骨膜源性细胞（hPDC）的 scRNA-seq 数据集。为了研究 TBX3 在成骨和血管生成中的作用，进行了细胞转染、qRT-PCR、western blot 和 CCK-8 细胞增殖检测。
发现 TCF7L2、TBX3、FLI1、NFKB2 和 EZH2 与相应细胞最密切相关，是转录因子（TFs）。然后可视化了这些 TF 的调控网络。我们的研究敲低了人成骨细胞系中 TBX3 的表达。在 TBX3 敲低组中，我们观察到 VEGFA、VEGFB 和 VEGFC 的表达降低。此外，Western blot 分析表明，下调 TBX3 导致 VEGFA 表达减少。并且 TBX3 在 CCK-8 测定中刺激成骨细胞增殖。
TBX3 调节 VEGFA 的表达，并促进骨骼微血管形成中的成骨细胞增殖。这些发现为研究 TBX3 在促进局部血管再生中的作用提供了理论基础。
