a State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing and the Ministry of Science and Technology, College of Biomedical Engineering, Chongqing Key Laboratory of Biomedical Engineering, Chongqing Collaborative Innovation Center for Minimally-invasive and Noninvasive Medicine , Chongqing Medical University , Chongqing , P.R.China.
b The Second Affiliated Hospital of Chongqing Medical University , Chongqing , P.R.China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):674-684. doi: 10.1080/21691401.2019.1576704.
In terms to investigate the effect of low-intensity pulsed ultrasound (LIPUS) for differentiation of bone marrow mesenchymal stem cells (BMSCs) and the feasibility of simultaneously inducing into osteoblasts and vascular endothelial cells within the cell culture medium in which two inductive agents are added at the same time with or without LIPUS. Cells were divided into a non-induced group, an osteoblast-induced group, a vascular endothelial-induced group, and a bidirectional differentiation-induced group. Each group was further subdivided into LIPUS and non-LIPUS groups. The cell proliferation in each group was measured by MTT assay. Cell morphological and ultrastructural changes were observed by inverted phase contrast microscopy and transmission electron microscopy. The differentiation of BMSCs was detected by confocal microscopy, flow cytometry and quantitative RT-PCR. Results demonstrated that both osteoblast and vascular endothelial cell differentiation markers were expressed in the bidirectional differentiation induction group and early osteogenesis and angiogenesis appeared. The cell proliferation, differentiation rate and expression of osteocalcin and vWF in the LIPUS groups were all significantly higher than those in the corresponding non-LIPUS group (p < .05), suggesting LIPUS treatment can promote the differentiation efficiency and rate of BMSCs, especially in the bidirectional differentiation induction group. This study suggests the combination of LIPUS and dual-inducing agents could induce and accelerate simultaneous differentiation of BMSCs to osteoblasts and vascular endothelial cells. These findings indicate the method could be applied to research on generating vascularized bone tissue with a shape and function that mimics natural bone to accelerate early osteogenesis and angiogenesis.
为了研究低强度脉冲超声（LIPUS）对骨髓间充质干细胞（BMSCs）分化的影响，以及在同时添加两种诱导剂的细胞培养物中，即在 LIPUS 或无 LIPUS 的情况下，同时将细胞诱导成成骨细胞和血管内皮细胞的可行性。细胞分为未诱导组、成骨细胞诱导组、血管内皮诱导组和双向分化诱导组。每组进一步分为 LIPUS 和非 LIPUS 组。通过 MTT 测定法测量每组细胞的增殖。通过倒置相差显微镜和透射电子显微镜观察细胞形态和超微结构变化。通过共聚焦显微镜、流式细胞术和定量 RT-PCR 检测 BMSCs 的分化。结果表明，双向分化诱导组中均表达成骨细胞和血管内皮细胞分化标志物，早期出现成骨和血管生成。LIPUS 组的细胞增殖、分化率以及骨钙素和 vWF 的表达均明显高于相应的非 LIPUS 组（p<0.05），提示 LIPUS 处理可以促进 BMSCs 的分化效率和速率，尤其是在双向分化诱导组中。本研究表明，LIPUS 与双诱导剂的联合应用可以诱导和加速 BMSCs 向成骨细胞和血管内皮细胞的同时分化。这些发现表明，该方法可应用于研究生成具有模拟天然骨形状和功能的血管化骨组织，以加速早期成骨和血管生成。
