Lohberger Birgit, Kaltenegger Heike, Stuendl Nicole, Payer Michael, Rinner Beate, Leithner Andreas
Department of Orthopaedic Surgery, Medical University of Graz, Auenbruggerplatz 5, 8036 Graz, Austria.
Department of Oral Surgery and Radiology, School of Dental Medicine, Medical University of Graz, Auenbruggerplatz 5, 8036 Graz, Austria.
Biomed Res Int. 2014;2014:189516. doi: 10.1155/2014/189516. Epub 2014 Apr 7.
We evaluated the effects of mechanical stimulation on the osteogenic differentiation of human intraoral mesenchymal stem and progenitor cells (MSPCs) using the Flexcell FX5K Tension System that mediated cyclic tensile stretch on the cells. MSPCs were isolated from human mandibular retromolar bones and characterized using flow cytometry. The positive expression of CD73, CD90, and CD105 and negativity for CD14, CD19, CD34, CD45, and HLA-DR confirmed the MSPC phenotype. Mean MSPC doubling time was 30.4 ± 2.1 hrs. The percentage of lactate dehydrogenase (LDH) release showed no significant difference between the mechanically stimulated groups and the unstimulated controls. Reverse transcription quantitative real-time PCR revealed that 10% continuous cyclic strain (0.5 Hz) for 7 and 14 days induced a significant increase in the mRNA expression of the osteogenesis-specific markers type-I collagen (Col1A1), osteonectin (SPARC), bone morphogenetic protein 2 (BMP2), osteopontin (SPP1), and osteocalcin (BGLAP) in osteogenic differentiated MSPCs. Furthermore, mechanically stimulated groups produced significantly higher amounts of calcium deposited into the cultures and alkaline phosphatase (ALP). These results will contribute to a better understanding of strain-induced bone remodelling and will form the basis for the correct choice of applied force in oral and maxillofacial surgery.
我们使用Flexcell FX5K张力系统评估了机械刺激对人口腔间充质干祖细胞(MSPCs)成骨分化的影响,该系统可介导细胞的周期性拉伸。MSPCs从人下颌第三磨牙骨中分离出来,并通过流式细胞术进行鉴定。CD73、CD90和CD105的阳性表达以及CD14、CD19、CD34、CD45和HLA-DR的阴性表达证实了MSPC的表型。MSPC的平均倍增时间为30.4±2.1小时。乳酸脱氢酶(LDH)释放百分比在机械刺激组和未刺激对照组之间无显著差异。逆转录定量实时PCR显示,在成骨分化的MSPCs中,10%的连续循环应变(0.5 Hz)持续7天和14天可显著增加成骨特异性标志物I型胶原蛋白(ColA))、骨连接蛋白(SPARC)、骨形态发生蛋白2(BMP2)、骨桥蛋白(SPP1)和骨钙素(BGLAP)的mRNA表达。此外,机械刺激组产生的沉积在培养物中的钙和碱性磷酸酶(ALP)量显著更高。这些结果将有助于更好地理解应变诱导的骨重塑,并将为口腔颌面外科中正确选择施加力奠定基础。