Schwarting Tim, Schenk Dano, Frink Michael, Benölken Michael, Steindor Friedrich, Oswald Martin, Ruchholtz Steffen, Lechler Philipp
a Center for Orthopaedics and Trauma Surgery , University of Giessen and Marburg , Marburg , Germany.
Connect Tissue Res. 2016;57(2):99-112. doi: 10.3109/03008207.2015.1087516. Epub 2015 Nov 11.
Preclinical studies have reported that bone morphogenetic protein (BMP)-2 promotes bone-tendon healing following anterior cruciate ligament reconstruction. We examined the region-specific effects of BMP-2 on osteoblast and fibroblast differentiation in a highly standardized murine in vitro co-culture model of bone-tendon integration.
We used quantitative PCR to measure the dose- and time-dependent influence of BMP-2 on the expression of alkaline phosphatase, osteocalcin, collagen type 1 (alpha 1 chain), runt-related transcription factor 2, osteopontin, collagen type 1 (alpha 2 chain), collagen type 5 (alpha 1 chain), decorin, fibromodulin, mohawk homeobox, bone morphogenetic protein receptor, type 1A, bone morphogenetic protein receptor, type 2, and Noggin in the osteoblast, interface, and fibroblast regions of a co-culture model of the murine preosteoblast cell line MC3T3-E1 and the fibroblast cell line 3T6.
Stimulation with BMP-2 resulted in a significant upregulation of alkaline phosphatase (p < 0.001), osteocalcin (p < 0.001), collagens (p < 0.001), runt-related transcription factor 2 (p < 0.05), and osteopontin (p < 0.001) expression in the osteoblast region. In the interface region, BMP-2 exposure led to dose- and time-dependent upregulation of alkaline phosphatase (p < 0.001), osteocalcin (p < 0.001), osteopontin (p < 0.001), runt-related transcription factor 2 (p < 0.001), and markers of extracellular matrix production (p < 0.001). Both BMP receptors showed a significant BMP-2-dependent upregulation at the interface region, and Noggin was downregulated at the osteoblast and interface region following BMP-2 exposure.
Exposure to BMP-2 upregulated the expression of genes associated with bone-tendon integration in vitro, suggesting the stimulation of transdifferentiation processes at the interface and fibroblast regions as well as the induction of positive feedback mechanisms. Further studies will be needed to establish BMP-2 dose and treatment algorithms following tendon reinsertion and reconstruction.
临床前研究报告称，骨形态发生蛋白（BMP）-2可促进前交叉韧带重建术后骨-肌腱愈合。我们在高度标准化的小鼠骨-肌腱整合体外共培养模型中，研究了BMP-2对成骨细胞和成纤维细胞分化的区域特异性影响。
我们使用定量PCR来测量BMP-2对小鼠前成骨细胞系MC3T3-E1和成纤维细胞系3T6共培养模型的成骨细胞、界面和成纤维细胞区域中碱性磷酸酶、骨钙素、1型胶原（α1链）、 runt相关转录因子2、骨桥蛋白、1型胶原（α2链）、5型胶原（α1链）、核心蛋白聚糖、纤调蛋白、莫霍克同源框、1A型骨形态发生蛋白受体、2型骨形态发生蛋白受体和Noggin表达的剂量和时间依赖性影响。
用BMP-2刺激导致成骨细胞区域中碱性磷酸酶（p < 0.001）、骨钙素（p < 0.001）、胶原（p < 0.001）、runt相关转录因子2（p < 0.05）和骨桥蛋白（p < 0.001）表达显著上调。在界面区域，暴露于BMP-2导致碱性磷酸酶（p < 0.001）、骨钙素（p < 0.001）、骨桥蛋白（p < 0.001）、runt相关转录因子2（p < 0.001）和细胞外基质产生标志物（p <
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