Murray Shannon J, Santangelo Kelly S, Bertone Alicia L
Comparative Orthopedic Molecular Medicine and Applied Research Laboratory, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA.
Am J Vet Res. 2010 Jan;71(1):103-14. doi: 10.2460/ajvr.71.1.103.
To evaluate early cellular influences of bone morphogenetic protein (BMP)12 and BMP2 on equine superficial digital flexor tenocytes (SDFTNs) and equine bone marrow-derived mesenchymal stem cells (BMDMSCs).
9 adult clinically normal horses.
BMDMSCs and SDFTNs were cultured in monolayer, either untreated or transduced with adenovirus encoding green fluorescent protein, adenovirus encoding BMP12, or adenovirus encoding BMP2. Cytomorphologic, cytochemical, immunocytochemical, and reverse transcriptase-quantitative PCR (RT-qPCR) analyses were performed on days 3 and 6. Genetic profiling for effects of BMP12 was evaluated by use of an equine gene expression microarray on day 6.
BMDMSCs and SDFTNs had high BMP12 gene expression and remained viable and healthy for at least 6 days. Type l collagen immunocytochemical staining for SDFTNs and tenocyte-like morphology for SDFTNs and BMDMSCs were greatest in BMP12 cells. Cartilage oligomeric matrix protein, as determined via RT-qPCR assay, and chondroitin sulfate, as determined via gene expression microarray analysis, were upregulated relative to control groups in SDFTN-BMP12 cells. The BMDMSCs and SDFTNs became mineralized with BMP2, but not BMP12. Superficial digital flexor tenocytes responded to BMP12 with upregulation of genes relevant to tendon healing and without mineralization as seen with BMP2.
Targeted equine SDFTNs may respond to BMP12 with improved tenocyte morphology and without mineralization, as seen with BMP2. Bone marrow-derived mesenchymal stem cells may be able to serve as a cell delivery method for BMP12.
评估骨形态发生蛋白（BMP）12和BMP2对马浅屈肌腱细胞（SDFTNs）和马骨髓间充质干细胞（BMDMSCs）的早期细胞影响。
9匹成年临床正常马匹。
将BMDMSCs和SDFTNs进行单层培养，分为未处理组，以及用编码绿色荧光蛋白的腺病毒、编码BMP12的腺病毒或编码BMP2的腺病毒转导的组。在第3天和第6天进行细胞形态学、细胞化学、免疫细胞化学和逆转录定量PCR（RT-qPCR）分析。在第6天使用马基因表达微阵列评估BMP12作用的基因谱分析。
BMDMSCs和SDFTNs具有较高的BMP12基因表达，并且至少6天内保持活力和健康状态。SDFTNs的I型胶原蛋白免疫细胞化学染色以及SDFTNs和BMDMSCs的腱细胞样形态在BMP12细胞中最为明显。通过RT-qPCR测定法测定的软骨寡聚基质蛋白以及通过基因表达微阵列分析测定的硫酸软骨素，相对于SDFTN-BMP12细胞中的对照组均上调。BMDMSCs和SDFTNs用BMP2处理后发生矿化，但用BMP12处理则未发生矿化。浅屈肌腱细胞对BMP12有反应，与肌腱愈合相关的基因上调，且未出现BMP2所导致的矿化现象。
靶向的马SDFTNs可能对BMP12有反应，表现为腱细胞形态改善且无矿化现象，这与BMP2所见不同。骨髓间充质干细胞可能能够作为BMP12的一种细胞递送方法。
