Evaluation of early cellular influences of bone morphogenetic proteins 12 and 2 on equine superficial digital flexor tenocytes and bone marrow-derived mesenchymal stem cells in vitro.
OBJECTIVE
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).


METHODS
9 adult clinically normal horses.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Murray
Shannon J
SJ
Comparative Orthopedic Molecular Medicine and Applied Research Laboratory, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA.
Santangelo
Kelly S
KS
Bertone
Alicia L
AL
D000818:Animals; D001853:Bone Marrow; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006736:Horses; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D040081:Protein Array Analysis; D013710:Tendons / Q000166:cytology*
D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; C086559:growth differentiation factor 7