Adenovirus mediated BMP-13 gene transfer induces chondrogenic differentiation of murine mesenchymal progenitor cells.
Chondrogenic/osteogenic differentiation of a mesenchymal progenitor stimulated by BMP-13 (CDMP-2) was studied. C3H10T1/2 cells were transduced by an adenoviral construct containing BMP-13 or BMP-2. BMP-13 supported chondrogenesis but not terminal differentiation, whereas BMP-2 stimulated endochondral ossification. The studies show that BMP-13 may fail to support terminal chondrocyte differentiation.


BACKGROUND
Bone morphogenetic protein (BMP)-13 is a member of the transforming growth factor beta (TGF-beta) superfamily of growth factors. Although the biological functions of BMP-13 remain poorly understood, continued postnatal expression of BMP-13 in articular cartilage suggests that this protein may function in an autocrine/paracrine fashion to regulate growth and maintenance of articular cartilage. The purpose of this study was to elucidate the role of BMP-13 in chondrogenic differentiation.


METHODS
Replication-deficient adenoviruses carrying human BMP-13 (Adv-hBMP13), bacterial beta-galactosidase (Adv-beta gal), and human BMP-2 (Adv-hBMP2) were constructed. Murine mesenchymal progenitor cells (C3H10T1/2) were transduced with these vectors, and differentiation to the chondrogenic lineage was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR), biochemical, and histological analyses.


CONCLUSIONS
Our findings revealed that hBMP-13 transduced cells differentiated into round cells that stained with Alcian blue. Analysis of gene expression in hBMP-13-transduced cells demonstrated presence of cartilage-specific markers, absence of hypertrophic chondrocyte specific markers, and upregulation of proteoglycan biosynthesis. In particular, hBMP-13-transduced cells had significantly less and delayed expression of alkaline phosphatase activity and calcium mineral accumulation than hBMP-2-transduced cells. Except for BMPR-IB/ALK-6, expression of BMP receptors was identified constitutively in C3H10T1/2 cells and was not affected by the presence of either of the BMPs. In summary, hBMP-13, while stimulating chondrogenesis, failed to support differentiation to hypertrophic chondrocytes and endochondral ossification similar to hBMP-2. Thus, this may prove to be a useful strategy for cell-based regeneration of articular cartilage.
Nochi
Hitoshi
H
Department of Orthopaedic Surgery and Division of Renal Diseases, Washington University School of Medicine, St Louis, Missouri 63110, USA.
Sung
Jin Hyung
JH
Lou
Jueren
J
Adkisson
H Davis
HD
Maloney
William J
WJ
Hruska
Keith A
KA
D000256:Adenoviridae / Q000235:genetics; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D053674:Aggrecans; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2; D052004:Bone Morphogenetic Protein Receptors; D019485:Bone Morphogenetic Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D024043:Collagen Type II / Q000235:genetics; D024121:Collagen Type X / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D006025:Glycosaminoglycans / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000235:genetics; D006801:Humans; D037181:Lectins, C-Type; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D015675:Osteocalcin / Q000235:genetics; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000235:genetics; D055758:SOX9 Transcription Factor; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection; D016212:Transforming Growth Factor beta; D015854:Up-Regulation / Q000502:physiology
C508320:Acan protein, mouse; D053674:Aggrecans; C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II; D024121:Collagen Type X; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D006609:High Mobility Group Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D017978:Receptors, Growth Factor; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; C527933:Sox9 protein, mouse; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; C090394:cartilage-derived-morphogenetic protein-2; D015675:Osteocalcin; D052004:Bone Morphogenetic Protein Receptors; D000469:Alkaline Phosphatase; D002118:Calcium