|
|
|
| Effects of Cartilage Oligomeric Matrix Protein Overexpression on BMP-2 Induced Cell Differentiation of Bone Marrow Mesenchymal Stem Cells |
| SHEN Peng-fei, WANG Bin, XIE Zi-kang, ZHENG Chong, QU Yu-xing |
| First Department of Orthopaedics, Changzhou TCM Hospital, Changzhou 213000, China |
|
|
|
Abstract Objective: To study the effects of cartilage oligomeric matrix protein (COMP) overexpression on bone morphogenetic protein 2 (BMP-2) induced cell osteogenic and chondrogenic differentiation of bone marrow mesenchymal stem cells (MSCs). Methods: MSCs, transfected with plasmid DNA encoding recombinant human COMP, were induced to differentiate into osteocytes and chondrocytes by BMP-2. Western blotting and real-time PCR were used to detect the overexpression of COMP, the expression level changes of osteogenesis related genes collagen type Ⅰ, RUNX2, osteocalcin and chondrogenesis related genes collagen type Ⅱ, SOX9, aggrecan, collagen type X, respectively. Alizarin red staining for osteogenic differentiation and alcian blue staining for chondrogenic differentiation were conducted to evaluate the tendency of cell differentiation. Results: Results of RT-PCR and Western blotting showed that the expression levels of COMP mRNA and protein were significantly increased in MSCs, which were transfected with plasmid DNA encoding recombinant human COMP (P<0.05). The mRNA expression levels of RUNX2, Col1a1 and the protein expression levels of RUNX2, osteocalcin in overexpressed group all decreased significantly compared with that of control group (P<0.05). However, the mRNA expression levels of SOX9, aggrecan and the protein expression levels of SOX9, Col2a1 in overexpressed group all increased significantly compared with that of control group (P<0.05). Alizarin red staining were weakened while alcian blue staining was enhanced. The expression level of Col10a1 gene in overexpressed group was higher than that in control group (P<0.05). Conclusion: Overexpression of COMP gene in MSCs could inhibit BMP-2 induced osteogenic differentiation, promote BMP-2 induced chondrogenic differentiation and supress the chondrocyte hypertrophy and maturation, which may provide new insight for cartilage tissue engineering.
|
|
Received: 05 April 2016
Published: 25 October 2016
|
|
|
|
|
| [1] 虞冀哲, 董学海, 陈海丹, 等. 间充质干细胞应用于软骨组织工程的研究进展. 生物骨科材料与临床研究, 2015, 12(4): 60-64. Yu J Z, Dong X H, Chen H D, et al. Reviewed the research progress of cartilage tissue engineering using mesenchymal stem cells. Orthop Biomech Mater Clin Study, 2015, 12(4): 60-64.
[2] Liang Y, Fu Y, Qi R, et al. Cartilage oligomeric matrix protein is a natural inhibitor of thrombin. Blood, 2015, 126(7): 905-914.
[3] Kluzek S, Bay-Jensen A C, Spector T, et al. Higher serum levels of cartilage oligomeric matrix protein (comp) are associated with self-reported knee pain. Osteoarthr Cartilage, 2014, 22(3): S74.
[4] Kluzek S, Bay-Jensen A C, Judge A, et al. Serum cartilage oligomeric matrix protein and development of radiographic and painful knee osteoarthritis. A community-based cohort of middle-aged women. Biomarkers, 2015, 20(8): 557-564.
[5] Kim M J, Lee B, Yang K, et al. BMP-2 peptide-functionalized nanopatterned substrates for enhanced osteogenic differentiation of human mesenchymal stem cells. Biomaterials, 2013, 34(30): 7236-7246.
[6] Barati D, Shariati S R, Moeinzadeh S, et al. Spatiotemporal release of BMP-2 and VEGF enhances osteogenic and vasculogenic differentiation of human mesenchymal stem cells and endothelial colony-forming cells co-encapsulated in a patterned hydrogel. J Controll Release, 2016, 223: 126-136.
[7] Guzzo R M, Gibson J, Xu R H, et al. Efficient differentiation of human iPSC-derived mesenchymal stem cells to chondroprogenitor cells. J Cell Biochem, 2013, 114(2): 480-490.
[8] 朱瑜琪, 王金荣, 王智耀. 间充质干细胞促进关节软骨的修复与再生. 中国组织工程研究, 2015, 19(50): 8195-8200. Zhu Y Q, Wang J R, Wang Z Y. Mesenchymal stem cells promote articular cartilage repair and regeneration. Chin J Tissue Eng Res, 2015, 19(50): 8195-8200.
[9] 曲峰, 袁邦拓, 齐玮, 等. Wnt3a 对骨髓间充质干细胞成软骨分化的影响. 中国矫形外科杂志, 2016, 24(2): 155-159. Qu F, Yuan B T, Qi W, et al. Effect of Wnt3a on chondrogenic differentiation of nesenchymal stem cells. Orthop J China, 2016, 24(2): 155-159.
[10] Dharmavaram R M, Liu G, Tuan R S, et al. Stable transfection of human fetal chondrocytes with a type II procollagen minigene: expression of the mutant protein and alterations in the structure of the extracellular matrix in vitro. Arthritis Rheum, 1999, 42(7): 1433-1442.
[11] Du Y, Wang Y, Wang L, et al. Cartilage oligomeric matrix protein inhibits vascular smooth muscle calcification by interacting with bone morphogenetic protein-2. Circ Res, 2011, 108(8): 917-928.
[12] Ishida K, Acharya C, Christiansen B A, et al. Cartilage oligomeric matrix protein enhances osteogenesis by directly binding and activating bone morphogenetic protein-2. Bone, 2013, 55(1): 23-35.
[13] Guo P, Shi Z L, Liu A, et al. Effects of cartilage oligomeric matrix protein on bone morphogenetic protein-2-induced differentiation of mesenchymal stem cells. Orthop Surg, 2014, 6(4): 280-287.
[14] Wang W J, Sun C, Liu Z, et al. Transcription factor Runx2 in the low bone mineral density of girls with adolescent idiopathic scoliosis. Orthop Surg, 2014, 6(1): 8-14.
[15] Ge C, Cawthorn W P, Li Y, et al. Reciprocal control of osteogenic and adipogenic differentiation by ERK/MAP kinase phosphorylation of Runx2 and PPARγ transcription factors. J Cell Physiol, 2016, 231(3): 587-596.
[16] Liu C F, Lefebvre V. The transcription factors SOX9 and SOX5/SOX6 cooperate genome-wide through super-enhancers to drive chondrogenesis. Nucleic Acids Res, 2015, 43(17): 8183-8203.
[17] Gurusinghe S, Young P, Michelsen J, et al. Suppression of differentiation and hypertrophy in canine chondrocytes through lentiviral vector expression of Sox9 and induced pluripotency stem cell factors. Biotechnol Lett, 2015, 37(7): 1495-1504.
[18] 王冠, 陈星星, 薛鑫, 等. Ⅱ 型胶原体外促进大鼠骨髓间充质干细胞向软骨细胞分化. 第三军医大学学报, 2015, 37(9): 886-890. Wang G, Chen X X, Xue X, et al. Collagen Ⅱ promotes differentiation of rat bone marrow mesenchymal stem cells into chondrocytes in vitro. J Third Mil Med Univ, 2015, 37(9): 886-890.
[19] Lu Y, Ding M, Li N, et al. Col10a1-Runx2 transgenic mice with delayed chondrocyte maturation are less susceptible to developing osteoarthritis. Am J Transl Res, 2014, 6(6): 736-745.
[20] Gu J, Lu Y, Li F, et al. Identification and characterization of the novel Col10a1 regulatory mechanism during chondrocyte hypertrophic differentiation. Cell Death Dis, 2014, 5(10): e1469. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
