Bone morphogenetic protein 2 stimulated osteo-chondrogenic differentiation of patellar tendon-derived stem cells isolated from a failed tendon-healing animal model of tendinopathy

doi:10.3969/j.issn.2095-4344.2014.19.020

Abstract

Abstract:

BACKGROUND: The pathogenesis of tendinopathy remains unclear and hence treatment of tendinopathy is usually palliative.
OBJECTIVE: To investigate the effects of bone morphogenetic protein 2 on the osteogenic and chondrogenic differentiation of patellar tendon-derived stem cells isolated from collagenase-induced tendinopathy rats in vitro.
METHODS: Patellar tendon-derived stem cells were isolated from patellar tendons of collagenase-induced tendinopathy rats. The multi-differentiation potential of patellar tendon-derived stem cells at passage 3 was identified by osteogenic, adipogenic and chondrogenic differentiation assays. The patellar tendon-derived stem cells were cultured to the 3rd passage in complete culture medium, and then the cells were divided into two groups with (experimental group) or without recombinant human bone morphogenetic protein 2 (control group) until the cells reached confluence for 7 days. Their osteogenic response to bone morphogenetic protein 2 in vitro was examined by alizarin red S staining of calciumnodule formation and quantification assay. The patellar tendon-derived stem cell pellets were cultured in complete culture medium with (experimental group) or without bone morphogenetic protein 2 (control grup) for 21 days. Chondrogenic differentiation of the cell pellets was evaluated by hematoxylin-eosin staining, alcian blue staining, immunohistochemical staining for Sox9 and collagen type II.
RESULTS AND CONCLUSION: Primary patellar tendon-derived stem cells from the tendinopathy rats cultured in vitro showed clonal growth; after passage, spindle fibroblast-like and flat-like cells were detectable. The cells were positive for oil red O staining at 10 days after adipogenic induction, positive for alizarin red staining at 7 days after osteogenic induction, and positive for hematoxylin-eosin staining and immunohistochemical staining of collagen type II at 14 days after chondrogenic induction. After patellar tendon-derived stem cells were induced with recombinant human bone morphogenetic protein 2 for 7 days, the result of alizarin red staining was positive in the experimental group, but negative in the control group without recombinant human bone morphogenetic protein 2. The difference in the result of alizarin red staining between the two groups was statistically significant. After patellar tendon-derived stem cells were induced with recombinant human bone morphogenetic protein 2 for 21 days, the results of hematoxylin-eosin staining, alcian blue staining, immunohistochemical staining for Sox9 and collagen type II were all positive. In conclusion, bone morphogenetic protein 2 could stimulate the osteogenic and chondrogenic differentiation of patellar tendon-derived stem cells isolated from collagenase-induced tendinopathy rats in vitro, which can help to better understand the pathogenesis of tendinopathy.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, tendinopathy, bone morphogenetic proteins, osteoblasts, chondrocytes, cell differentiation

CLC Number:

R394.2

Lin Yu-cheng, Wang Chen, Rui Yun-feng, Cheng Xin-kun, Ma Liang-yu. Bone morphogenetic protein 2 stimulated osteo-chondrogenic differentiation of patellar tendon-derived stem cells isolated from a failed tendon-healing animal model of tendinopathy[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(19): 3075-3081.

Figures/Tables 3

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