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Morphology and Phenotype Expression of Types I, II, III, and X Collagen and MMP-13 of Chondrocytes Cultured from Articular Cartilage of Kashin-Beck Disease
WEIZHUO WANG, XIONG GUO, JUNCHANG CHEN, PENG XU, and MIKKO J. LAMMI
ABSTRACT. Methods. Samples of articular cartilage were divided into 2 groups: KBD group (8 samples, 8 cases) and the control (8 samples, 8 cases). KBD patients were diagnosed according to "Pathological Criteria to Diagnose KBD in China." Hyaline cartilage was digested with collagenase into cell suspensions and cultured in monolayers. Chondrocyte ultrastructure was observed by electron microscope at 10th day in vitro. Primary articular chondrocytes were seeded on microscope slides and immunostained on 12th day of cultivation for types I, II, III, and X collagens and MMP-13. Positive findings were counted by light microscopy and confirmed by flow cytometric analyses. Results. Considerable amounts of vacuoles and distorted nuclei, as well as thickening and irregular arrangement of collagen fibrils, were seen in the KBD samples by electron microscopy. Types I, III, and X collagen were stained in the KBD, but not in the control cultures. The percentages of positive staining for type II collagen were significantly lower in KBD than those in controls (tcol II = -5.54, p < 0.001), and for MMP-13 in the KBD group were significantly higher (tMMP-13 = 3.70, p < 0.01). Conclusion. Phenotype expressions of types I, II, III, and X collagen and MMP-13 in chondrocytes cultured in vitro were significantly different between the KBD and control cultures, indicating degenerative and hypertrophic changes in chondrocytes of KBD articular cartilage. (J Rheumatol First Release Mar 1 2008) Key Indexing Terms:
KASHIN-BECK DISEASE
From the Key Laboratory of Environment and Gene Related Diseases, Xi'an Jiaotong University, Ministry of Education; the Department of Orthopedics Surgery, Second Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China; the Institute of Biomedicine, Anatomy, University of Kuopio; and the Department of Biosciences, Applied Biotechnology, University of Kuopio, Kuopio, Finland. Supported by the Ministry of Science and Technology (2006 DFA33610), The Natural Scientific Fund of China (No. 30630058), and The International Co-operative Fund in Shaanxi (2005KW-13). W.Z. Wang, MD, PhD; J.C. Chen, MD, Department of Orthopedic Surgery, Second Hospital of Xi'an Jiaotong University; X. Guo, Professor, MD; P. Xu, Professor, MD, PhD, Key Laboratory of Environment and Genes Related to Diseases, Xi'an Jiaotong University; M.J. Lammi, PhD, Professor, Department of Anatomy, Institute of Biomedicine, University of Kuopio, and Department of Biosciences, Applied Biotechnology, University of Kuopio. Address reprint requests to Dr. X. Guo, Medical School of Xi'an Jiaotong University, No. 76 West Road of Yanta, Xi'an 710061, China. E-mail: guox@mail.xjtu.edu.cn Accepted for publication November 23, 2007.
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