Co-culture of subchondral osteoblasts and chondrocytes from human knee joint

doi:10.3969/j.issn.2095-4344.2013.11.002

Abstract

Abstract:

BACKGROUND: Cartilage tissue engineering provides a potential method for the treatment of osteoarthritis. The relation and mechanism of interaction that subchondral osteoblasts act on chondrocytes are worthy of further research.
OBJECTIVE: To co-culture subchondral osteoblasts and chondrocytes from human knee joint, thereby constructing a co-culture system of subchondral osteoblasts and chondrocytes to investigate the reaction between osteoblasts and chondrocytes.
METHODS: Subchondral bone specimens were obtained from patients undergoing total knee replacement for primary osteoarthritis and those receiving amputation surgeries for serious limb injuries. Primary osteoblasts were isolated from subchondral bone specimens using sequential digestion method, and identified using type Ⅰ collagen immunochemistry. Primary chondrocytes were isolated from articular specimens after amputation and identified using type Ⅱ collagen immunochemistry. Then, a three-dimensional co-culture system of osteoblasts and chondrocytes was constructed using cell culture inserts.
RESULTS AND CONCLUSION: Osteoblasts and chondrocytes were identified successfully using immunochemistry, NBT/BCIP staining, and alizarin red staining. Real-time quantitative reverse transcription-PCR results exhibited that expression of type Ⅱ collagen and proteoglycans significantly decreased in chondrocytes co-cultured with subchondral osteoblasts from osteoarthritis patients. These findings show that osteoblasts from osteoarthritis patients can initiate chondrocyte phenotypes shifting towards degenerative differentiation.

Key words: tissue construction, cartilage tissue construction, subchondral bone, osteoblasts, chondrocytes, osteoarthritis, co-culture, real-time quantitative reverse transcription-PCR, type Ⅱ collagen, proteoglycans, type Ⅰ collagen, alkaline phosphatase, the National Natural Science Foundation of China, tissue construction photographs-containing paper

CLC Number:

R318

Zhang Li-zhi, Jiang Yao, Mao Yan-jie, Hang Cheng-long. Co-culture of subchondral osteoblasts and chondrocytes from human knee joint[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(11): 1909-1916.

Figures/Tables 6

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