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Chinese Journal of Joint Surgery(Electronic Edition) ›› 2019, Vol. 13 ›› Issue (06): 693-698. doi: 10.3877/cma.j.issn.1674-134X.2019.06.008

Special Issue:

• Basic Research • Previous Articles    

Experimental study of three-dimensional co-culture system promoting dedifferentiated chondrocyte redifferentiation

Tingshuai Zhang1, Jianyu Zou1, Hanzheng Chen1, Rixu Liu1, Shicong Zheng1, Yi Chen1, Shujiang Zhang1, Yongchang Yao1,()   

  1. 1. Department of Joint Surgery, 1st Affiliated Hospital of Guangzhou Medical University; Guangdong Key Laboratory of Orthopaedic Technology and Implant Materials, Guangzhou 510120, China
  • Received:2019-09-06 Online:2019-12-01 Published:2019-12-01
  • Contact: Yongchang Yao
  • About author:
    Corresponding author: Yao Yongchang, Email:

Abstract:

Objective

To investigate the effect of alginate hydrogel 3D co-culture on redifferentiation of dedifferentiated chondrocytes.

Methods

The first generation (P1) chondrocytes and the fourth generation (P4) chondrocytes were co-cultured with ATDC5 at the ratio of 3: 1, and wee named as AP1 and AP4 respectively. Another 3D culture system consisting of single P1, P4 and ATDC5 was established, and was named as P1, P4 and ATDC, respectively. All the groups were cultured in the chondrocyte-inducing medium with the supply of TGF-β3. After 28 days, the expression of collagen typeⅠ (ColⅠ), collagen typeⅡ (ColⅡ) and aggrecan (Acan) genes was assayed by Q-PCR. Histological staining was performed. The cartilage-related genes (Col Ⅱ and Acan) of each group were compared. The results were analyzed by Bonferroni test.

Results

The expression of ColⅡ and Acan in AP4 group was significantly up-regulated compared to P1 and P4 groups (ColⅡ: F=38.41, P<0.01, Acan: F=5, P<0.01 ), and the deposition of relative protein products could also be clearly observed on histological staining.

Conclusion

The 3D co-culture system can make the dedifferentiated chondrocytes regain their unique phenotype, the mechanism of which involves cellular factors and the cell-cell connection.

Key words: Transforming growth factor beta3, Hydrogel, Collagen type Ⅰ, Collagen type Ⅱ, Biocompatible materials, Chondrocyte, Dedifferentiation

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