Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (23): 3621-3626.doi: 10.3969/j.issn.2095-4344.2724

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Basic fibroblast growth factors protect chondrocytes by antagonizing extracellular inflammatory factors

Yang Fan1, 2, Liu Baoyi2, Cao Meng2, Zhu Xiaoshu3, Zhang Yu2, Qin Kairong1, Zhao Dewei1, 2   

  1. 1Department of Biomedical Engineering, Dalian University of Technology, Dalian 116024, Liaoning Province, China; 2Department of Orthopedics, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, Liaoning Province, China; 3The Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen 518033, Guangdong Province, China
  • Received:2019-08-14 Revised:2019-08-16 Accepted:2019-10-15 Online:2020-08-18 Published:2020-04-25
  • Contact: Zhao Dewei, MD, Chief physician, Professor, Department of Biomedical Engineering, Dalian University of Technology, Dalian 116024, Liaoning Province, China; Department of Orthopedics, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, Liaoning Province, China
  • About author:Yang Fan, MD candidate, Physician, Department of Biomedical Engineering, Dalian University of Technology, Dalian 116024, Liaoning Province, China; Department of Orthopedics, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, Liaoning Province, China
  • Supported by:
    the Postdoctoral Science Foundation of China (General Project), No. 2017M621116; Dalian High-Level Talent Innovation Support Program-“Science and Technology Star” Project, No. 2017RQ154; Liaoning Provincial Doctoral Start-up Foundation, No. 201601299; General Research Project of Liaoning Education Department, No. L2015021

Abstract:

BACKGROUND: Nowadays, there are many studies addressing basic fibroblast growth factor (bFGF) that promotes chondrocyte differentiation and inflammatory factors that damage chondrocyte structure, such as interleukin-1, but there are no reports on the combined effect of the two factors on chondrocytes.

OBJECTIVE: To observe the effect of bFGF and inflammatory cytokines on growth characteristics and differentiation potential of chondrocytes.

METHODS: Passage 3 chondrocytes from Sprague-Dawley rats cultured in vitro were divided into four groups: a blank control group in which chondrocytes were cultured alone, a negative control group in which chondrocytes were cultured with serum-free DMEM/F12 containing tumor necrosis factor-α, interleukin-1 and interleukin-6, a positive control group in which chondrocytes were cultured with serum-free DMEM/F12 containing bFGF, and an experimental group in which chondrocytes were cultured with serum-free DMEM/F12 containing bFGF, tumor necrosis factor-α, interleukin-1 and interleukin-6. At 3, 5, and 7 days after culture, the proliferative activity of chondrocytes was detected by cell counting kit-8 method; the levels of inflammatory cytokines were determined by ELISA method; and mRNA expression of collagen type II and proteoglycan in chondrocytes was measured by real-time PCR. Protein levels of collagen type II and proteoglycan were assessed by immunofluorescence assay at 7 days after culture.

RESULTS AND CONCLUSION: At 3, 5, 7 days after culture, the proliferation of chondrocytes ranked as follows: positive control group > experimental group > blank control group > negative control group. Compared with the blank control group, the expressions of interleukin-1, interleukin-6 and tumor necrosis factor-α were significantly increased in the negative control group. However, compared with the negative control group, the expressions of the above three inflammatory cytokines in the experimental group showed a significant decline (P < 0.05). There was positive expression of collagen type II and proteoglycan in the blank control, positive control and experimental groups, especially in the negative control group. Compared with the blank control group, the expressions of collagen type II and proteoglycan were up-regulated significantly in the positive control and experimental groups. To conclude, the rationale use of bFGF can maintain the phenotype of chondrocytes, inhibit dedifferentiation and promote cell proliferation.

Key words: chondrocytes, cell proliferation, cell differentiation, basic fibroblast growth factor; interleukin-1, interleukin-6, tumor necrosis factor-

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