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. 2010 Mar;11(3):200-8.
doi: 10.1631/jzus.B0900074.

Effects of moniliformin and selenium on human articular cartilage metabolism and their potential relationships to the pathogenesis of Kashin-Beck disease

An Zhang  1 , Jun-ling Cao, Bo Yang, Jing-hong Chen, Zeng-tie Zhang, Si-yuan Li, Qiang Fu, Clare E Hugnes, Bruce Caterson
Affiliations

Effects of moniliformin and selenium on human articular cartilage metabolism and their potential relationships to the pathogenesis of Kashin-Beck disease

An Zhang et al. J Zhejiang Univ Sci B. 2010 Mar.

Abstract

Objective: To investigate the effects of mycotoxin moniliformin (MON) on the metabolism of aggrecan and type II collagen in human chondrocytes in vitro and the relationship between MON and Kashin-Beck disease (KBD).

Methods: Human chondrocytes were isolated and cultured on bone matrix gelatin to form an artificial cartilage model in vitro with or without MON toxin. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of aggrecan and type II collagen in the cartilage was determined using immunocytochemical staining.

Results: MON toxin inhibited chondrocyte viability in dose-dependent and time-dependent manners. MON reduced aggrecan and type II collagen syntheses in the tissue-engineered cartilage. MON also increased the expression of matrix metalloproteinase-1 (MMP-1), MMP-13, BC4 epitopes, and CD44 in cartilages. However, the expression of 3B3(-) epitopes in cartilages was inhibited by MON. Selenium partially alleviated the damage of aggrecan induced by MON toxin.

Conclusion: MON toxin promoted the catabolism of aggrecan and type II collagen in human chondrocytes.

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Figures

Fig. 1
Fig. 1
Effects of different concentrations of MON on the cell viability of chondrocytes Human chondrocytes cultured as monolayer were incubated with different concentrations of MON toxin for 5 d. MTT assay was done to determine the cell viability. The cellular viability of control is regarded as 1. Data are the mean of the experiment (n=5). * P≤0.05 when compared with the control group
Fig. 2
Fig. 2
H & E staining of the chondrocytes cultured on BMG Human chondrocytes were cultured on BMG for 16 d. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group. Black arrow: cell necrosis
Fig. 2
Fig. 2
H & E staining of the chondrocytes cultured on BMG Human chondrocytes were cultured on BMG for 16 d. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group. Black arrow: cell necrosis
Fig. 2
Fig. 2
H & E staining of the chondrocytes cultured on BMG Human chondrocytes were cultured on BMG for 16 d. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group. Black arrow: cell necrosis
Fig. 2
Fig. 2
H & E staining of the chondrocytes cultured on BMG Human chondrocytes were cultured on BMG for 16 d. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group. Black arrow: cell necrosis
Fig. 3
Fig. 3
TB staining of the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. Proteoglycan were stained with purple and the nuclei were stained with blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 3
Fig. 3
TB staining of the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. Proteoglycan were stained with purple and the nuclei were stained with blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 3
Fig. 3
TB staining of the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. Proteoglycan were stained with purple and the nuclei were stained with blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 3
Fig. 3
TB staining of the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. Proteoglycan were stained with purple and the nuclei were stained with blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 4
Fig. 4
Immunostaining of type II collagen in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The type II collagen was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 4
Fig. 4
Immunostaining of type II collagen in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The type II collagen was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 4
Fig. 4
Immunostaining of type II collagen in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The type II collagen was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 4
Fig. 4
Immunostaining of type II collagen in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The type II collagen was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 5
Fig. 5
Immunostaining of 3B3(−) in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The epitopes of 3B3(−) were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 5
Fig. 5
Immunostaining of 3B3(−) in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The epitopes of 3B3(−) were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 5
Fig. 5
Immunostaining of 3B3(−) in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The epitopes of 3B3(−) were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 5
Fig. 5
Immunostaining of 3B3(−) in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. The epitopes of 3B3(−) were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 6
Fig. 6
Immunostaining of BC4 epitopes in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. The epitopes of BC4 were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 6
Fig. 6
Immunostaining of BC4 epitopes in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. The epitopes of BC4 were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 6
Fig. 6
Immunostaining of BC4 epitopes in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. The epitopes of BC4 were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 6
Fig. 6
Immunostaining of BC4 epitopes in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d after seeded on BMG. The epitopes of BC4 were green and the nuclei were red. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 7
Fig. 7
Immunostaining of MMP-1 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-1 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 7
Fig. 7
Immunostaining of MMP-1 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-1 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 7
Fig. 7
Immunostaining of MMP-1 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-1 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 7
Fig. 7
Immunostaining of MMP-1 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-1 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 8
Fig. 8
Immunostaining of MMP13 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-13 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 8
Fig. 8
Immunostaining of MMP13 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-13 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 8
Fig. 8
Immunostaining of MMP13 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-13 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 8
Fig. 8
Immunostaining of MMP13 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. MMP-13 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 9
Fig. 9
Immunostaining of CD44 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. CD44 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 9
Fig. 9
Immunostaining of CD44 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. CD44 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 9
Fig. 9
Immunostaining of CD44 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. CD44 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
Fig. 9
Fig. 9
Immunostaining of CD44 in the cartilage reconstructed in vitro Chondrocytes were cultured for 16 d before the immunostaining. CD44 was deep brownish red and the nuclei were blue. (a) Control group; (b) MON group; (c) Control+Se group; (d) MON+Se group
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