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Glucan HBP-A increase type II collagen expression of chondrocytes in vitro and tissue engineered cartilage in vivo

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Abstract

Objective

Although chondroprotective activities have been documented for polysaccharides, the potential target of different polysaccharide may differ. The study was aimed to explore the effect of glucan HBP-A in chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs in vivo, especially on the expression of type II collagen.

Methods

Chondrocytes isolated from rabbit articular cartilage were cultured and verified by immunocytochemical staining of type II collagen. Chondrocyte viability was assessed after being treated with HBP-A in different concentrations. Morphological status of chondrocytes-alginate hydrogel constructs in vitro was observed by scanning electron microscope (SEM). The constructs were treated with HBP-A and then injected to nude mice subcutaneously. Six weeks after transplantation, the specimens were observed through transmission electron microscopy (TEM). The mRNA expressions of disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTs-5), aggrecan and type II collagen in both monolayer culture and constructs were determined by real time polymerase chain reaction (PCR). The expression of type II collagen and matrix metalloproteinases-3 (MMP-3) in chondrocyte monolayer culture was also tested through Western blot and enzyme linked immunosorbent assay (ELISA), respectively.

Results

MMP-3 secretion and ADAMTs-5 mRNA expression in vitro were inhibited by HBP-A at 0.3 mg/mL concentration. In morphological study, there were significant appearance of collagen in those constructs treated by HBP-A. Accordingly, in both chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs, the expression of type II collagen was increased significantly in HBP-A group when compared with control group (P<0.001).

Conclusions

The study documented that the potential pharmacological target of glucan HBP-A in chondrocytes monolayer culture and tissue engineered cartilage in vivo may be concerned with the inhibition of catabolic enzymes MMP-3, ADAMTs-5, and increasing of type II collagen expression.

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Authors and Affiliations

  1. Research Institute of Orthopaedics, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Yue-long Cao  (曹月龙), Ting Liu  (刘 婷), Jian Pang  (庞 坚), Ning-yang Gao  (高宁阳), Hong-sheng Zhan  (詹红生), Yin-yu Shi  (石印玉) & Xiang Wang  (王 翔)

  2. Institute of Chinese Materia Medica of Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Shun-chun Wang  (王顺春)

Authors
  1. Yue-long Cao  (曹月龙)
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  2. Ting Liu  (刘 婷)
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  3. Jian Pang  (庞 坚)
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  4. Ning-yang Gao  (高宁阳)
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  5. Hong-sheng Zhan  (詹红生)
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  6. Yin-yu Shi  (石印玉)
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  7. Xiang Wang  (王 翔)
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  8. Shun-chun Wang  (王顺春)
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Corresponding author

Correspondence to Yue-long Cao  (曹月龙).

Additional information

Supported by the National Natural Science Foundation of China (No. 30300459, 81072830), Shanghai Leading Academic Discipline Project (No. T0303) and the Shanghai Youth Phospherus Project (No. 08QA14063)

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Cao, Yl., Liu, T., Pang, J. et al. Glucan HBP-A increase type II collagen expression of chondrocytes in vitro and tissue engineered cartilage in vivo . Chin. J. Integr. Med. 21, 196–203 (2015). https://doi.org/10.1007/s11655-013-1511-x

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  • DOI: https://doi.org/10.1007/s11655-013-1511-x

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