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Chondrocyte-alginate constructs with or without TGF-β1 produces superior extracellular matrix expression than monolayer cultures

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Abstract

Tissue engineering approaches often require expansion of cell numbers in vitro to accelerate tissue regenerative processes. Although several studies have used this technique for therapeutic purposes, a major concern involving the use of isolated chondrocyte culture is the reduction of extracellular matrix (ECM) protein expressed due to the transfer of cells from the normal physiological milieu to the artificial 2D environment provided by the cell culture flasks. To overcome this issue, the use of alginate hydrogel beads as a substrate in chondrocyte cultures has been suggested. However, the resultant characteristics of cells embedded in this bead is elusive. To elucidate this, a study using chondrocytes isolated from rabbit knee articular cartilage expanded in vitro as monolayer and chondrocyte-alginate constructs was conducted. Immunohistochemical evaluation and ECM distribution was examined with or without transforming growth factor (TGF-β1) supplement to determine the ability of cells to express major chondrogenic proteins in these environments. Histological examination followed by transmission electron microscopy and scanning electron microscopy was performed to determine the morphology and the ultrastructural characteristics of these cells. Results demonstrated a significant increase in glycosaminoglycan/mg protein levels in chondrocyte cultures grown in alginate construct than in monolayer cultures. In addition, an abundance of ECM protein distribution surrounding chondrocytes cultured in alginate hydrogel was observed. In conclusion, the current study demonstrates that the use of alginate hydrogel beads in chondrocyte cultures with or without TGF-β1 supplement provided superior ECM expression than monolayer cultures.

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Acknowledgments

This project was supported by HIR-MOHE research grant and University of Malaya Research Grant (RG144/09HTM)

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Authors declare there is no conflict of interest

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Authors and Affiliations

  1. Faculty of Medicine, Universiti Teknologi MARA, 40450, Shah Alam, Selangor, Malaysia

    Sharaniza Ab-Rahim

  2. School of Medicine and Health Sciences, Monash University, Sunway Campus, 46150, Subang Jaya, Selangor, Malaysia

    Lakshmi Selvaratnam

  3. Tissue Engineering Group, National Orthopaedic Centre of Excellence in Research and Learning (NOCERAL), Department of Orthopaedic Surgery, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Hanumantha Rao Balaji Raghavendran & Tunku Kamarul

Authors
  1. Sharaniza Ab-Rahim
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  2. Lakshmi Selvaratnam
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  3. Hanumantha Rao Balaji Raghavendran
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  4. Tunku Kamarul
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Correspondence to Tunku Kamarul.

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Ab-Rahim, S., Selvaratnam, L., Raghavendran, H.R.B. et al. Chondrocyte-alginate constructs with or without TGF-β1 produces superior extracellular matrix expression than monolayer cultures. Mol Cell Biochem 376, 11–20 (2013). https://doi.org/10.1007/s11010-012-1543-0

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  • DOI: https://doi.org/10.1007/s11010-012-1543-0

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