The effect of self-designed bifunctional RGD-containing fusion protein on the behavior of human keratinocytes and dermal fibroblasts

  1. Tzu-Wei Wang1,
  2. Hsi-Chin Wu1,
  3. Yi-Chau Huang1,
  4. Jui-Sheng Sun2,3,
  5. Feng-Huei Lin1,*

Article first published online: 28 APR 2006

DOI: 10.1002/jbm.b.30552

Issue

Journal of Biomedical Materials Research Part B: Applied Biomaterials

Journal of Biomedical Materials Research Part B: Applied Biomaterials

Volume 79B, Issue 2, pages 379–387, November 2006

Additional Information

How to Cite

Wang, T.-W., Wu, H.-C., Huang, Y.-C., Sun, J.-S. and Lin, F.-H. (2006), The effect of self-designed bifunctional RGD-containing fusion protein on the behavior of human keratinocytes and dermal fibroblasts. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 79B: 379–387. doi: 10.1002/jbm.b.30552

Author Information

  1. 1

    Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan

  2. 2

    Institute of Rehabilitation Science and Technology, National Yang-Ming University, Taipei, Taiwan

  3. 3

    Department of Orthopedic Surgery, Taipei City Hospital, Yang-Ming Branch, Taipei, Taiwan

*Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan

Publication History

  1. Issue published online: 6 OCT 2006
  2. Article first published online: 28 APR 2006
  3. Manuscript Accepted: 27 DEC 2005
  4. Manuscript Revised: 14 OCT 2005
  5. Manuscript Received: 2 SEP 2005

Funded by

  • National Science Council

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Keywords:

  • bifunctional RGD-containing fusion protein;
  • attachment;
  • cytotoxicity;
  • keratinocytes;
  • dermal fibroblasts

Abstract

In this study, self-designed bifunctional RGD-containing fusion protein (BFP) was grafted on the petri dish to evaluate its cytotoxicity and attachment efficiency on primary cultured keratinocytes and dermal fibroblasts. Two lengths of the GRGDS sequences were separately fused to the N-terminus and C-terminus of the Trichoderma koningii cellobiohydrolase I gene cellulose-binding domain, to serve as linking molecule between the cell and the substrate. The grafting procedure was no more labor-intensive and could be done just in aqueous condition itself. The epidermal keratinocytes and dermal fibroblasts, harvested and separated from human foreskin, were cultured in serum-free keratinocyte culture medium and DMEM, respectively. The BFP was dissolved in double-deionized water, and was prepared at different concentrations. The BFP solution was subsequently added into the petri dish for grafting. MTT assay, total DNA measurement, and lactate dehydrogenase analysis were used to evaluate the cell viability, cell proliferation, and cytotoxicity. The immunochemical stain and SEM examination were chosen to make sure that the cultured cells still kept in phenotype. The results showed that the self-designed BFP was successfully coated on the petri dish to improve the cells' adhesion. The whole coating procedure was just done in aqueous solution without any organic solvent being involved. This method was much simpler than the traditional one, and there was no possibility to damage the immobilized biomolecules. From the results of the study, BFP could enhance attachment of keratinocytes and dermal fibroblasts without losing normal cell morphology and keep keratinocytes on the desired differentiation pathway. We believe that coating BFP on petri dish not only enhanced the keratinocyte attachment but also promoted keratinocytes proliferation. We suggest that the self-designed BFP has a great potential to apply on surface modification for the tissue-engineering scaffolds in the future. © 2006 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006

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