May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Characteristics of a gelatin–chitosan biomembrane for engineering the conjunctiva
Author Affiliations & Notes
  • X. Zhu
    Singapore Eye Research Institute, Singapore
  • R.W. Beuerman
    Singapore Eye Research Institute, Singapore
    Dept. Ophthalmology, National University of Singapore, Singapore, Singapore
  • J.Y. Seah
    Singapore Eye Research Institute, Singapore
  • S. Nambiar
    Singapore Eye Research Institute, Singapore
  • D.T. H. Tan
    Singapore Eye Research Institute, Singapore
    Dept. Ophthalmology, National University of Singapore, Singapore, Singapore
  • Footnotes
    Commercial Relationships  X. Zhu, None; R.W. Beuerman, None; J.Y. Seah, None; S. Nambiar, None; D.T.H. Tan, None.
  • Footnotes
    Support  NMRC IBG and NUS R–191–000–004–112
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 3931. doi:
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      X. Zhu, R.W. Beuerman, J.Y. Seah, S. Nambiar, D.T. H. Tan; Characteristics of a gelatin–chitosan biomembrane for engineering the conjunctiva . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3931.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: To develop, fabricate and evaluate a novel gelatin–chitosan (GC) biomembrane which could be used as a substrate for the clinical engineering of ocular surface tissues. Methods:The GC membrane was fabricated by casting the solution onto a metal mold, evaporating the solvent in vacuum oven and neutralizing the membrane in NaOH solution. Rabbit conjunctiva explants were grown on the GC membrane to form an epithelial layer in the presence of 1:1 DMEM–F12 culture medium supplemented by 10% FBS and growth factors. Rabbit conjunctiva fibroblasts were also cultured on this biomembrane in the presence of DMEM culture medium containing 10% FBS. Circular 4–5 mm diameter wounds were created on an upper quadrant of the conjunctiva and prototype GC membranes were sutured on the wound surface and healing was monitored by slit–lamp with fluorescein staining. Results: We tested several ratios (ranging from 1:1 to 1:4) of gelatin to chitosan to determine the optimal membrane properties. Transparency and elasticity of the GC membrane varied according to the ratio of gelatin and chitosan. Increasing the amount of gelatin resulted in an increase in transparency and elasticity, but lowered the tensile strength of the membrane. We also optimized the G:C ratio for conjunctival fibroblast proliferation. Although the conjunctival fibroblasts showed good attachment and spreading on the GC biomembranes, cell proliferation studies using the MTT assay showed that conjunctival fibroblasts significantly decreased from 0.177±0.011 to 0.068±0.017 (p=0.014, n=3) when the chitosan increased from 50% (1:1) to 80% (1:4). Additional culture studies on conjunctival epithelial cells showed that conjunctival tissues explanted on the GC membrane formed cell colonies. The cell behavior and morphology were similar to that observed from explants grown on culture dishes without GC membrane. Thus, both epithelial cells and fibroblasts grew to confluence illustrating good biocompatibility of the GC membrane and good cell viability. Membranes have been successfully sutured onto the conjunctiva in our rabbit animal model, and appear to be viable in the short–term. Conclusion:The properties of the GC biomembrane suggest that it may be a useful potential candidate, as a cellular carrier and/or scaffold, for reconstructive tissue engineering of the conjunctiva. Supported by: NMRC IBG and NUS R–191–000–004–112

Keywords: conjunctiva • extracellular matrix • regeneration 
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