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Sodium hyaluronate (hyaluronic acid) promotes migration of human corneal epithelial cells in vitro.

ABSTRACT: PURPOSE: Sodium hyaluronate (hyaluronic acid) is known to promote corneal epithelial wound healing in vivo and in vitro, in animal experiments. Sodium hyaluronate is the ligand for CD44, a cell surface adhesion molecule which has been found on normal human corneal epithelial cells. The purpose of this study was to investigate the effect of sodium hyaluronate on human corneal epithelial cell migration, proliferation, and CD44 receptor expression. METHODS: Human corneal epithelial cell cultures were established from 32 donor corneoscleral rims and maintained separately in three different culture conditions: (1) culture medium only, (2) sodium hyaluronate enriched (0.6 mg/ml) medium, and (3) hydroxypropylmethylcellulose enriched (2.5 mg/ml) medium. The total area of migrating epithelial cell sheets in each case was measured by planimetry on days 4, 8, 12, and 16. Cytospin preparations of cells cultured in the different culture conditions were examined immunohistochemically for proliferation and CD44 receptor expression using antibodies directed against Ki67 and CD44 respectively. RESULTS: Cells cultured in the presence of sodium hyaluronate showed significantly increased migration at days 12 and 16 (Friedmen test: p = 0.0012, day 16; p = <0.001, day 12) compared with cells cultured in the other media. There was no difference in cell proliferation (Ki67) or CD44 expression on cells cultured in the different culture conditions. CONCLUSIONS: Sodium hyaluronate promotes migration but not proliferation or CD44 expression on human corneal epithelial cells in vitro. The beneficial effect of sodium hyaluronate in corneal wound healing is likely to be related to rapid migration of cells leading to rapid wound closure. This may be facilitated by the adhesion between CD44 on the cells and hyaluronic acid, which coats the surface of the denuded cornea.

SUBMITTER: Gomes JA

PROVIDER: S-EPMC1772195 | biostudies-other | 2004 Jun

REPOSITORIES: biostudies-other

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Project description:Stem cells with enhanced resistance to oxidative stress after in vitro expansion have been shown to have improved engraftment and regenerative capacities. Such cells can be generated by preconditioning them with exposure to an antioxidant. In this study we evaluated the effects of Tualang honey (TH), an antioxidant-containing honey, on human corneal epithelial progenitor (HCEP) cells in culture. Cytotoxicity, gene expression, migration, and cellular resistance to oxidative stress were evaluated. Immunofluorescence staining revealed that HCEP cells were holoclonal and expressed epithelial stem cell marker p63 without corneal cytokeratin 3. Cell viability remained unchanged after cells were cultured with 0.004, 0.04, and 0.4% TH in the medium, but it was significantly reduced when the concentration was increased to 3.33%. Cell migration, tested using scratch migration assay, was significantly enhanced when cells were cultured with TH at 0.04% and 0.4%. We also found that TH has hydrogen peroxide (H2O2) scavenging ability, although a trace level of H2O2 was detected in the honey in its native form. Preconditioning HCEP cells with 0.4% TH for 48 h showed better survival following H2O2-induced oxidative stress at 50 µM than untreated group, with a significantly lower number of dead cells (15.3 ± 0.4%) were observed compared to the untreated population (20.5 ± 0.9%, p<0.01). Both TH and ascorbic acid improved HCEP viability following induction of 100 µM H2O2, but the benefit was greater with TH treatment than with ascorbic acid. However, no significant advantage was demonstrated using 5-hydroxymethyl-2-furancarboxaldehyde, a compound that was found abundant in TH using GC/MS analysis. This suggests that the cellular anti-oxidative capacity in HCEP cells was augmented by native TH and was attributed to its antioxidant properties. In conclusion, TH possesses antioxidant properties and can improve cell migration and cellular resistance to oxidative stress in HCEP cells in vitro.

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Sodium hyaluronate (hyaluronic acid) promotes migration of human corneal epithelial cells in vitro.

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