Figure 5.
Effects of HA of different molecular weights and HA-containing OTC eye drops on corneal epithelial wound healing were investigated using a scratch assay. Confluent hTCEpi were subjected to scratch wounds and the cells allowed to migrate into the cell free area for 24 hours in presence of HMWHA, mid-MWHA, and LMWHA at a concentration of 0.05 mg/mL or various HA-containing OTC eye products diluted 1:10 in media. Images were captured at zero, six, 18, and 24 hours under an EVOS microscope or an LSM 800 confocal microscope (Carl Zeiss Microscopy LLC), and the cell-free area was calculated using the plugin in Image J designed by Suarez-Arnedo et al.67 that is specific for in vitro wound healing assays. The cell-free area was calculated at each timepoint, and the wounded area was calculated as a percentage compared to zero hours. (A) Indicates the migration profile of corneal epithelial cells in the presence of HMWHA, mid-MWHA, and LMWHA compared to PBS control. (B) Indicates the migration profile of corneal epithelial cells in the presence of HA-containing eye drops that contained HA primarily in the HMWHA range compared to the HMWHA standard. (C) Indicates the migration profile of corneal epithelial cells in the presence of HA-containing eye drops that contained HA primarily in the mid-MWHA range compared to the mid-MWHA standard. (D) Indicates the migration profile of corneal epithelial cells in the presence of HA-containing eye drops that contained HA primarily in the LMWHA range compared to the LMWHA standard.