Abstract:

Objective

The aim of this study is to investigate the effects and mechanisms of polyvinylidene fluoride（PVDF）on the migration and proliferation of human corneal epithelial cell（HCEC）.

Methods

PVDF films were prepared using solution casting and electrospinning methods.Scanning electron microscopy，X-ray diffraction，fourier-transform infrared spectroscopy，differential scanning calorimetry，and UV-Vis spectrophotometry were employed to select the optimal process from the aspect of the surface morphology，crystallinity，and visible light transmittance for two fabrication methods，and used for subsequent cell experiments.PVDF was divided into non-polarized and corona-poled groups with or without spontaneous polarization.Biocompatibility was assessed using cell counting kit-8.Scratch assay，transwell migration assay，and EdU proliferation assay were used to evaluate the effects of PVDF on cell migration and proliferation.Immunofluorescence and high-content cellular texture analysis were applied to assess cytoskeletal changes.Migration rates，migrated cell counts，proliferation rates，cytoskeletal length，and width met homogeneity of variance and normal distribution，were expressed as ±s and compared by one-way ANOVA with Bonferroni post hoc tests for intergroup.

Results

Solution-cast PVDF films exhibited superior physicochemical properties compared to electrospun films and were selected for cell experiments.PVDF significantly promoted HCEC migration and proliferation.In the scratch assay，the migration rates of the control group，non-polarized PVDF group，and polarized PVDF group were（61.43±2.86）%，（70.36±5.87）%，and（96.12±0.48）%，respectively，with statistically significant differences（F＝75.91，P＜0.05）.In the Transwell assay，the numbers of migrating cells in the control group，non-polarized PVDF group，and polarized PVDF group were（1643.11±51.17），（1827.44±51.81），and（2392.89±57.99），respectively，with statistically significant intergroup differences（F＝105.69，P＜0.05）.The cell proliferation rates in the control group，non-polarized PVDF group，and polarized PVDF group were（27.83±1.62）%，（30.58±1.46）%，and（41.20±1.89）%，respectively，with statistically significant intergroup differences（F＝115.13，P＜0.05）.The microtubule lengths and widths of HCECin the control group，non-polarized PVDF group，and polarized PVDF group were（1.41±0.06）μm，（0.41±0.01）μm，（1.52±0.02）μm，（0.44±0.01）μm，（1.72±0.03）μm，and（0.44±0.01）μm，respectively.Statistically significant differences were observed among the three groups in both microtubule length and width（F＝29.76，9.67；P＜0.05）.The microfilament lengths and widths were（1.24±0.01）μm，（0.38±0.01）μm，（1.31±0.01）μm，（0.40±0.01）μm，（1.40±0.03）μm，and（0.40±0.01）μm，respectively，with statistically significant intergroup differences in both microfilament length and width（F＝42.70，62.70；P＜0.05）.

Conclusions

Solution-cast PVDF demonstrates excellent physicochemical properties and biocompatibility.It promotes HCECmigration and proliferation by regulating cytoskeletal reorganization via an electrical microenvironment generated by spontaneous polarization，with corona poling significantly enhancing this effect，which providing a novel strategy for corneal regenerative medicine.

Key words: Polyvinylidene fluoride, Corneal epithelial cells, Migration, Proliferation, Cytoskeleton