Abstract

The objective of this in vitro study was to evaluate the possible regenerative effects of dehydrated human amnion chorion membrane (ACM) on cell proliferation, migration, and differentiation/mineralization of bone marrow-derived stem cells (BMSCs). The dehydrated human ACM has been commonly used in periodontal regeneration and ridge augmentation procedures due to its membrane properties and growth factors. For the study, commercially available bone marrow-derived stem cells were thawed, cultured, and exposed to different concentrations of amnion-chorion membrane extract (50, 100, 200, 500, and 1000 μg/mL) in mesenchymal stem cell basal media. Cell proliferation was analyzed using the WST-1 assay, migration with the scratch-wound assay, and differentiation/mineralization with Alizarin Red S staining. Two-way ANOVA was used to assess the effects of concentration and time, with pair-wise comparisons made using Fisher's Protected Least Significant Difference at a 5% significance level. Results showed that the control, 50 μg/mL, and 100 μg/mL groups exhibited lower cell proliferation compared to the 500 μg/mL and 1000 μg/mL groups. Lower concentrations also displayed reduced cell migration compared to the higher concentrations. In the mineralization assay, the control, 200 μg/mL, and 500 μg/mL groups exhibited lower cell mineralization compared to 1000 μg/mL. Temporally, shorter durations showed higher levels of cell migration and lower levels of cell mineralization. Overall, the use of amnion-chorion membrane positively affected cell proliferation, migration, and mineralization, with increased concentrations of amnion-chorion membrane extract yielding greater benefits. This study indicates that amnion-chorion membrane may enhance stem cell activities due to the presence of growth factors in the membrane. These findings are preliminary and must be interpreted cautiously. Further in vivo and clinical studies are necessary to validate these observations.