(59) The effect of Amnion-Chorion Membrane on Bone Marrow Derived Stem Cells

Introduction:

Background: Periodontal regeneration is defined as the reestablishment of lost periodontium, including alveolar bone, cementum, PDL, and gingival tissue. The dehydrated human amnion-chorion membrane (ACM) has been commonly used in daily practice to aim periodontal regenerations and ridge augmentations. ACM has been used successfully as membrane and sources of growth factor during periodontal regeneration procedures. However, to the best of our knowledge, there are no studies to evaluate ACM regarding stem cell proliferation.


Aim: The aim of this study was to evaluate the effects of ACM on cell proliferation of the bone marrow derived stem cell (BMSCs).

Methods: Material and

Methods: Commercially available BMSCs was utilized and following the media preparation, the cells were thawed and cultured. 10x20mm (10mg) ACM was used and pulverized in 10ml of PBS (1mg/ml). The medium was diluted to ACM extract of 1000 µg/ml, 500 µg/ml, 200 µg/ml, 100 µg/ml, and 50 µg/ml. BMSCs were seeded on a 96-well plate and were cultured for 3 days and then treated with ACM at different concentrations for 48 hours. At the end of 48 hours, WST-1 assay was used to analyze cell proliferation of BMSCs.

Results:

Results: BMSCs were seeded on three 96-well plates (18 well/concentration). A one way analysis of variance (ANOVA) with ACM extract level (control, 0.05mg, 0.1mg, 0.2mg,0.5mg, and 1mg) as the independent variable was employed to determine whether there are statistically significant cell proliferation levels among these 5 ACM levels with one control group over a 48-hour time period. The results indicated a statistically significant effect of ACM level on cell proliferation, F5, 102 = 102.40, p < .001. Because Levene’s test turned out to be statistically significant thereby suggesting that homogeneity of variance was not established and the sample size was relatively smaller, a conservative post-hoc test, Dunnett C test was run to detect which levels of ACM were different from each other. The results of this test revealed that 0.5mg and 1mg levels were the only ones that led to cell proliferation statistically significantly higher than the control group in this study, p’s < .001. Likewise, 1mg ACM level resulted in statistically significantly higher cell proliferation compared to 0.05mg, 0.1, 0.2mg, and 0.5mg levels, p’s <.001, thus leading to the largest cell proliferation. Finally, the 0.5mg ACM level was also associated with a statistically significantly higher level of cell proliferation than 0.05mg and 0.1mg levels of ACM only, p’s < .001 (not 0.2mg level).

Conclusions:

Conclusion: ACM significantly increased BMSC proliferation and higher concentrations of ACM (0.5 mg/ml and 1 mg/ml) showed higher results when compared to lower concentrations.

Lead Author’s Contributions to the Research Project: The lead author was responsible to perform experiments, for data collection, and composition of the abstract.