Rahadian Indarto Susilo, Rahadian and Joni Wahyuhadi, Joni and I Ketut Sudiana, I Ketut and Fedik Abdul Rantam, Fedik Cytotoxicity test for the use of freeze-dried amniotic membranes against viability, proliferation, and apoptosis on brain cell culture: An in vitro study. Interdisciplinary Neurosurgery: Advanced Techniques and Case Management, 23. ISSN 2214-7519

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Abstract

Abstract Objective This study aimed to conduct a cytotoxicity test in determining biocompatibility of freeze-dried amniotic membranes on brain cell culture. Methods An in vitro study was carried out on rat’s brain cell culture. Samples were divided into three groups: control, conditioned medium, and direct amniotic membrane exposure. Each group was stained with MTT, DAPI and Annexin-V for its viability, proliferation, and apoptosis, respectively. Statistical analysis was conducted using ANOVA, post-hoc, or the Kruskal-Wallis test with CI 95%. Results Viability test using MTT staining showed viable cell in conditioned medium group of 76.99 ± 10.19, and direct amniotic membrane exposure group of 90.36 ± 23.31 (p = 0.001). DAPI staining showed a median value of 5.32 (1.97-16.53) for control group, 6.00 (2.42-16.62) for conditioned medium group, and 3.53 (1.32-8.69) for direct amniotic membrane exposure. Annexin-V single staining showed mean value of 10.28 ± 2.43 (control group), 10.07 ± 0.97 (conditioned medium), and 10.42 ± 4.07 (direct amniotic membrane; p = 0.969). Double staining by Annexin V + DAPI showed mean value of 10.43 ± 1.82 (control), 10.01 ± 1.07 (conditioned medium), and 12.40 ± 3.67 (direct amniotic membrane exposure; p = 0.148). Conclusion Amniotic membrane exposure affected rat’s brain cell culture viability in tolerable limit, while proliferation and apoptosis do not get affected

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