Cytotoxicity test for the use of freeze-dried amniotic membranes against viability, proliferation, and apoptosis on brain cell culture: An in vitro study

• Published in: Interdisciplinary neurosurgery : Advanced techniques and case management 2021-03, Vol.23, p.100947, Article 100947
• Main Authors: Susilo, Rahadian Indarto, Wahyuhadi, Joni, Ketut Sudiana, I, Rantam, Fedik Abdul
• Format: Article
• Language: English
• Subjects: Cytotoxicity; Dura mater defect; Freeze-dried amniotic membrane; Viability
• ISSN: 2214-7519; 2214-7519
• DOI: 10.1016/j.inat.2020.100947

This study aimed to conduct a cytotoxicity test in determining biocompatibility of freeze-dried amniotic membranes on brain cell culture. 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%. 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). Amniotic membrane exposure affected rat’s brain cell culture viability in tolerable limit, while proliferation and apoptosis do not get affected.