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

• Main Authors: Rahadian Indarto Susilo, Rahadian, Joni Wahyuhadi, Joni, I Ketut Sudiana, I Ketut, Fedik Abdul Rantam, Fedik
• Format: Article PeerReviewed
• Language: English; English; English; English
• Published: Elsevier
• Subjects: R5-920 Medicine (General)
• Online Access: https://repository.unair.ac.id/120038/1/08.%20Cytotoxicity%20Test%20for%20the%20Use%20of%20Freeze-Dried.pdf; https://repository.unair.ac.id/120038/2/08.%20Artikel_compressed.pdf; https://repository.unair.ac.id/120038/3/08.%20turnitin.pdf; https://repository.unair.ac.id/120038/8/08.%20karil.pdf; https://repository.unair.ac.id/120038/; https://www.sciencedirect.com/science/article/pii/S2214751920305089
• Institution: Universitas Airlangga

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