Cytotoxicity of Ethanol Extract of Borassus Flabellifer L. Fruit Seed Skin (Evaluation of Nuclear Fragmentation and Reduction on HSC-3 Cell Line) Raynita Rosalinda Angeline, Janti Sudiono
Department of Oral Pathology, Faculty of Dentistry, Universitas Trisakti
Abstract
Background: Longstanding cancer treatment causes side effects, necessitating alternative herbal treatment development like Borassus flabellifer L., with flavonoid and phenols as phytochemical compounds that can induce cancer cell apoptosis which can be qualitatively observed via cell morphological changes. Objectives: Determine the cytotoxicity of ethanol extract of B.flabellifer L. fruit seed skin on HSC3 cell line in the form of nuclear fragmentation and reduction. Method: Seven groups were used: negative control, positive control (doxorubicin 3micromole), and five B.flabellifer L. seed skin extract groups with different concentrations, applied to HSC3 cells planted in chamber slides, then incubated for 24, 48, and 72hours and microscopically observed after hematoxylin eosin staining. Three of four fields of view taken for each group were analyzed, considering nuclear fragmentation present with at least one fragmented nucleus. For nuclear reduction, surface area of all nucleus in each field of view are measured in ImageJ, calibrated with a scale bar, then recording the mean, standard deviation, minimum, and maximum values. Statistical analysis used Chisquare test for nuclear fragmentation and KruskalWallis test for nuclear reduction. Results: Nuclear fragmentation occurs in all extract groups at 24, 48 and 72hours, showing significant differences with the negative control (p=0.00). Significant differences in nuclear reduction between the negative control and extract groups of 750microgram/mL (p=0.02), 1500microgram/mL (p=0.01) (24hours), 375microgram/mL (p=0.00), 750microgram/mL (p=0.015) (48hours), 375microgram/mL (p=0.01), 750microgram/mL (p=0.013), 1500microgram/mL (p=0.022) (72hours). Conclusion: Ethanol extract of B.flabellifer L. fruit seed skin has cytotoxic effects on HSC-3 cell line in the form of nuclear fragmentation and reduction.
