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Traditional herbal remedies are naturally occurring, plant-derived substances with limited to no chemical processing and have been used in local or national healing traditions to treat illness. In global health debates, traditional herbal medicines are gaining considerable attention. Many hope new research into herbal medicine will play a vital role in global health. Countries like China, India, Nigeria, USA, and also the World Health Organization (WHO) made large investments in ancient herbal medicines. Currently, the use of the Indonesian plant has dramatically improved the medical and dentistry field. The dental and oral care is often related to wounds, and the Ambonese banana stem has been proven as an effective treatment for these injuries. This study aimed to evaluate the effectiveness and safety of the Ambonese banana stem sap through cytotoxicity tests on the fibroblast cell culture of Baby Hamster Kidney-21 (BHK-21). This study was carried out on three BHK-21 fibroblast cell culture, namely, the media and cell control, and the Ambonese banana stem sap with a concentration of 10%, 20%, 30%, 40%, 50%, 60 %, 70%, 80%, 90%, and 100% incubated for 24 hours at 37ºC and 5% CO2. Then, MTT was evenly dispersed on the media to obtain accurate optical density value. All quantitative data were statistically analyzed using one-way ANOVA and Tukey's HSD Test. The result showed significant differences in optical density values between groups with p = 0.000 (p <0.05). There was no significant difference between the cell control and the Ambonese banana stem sap group with concentrations of 90%, 80%, 70%, 60%, 50%, 30%, 20%, and 10%. It was also observed that the Ambonese banana stem sap is nontoxic to fibroblast cells since its viability value was more than 60%. [ABSTRACT FROM AUTHOR]