Introduction: Herbal drugs have healing properties to prevent and treat various health conditions, including topical treatments, which contain therapeutic active ingredients like Hibiscus rosa-sinensis L. leaf extract combined with a carrier substance such as bovine gelatin hydrogel. Hibiscus rosa-sinensis L. extract also exhibits other pharmacological effects including antioxidant, antibacterial, antifungal, anti- inflammatory, antidiabetic, and anticancer properties. Bovine gelatin-based hydrogels have non-immunogenic properties derived from natural materials, with controlled drug release, as well as good biodegradability and biocompatibility, making them non-toxic. The combination of bovine gelatin hydrogel with Hibiscus rosa-sinensis L. leaf extract has been studied to develop a new topical drug. This research focuses on assessing the safety of this combination through cytotoxicity testing. Materials and Methods: This in vitro experimental study, utilized a post-test only control group design. The control group utilizes a bovine gelatin-based hydrogel, while the treatment group uses the same hydrogel combined with Hibiscus rosa-sinensis L. leaf extract on BHK-21 fibroblast cell cultures. Preclinical testing using the MTT assay served as a cytotoxicity test to validate the safety of this topical drug. Comparative study using a T-test with a significance level of p<0.05. Data analysis was carried out using Statistical Product and Service Solution (SPSS) Software for Mac version 29. Results: The cytotoxicity test results for bovine gelatin hydrogel and the combination with the extract of Hibiscus rosa-sinensis L. leaves were shown in the percentage of viable cells of 92.3% and 96.4%. The parametric T-test revealed no significant difference between the two groups (p = 0.754). Conclusions: The combination of bovine gelatin hydrogel with the extract of Hibiscus rosa-sinensis L. leaves in the development of herbal drugs are non-toxic
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