Formulation of Cantigi (Vaccinium varingiaefolium Blume Miq.) Leaf Extract-Loaded Gelatin Nanoparticles as Anti-Breast Cancer In vitro, Edition 1

Background: Indonesia, an archipelagic country with numerous volcanoes, is home to Cantigi (Vaccinium varingiaefolium Blume Miq.), which thrives around the volcanic craters, including on Mount Tangkuban Parahu in North Bandung. Empirically, the local community uses it for salad, cosmetics, and health. Previous studies have demonstrated that Cantigi leaf extract exhibits significant cytotoxic activity against L1210 leukemia cells, though its effects on breast cancer cells have yet to be studied.

Objectives: To obtain simplicia and extracts of Cantigi leaves that meet the quality standards of simplicia and extracts; to determine the toxicity data of Cantigi leaf extract using BSLT and MTT methods on T47D and MCF-7 breast cancer cells, and on normal Vero cells, and determine their selectivity index; to obtain data on volatile bioactive compounds using the GC-MS method and non-volatile bioactive compounds using the LC-MS/MS method; to formulate and obtain physicochemical data and optimum cytotoxic activity of Cantigi leaf extract-loaded gelatin nanoparticles.

Methodology: Cantigi leaves of Mount Tangkuban Parahu were made into simplicia using an oven, and the extract using maceration in stages with hexane, ethyl acetate, and ethanol. The simplicia and extracts were tested and standardized against specific and non-specific parameters. The extract cytotoxic activity was prepared and tested by BSLT (Artemia salina larva) and MTT (T47D, MCF-7, and Vero cells) methods, and the selectivity index was calculated and compared. Extract phytochemicals were analyzed using GC-MS and LC-MS/MS methods. The extract was prepared and synthesized into gelatin nanoparticles by varying two process variables and three material concentrations and characterized by particle size, polydispersity index, zeta potential, entrapment efficiency, IC50 (T47D cells), FTIR, morphology, and release profile.

Results: The simplicia and extract met the specific and non-specific quality standards. The ethyl acetate extract was toxic on the BSLT test (LC50 of 320,63 ppm) and had the highest cytotoxic activity ((IC50 of 75.16+3.05 ppm, the MTT test on T47D cells). GC-MS identified two main compounds (palmitic acid and stearic acid), and LC-MS/MS identified three main compounds (bufalin, trametenolic acid, and beta-sitosterol-3-O-beta-glucopyranoside), and all had cytotoxic activities on breast cancer cells. Gelatin nanoparticles can be prepared and characterized. The most optimum gelatin nanoparticles have the following characteristics: Particle size of 128.7+28.8 nm, polydispersity index of 0.293+0.00, zeta potential of 9.53+0.00 mV, entrapment efficiency of 74.87+0.00 %, and cytotoxic activity (IC50) of 16.88+1.42 ppm. In general, the gelatin nanoparticles met the nanoparticle criteria.

Conclusion: The formulation of gelatin nanoparticles from Cantigi leaf extract (Vaccinium varingiaefolium Blume Miq.) as an in vitro breast cancer agent can produce gelatin nanoparticles that have optimum cytotoxic activity with IC50 of 16.88 ppm, and this activity is better than the cytotoxic activity of the crude extract with IC50 of 75.16 ppm.