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The recent developments in nanotechnology have driven researchers towards the application of latex extracts for the green synthesis of silver nanoparticles (AgNPs). In this study, AgNPs were biologically synthesized using latex extracts from <i>Tabernaemontana ventricosa</i>, characterized, and their respective biological activities were assessed. Our results showed prominent silver (Ag) peaks at 410 nm confirmed by UV-vis while the elemental percentage composition (3.89 ± 0.16%) of the AgNPs was verified by EDX. The SEM and HRTEM analysis revealed spherical, ovate, and triangular AgNPs, with diameters ranging from 5.00 nm–17.50 nm; however, larger hydrodynamic diameters were revealed by NTA analysis. The FTIR spectra results displayed several peaks of bending and stretching associated with various functional groups such as alcohols, alkanes, amines, proteins, enzymes, and other biomolecules possibly responsible for the capping, reduction, and functionalization of AgNPs. In addition, the AgNPs showed strong antibacterial activity (diameter of the zone of inhibition) against <i>Escherichia coli</i> (12.67 ± 1.15 mm), <i>Staphylococcus aureus</i> (11.67 ± 0.58 mm), and <i>Pseudomonas aeruginosa</i> (11.33 ± 0.58 mm), with significant cytotoxic activity noted in the HeLa cells (10.52 µg/mL). The study confirmed the successful production of AgNPs and recommends <i>T. ventricosa</i> latex extracts as effective capping agents of nanoparticles (NPs).