The biocompatibility assessment of medical devices is a vital component of preclinical safety evaluation to ensure their compatibility with biological systems before clinical application. The present study assesses both the Cytotoxicity (in vitro) and acute systemic toxicity (in vivo) potential of the Venous Implantable Port in accordance with ISO 10993 series standards and OECD principles of Good Laboratory Practice (GLP). The in vitro cytotoxicity test was conducted using the L929 mouse fibroblast cell line as per ISO 10993-5:2009/EN ISO 10993-5:2009 and US FDA (FR 2-245):2016 guidelines. Cytotoxic potential was evaluated by morphological observation and the MTT colorimetric assay. The study revealed normal cell morphology and cell viability above the acceptance threshold, indicating the absence of cytotoxicity. The in vivo acute systemic toxicity evaluation was performed in Swiss albino mice as per ISO 10993-11:2017/EN ISO 10993-11:2018 and ISO 10993-12:2021 guidelines. Appropriate extracts of the test item were administered via different dosing routes. Throughout the observation period, no mortality, morbidity, or abnormal clinical signs were observed and gross pathological examination showed no visible tissue or organ abnormalities. Overall, the Venous Implantable Port was found to be non-cytotoxic in cytotoxicity study, and non-toxic in acute systemic toxicity study, satisfying the biological safety requirements and toxicological end points for the same as per the ISO 10993 standard.