Aerides odoratum Lour., an epiphytic medicinal orchid native to Bangladesh, holds significant therapeutic and ornamental value but faces severe population decline due to overharvesting and habitat destruction. This study established an optimized in vitro propagation protocol and comprehensively evaluated quantitative phytochemical profiles, antioxidant, anti-inflammatory and cytotoxic activities across its naturally grown leaf, stem and root extracts. A robust propagation system utilizing varied basal medium supplemented with plant growth regulators (PGRs) successfully promoted seed germination, protocorm-like bodies (PLBs) proliferation and shoot-root differentiation and ex vitro acclimatization. Qualitative phytochemical screening revealed abundant secondary metabolites, including alkaloids, flavonoids, tannins, terpenoids and quinones, distributed differentially across organs. Biological efficacies demonstrated concentration-dependent responses. In the DPPH free radical scavenging assay, the leaf extract exhibited superior antioxidant activity with an IC50 of 181.25 µg/ml compared to stem (IC50 = 135.05 µg/ml) and root (IC50 = 57.138 µg/ml) extracts (standard ascorbic acid IC50 = 26.08 µg/ml). Anti-inflammatory evaluation via egg albumin denaturation inhibition revealed exceptional potency in the leaf extract (IC50 = 89.81 µg/ml), root extract (IC50 = 82.80 µg/ml), stem extract (IC50 = 40.56 µg/ml) and relative to standard acetylsalicylic acid (IC50 = 30.24 µg/ml). Cytotoxic evaluation using the brine shrimp lethality bioassay indicated moderate safety thresholds with median lethal concentration (LC50) values of 234.84 µg/ml (stem), 143.05 µg/ml (leaf) and 292.88 µg/ml (root) and relative to standard (LC50 = 43.50 µg/ml). These quantitative findings scientifically validate the traditional ethno-medicinal uses of A. odoratum and confirm its pharmacological potential, while the established tissue culture protocol offers a sustainable conservation framework.