Crassula "Buddha's Temple" aqueous extract (BTAE), prepared via an optimized, reproducible hydrothermal extraction, exhibits protective antioxidant and lipid regulatory effects in adipocyte models subjected to oxidative stress. The phytochemical profile of BTAE revealed a chemically diverse composition enriched in polyphenols (chlorogenic acid, quercetin, kaempferol, catechin) alongside sulfur-containing metabolites, tricarboxylic acid cycle intermediates, nucleotides, and peptide-like compounds, indicating multifaceted biochemical activity. In both 2D monolayer and advanced 3D bioprinted 3T3-L1 adipocyte cultures exposed to tert-butyl hydroperoxide (tBHP), BTAE pretreatment (5.7 ng/mL) significantly attenuated cytotoxicity, reduced intracellular and mitochondrial reactive oxygen species accumulation, preserved mitochondrial membrane potential, and suppressed lipid peroxidation. Concurrently, BTAE modulated gene expression by downregulating proapoptotic (Bax, CASP3) and lipogenic genes (GPAT1, ABHD5) while upregulating the antiapoptotic gene Bcl2. Notably, BTAE's efficacy matched or exceeded that of N-acetylcysteine, a standard antioxidant control. Enhanced bioactivity of BTAE in 3D adipocyte constructs suggests physiological microenvironment-dependent potentiation of its cytoprotective and metabolic regulatory functions. These findings support BTAE's potential as a natural, safe, and cost-effective candidate for mitigating oxidative stress and lipid dysregulation in adipose tissues, with implications for metabolic disease therapeutics including nonalcoholic fatty liver disease.
Keywords: 3D bioprinting; adipocytes; lipid metabolism; mitochondrial function; nonalcoholic fatty liver disease; oxidative stress.
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