| Summary: | This study demonstrates that the fermentation of <i>Tetragonia tetragonioides</i> (<i>T. tetragonioides</i>) by <i>Bacillus subtilis</i> (<i>B. subtilis</i>) subsp. <i>spizizenii</i> enhances its antioxidant, anti-inflammatory, and skin-moisturizing activities. Fermented <i>T. tetragonioides</i> extracts (FTEs) showed a significant increase (<i>p</i> < 0.05) of approximately 1.3 to 3.07 times in their total polyphenol content (TPC), total flavonoid content (TFC), and vanillic acid content compared to pre-fermentation <i>T. tetragonioides</i> extracts (TEs). Additionally, the 2,2-diphenyl-1-picrylhydrazyl (DPPH)- and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging activities were significantly higher (<i>p</i> < 0.05) in FTEs than in TEs, showing an increase of about 1.25 to 1.64 times. The anti-inflammatory effects, evaluated through the inhibition of nitric oxide (NO) in RAW 264.7 cells stimulated by lipopolysaccharide (LPS), revealed that FTEs exhibited significant (<i>p</i> < 0.05) NO inhibition activity at less than half the concentration of TEs. It is particularly noteworthy that the FTE at 200 μg/mL significantly suppressed the expression of the cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and tumor necrosis factor-alpha (TNF-α) proteins. In HaCaT cells, FTEs substantially (<i>p</i> < 0.001) increased the mRNA expression of filaggrin (FLG), hyaluronan synthase (HAS)-1, and HAS-3, indicating improved skin protection and moisturization. In conclusion, this study confirms that <i>T. tetragonioides</i>’ antioxidant, anti-inflammatory, and skin-moisturizing activities are enhanced by <i>B. subtilis</i> fermentation, suggesting the potential of FTEs as a cosmeceutical ingredient.
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