Osteoporosis is a disease characterized by decreased bone strength, decreased bone mass, and bone deterioration. The current study investigates the effects of Gloiopeltis furcata ethanol extract (GFE) on antioxidant activity and osteoblast differentiation in MC3T3-E1 osteoblastic cells. Antioxidant activities of GFE were examined by evaluating 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, superoxide dismutase (SOD) activity, and total polyphenol content. Results showed that GFE contains 3.04 mg gallic acid equivalents/mL. GFE exhibited significantly strong scavenging activity in the DPPH assay, and 54.26% SOD activity was exhibited at 1,000 mg/mL concentration. Furthermore, GFE significantly increased the viability and proliferation of the MC3T3-E1 osteoblastic cells. Exposure to GFE promoted alkaline phosphatase (ALP) activity and mineralized nodules in MC3T3-E1 cells. Moreover, a dose-dependent increase in the expression of proteins associated with osteoblast growth and differentiation, such as Runx2, ALP, and osterix, was observed in the GFE-treated cells. Taken together, these results indicate that GFE encompasses both antioxidant potential and osteoblast differentiation properties. Thus, we conclude that GFE can potentially be applied as an alternate therapy for the prevention and treatment of osteoporosis.