This study investigates the influence of the thickness of the oxide film formed on the surface of TC4 titanium alloy after anodization on the laser transmission welding between the titanium alloy and high borosilicate glass. The effects of oxidation time on the morphology and thickness of the oxide film were examined, and the elemental and phase composition of the oxide film were characterized. The differences in welding strength, weld morphology, and fracture surface between the titanium alloy before and after oxidation treatment and high borosilicate glass welding were compared, and analyzed the welding seam formation mechanism. Within our research scope, the results indicate that with an increase in oxide film thickness, the surface roughness gradually increases and the internal voids gradually increase, resulting in a decrease trend in the bonding strength of the welded joints with the increase in oxide film thickness. The interpenetrating structure formed by the mutual infiltration of glass and metal in the molten pool is identified as the main reason for their connection.