This study evaluated the effect of acid etching on surface characteristics, flexural strength and osteoblast cell response of glass-infiltrated zirconia. Zirconia specimens were divided into six groups: untreated zirconia (Z); glass-infiltrated zirconia (ZG); glass-infiltrated and sandblasted zirconia (ZGS); glass-infiltrated, sandblasted and 5 min acid-etched zirconia (ZGS-E5); glassinfiltrated, sandblasted and 15 min acid-etched zirconia (ZGS-E15); glass-infiltrated, sandblasted and 25 min acid-etched zirconia (ZGS-E25). Surface roughness, biaxial flexural strength and MC3T3-E1 cell proliferation were evaluated. When increasing etching time, surface roughness significantly increased while flexural strength decreased. Cell proliferation rate at day 3 on group ZGS-E15 and ZGS-E25 was significantly higher than that of other groups. Surface roughness and flexural strength of glass-infiltrated zirconia can be controlled by adjusting etching time. Rough surface made by acid etching following glass infiltration significantly enhanced osteoblast cell response. Glass infiltration improved strength of zirconia but severe acid etching slightly reduced strength of zirconia.