Previous studies have shown that bioactive glasses can support osteoblastic growth and differentiation in vitro as well as in vivo. More recently, a new category of sol-gel glasses has been developed with enhanced bioactivity and open pores enclosed in a mesoporous matrix. In our study, we investigated the effect of 58S sol-gel glasses on the growth and differentiation of mouse calvaria osteoblasts. Two types of granules were used: 58S sol-gel granules and 60S inert glasses used as control. Phase contrast microscopy showed that cells proliferated and formed mineralized bone nodules in both cultures. However, this phenomenon occurred earlier and to a higher degree in cultures with 58S sol-gel glasses. Northern blot analysis of the expression of osteoblastic markers revealed that osteoblasts retained their phenotype in both types of cultures. Interestingly, stimulation of alkaline phosphatase, bone sialoprotein, and osteocalcin was noticed at day 18 in sol-gel cultures when compared with that in control. These data confirm that 58S bioactive glasses are capable of supporting the growth and maturation of primary mouse osteoblasts. In addition, it was shown that 58S glasses affected the gene-expression profile, causing an up-regulation of the major bone markers. These results indicated that 58S sol-gel glasses appeared as suitable candidates for osteoblast scaffolds in the field of bone tissue engineering.