Figure 4: Effects of Li⁺ in extracts on proliferation and differentiation of MC3T3-E1 cells.

(A) Li⁺ were released linearly from cements after immersion in medium for the same time, and most Li⁺ were released within 1day, only a small amount was released after 7 days (n = 5). (B) Cell proliferation on Li/CPC-50 and Li/CPC-100 was better at 3, 5 and 7 days than that on Li/CPC-200 and CPC (n = 5). (C) ALP activity at 7 days was better in Li/CPC-50 and Li/CPC-100 than for Li/CPC-200 and CPC (n = 5). (D,E) Osteogenic differentiation (ALP staining) and mineralisation (alizarin-red staining) of Li/CPC-50 and Li/CPC-100 were more obvious than those on Li/CPC-200 and CPC (n = 3). (F) Cells attached on Li/CPC-50 and Li/CPC-100 showed more spread and superior extension of filopodia, as well as greater focal adhesion via well-organized F-actin stress fibres (n = 3). Statistical analyses were done using Students’t-test. *p < 0.05 was considered significant.