Bioactive Silicate Nanoplatelets for Osteogenic Differentiation of Human Mesenchymal Stem Cells
Akhilesh K. Gaharwar, Silvia M. Mihăilă, Archana Swami, Alpesh Patel, Shilpa Sant, Rui L. Reis, Alexandra P. Marques, Manuela E. Gomes, Ali Khademhosseini
Abstract
Figure 2b. Effect of silicate nanoplatelets on hMSCs differentiation (Day 21).hMSCs were cultured in presence or absence of silicate nanoparticles (100 µg mL -1 ) in either normal media (NM), osteoconductive (OC) media, or osteoinductive (OI) medium for 21 days.The addition of silicate significantly enhances production of extracellular matrix after 21 days.An increase in runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN) was observed due to the addition of silicates (scale bar = 100 µm).Cells in normal media without silicate particles act as a negative control whereas cells in OI serve as a positive control.Cell nuclei were counterstained with DAPI (yellow).c) The protein production was quantified using image analysis from the fluorescence images.The intensity of protein per cell was qualified and later normalized by the control (hMSCs in normal growth media with no silicate particles) to obtain fold increase in the production of protein.The addition of silicate (100 µg mL -1 ) results in the production of RUNX2, OCN, and OPN in normal media, which indicates a strong bioactive character of silicate nanoplatelets.The addition of OC and OI media further enhances the production of osteoblast-related proteins.The results indicate that the addition of silicate promote osteogenesis of hMSCs.(one-way ANOVA followed by Tukey post-hoc, * P < 0.05, ** P < 0.01,