Fig. 3.

Biological evaluation of the microfluidic chip substrate. (A) Live/dead staining of hFOB cells cultured for 1, 4 and 7 days. Live cells are stained green, and dead cells are stained red. Scale bar = 200 μm. (B) CCK-8 assay of hFOB cells cultured on different materials for 1, 4 and 7 days. (C) qRT-PCR analysis of mRNA levels of osteogenic genes. Relative gene expression levels of RUNX2, ALP, COL-1, OCN and OPN in hFOB cells after 7 days of culture. Immunofluorescence staining of hFOB cells cultured on different materials for 7 days: (D) OCN, (E) OPN, (F) ALP, (G) RUNX2, and (H) COL-1. Scale bar = 100 μm. (I) Area statistics of specific red fluorescence per cell, obtained with the help of ImageJ software (total red fluorescence area divided by the number of cells). Data are presented as the mean ± standard error, *p < 0.05, **p < 0.01, ***p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)