Cell morphology in response to borosilicate glasses.

a. Phalloidin staining (red) of actin cytoskeleton of hASCs cultured on glass discs for 7d. b. Phalloidin staining (red) of actin cytoskeleton of hASCs cultured in glass extracts for 7d. Nuclei were stained blue with DAPI. Scale bars 200 μm. BM = basic medium, OM = osteogenic medium. B1 = undiluted extract, B2 = 1:10 dilution, B3 = 1:100 dilution.</p

Ion concentrations within BaG-free DMEM/F-12 and BaG extract media.

The extract media were prepared from B25 glass granules. In addition to undiluted extract (B1), dilutions of 1:10 (B2) and 1:100 (B3) were evaluated.</p

Borosilicate glass-induced early osteogenic commitment of hASCs.

a. ALP activity (n = 9) and expression of osteogenic marker genes RUNX2a (b), OSTERIX (c), DLX5 (d), OSTEOPONTIN (e) and OSTEOCALCIN (f) (n = 2–4) in hASCs cultured on glass discs. g. ALP activity (n = 12) and the expression of osteogenic marker genes RUNX2a (h), OSTERIX (i), DLX5 (j), OSTEOPONTIN (k) and OSTEOCALCIN (l) (n = 3–4) in hASCs cultured in glass extracts. * p<0.05 between the indicated group and the control (ctrl) group at the same time point. BM = basic medium, OM = osteogenic medium. B1 = undiluted extract, B2 = 1:10 dilution, B3 = 1:100 dilution.</p

Characterization of glass dissolution and reaction layer formation.

a-d. Si, B, Ca and P concentrations, respectively, in SBF as a function of immersion time. e. The pH of SBF solution as a function of immersion time. f. SEM images of S53P4, B25 and B50 glass particles immersed for 7d in SBF. Scale bar 20 μm. g. SEM images of S53P4, B25 and B50 glass disc cross-sections, after immersion for 21 days in cell culture medium. Scale bar 10 μm.</p

The nominal compositions of the BaGs used in this study.

The nominal compositions of the BaGs used in this study.</p

Borosilicate glass-induced late osteogenic differentiation.

a. Immunocytochemical staining of osteocalcin (green) in hASCs cultured on glass discs for 21d. b. Immunocytochemical staining of osteocalcin (green) in hASCs cultured in borosilicate glass extracts for 19d. Nuclei were stained blue with DAPI and actin cytoskeleton red with phalloidin. Scale bars 200 μm. c. Alizarin red S staining of CaP mineral in borosilicate glass extracts after 19d of culture. Mineral is stained red. Each image represents the whole well of a 24-well plate. d. Quantified Alizarin red S staining at 19d. n = 6. * p<0.05 between the indicated group and the control (ctrl) group at the same time point. BM = basic medium, OM = osteogenic medium. B1 = undiluted extract, B2 = 1:10 dilution, B3 = 1:100 dilution.</p

The primer sequences and the accession numbers of the mRNAs studied with qRT-PCR.

The primer sequences and the accession numbers of the mRNAs studied with qRT-PCR.</p

Cell viability and proliferation in response to borosilicate glasses.

a. Cell viability on bioactive glass discs at 21d. b. Cell viability in borosilicate glass extracts at 19d. Viability was analyzed with Live/dead staining. Scale bars 1.0 mm. c. Cell proliferation on glass discs (n = 9). d. Cell proliferation in glass extracts (n = 12). Proliferation was analyzed with CyQUANT assay. * p<0.05 between the indicated group and the control (ctrl) group at the same time point. BM = basic medium, OM = osteogenic medium. B1 = undiluted extract, B2 = 1:10 dilution, B3 = 1:100 dilution.</p

The surface marker expression of undifferentiated hASCs at passage 1, cultured in BM.

The surface marker expression of undifferentiated hASCs at passage 1, cultured in BM.</p

Collagen-I production in response to borosilicate glasses.

a. Immunocytochemical staining of collagen-1 (green) in hASCs cultured on glass discs for 21d. b. Immunocytochemical staining of collagen-1 (green) in hASCs cultured in borosilicate glass extracts for 18d. Nuclei were stained blue with DAPI and actin cytoskeleton red with phalloidin. Scale bars 200 μm. BM = basic medium, OM = osteogenic medium. B1 = undiluted extract, B2 = 1:10 dilution, B3 = 1:100 dilution.</p