Bioactive glass-induced osteoblast differentiation: a noninvasive spectroscopic study.
Here, we report on a rapid, noninvasive biophotonics system using Raman spectroscopy to detect real-time biochemical changes in foetal osteoblasts (FOBs) following exposure to 45S5 Bioglass (BG)-conditioned media. Bio-Raman spectroscopy, combined with multivariate statistical analysis techniques (principal component analysis and least squares analysis), was able to noninvasively identify biochemical differences in FOBs cultured for different time periods and between FOBs exposed/or not to BG-conditioned media. Gene and protein expression studies were also performed for known markers of osteoblastic differentiation, namely, alkaline phosphatase, bone sialoprotein, and collagen type I. Quantitative RT-PCR confirmed upregulation of genes associated with osteoblast differentiation after exposure to BG-conditioned media. These results suggest that Raman spectroscopy can noninvasively detect biochemical changes in FOBs associated with differentiation. This technique could have important applications in the field of regenerative medicine by enabling rapid characterization of cell or organoid behavior on novel bioactive scaffolds without damage to either cell or biomaterial.
Jell
G
G
Department of Materials, Imperial College London, London, SW7 2AZ, United Kingdom.
Notingher
I
I
Tsigkou
O
O
Notingher
P
P
Polak
J M
JM
Hench
L L
LL
Stevens
M M
MM
D000469:Alkaline Phosphatase / Q000378:metabolism; D001672:Biocompatible Materials / Q000737:chemistry*; D002454:Cell Differentiation; D002478:Cells, Cultured; D002516:Ceramics; D017077:Culture Media, Conditioned / Q000378:metabolism; D005898:Glass / Q000737:chemistry*; D006801:Humans; D058574:Integrin-Binding Sialoprotein; D016018:Least-Squares Analysis; D015233:Models, Statistical; D010006:Osteoblasts / Q000166:cytology*; D017785:Photons; D025341:Principal Component Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000378:metabolism; D013059:Spectrum Analysis, Raman / Q000379:methods*
D001672:Biocompatible Materials; D017077:Culture Media, Conditioned; C546871:IBSP protein, human; D058574:Integrin-Binding Sialoprotein; D012795:Sialoglycoproteins; C425330:bioactive glass 45S5; D000469:Alkaline Phosphatase