Bioactive glass‐induced osteoblast differentiation: A noninvasive spectroscopic study

Abstract

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.

Twenty-first century challenges for biomaterials

“…The bio-Raman system offers the potential of monitoring and maintaining cell phenotype by in situ spectroscopic analysis of the cells Jones et al 2005;Jell et al 2008;Swain et al 2008a;Gentleman et al 2009). Notingher et al show that two statistical-based spectroscopic data management programs, called principal component analysis and linear discriminant analysis (LDA), are powerful tools for distinguishing cell phenotypes in real time.…”

“…Notingher et al show that two statistical-based spectroscopic data management programs, called principal component analysis and linear discriminant analysis (LDA), are powerful tools for distinguishing cell phenotypes in real time. Bio-Raman spectroscopic analyses were made of MG63 immortal human osteosarcoma-derived osteoblasts and compared with primary human osteoblasts, obtained from surgically excised femoral heads ( Jell et al 2008). An LDA model prepared from the data showed high cross-validation sensitivity (100%) and high specificity (95%) for discriminating the MG63 cells from the primary cells, with 96 per cent of the cells being correctly classified either as tumour-derived or non-tumourderived cells.…”

Bioactive glass induced osteogenic differentiation of human adipose stem cells is dependent on cell attachment mechanism and mitogen-activated protein kinases

“…Cells adhere to the growth substrate by means of FAs, which, upon maturation, are characterised by the presence of vinculin (Burridge and ChrzanowskaWodnicka, 1996). Surfaces that induce the formation of prominent FAs support osteogenic differentiation of stem and progenitor cells, whereas small and evenly spread FAs are linked to a decrease in osteogenic and an increase in adipogenic commitment (Abagnale et al, 2015;Biggs et al, 2009;Kilian et al, 2010). In contrast to these studies, conducted with various surface topographies and micropatterns created in polystyrene and polyimide, the results presented here indicated that the mature FAs of hASCs on S53P4 and 1-06 were small and evenly dispersed despite the evident osteogenic potential of both BaG types.…”

Bioactive glass and glass-ceramic scaffolds for bone tissue engineering

“…Key mechanisms leading to enhanced new bone growth are now known to be related to the controlled release of ionic dissolution products from the degrading bioactive glass, especially critical concentrations of biologically active, soluble silica and calcium ions [8,191]. Recent studies have shown that bioactive (partially) resorbable glasses and their ionic dissolution products enhance osteogenesis by regulating osteoblast proliferation, differentiation, and gene expression [7,41,51,[191][192][193][194][195][196][197][198].…”