Application of Raman Spectroscopic Imaging to Assess the Structural Changes at Cell-Scaffold Interface

Abstract

Raman spectroscopic imaging and mapping were applied to characterise three-compound ceramic composite biomaterial consisting of chitosan, β-1,3-d-glucan (curdlan) and hydroxyapatite (HA) developed as a bone tissue engineering product (TEP). In this rapidly advancing domain of medical science, the urge for quick, reliable and specific method for products evaluation and tissue–implant interaction, in this case bone formation process, is constantly present. Two types of stem cells, adipose-derived stem cells (ADSCs) and bone marrow-derived stem cells (BMDSCs), were cultured on composite surface. Raman spectroscopic imaging provided advantageous information on molecular differences and spatial distribution of compounds within and between the cell-seeded and untreated samples at a microscopic level. With the use of this, it was possible to confirm composite biocompatibility and bioactivity in vitro. Deposition of HA and changes in its crystallinity along with protein adsorption proved new bone tissue formation in both mesenchymal stem cell samples, where the cells proliferated, differentiated and produced biomineralised extracellular matrix (ECM). The usefulness of spectroscopic Raman imaging was confirmed in tissue engineering in terms of both the organic and inorganic components considering composite–cells interaction.

Effectiveness of the production of tissue-engineered living bone graft: a comparative study using perfusion and rotating bioreactor systems

“…For better comparison spectra were normalized to the highest band at 960 cm −1 , assigned to ν 1 stretching of P–O bands in hydroxyapatite crystals’ PO 4 groups 24 . Spectra had characteristic features ascribed to hydroxyapatite, with band maxima at 430, 590, 960, 1045, and 1074 cm −1 corresponding to phosphate group vibrations: ν 2 , ν 4 , ν 1 , ν 3, and (CO 3 ) 2− , respectively 25 . The untreated biomaterial (native) was the reference sample for cell-seeded biomaterials maintained in static conditions (mat_S), in bioreactors (mat_P and mat_R) and for non-seeded biomaterial incubated in the culture medium (sample marked as control).…”

Raman Spectroscopy in Skeletal Tissue Disorders and Tissue Engineering: Present and Prospective

“… 49 , 114–116 In a recent study, by combining classical RS spectra analysis, Raman spectral imaging, and mapping, Kalisz et al could have access not only to the biochemical changes at the interface between stem cells and osteo-inductive scaffolds, but also to the three-dimensional deposition of the ECM components and mineralization at the cell-material interface. 121 In their study, the authors first measured the Raman profiles of the three raw components (Chitosan [CHI], beta 1,3 glucan polysaccharide, and HA) that they used in the fabrication of theirs scaffolds, and they monitored the matrix deposited by MSCs from adipose tissue and bone marrow ( Fig. 7 ).…”

“…Figures are reproduced with permission from Kalisz et al (doi:10.3390/ijms22020485). 121 ECM, extracellular matrix. Color images are available online.…”