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

Grzegorz Kalisz,Michal Jedrek,Anna Sroka-Bartnicka,Agata Przekora,Barbara Gieroba,Paulina Kazimierczak,Wojciech Grudzinski,Wieslaw I Gruszecki,Grazyna Ginalska

doi:10.3390/ijms22020485

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.

Highlights

Modern methods for bone lesion treatment are developing in a wide range of approaches, with meticulous analysis of biological and physical properties of connective tissue
Recent reports have indicated that mesenchymal stem cells acquired from adipose tissue have comparable morphology, phenotype and differentiation potential to bone marrow-derived stem cells (BMDSCs)
It was reported that both adipose-derived stem cells (ADSCs) and BMDSCs have the ability to proliferate and differentiate on the biomaterial scaffold, the process is influenced by multiple features of the scaffold [3]

Summary

Introduction

Modern methods for bone lesion treatment are developing in a wide range of approaches, with meticulous analysis of biological and physical properties of connective tissue. Autologous bone grafts are considered to be a close future gold standard for reconstructive applications in the biomedical sciences Nowadays, this approach encounters issues associated with limited availability of autografts, chronic pain, morbidities and infections [3]. Recent reports have indicated that mesenchymal stem cells acquired from adipose tissue have comparable morphology, phenotype and differentiation potential to BMDSCs. ADSCs (adipose-derived stem cells) seem to be promising source of cells for applications in tissue engineered biomaterials [10]. Structure based on hydroxyapatite (HA) and chitosan (CHIT) mixture with addition of 1,3-beta-D-glucan (GLU, curdlan) was converted to be more elastic with maintaining mechanical properties of bioceramics and physicochemical properties of the surface [12,13] It has huge potential in biomedical applications [14]

Methods

Results

Discussion

Conclusion