Nanostructured TiC Layer is Highly Suitable Surface for Adhesion, Proliferation and Spreading of Cells

Mariangela Lopreiato,Anna Scotto D’Abusco,Pietro Dalla Vedova,Alessia Mariano,Roberto Scandurra,Rossana Cocchiola,Giovanni Longo

doi:10.3390/condmat5020029

Abstract

Cell culture is usually performed in 2D polymer surfaces; however, several studies are conducted with the aim to screen functional coating molecules to find substrates more suitable for cell adhesion and proliferation. The aim of this manuscript is to compare the cell adhesion and cytoskeleton organization of different cell types on different surfaces. Human primary fibroblasts, chondrocytes and osteoblasts isolated from patients undergoing surgery were seeded on polystyrene, poly-d-lysine-coated glass and titanium carbide slides and left to grow for several days. Then their cytoskeleton was analyzed, both by staining cells with phalloidin, which highlights actin fibers, and using Atomic Force Microscopy. We also monitored the production of Fibroblast Growth Factor-2, Bone Morphogenetic Protein-2 and Osteocalcin, using ELISA, and we highlighted production of Collagen type I in fibroblasts and osteoblasts and Collagen type II in chondrocytes by immunofluorescences. Fibroblasts, chondrocytes and osteoblasts showed both an improved proliferative activity and a good adhesion ability when cultured on titanium carbide slides, compared to polystyrene and poly-d-lysine-coated glass. In conclusion, we propose titanium carbide as a suitable surface to cultivate cells such as fibroblasts, chondrocytes and osteoblasts, allowing the preservation of their differentiated state and good adhesion properties.

Highlights

In the last years, the in vitro cell culture fields have undergone significant developments, especially in their application to a wide range of research areas, such as screening of molecules to be used in the treatments of pathologies, toxicology and disease studies [1]
We monitored the production of Fibroblast Growth Factor-2, Bone Morphogenetic Protein-2 and Osteocalcin, using ELISA, and we highlighted production of Collagen type I in fibroblasts and osteoblasts and Collagen type II in chondrocytes by immunofluorescences
Human Primary Chondrocytes (HPCs) cultured on poly-d-Lys and txaFnOiRuPmEERcaRrEbViIdEWe layer (TiC) produced a higher amount of Bone Morphogenetic Protein-2 (BMP-2) compared to polystyrene; considering that the chondrocytes we used are from adult tissue, the production of BMP-2 by these cells is very promising

Summary

Introduction

The in vitro cell culture fields have undergone significant developments, especially in their application to a wide range of research areas, such as screening of molecules to be used in the treatments of pathologies, toxicology and disease studies [1]. The hOBs showed on TiC an arrangement which is much more resembling to that commonly found in bone: an increased production of collagens and other ECM components and subsequently an improved differentiation in osteocytes, that have several filaments used to stay in contact with other cells and with the bone microenvironment Both of these features were more present in the hOBs grown on TiC, compared to the cells cultivated on poly-d-Lysine and polystyrene (Figures 2 and 4). HPCs cultured on poly-d-Lys and TiC produced a higher amount of BMP-2 compared to polystyrene; considering that the chondrocytes we used are from adult tissue, the production of BMP-2 by these cells is very promising. Osteoblasts cultivated on TiC produced a statistically significant higher amount of OC compared to polystyrene and poly-d-Lys, and a higher amount of Coll I, a collagen very abundant in bone matrix (Figures 2 and 3)

Materials and Methods

Findings

Conclusions