Interdisciplinary approach to cell–biomaterial interactions: biocompatibility and cell friendly characteristics of RKKP glass–ceramic coatings on titaniumby Mario Ledda, Angela De Bonis, Francesca Romana Bertani, Ilaria Cacciotti, Roberto Teghil, Maria Grazia Lolli, Antonio Ravaglioli, Antonella Lisi, Julietta V Rau

Biomed. Mater.


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Interdisciplinary approach to cell–biomaterial interactions: biocompatibility and cell friendly characteristics of RKKP glass–ceramic coatings on titanium

View the table of contents for this issue, or go to the journal homepage for more 2015 Biomed. Mater. 10 035005 (

Home Search Collections Journals About Contact us My IOPscience © 2015 IOP Publishing Ltd 1. Introduction

Current and forthcoming future regenerative medicine requires new smart materials for tissue repair and engineering. Among these, bioactive glasses are known as a promising class of materials due to their ability to bind to biological hard and soft tissues [1–3]. Since the discovery of bioglass in the late 1960s by Hench, this material underwent a significant development from the point of view of composition [4]. It is known that various additions to the original composition (i.e. 45S5) may significantly change and vary material properties. Furthermore, recent studies demonstrated that a new generation of ‘smart’ bioglasses are able to stimulate cell adhesion, proliferation, and angiogenesis processes, triggering signalling mechanisms through the gene expression involved in new tissue development [5, 6]. A glass–ceramic material named RKKP (stands for Ravaglioli, Krajewski, Kirsch, Piancastelli) and developed at ISTEC-CNR (Faenza, Italy) [7, 8] showed that the addition of a couple of ions, such as La3+/Ta5+, further improved the material properties, supplying the surface with a suitable Z-potential, regulating adsorption of some proteins [9]. This first RKKP glass– ceramic was prepared via a traditional melt-processing route. Successively, we applied sol–gel methodology for the synthesis of RKKP composition [10] and used the obtained powder for the bulk RKKP material

M Ledda et al

Printed in the UK 035005 bMM © 2015 IOP Publishing Ltd 2015 10 biomed. Mater. bMM 1748-6041 10.1088/1748-6041/10/3/035005 3 00 00 biomedical Materials


June 2015

Interdisciplinary approach to cell–biomaterial interactions: biocompatibility and cell friendly characteristics of RKKP glass–ceramic coatings on titanium

Mario Ledda1,9, Angela De Bonis2,9, Francesca Romana Bertani3, Ilaria Cacciotti4,5,6, Roberto Teghil2,

Maria Grazia Lolli1, Antonio Ravaglioli7, Antonella Lisi1,9 and Julietta V Rau8,9 1 Institute of Translational Pharmacology (IFT-CNR), Via del Fosso del Cavaliere, 100, 00133 Rome, Italy 2 Dipartimento di Scienze, Università della Basilicata, Via dell’Ateneo Lucano 10, 85100 Potenza, Italy 3 Istituto di Sistemi Complessi (ISC-CNR),Via del Fosso del Cavaliere, 100, 00133 Rome, Italy 4 Università di Roma ‘Niccolò Cusano’, Via Don Carlo Gnocchi, 3, 00166 Rome, Italy 5 Università di Roma ‘Tor Vergata’, Dipartimento di Ingegneria dell’Impresa, Via del Politecnico, 1, 00133 Rome, Italy 6 UdR INSTM-‘Roma Tor Vergata’, Via del Politecnico, 1, 00133 Rome, Italy 7 Ceramic Science and Technology, Via Granarolo, 48018 Faenza, Italy 8 Istituto di Struttura della Materia (ISM-CNR), Via del Fosso del Cavaliere, 100, 00133 Rome, Italy

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Keywords: glass–ceramics, RKKP films, CaCo-2 cell line, pulsed laser deposition, regenerative medicine, tissue engineering


In this work, titanium (Ti) supports have been coated with glass–ceramic films for possible applications as biomedical implant materials in regenerative medicine. For the film preparation, a pulsed laser deposition (PLD) technique has been applied. The RKKP glass–ceramic material, used for coating deposition, was a sol–gel derived target of the following composition: Ca-19.4,

P-4.6, Si-17.2, O-43.5, Na-1.7, Mg-1.3, F-7.2, K-0.2, La-0.8, Ta-4.1 (all in wt%). The prepared coatings were compact and uniform, characterised by a nanometric average surface roughness.

The biocompatibility and cell-friendly properties of the RKKP glass–ceramic material have been tested. Cell metabolic activity and proliferation of human colon carcinoma CaCo-2 cells seeded on RKKP films showed the same exponential trend found in the control plastic substrates. By the phalloidin fluorescence analysis, no significant modifications in the actin distribution were revealed in cells grown on RKKP films. Moreover, in these cells a high mRNA expression of markers involved in protein synthesis, proliferation and differentiation, such as villin (VIL1), alkaline phosphatase (ALP1), β-actin (β-ACT), Ki67 and RPL34, was recorded. In conclusion, the findings, for the first time, demonstrated that the RKKP glass–ceramic material allows the adhesion, growth and differentiation of the CaCo-2 cell line.

PaPer 9 These authors contributed equally to this work. received 18 November 2014 revised 16 February 2015 accePted for Publication 8 April 2015

Published 4 June 2015 doi:10.1088/1748-6041/10/3/035005Biomed. Mater. 10 (2015) 035005 2M Ledda et al preparation [11]. As a promising tailoring tool for third generation biomaterials, the sol–gel methodology was used with the aim of introducing other functional ions and to prepare multidoped glass–ceramic materials, adjustable for specific biomedical applications, like orthopaedics, odontoiatrics and cardiosurgery.

Compared to the conventional high temperature melt-processing route often used for glass preparation, the sol–gel process allows compositional limits to be overcome, to significantly extend the additives number, and to control such parameters as components quantity, material’s homogeneity, porosity, purity, etc [12].

It should be noted that titanium (Ti), an exogenous material for the human body, is currently used in medicine in its native form. When coated with a more bioactive material, Ti could significantly improve its performances and could be used also for tissue engineering applications. Recently, RKKP glass–ceramic films (RKKP films) on Ti were successfully prepared for potential use as implant coatings for orthopaedics and dentistry, applying the pulsed laser deposition (PLD) technique [11]. Physico-chemical properties of films were investigated, proving that the prepared coatings were compact and uniformly deposited, characterised by the nanometric average surface roughness and enhanced mechanical properties.