Stefania Cometa

Polycaprolactone Scaffolds Fabricated via Bioextrusion for Tissue Engineering Applications

The most promising approach in Tissue Engineering involves the seeding of porous, biocompatible/biodegradable scaffolds, with donor cells to promote tissue regeneration. Additive biomanufacturing processes are increasingly recognized as ideal techniques to produce 3D structures with optimal pore size and spatial distribution, providing an adequate mechanical support for tissue regeneration while shaping in-growing tissues. This paper presents a novel extrusion-based system to produce 3D scaffolds with controlled internal/external geometry for TE applications.The BioExtruder is a low-cost system that uses a proper fabrication code based on the ISO programming language enabling the fabrication of multimaterial scaffolds. Poly(ε-caprolactone) was the material chosen to produce porous scaffolds, made by layers of directionally aligned microfilaments. Chemical, morphological, and in vitro biological evaluation performed on the polymeric constructs revealed a high potential of the BioExtruder to produce 3D scaffolds with regular and reproducible macropore architecture, without inducing relevant chemical and biocompatibility alterations of the material.

An innovative, easily fabricated, silver nanoparticle-based titanium implant coating: development and analytical characterization

AbstractMicrobial colonization and biofilm formation on implanted devices represent an important complication in orthopaedic and dental surgery and may result in implant failure. Controlled release of antibacterial agents directly at the implant site may represent an effective approach to treat these chronic complications. Resistance to conventional antibiotics by pathogenic bacteria has emerged in recent years as a major problem of public health. In order to overcome this problem, non-conventional antimicrobial agents have been under investigation. In this study, polyacrylate-based hydrogel thin coatings have been electrosynthesised on titanium substrates starting from poly(ethylene glycol diacrylate)–co–acrylic acid. Silver nanoparticles (AgNPs) with a narrow size distribution have been synthesized using a “green” procedure and immobilized on Ti implant surfaces exploiting hydrogel coatings’ swelling capabilities. The coatings have been characterized by XPS and SEM/EDX, while their silver release performances have been monitored by ICP–MS. The antibacterial activity of these AgNP-modified hydrogel coatings was tested evaluating in vitro inhibition growth of Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli, among the most common pathogens in orthopaedic infections. Moreover, a preliminary investigation of the biocompatibility of silver-loaded coatings versus MG63 human osteoblast-like cells has been performed. An important point of strength of this paper, in fact, is the concern about the effect of silver species on the surrounding cell system in implanted medical devices. Silver ion release has been properly tuned in order to assure antibacterial activity while preserving osteoblasts’ response at the implant interface. FigureSilver nanoparticles-loaded PEGDA-AA hydrogel coatings for inhibition of titanium implants associated infections

Characterization and cytocompatibility of an antibiotic/chitosan/ cyclodextrins nanocoating on titanium implants

A novel ciprofloxacin loaded chitosan nanoparticle-based coating onto titanium substrates has been developed and characterized to obtain an orthopaedic implant surface able to in situ release the antibiotic for the prevention of post-operative infections. Ciprofloxacin loaded chitosan nanoparticles were obtained using the combination of sulfobutyl ether-beta-cyclodextrin and gamma-cyclodextrin. The resulting nanoparticulate system was characterized by TEM, HPLC and XPS. Particle size was in the range 426-552 nm and zeta potential values were around +30 mV. This antibacterial coating was able to in vitro inhibit two nosocomial Staphylococcus aureus strains growth, with a reduction of about 20 times compared to controls. No impairment in MG63 osteoblast-like cells viability, adhesion and gene expression were detected at 48 h, 7 and 14 days of culture. Overall, the investigated coating represents a promising candidate for the development of a new antibiotic carrier for titanium implants.

Ciprofloxacin-modified electrosynthesized hydrogel coatings to prevent titanium-implant-associated infections

New promising and versatile materials for the development of in situ sustained release systems consisting of thin films of either poly(2-hydroxyethyl methacrylate) or a copolymer based on poly(ethylene-glycol diacrylate) and acrylic acid were investigated. These polymers were electrosynthesized directly on titanium substrates and loaded with ciprofloxacin (CIP) either during or after the synthesis step. X-ray photoelectron spectroscopy was used to check the CIP entrapment efficiency as well as its surface availability in the hydrogel films, while high-performance liquid chromatography was employed to assess the release property of the films and to quantify the amount of CIP released by the coatings. These systems were then tested to evaluate the in vitro inhibition of methicillin-resistant Staphylococcus aureus (MRSA) growth. Moreover, a model equation is proposed which can easily correlate the diameter of the inhibition haloes with the amount of antibiotic released. Finally, MG63 human osteoblast-like cells were employed to assess the biocompatibility of CIP-modified hydrogel coatings.

Biocompatibility of Poly(Acrylic Acid) Thin Coatings Electro-synthesized onto TiAlV-based Implants

The protection of metal orthopedic implants against corrosion is a crucial medical problem. It was found that electrochemical polymerization of thin, passive poly(acrylic acid) (PAA) films on titanium and TiAlV substrates provides good anti-corrosion properties. In this work, an investigation of anti-corrosion features was carried out to clarify the hypothesis of the presence of an electrostatic contribution to the performance of a PAA coating. Ion release tests were performed at three different pHs; the pH dependence of the polymer swelling was examined by quartz crystal microbalance with dissipation monitoring, to establish the role of this phenomenon on the polymer barrier properties. The potential application of these PAA thin films as biocompatible protective coatings for metal implants and compatibility towards MG-63 human osteoblast-like cells was assessed.

Development and characterization of rhVEGF-loaded poly(HEMA-MOEP) coatings electrosynthesized on titanium to enhance bone mineralization and angiogenesis.

Osteointegration of titanium implants could be significantly improved by coatings capable of promoting both mineralization and angiogenesis. In the present study, a copolymeric hydrogel coating, poly-2-hydroxyethyl methacrylate-2-methacryloyloxyethyl phosphate (P(HEMA-MOEP)), devised to enhance calcification in body fluids and to entrap and release growth factors, was electrosynthesized for the first time on titanium substrates and compared to poly-2-hydroxyethyl methacrylate (PHEMA), used as a blank reference. Polymers exhibiting negatively charged groups, such as P(HEMA-MOEP), help to enhance implant calcification. The electrosynthesized coatings were characterized by X-ray photoelectron spectroscopy and atomic force microscopy. MG-63 human osteoblast-like cell behaviour on the coated specimens was investigated by scanning electron microscopy, MTT viability test and osteocalcin mRNA detection. The ability of negatively charged phosphate groups to promote hydroxyapatite-like calcium phosphate deposition on the implants was explored by immersing them in simulated body fluid. Similar biological responses were observed in both coated specimens, while calcium-phosphorus globules were detected only on P(HEMA-MOEP) surfaces pretreated with alkaline solution. Testing of the ability of P(HEMA-MOEP) hydrogels to entrap and release human recombinant vascular endothelial growth factor, to tackle the problem of insufficient oxygen and nutrient delivery, suggested that P(HEMA-MOEP)-coated titanium prostheses could represent a multifunctional material suitable for bone restoration applications.

Evaluation of in vitro degradation of PCL scaffolds fabricated via BioExtrusion. Part 1: Influence of the degradation environment

One of the most promising approaches in tissue engineering (TE) comprises the development of 3D porous scaffolds which are able to promote tissue regeneration. Biocompatible and biodegradable poly(ϵ-caprolactone) (PCL) structures are increasingly used as temporary extra-cellular matrices for bone tissue engineering. To ensure an appropriate bone restoration over the long term, the selected material must have a degradation rate that match the in-growth of new bone. The in vivo process, by which the scaffold degrades and is resorbed transferring the load and function back to the host tissue, is complex. Consequently, an appropriate preliminary in vitro study is required. A novel extrusion-based technology called BioExtruder was used to produce PCL porous scaffolds made with layers of directionally aligned microfilaments. The in vitro degradation behaviour in both simulated body fluid (SBF) and phosphate buffer solution (PBS) were investigated over 6 months. The characterization of the degradation behaviour of the structures was performed at specific times by evaluating changes in the average molecular weight, the weight loss and its thermal properties. Morphological and surface chemical analyses were also performed using a Scanning Electron Microscopy (SEM) and an X-ray Photoelectron Spectroscopy (XPS), respectively.

Analytical characterization and antimicrobial properties of novel copper nanoparticle–loaded electrosynthesized hydrogel coatings

In this study, a novel antimicrobial coating was developed to avoid infections and to provide sterile conditions for stainless steel devices. Poly(ethylene glycol diacrylate) hydrogel thin films were modified with copper-based nanoparticles, following two different entrapment procedures. These coatings were firmly attached on metal substrates by means of a simple and fast electrochemical polymerization technique. The surface composition of the Cu nanoparticles–modified hydrogel coatings and their bactericidal effect against Staphylococcus aureus and Escherichia coli was studied, and the efficacy of such systems in preventing bacterial infections demonstrated.

PHEMA-based thin hydrogel films for biomedical applications

Poly(2-hydroxyethyl methacrylate) based thin coatings were electro-synthesized by cyclic voltammetry on Au-coated quartz crystal surfaces to study different solid—liquid interfacial processes. By varying the electrochemical parameters and the presence or not of a crosslinking agent, films were obtained with thicknesses ranging from 5 to 90 nm. Surface characterization was performed by X-ray photoelectron spectroscopy, atomic force microscopy, and static contact angle measurements. Using quartz crystal microbalance with dissipation monitoring to investigate the relationship between the film thickness and the swelling behavior, it was found that these characteristics can be modulated by varying either the number of voltammetric cycles or the presence of the crosslinker. Cell adhesion and biocompatibility tests indicate that these film coatings were suitable for biomedical applications.

Insight Into Halloysite Nanotubes-Loaded Gellan Gum Hydrogels For Soft Tissue Engineering Applications

A tri-component hydrogel, based on gellan gum (GG), glycerol (Gly) and halloysite nanotubes (HNT), is proposed in this work for soft tissue engineering applications. The FDA-approved GG polysaccharide has been recently exploited as biomaterial because its biomimetic features. Gly is added as molecular spacer to improve hydrogel viscosity and mechanical properties. HNT incorporation within the hydrogel offers the versatility to improve the GG-Gly biocompatibility with potential incorporation of target biomolecules. In this work, hydrogels with different composition ratios are physically crosslinked for tuning physico-mechanical properties. An accurate physico-chemical characterization is reported. HNT addition leads to a water uptake decrease of 30-35% and tuneable mechanical properties with a compressive Youngs modulus ranging between 20 and 75kPa. Finally, in vitro study with human fibroblasts on GG-Gly hydrogels loaded with 25% HNT offered the higher metabolic activities and cell survival up to 7days of incubation.