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Considerable attention has been given to the use of chitosan (CS)-based materials reinforced with inorganic bioactive signals such as hydroxyapatite (HA) to treat bone defects and tissue loss. It is well known that CS/HA based materials possess minimal foreign body reactions, good biocompatibility, controlled biodegradability and antibacterial property. Herein, the bioactivity of these composite systems was analyzed on in vitro bone cell models for their applications in the field of bone tissue engineering (BTE). The combination of sol-gel approach and freeze-drying technology was used to obtain CS/HA scaffolds with three-dimensional (3D) porous structure suitable for cell in-growth. Specifically, our aim was to investigate the influence of bioactive composite scaffolds on cellular behavior in terms of osteoinductivity and anti-inflammatory effects for treating bone defects. The results obtained have demonstrated that by increasing inorganic component concentration, CS/HA (60 and 70% v/v) scaffolds induced a good biological response in terms of osteogenic differentiation of human mesenchymal stem cells (hMSC) towards osteoblast phenotype. Furthermore, the scaffolds with higher concentration of inorganic fillers are able to modulate the production of pro-inflammatory (TGF-β) and anti-inflammatory (IL-4, IL-10) cytokines. Our results highlight the possibility of achieving smart CS/HA based composites able to promote a great osteogenic differentiation of hMSC by increasing the amount of HA nanoparticles used as bioactive inorganic signal. Contemporarily, these materials allow avoiding the induction of a pro-inflammatory response in bone implant site.

Il presente lavoro è una overview delle tecniche di fabbricazione degli scaffold per la rigenerazione del tessuto osseo e cartilagineo

Magnesium (Mg)- and silicon (Si)-substituted hydroxyapatite (HA) scaffolds were synthesized using the sponge replica method. The influence of Mg2+ and SiO44− ion substitution on the microstructural, mechanical and biological properties of HA scaffolds was evaluated. All synthesized scaffolds exhibited porosity &gt, 92%, with interconnected pores and pore sizes ranging between 200 and 800 μm. X-ray diffraction analysis showed that β-TCP was formed in the case of Mg substitution. X-ray fluorescence mapping showed a homogeneous distribution of Mg and Si ions in the respective scaffolds. Compared to the pure HA scaffold, a reduced grain size was observed in the Mg- and Si-substituted scaffolds, which greatly influenced the mechanical properties of the scaffolds. Mechanical tests revealed better performance in HA-Mg (0.44 ± 0.05 MPa), HA-Si (0.64 ± 0.02 MPa) and HA-MgSi (0.53 ± 0.01 MPa) samples compared to pure HA (0.2 ± 0.01 MPa). During biodegradability tests in Tris-HCl, slight weight loss and a substantial reduction in mechanical performances of the scaffolds were observed. Cell proliferation determined by the MTT assay using hBMSC showed that all scaffolds were biocompatible, and the HA-MgSi scaffold seemed the most effective for cell adhesion and proliferation. Furthermore, ALP activity and osteogenic marker expression analysis revealed the ability of HA-Si and HA-MgSi scaffolds to promote osteoblast differentiation.

The advances in polymer chemistry have allowed the preparation of biomedical polymers using human metabolites as monomers that can hold unique properties beyond the required biodegradability and biocompatibility. Herein, we demonstrate the use of endogenous human metabolites (succinic and dilinoleic acids) as monomeric building blocks to develop a new series of renewable resource-based biodegradable and biocompatible copolyesters. The novel copolyesters were characterized in detail employing several standard techniques, namely 1H NMR, 13C NMR, and FTIR spectroscopy and SEC, followed by an in-depth thermomechanical and surface characterization of their resulting thin films (DSC, TGA, DMTA, tensile tests, AFM, and contact angle measurements). Also, their anti-fungal biofilm properties were assessed via an anti-fungal biofilm assay and the biological properties were evaluated in vitro using relevant human-derived cells (human mesenchymal stem cells and normal human dermal fibroblasts). These novel highly biocompatible polymers are simple and cheap to prepare, and their synthesis can be easily scaled-up. They presented good mechanical, thermal and anti-fungal biofilm properties while also promoting cell attachment and proliferation, outperforming well-known polymers used for biomedical applications (e.g. PVC, PLGA, and PCL). Moreover, they induced morphological changes in the cells, which were dependent on the structural characteristics of the polymers. In addition, the obtained physicochemical and biological properties can be design-tuned by the synthesis of homo- and -copolymers through the selection of the diol moiety (ES, PS, or BS) and by the addition of a co-monomer, DLA. Consequently, the copolyesters presented herein have high application potential as renewable and cost-effective biopolymers for various biomedical applications.

Over the past few years, different in vitro and in vivo studies have been highlighting the great potentiality of hyaluronic acid (HA) as a biomaterial in wound healing treatment thanks to its good capability to induce mesenchymal and epithelial cell growth and differentiation, angiogenesis, and collagen deposition. However, the need to improve its mechanical properties as well as its residence time has led scientists to study new functionalization strategies. In this work, chemically modified HA-based hydrogels were obtained by methacrylic and maleic functionalization. Methacrylated (MEHA) and maleated HA (MAHA) hydrogels have shown important physico-chemical properties. The present study provides a deeper insight into the biocompatibility of both synthesized materials and their effects on tissue inflammation using in vitro and in vivo models. To this aim, different cell lines involved in wound healing, human dermal fibroblasts, human adipose-derived stem cells and human umbilical vein endothelial cells, were seeded on MEHA and MAHA hydrogels. Furthermore, an inflammation study was carried out on a murine macrophage cell line to assess the effects of both hydrogels on inflammatory and anti-inflammatory interleukin production. The results showed that both MAHA and MEHA supported cell proliferation with anti-inflammation ability as highlighted by the increased levels of IL-10 (57.92 ± 9.87 pg mL-1 and 68.08 ± 13.94 pg mL-1, for MEHA and MAHA, respectively). To investigate the inflammatory response at tissue/implant interfaces, an in vivo study was also performed by subcutaneous implantation of the materials in BALB/c mice for up to 28 days. In these analyses, no significant chronic inflammation reaction was demonstrated in either MEHA or MAHA in the long-term implantation.

Bone cancer is a demanding challenge for contemporary medicine due to its high frequency of presentation and significant heterogeneity of malignant lesions developing within the bone. To date, available treatments are rarely curative and are primarily aimed at prolonging patients’ survival and ameliorating their quality of life. Furthermore, both pharmacological and surgical therapies are aggravated by a consistent burden of adverse events and subsequent disability due to the loss of healthy bone structural and functional properties. Therefore, great research efforts are being made to develop innovative biomaterials able to selectively inhibit bone cancer progression while reducing the loss of bone structural properties secondary to local tissue invasion. In this review, we describe the state of the art of innovative biomaterials for the treatment of bone cancer. Along with physiological bone remodeling, the development of bone metastasis and osteosarcoma will be depicted. Subsequently, recent advances on nanocarrier-based drug delivery systems, as well as the application of novel, multifunctional biomaterials for the treatment of bone cancer will be discussed. Eventually, actual limitations and promising future perspectives regarding the employment of such approaches in the clinical scenario will be debated.

Over the years, there has been an increasing number of cardiac and orthopaedic implanted medical devices, which has caused an increased incidence of device-associated infections. The surfaces of these indwelling devices are preferred sites for the development of biofilms that are potentially lethal for patients. Device-related infections form a large proportion of hospital-acquired infections and have a bearing on both morbidity and mortality. Treatment of these infections is limited to the use of systemic antibiotics with invasive revision surgeries, which had implications on healthcare burdens. The purpose of this review is to describe the main causes that lead to the onset of infection, highlighting both the biological and clinical pathophysiology. Both passive and active surface treatments have been used in the field of biomaterials to reduce the impact of these infections. This includes the use of antimicrobial peptides and ionic liquids in the preventive treatment of antibiotic-resistant biofilms. Thus far, multiple in vivo studies have shown efficacious effects against the antibiotic-resistant biofilm. However, this has yet to materialize in clinical medicine.

Electrospun porous bone scaffolds are known to imitate the extracellular matrix very well and provide an environment through which the tissue formation is enhanced. Although polymeric scaffolds have a great potential in bone tissue regeneration, their weak bioactivity (bone bonding ability) and mechanical properties have left room for improvement. Therefore, the present study focused on the developing a ternary multifunctional platform composed of polycaprolactone (PCL)/silk fibroin (SF)/Zn-substituted Mg

Cellulose represents a low cost, abundant, and renewable polysaccharide with great versatility, it has a hierarchical structure composed of nanofibers with high aspect ratio (3–4 nm wide, hundreds of μm long). TEMPO-mediated oxidation represents one of the most diffused methods to obtain cellulose nanofibers (CNFs): It is possible to obtain physically crosslinked hydrogels by means of divalent cation addition. The presence of inorganic components, such as calcium phosphates (CaP), can improve not only their mechanical properties but also the bioactivity of the gels. The aim of this work is to design and characterize a TEMPO-oxidized cellulose nanofibers (TOCNFs) injectable hydrogel embedded with inorganic particles, CaP and CaP-GO, for bone tissue regeneration. Inorganic particles act as physical crosslinkers, as proven by rheological characterization, which reported an increase in mechanical properties. The average load value registered in injection tests was in the range of 1.5–4.4 N, far below 30 N, considered a reasonable injection force upper limit. Samples were stable for up to 28 days and both CaP and CaP-GO accelerate mineralization as suggested by SEM and XRD analysis. No cytotoxic effects were shown on SAOS-2 cells cultured with eluates. This work demonstrated that the physicochemical properties of TOCNFs-based dispersions could be enhanced and modulated through the addition of the inorganic phases, maintaining the injectability and bioactivity of the hydrogels.

Ti6Al4V alloy is still attracting great interest because of its application as an implant material for hard tissue repair. This research aims to produce and investigate in-situ chitosan/hydroxyapatite (CS/HA) nanocomposite coatings based on different amounts of HA (10, 50 and 60 wt.%) on alkali-treated Ti6Al4V substrate through the sol-gel process to enhance in vitro bioactivity. The influence of different contents of HA on the morphology, contact angle, roughness, adhesion strength, and in vitro bioactivity of the CS/HA coatings was studied. Results confirmed that, with increasing the HA content, the surface morphology of crack-free CS/HA coatings changed for nucleation modification and HA nanocrystals growth, and consequently, the surface roughness of the coatings increased. Furthermore, the bioactivity of the CS/HA nanocomposite coatings enhanced bone-like apatite layer formation on the material surface with increasing HA content. Moreover, CS/HA nanocomposite coatings were biocompatible and, in particular, CS/10 wt.% HA composition significantly promoted human mesenchymal stem cells (hMSCs) proliferation. In particular, these results demonstrate that the treatment strategy used during the bioprocess was able to improve in vitro properties enough to meet the clinical performance. Indeed, it is predicted that the dense and crack-free CS/HA nanocomposite coatings suggest good potential application as dental implants.

Hyaluronic acid (HA)-based hydrogels are one of the most promising naturally derived biomaterials for tissue engineering applications, as they can play an important role in many key cellular processes. In this study, HA was chemically functionalized with photo-cross-linkable motifs by reacting with methacrylic anhydride (MA) to obtain methacrylated hyaluronic acid (MeHA). A range of MA/HA molar ratios was used to obtain different degrees of substitution (DS) ranging from 3.5% to 74.5%, as showed by nuclear magnetic resonance and attenuated total reflection spectroscopy. By fine tuning the DS, the chemical reaction parameters, and the polymer concentration, it was demonstrated the possibility to tailor their mechanical features. Double network (DN) hydrogels were prepared through the synergic use of MeHA and polyethylene glycole diacrylate (PEGDA). To improve the biological properties of DN hydrogels, bioactive solid signals such as hydroxyapatite nanoparticles (HAp) prepared by sol-gel approach were used in combination with DN hydrogels to obtain an advanced composite material with dual function in terms of mechanical and biological support for soft/hard tissue formation. The results highlighted that composite-DN hydrogels showed a 10-time increase of the storage modulus, if compared to neat MeHA, and an early alkaline phosphatase expression from human mesenchymal stem cells in basal medium. This work can be considered a first systematic approach for the designing of photo-cross-linkable hydrogels, based on a combination of natural/synthetic polymers and HAp, that could be applied in three-dimensional additive manufacturing techniques such as stereolithography. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 3079-3089, 2018.

The aim of this work is to investigate properties of interfacial layer in chitosan-hydroxyapatite nanoparticles (CS-nHAp) films. Simultaneous mechanical and dielectric percolation phenomena in films are observed for 30–40% of nHAp loadings. Structure, electrical and mechanical properties of composites and their dependencies on the hydroxyapatite concentrations are investigated by SEM, thermogravimetry, FTIR spectroscopy, nanoindentation and dielectric spectroscopy measurements. Hardness, reduced elastic modulus, conductivity and dielectric constant exhibit a strong dependence on nanoparticle concentration such that maximum values of referred properties are obtained at 30–40 wt % of nHAp. A three-phase model is used to include chitosan matrix, nanoparticles of hydroxyapatite and interfacial layer with dielectric constant higher than that of chitosan and hydroxyapatite. This layer between nanoparticles and matrix is due to strong interactions between chitosan's side groups with PO43− of hydroxyapatite. By performing simulations of spectroscopy measurements in the Hz-GHz frequency range we show that a maximum in all properties is traceable to the existence of overlapping and decreasing of thickness of interfacial layer. Our study proposes a systematic approach to design smart materials for bone tissue engineering.

This study reports on the development of a scaffold with a gradient of bioactive solid signal embedded in the biodegradable polymer matrix by combining a sol-gel approach and freeze-drying technology. The chemical approach based on the sol-gel transition of calcium phosphates ensures the particles dispersion into the gelatin matrix and a direct control of interaction among COOHgelatin /Ca2+ ions. Morphological analysis demonstrated that on the basis of the amount of inorganic component and by using specific process conditions, it is possible to control the spatial distribution of nanoparticles around the gelatin helix. In fact, methodology and formulations were able to discriminate between the different hydroxyapatite concentrations and their respective morphology. The good biological response represented by good cell attachment, proliferation and increased levels of alkaline phosphatase as an indicator of osteoblastic differentiation of human mesenchymal stem cells toward the osteogenic lineage, demonstrating the effect of bioactive solid signals on cellular behavior. Furthermore, the inhibition of reactive oxygen species production by composite materials predicted potential anti-inflammatory properties of scaffolds thus confirming their biocompatibility. Indeed, these interesting biological results suggest good potential application of this scaffold as filler to repair bone defects. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2007-2019, 2018.

The control of the inflammatory response induced by the implantation of foreign biomaterials is fundamental in determining tissue healing. It has been shown that the activation of specific macrophage pathways upon contact with a biomaterial can lead either to a chronic inflammation preventing a physiological tissue repair or to an improved tissue healing. In the case of bone repair, calcium phosphate cements with good osteoconductivity properties have been successfully applied in bone defect filling and repair, but poor handling properties, insufficient viscous flow and unmatched degradation rate are still problems that remain unsolved. In this study, a strontium-doped hydroxyapatite (HA) gel was modified by integrating branched poly(epsilon-lysine) dendrons with third-generation branches exposing phosphoserine (SrHA/G3-K PS). The interaction of this material with macrophages was investigated in vitro, focusing on the secretion and gene expression of specific pro-inflammatory cytokines. Our results showed that the addition of strontium and G3-K PS to HA sol-gel could down-regulate the gene expression of inflammatory factors such as IL-1 beta, TNF-alpha and MCP-1, while increasing the gene expression of IL-6, a cytokine known for its osteogenic effect. These results were further confirmed by ELISA test of the respective protein concentrations. When exposed to supernatants of macrophage culture in the presence of strontium and G3-K PS, osteoblast viability was promoted with elevated osteogenic gene markers, in terms of OPG, ALP, OCN and COL-I. In vivo implantation experiments using an osteoporotic rat model with bone defect further confirmed that the addition of G3-K PS to HA could dramatically promote new bone regeneration. Although the introduction of strontium improved the degradation properties of the injectable materials, no positive effect on promoting in vivo bone regeneration was observed.

Within the framework of neurodegenerative disorder therapies, the fabrication of 3D eumelanin architectures represents a novel strategy to realize tissue-engineering scaffolds for neuronal cell growth and control by providing both mechanical support and biological signals. Here, an appropriate procedure combining electrospinning, spin coating and solid-state polymerization process is established to realize the scaffolds. For biological analysis, a human derived cell line SH-SY5Y from neuroblastoma is used. Cell maturation on eumelanin microfibers, random and aligned, is evaluated by using confocal analysis and specific markers of differentiating neurons (beta III tubulin and GAP-43 expression). Cell morphology is tested by SEM analysis and immunofluorescence techniques. As results, eumelanin coated microfibers prove capable to support biological response in terms of cell survival, adhesion and spreading and to promote cell differentiation toward a more mature neuronal phenotype as confirmed by GAP-43 expression over the culture.

Injectable biphasic calcium phosphates have been proposed as a solution in the treatment of a range of clinical applications including as fillers in the augmentation of osteoporotic bone. To date, various biodegradable natural or synthetic organics have been used as a polymer component of bone materials to increase their cohesiveness. Herein, a novel bone material was developed combining osteoconductive biphasic calcium phosphate (BCP) nanoparticles with phosphoserine-tethered generation 3 poly(epsilon-lysine) dendron (G3-K PS), a class of hyperbranched peptides previously shown to induce biomineralization and stem cell osteogenic differentiation. Strontium was also incorporated into the BCP nanocrystals (SrBCP) to prevent bone resorption. Within 24 h, an antiwashout behavior was observed in G3-K PS-integrated pure BCP group (BCPG3). Moreover, both in vitro tests by relevant cell phenotypes and an in vivo tissue regeneration study by an osteoporotic animal bone implantation showed that the integration of G3-K PS would downregulate Cxcl9 gene and protein expressions, thus enhancing bone regeneration measured as bone mineral density, new bone volume ratio, and trabecular microarchitectural parameters. However, no synergistic effect was found when Sr was incorporated into the BCPG3 bone pastes. Notably, results indicated a concomitant reduction of bone regeneration potential assessed as reduced Runx2 and PINP expression when bone resorptive RANKL and CTX-I levels were reduced by Sr supplementation. Altogether, the results suggest the potential of injectable BCPG3 bone materials in the treatment of osteoporotic bone defects.

Injectable materials represent very attractive ready-to-use biomaterials for application in minimally invasive surgical procedures. It is shown that this approach to treat, for example, vertebral fracture, craniofacial defects, or tumor resection has significant clinical potential in the biomedical field. In the last four decades, calcium phosphate cements have been widely used as injectable materials for orthopedic surgery due to their excellent properties in terms of biocompatibility and osteoconductivity. However, few clinical studies have demonstrated certain weaknesses of these cements, which include high viscosity, long degradation time, and difficulties being manipulated. To overcome these limitations, the use of sol-gel technology has been investigated, which has shown good results for synthesis of injectable calcium phosphate-based materials. In the last few decades, injectable hydrogels have gained increasing attention owing to their structural similarities with the extracellular matrix, easy process conditions, and potential applications in minimally invasive surgery. However, the need to protect cells during injection leads to the development of double network injectable hydrogels that are capable of being cross-linked in situ. This review will provide the current state of the art and recent advances in the field of injectable biomaterials for minimally invasive surgery.

Current bone implants based on new biomaterials may cause a foreign body reaction (FBR) around the implant itself thus prolonging the healing time following bone fractures. In this paper, biomimetic chitosan-based scaffolds promoting bone tissue regeneration and controlling inflammatory response are proposed. First, the anti-inflammatory potential of scaffolds on hMSCs stimulated by lipopolysaccharide (LPS) was investigated by dosing the levels of some interleukins and oxidative stress metabolites (IL-1?, IL-10 and nitrites) involved in immune response. Then, to mimic the inflammation process at osteoporotic site, the effect of scaffolds was evaluated on in vitro co-culture model based on osteoblasts and macrophages stimulated by LPS. Results demonstrated that bioactivated scaffolds are able to i) inhibit synthesis of inflammatory mediators such as IL-1?; ii) reduce oxidative stress metabolites; and iii) promote anti-inflammatory markers generation (IL-10) in hMSCs. Finally, bioactivated scaffolds show an anti-inflammatory activity also on in vitro co-cultures, which better mimic in vivo damaged bone microenvironment.

Superabsorbent hydrogels are macromolecular networks able to absorb and retain large amounts of water solutions within their fine mesh-like structure. More importantly, they are capable of multiple swelling/shrinking transitions in response to specific environmental cues (e.g., pH, ionic strength, temperature, presence of given electrolytes), thus exhibiting a stimuli-sensitive behavior, which makes them appealing for the design of smart devices in a number of technological fields. In particular, in the last two decades, cellulose-based superabsorbent hydrogels have proven to be an environmentally friendly and cost-effective alternative to acrylamide-based products. This chapter reviews the relationship between the molecular structure of cellulose-based hydrogels and their physicochemical properties. First, the network formation through the use of different cellulose derivatives and chemical or physical crosslinking agents is presented. Successively, the smart swelling capability of the hydrogels as a function of composition and structure is thoroughly discussed. Finally, several approaches to the hydrogel characterization are reviewed, with focus on the assessment of key mechanical, thermal and morphological properties.

Electrospun porous bone scaffolds are known to imitate the extracellular matrix very well and provide an environment through which the tissue formation is enhanced. Although polymeric scaffolds have a great potential in bone tissue regeneration, their weak bioactivity (bone bonding ability) and mechanical properties have left room for improvement. Therefore, the present study focused on the developing a ternary multifunctional platform composed of polycaprolactone (PCL)/silk fibroin (SF)/Zn-substituted Mg2SiO4 nanoparticles for bone tissue regeneration. This study is composed of two connected sections including synthesis and characterization of Mg(2-x)ZnxSiO4, x = 0, 0.5, 1, 1.5, 2 through surfactant-assisted sol-gel technique followed by incorporation of the nanoparticles into PCL/SF hybrid scaffold via electrospinning technique. The weight ratios of polymers and ceramic nanoparticles were optimized to reach desirable textural—porosity, pore size, and fiber diameter—and mechanical properties. Having optimized the ternary scaffold, it was then undergone different physical, chemical, and biological tests in vitro. A precise comparison study between the ternary (PCL/SF/ceramic nanoparticles), binary (PCL/SF), and pure PCL was made to shed light on the effect of each composition on the applicability of ternary scaffold. The overall results confirmed that the Mg1Zn1SiO4 nanoparticles-incorporated PCL/SF scaffold with fluorescence property was the one yielding the highest Young's modulus and desirable textural properties. The ternary scaffold showed improved biological properties making it a promising candidate for further studies towards bone tissue regeneration.

Herein, we demonstrate the effect of imidazolium ionic liquids (IL) applied as additives in the in situ formation of high amorphous vinyl alcohol (HAVOH)-silica bionanocomposites, using a simple sol-gel process approach. A complementary set of alkyl-, ether-, and carboxy-functionalized IL was used, allowing silica structure control and polymer-silica interphase tuning. Consequently, hybrids with diverse morphologies, as well as improved thermo-mechanical and barrier properties, were obtained. This diversity also highlighted the systems' dependency on the IL's molecular structure, where both the cation and anion influenced the hybrids' final properties. This could be evidenced as the polar group functionalized-IL (ether- and carboxy-functionalized IL) allowed the formation of multiple hydrogen bonds at the organic-inorganic interphase, inducing a fine hybrid morphology with well-dispersed silica nanodomains. This significantly increased the storage (~50%) and tensile moduli (~20%), extensibility (up to 300%), and glass transition temperature (>20 °C) and decreased the water vapor permeability (~50%), which are desirable characteristics for potential food and medical packaging.

In this study, a new foaming method, based on physical foaming combined with microwave-induced curing, is proposed in combination with a surface bioactivation to develop scaffold for bone tissue regeneration. In the first step of the process, a stable physical foaming was induced using a surfactant (Pluronic) as blowing agent of a homogeneous blend of Chitosan and polyethylene glycol diacrylate (PEGDA700) solutions. In the second step, the porous structure of the foaming was chemically stabilized by radical polymerization induced by homogeneous heating of the sample in a microwave reactor. In this step, 2,2-azobis[2-(2-imidazolin-2yl)propane]dihydrochloride was used as thermoinitiator (TI). Chitosan and PEGDA were mixed in different blends to investigate the influence of the composition on the final properties of the material. The chemical properties of each sample were evaluated by infrared attenuated total reflectance analysis, before and after curing in order to maximize reaction yield and optimize kinetic parameters (i.e. time curing, microwave power). Absorption capacity, elastic modulus, porosity and morphology of the porous structure were measured for each sample. The stability of materials was evaluated in vitro by degradation test in phosphate-buffered saline. To improve the bioactivity and biological properties of chitosan scaffold, a biomineralization process was used. Biological characterization was carried out with the aim to prove the effect of biomineralization scaffold on human mesenchymal stem cells behaviour. Copyright © 2016 John Wiley & Sons, Ltd.

Candida species have great ability to colonize and form biofilms on medical devices, causing infections in human hosts. In this study, poly(l-lactide) films with different imidazolium salt (1-n-hexadecyl-3-methylimidazolium chloride (C16MImCl) and 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16MImMeS)) contents were prepared, using the solvent casting process. Poly(l-lactide)-imidazolium salt films were obtained with different surface morphologies (spherical and directional), and the presence of the imidazolium salt in the surface was confirmed. These films with different concentrations of the imidazolium salts C16MImCl and C16MImMeS presented antibiofilm activity against isolates of Candida tropicalis, Candida parapsilosis, and Candida albicans. The minor antibiofilm concentration assay enabled one to determine that an increasing imidazolium salt content promoted, in general, an increase in the inhibition percentage of biofilm formation. Scanning electron microscopy micrographs confirmed the effective prevention of biofilm formation on the imidazolium salt containing biomaterials. Lower concentrations of the imidazolium salts showed no cytotoxicity, and the poly(l-lactide)-imidazolium salt films presented good cell adhesion and proliferation percentages with human mesenchymal stem cells. Furthermore, no acute microscopic lesions were identified in the histopathological evaluation after contact between the films and pig ear skin. In combination with the good morphological, physicochemical, and mechanical properties, these poly(l-lactide)-based materials with imidazolium salt additives can be considered as promising biomaterials for use in the manufacturing of medical devices.

In the current work, our purpose was based on the assessment of bioactive chitosan (CS)/Poly(ethylene glycol) diacrylate (PEGDA) based scaffolds ability to stimulate in vitro angiogenesis process. The bioactivation of the scaffolds was accomplished by using organic (BMP-2 peptide) and inorganic (hydroxyapatite nanoparticles) cues. In particular, the properties of the materials in terms of biological response promotion on human umbilical vein endothelial cells (HUVECs) were studied by using in vitro angiogenesis tests based on cell growth and proliferation. Furthermore, our interest was to examine the scaffolds capability to modulate two important steps involved in angiogenesis process: migration and tube formation of cells. Our data underlined that bioactive signals on CS/PEGDA scaffolds surface induce a desirable effect on angiogenic response concerning angiogenic marker expression (CD-31) and endothelial tissue formation (tube formation). Taken together, the results emphasized the concept that bioactive CS/PEGDA scaffolds may be novel implants for stimulating neovascularization of tissue-engineered constructs in regenerative medicine field.

In this study, polycaprolactone (PCL) and polycaprolactone/fluoride substituted-hydroxyapatite (PCL/FHA) nanocomposite coatings at different composition (10, 20 and 30 wt.% of FHA) are deposited on alkali-treated Ti6Al4V substrate via in-situ sol-gel method to improve its corrosion behavior and in vitro bioactivity. The PCL and PCL/FHA nanocomposite coatings are characterized by scanning electron microscopy (SEM) equipped with X-ray energy dispersive spectrometer (EDS), X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR). The in vitro bioactivity and corrosion behavior of the coatings are also investigated to show their potential application for dental implants. The results indicate that the in-situ sol-gel process is capable of forming compact and crack-free coatings with a homogeneous dispersion of FHA as a ceramic phase in the polymeric matrix. By increasing the FHA content, the hydrophilicity, adhesion strength and surface roughness were increased. Furthermore, in vitro bioactivity of the PCL/FHA nanocomposite coatings was improved by increasing the FHA content and the incubation time in simulated body fluid (SBF). The corrosion resistance of the PCL/10 wt.% FHA coatings considerably increased as compared to the pure PCL and PCL/FHA coatings with 20 and 30 wt.% of FHA. Additionally, the MTT assay confirmed that the coatings have no considerable cytotoxicity effect and PCL/20 wt.% FHA coating promoted the proliferation and spreading of MG63 cells. Indeed, the results showed that the PCL/FHA nanocomposite coatings on alkali-treated Ti6Al4V substrate may be a promising candidate for dental implant applications.

Nowadays chemotherapy is the main treatment for osteosarcoma disease, even if limited by the lack of selectivity between healthy and cancer cells during the inhibition of cell division. Herein, we propose the use of few-layer two-dimensional black phosphorous (2D bP) as an alternative tool for osteosarcoma treatment and report how 2D bP can stimulate newly forming bone tissue generation after osteosarcoma resection. In our study, we have developed an in vitro model to evaluate the efficacy of 2D bP material with and without near-infrared light irradiation treatment, and we have demonstrated that the presence of 2D bP without treatment inhibits the metabolic activity of osteosarcoma cells (SAOS-2) while inducing both the proliferation and the osteogenic differentiation of human preosteoblast cells (HOb) and mesenchymal stem cells. Furthermore, we also propose an in vitro coculture model (SAOS-2 and HOb cell lines) in order to study the effect of 2D bP on inflammatory response related to cancer. On this coculture model, 2D bP may increase anti-inflammatory cytokine generation (i.e., interleukin-10) and inhibit proinflammatory mediators synthesis (i.e., interleukin-6), thus suggesting the opportunity to prevent cancer-related inflammation. Finally, we have demonstrated that 2D bP represents a promising candidate for future regenerative medicine and anticancer applications.

As a minimally invasive surgery, percutaneous cement discoplasty (PCD) is now contemplated to treat lumbar disc degeneration disease in elder population. Here, we investigated whether the osteogenic mineralized collagen (MC) modified polymethylmethacrylate (PMMA) cement could be a suitable material in PCD surgery. Injectability, hydrophilicity and mechanical properties of the MC-modified PMMA (PMMA-MC) was characterized. The introduction of MC did not change the application and setting time of PMMA and was easy to be handled in minimally invasive operation. Hydrophilicity of PMMA-MC was greatly improved and its elastic modulus was tailored to complement mechanical performance of bone under dynamic stress. Then, PCD surgery in a goat model with induced disc degeneration was performed with implantation of PMMA-MC or PMMA. Three months after implantation, micro-computed tomography analysis revealed a 36.4% higher circumferential contact index between PMMA-MC and bone, as compared to PMMA alone. Histological staining confirmed that the surface of PMMA-MC was in direct contact with new bone, while the PMMA was covered by fibrous tissue. The observed gathering of macrophages around the implant was suspected to be the cause of fibrous encapsulation. Therefore, the interactions of PMMA and PMMA-MC with macrophages were investigated in vitro. We discovered that the addition of MC could hinder the proliferation and fusion of the macrophages. Moreover, expressions of fibroblast-stimulating growth factors, insulin-like growth factor, basic fibroblast growth factor and tumor necrosis factor-β were significantly down-regulated in the macrophages cocultured with PMMA-MC. Together, the promoted osteointegration and reduced fibrous tissue formation observed with PMMA-MC material makes it a promising candidate for PCD surgery.

The main driving idea of the present study was the comparison between two different chemical modifications of hyaluronic acid (HA) followed by the development of nanocomposite hydrogels directly in situ by biomineralization of photocrosslinkable HA polymers through sol-gel synthesis. In this way, it has been possible to overcome some limitations due to classical approaches based on the physical blending of inorganic fillers into polymer matrix. To this aim, methacrylated and maleated HA, synthesized with similar degree of substitution (DS) were compared in terms of mechanical and physico-chemical properties. The success of in situ biomineralization was highlighted by reflect Fourier transform infrared spectroscopy and thermogravimetric analysis. Furthermore, mechanical characterization demonstrated the reinforcing effect of inorganic fillers evidencing a strong correlation with DS. The swelling behavior resulted to be correlated with filler concentration. Finally, the cytotoxicity tests revealed the absence of toxic components and an increase of cell proliferation over culture time was observed, highlighting these bio-nanocomposite hyaluronan derivatives as biocompatible hydrogel with tunable properties.

Bioactive glasses are well-known materials suitable for bone-related applications thanks to their biocompatibility and osteoconductivity. In order to improve their in vivo performance, the modification of the glass composition by adding ions with specific biological functions is required. As copper (Cu) possesses antibacterial properties, in this study, 5 wt % of CuO has been added to the 45S5 bioactive glass composition. The investigation of the effect of the Cu-containing bioactive glass on cellular behavior has revealed that the presence of Cu induces an early differentiation of human mesenchymal stem cells through osteoblast phenotype, Target la earn promotes the expression of anti-inflammatory interleukin, and reduces proinflammatory interleukin expression. With the aim to produce coatings with antibacterial properties, the Cu-containing bioactive glass was used as the target material for the pulsed laser deposition (PLD) of bioactive thin films. PLD experiments were carried out at different substrate temperatures to study the effect on the film's characteristics. All of the films are compact, crack-free, and characterized by a rough morphology and good wettability. The in vitro bioactivity was demonstrated by the apatite growth on the coating surface, after soaking in simulated body fluid, revealed by Raman spectroscopy and scanning electron microscopy-energy dispersive X-ray analyses. The antibacterial study proved that the material showed more effective activity against three Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Salmonella enterica) rather than against Gram-positive bacteria (Staphylococcus aureus).