Bone and joint defects or diseases, coupled with edentation, pose challenging and commonly encountered clinical issues with a significant incidence in the medical domain [...]

Hydroxyapatite (HA) promotes excellent bone regeneration in bone-tissue engineering, due to its similarity to bone mineral and its ability to connect to living tissues. These factors promote the osteointegration process. This process can be enhanced by the presence of electrical charges, stored in the HA. Furthermore, several ions can be added to the HA structure to promote specific biological responses, such as magnesium ions. The main objective of this work was to extract hydroxyapatite from sheep femur bones and to study their structural and electrical properties by adding different amounts of magnesium oxide. The thermal and structural characterizations were performed using DTA, XRD, density, Raman spectroscopy and FTIR analysis. The morphology was studied using SEM, and the electrical measurements were registered as a function of frequency and temperature. Results show that: (i) an increase of MgO amount indicates that the solubility of MgO is below 5%wt for heat treatments at 600 °C; (ii) the rise of MgO content increases the capacity for electrical charge storage; (iii) sheep hydroxyapatite presents itself as a natural source of hydroxyapatite, environmentally sustainable and low cost, and promising for applications in regenerative medicine.

Bone tissue engineering is based on a comprehensive understanding of bone structure, bone mechanics, and biology. In order to create nanostructured hydroxyapatite powders with customized properties, many synthesis strategies such as wet chemical precipitation, sol-gel, hydrothermal, and biomimetic approaches have been intensively researched through the years. Calcium phosphate (CaP)-based ceramic nanoparticles, including hydroxyapatite (HAp), were synthesized by the chemical precipitation technique at pH ranges of 7 to 11 and different calcination temperatures of 600 to 1100 °C. The synthesized powders were characterized by several techniques, including scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), energy dispersive X-ray analysis (EDX), and in vitro cell culture assays. The particle size analysis and zeta potential of these powders were also carried out using the dynamic light scattering (DLS) and laser Doppler electrophoresis methods. The results showed that the pH levels of 9 to 11 range and calcination temperatures of 600 to 800 °C were adequate for appropriate nanohydroxyapatite powder production using this method. The particle size of the nanohydroxyapatite was approximately 55 nm, although they were agglomerated after calcination. The biocompatibility tests demonstrated that these nanohydroxyapatite (nHAp) powders produced have appropriate cytocompatibility and can be used for bone graft production and other biomedical applications.

Novel biomaterials with promising bone regeneration potential, derived from rich, renewable, and cheap sources, are reported. Thus, thin films were synthesized from marine-derived (i.e., from fish bones and seashells) hydroxyapatite (MdHA) by pulsed laser deposition (PLD) technique. Besides the physical–chemical and mechanical investigations, the deposited thin films were also evaluated in vitro using dedicated cytocompatibility and antimicrobial assays. The morphological examination of MdHA films revealed the fabrication of rough surfaces, which were shown to favor good cell adhesion, and furthermore could foster the in-situ anchorage of implants. The strong hydrophilic behavior of the thin films was evidenced by contact angle (CA) measurements, with values in the range of 15–18°. The inferred bonding strength adherence values were superior (i.e., ~49 MPa) to the threshold established by ISO regulation for high-load implant coatings. After immersion in biological fluids, the growth of an apatite-based layer was noted, which indicated the good mineralization capacity of the MdHA films. All PLD films exhibited low cytotoxicity on osteoblast, fibroblast, and epithelial cells. Moreover, a persistent protective effect against bacterial and fungal colonization (i.e., 1- to 3-log reduction of E. coli, E. faecalis, and C. albicans growth) was demonstrated after 48 h of incubation, with respect to the Ti control. The good cytocompatibility and effective antimicrobial activity, along with the reduced fabrication costs from sustainable sources (available in large quantities), should, therefore, recommend the MdHA materials proposed herein as innovative and viable solutions for the development of novel coatings for metallic dental implants.

In this study, an IPN based enzymatic levan-polyacrylamide hydrogel (EL-PA) was developed and charac-terized for its structural, morphological, rheological properties and swelling kinetics to underline hydrogel properties and its potential use in paper conservation. The addition of levan also led to changes in the viscoelastic behavior of the hydrogels, with the complex viscosity of EL-PA samples showing pronounced dependence on shear rate. The swelling and the overall surface area of the hydrogels were increased with the addition of levan into the polymer network. Source associated structural differences were found to be negligible such that both microbially produced linear and enzymatically produced branched forms of levan performed equally well. Solvent loaded hydrogels were then applied on an artifact, a 19th century book of Namik Kemal, and investigated using FTIR, SEM, XRD and colorimetric analysis. Old adhesive lay-ers were successfully removed, and hydrogels showed good compatibility and ease of application. This study has shown that levan has improved hydrogel properties and levan based systems bear high poten-tial in conservation science.(c) 2023 Consiglio Nazionale delle Ricerche (CNR). Published by Elsevier Masson SAS. All rights reserved.

Corneal ulcer, which is brought on by a breach in the epithelial barrier, is a dangerous infection of the avascular corneal stroma. New treatment strategies are needed, suppressing the aggressive nature of the disease and including a combination of different drugs. In this study, vancomycin (VAN) and fluconazole (FLU) dual-drug loaded dual-layered polyvinyl alcohol and gelatin (PVA/GEL) nanofibrous patches are produced by electrospinning. Scanning electron microscopy (SEM) images show smooth surfaces are obtained for both pure and drug-loaded nanofibrous patches. The tensile test results report that loading the FLU and VAN separately into the PVA/GEL patches decrease both the tensile strength and elongation at break and it is further reduced when combining two drug-loaded layers in one patch. According to drug release results, the FLU and VAN-loaded nanofibrous patches show a controlled release profile extending up to 96 h. Moreover, PVA/GEL/FLU, PVA/GEL/VAN, and PVA/GEL/FLU/VAN nanofibrous patches display significant antimicrobial activity against Candida albicans and Staphylococcus aureus. SEM, 4'-6diamidynofenyloindol (DAPI) staining, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay show that PVA/FLU and PVA/GEL/FLU/VAN nanofibrous patches have a superior effect on NIH3T3 cell spreading and proliferation. The novelty of this study lays in the development of a potential dual drug rapid treatment for corneal ulcers of aggressive nature.

We report on hydroxyapatite (HA) of biological-origin doped with lithium carbonate (LiC) and lithium phosphate (LiP) coatings synthesized by Pulsed laser deposition onto Ti6Al4V substrates fabricated by the Additive manufacturing technique. A detailed comparison from the structural, morphological, chemical composition, wetting behavior and bonding strength standpoints of as-deposited (NTT) and post-deposition thermal-treated (TT) coatings at temperatures ranging from 400 to 700 °C (i.e., TT400–TT700), was performed. Structural investigations indicated a complete crystallization of the initially amorphous HA-based layers at temperatures in excess of 500 °C. The morphological analyses emphasized the rough appearance of the film surfaces, consisting of particulates whose dimensions increased at higher temperatures, with an emphasis on LiC coatings. AFM investigations evidenced rough surfaces, with a clear tendency to increase in corrugation with the applied temperature, in the case of LiC coatings. A hydrophobic behavior was observed for control, NTT and TT400 samples, whilst a radical shift towards hydrophilicity was demonstrated for both types of structures at higher temperatures. In the case of TT500–TT700 coatings, the pull-out adherence values increased considerably compared to control ones. Taking into consideration the obtained results, the positive influence of post-deposition thermal treatments (performed at higher temperatures) on the physical–chemical and mechanical properties of LiC and LiP coatings was indicated. Alongside these improved characteristics observed at elevated temperatures, the sustainable nature of the used BioHA materials should recommend them as viable alternatives to synthetic HA ones for bone implant applications.

The marine species are specially used for the fabrication of bioceramic nano-powders with natural methods for their use in controlled drug delivery. However, there are only very limited studies regarding the production and synthesis of hydroxyapatite (HA)–based drug delivery systems from marine structures. In this study, poly (vinyl alcohol) (PVA) containing Rifampicin (RIF)–loaded Orange Spiny Oyster Seashell (Spondylus barbatus) hydroxyapatite (HA) composite is synthesized by an in situ ultrasound-assisted method. All samples were analyzed by X-ray diffraction (XRD), Fourier transforms infrared (FTIR) spectroscopy, and Scanning electron microscope (SEM), respectively. The in vitro drug release tests of the obtained samples were performed in a phosphate-buffered medium (PBS) at 37 °C. Drug release was evaluated according to five varying kinetic models. In vitro RIF release from HA/PVA composite in phosphate buffer (pH 7.4) showed prolonged sustained drug release. From the drug release kinetic models, Higuchi and Korsmeyer–Peppas were found to be the best model for the three ratios based on the correlation coefficient. The diffusion component is less than 0.5, which indicates quasi-fickian diffusion. From the kinetic study results, the RIF-loaded marine phase composite has potential use in drug delivery applications as it shows positive sustained drug release behavior.

Bioceramics are a fast-growing materials group, which are widely used in orthopedics, maxillofacial, dental, and reconstructive surgeries. They are produced using raw materials either from synthetic or natural sources. As naturally originated resources, the bones of sheep and cows are used after converting to calcium phosphates. Human-originated sources in the past were obtained from human cadaver bones, however now-a-days this has been discontinued. On the other hand, the "golden standard" in the reconstruction surgery has been using patients own bones, -i.e., autogenous bones, which heal better than other alternatives. Besides natural products, synthetic materials are produced from a range of inorganic raw and natural materials based on marine sources, such as corals, and other marine-derived materials (i.e., seashells, nacre). These are used to produce bioceramics and hence implants, devices, and bone grafts. Although during the last four decades a number of excellent books and book chapters have been published, no comprehensive review has been yet reported to cover the available marine materials and to indicate the related work and corresponding references to allow for both medical and ceramic scientists to access directly and open new avenues for further research on marine structures and their applications in orthopedic, maxillofacial, and reconstructive surgery areas. Hence, this review covers the general marine structures, their locations and availability in different countries and, current research on production methods of these unique structures that are difficult to fabricate synthetically. The authors are confident that this comprehensive review will be an excellent source not only for the ceramists, but also for the medical scientists.

Fungal keratitis is a severe corneal infection that causes irreversible damage to the cornea, for which conventional drug treatments may be insufficient. With the new generation of drug delivery systems, it is desired to ensure the ocular usability of drugs. In this study, two-layer polyvinyl alcohol and gelatin (PVA/GEL) nanofibers with high drug loading capacity were produced by the electrospinning method. Cinnamaldehyde (CA), an FDA-approved volatile molecule found in cinnamon essential oil, and fluconazole (FLU), an antifungal drug, were incorporated into PVA/GEL nanofibers to inhibit the growth and biofilm formation of Candida albicans, one of the pathogens that cause fungal keratitis. The morphology, chemical structures, and thermal transitions of the produced pure (PVA/GEL), FLU loaded (PVA/GEL/FLU), CA loaded (PVA/GEL/CA), and combined FLU and CA loaded (PVA/GEL/COM) nanofibers were analysed by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC), respectively. The mechanical analysis, swelling and degradation behaviour, and drug release kinetics of the nanofibers were investigated. PVA/GEL/FLU, PVA/GEL/CA and PVA/GEL/FLU/COM nanofibers were evaluated for their antifungal and antibiofilm activity against Candida albicans. Results showed that PVA/GEL/FLU and PVA/GEL/COM nanofibers have significant antifungal activity and inhibited biofilm formation by 37% and 49%, respectively. Furthermore, it was determined by MTT analysis using a human embryonic kidney (HEK) that the nanofibers were not cytotoxic. In the treatment of fungal keratitis, double-layer PVA/GEL/COM nanofiber with CA in the first layer and FLU in the second layer can create a new treatment approach as an alternative drug delivery system.

In recent years, scaffolds produced in 3D printing technology have become more widespread tool due to providing more advantages than traditional methods in tissue engineering applications. In this research, it was aimed to produce patches for the treatment of tympanic membrane perforations which caused significant hearing loss by using 3D printing method. Polylactic acid(PLA) scaffolds with Chitosan(CS) and Sodium Alginate(SA) added in various ratios were prepared for artificial eardrum patches. Different amounts of chitosan and sodium alginate added to PLA increased the biocompatibility of the produced scaffolds. The created patches were designed by mimicking the thickness of the natural tympanic membrane thanks to the precision provided by the 3D printed method. The produced scaffolds were analyzed separately for chemical, morphological, mechanical and biocompatibility properties. Scanning electron microscope (SEM), Fourier-transform infrared (FT-IR) spectroscopy was performed to observe the surface morphology and chemical structure of the scaffolds. Mechanical, thermal and physical properties, swelling and degradation behaviors were examined to fully analyze whole characteristic features of the samples. Cell culture study was also performed to demonstrate the biocompatibility properties of the fabricated scaffolds with human adipose tissue-derived mesenchymal stem cells (hAD-MSCs). 15 wt % PLA was selected as the control group and among all concentrations of CS and SA, groups containing 3 wt% CS and 3 wt% SA showed significantly superior and favorable features in printing quality. The research continued with these two scaffolds (3 wt% CS, and 3 wt% SA), which showed improved print quality when added to PLA. Overall, these results show that PLA/CS and PLA/SA 3D printed artificial patches have the potential to tissue engineering solutions to repair tympanic membrane perforation for people with hearing loss.

Vitamin D3, vitamin K2, and Mg (10%, 1.25%, and 5%, w/w, respectively)-loaded PLA (12%, w/v) (TCP (5%, w/ v))/PCL (12%, w/v) 1:1 (v/v) composite nanofibers (DKMF) were produced by electrospinning method (ES) and their osteoinductive effects were investigated in cell culture test. Neither pure nanofibers nor DKMF caused a significant cytotoxic effect in fibroblasts. The induction of the stem cell differentiation into osteogenic cells was observed in the cell culture with both DKMF and pure nanofibers, separately. Vitamin D3, vitamin K2, and magnesium demonstrated to support the osteogenic differentiation of mesenchymal stem cells by expressing Runx2, BMP2, and osteopontin and suppressing PPAR-gamma and Sox9. Therefore, the Wnt/beta-catenin signaling pathway was activated by DKMF. DKMF promoted large axonal sprouting and needle-like elongation of osteoblast cells and enhanced cellular functions such as migration, infiltration, proliferation, and differentiation after seven days of incubation using confocal laser scanning microscopy. The results showed that DKMF demonstrated sustained drug release for 144 h, tougher and stronger structure, higher tensile strength, increased water up-take capacity, decreased degradation ratio, and slightly lower Tm and Tg values compared to pure nanofibers. Consequently, DKMF is a promising treatment approach in bone tissue engineering due to its osteoinductive effects.

Tissue engineering is considered a promising alternative due to the limitations of conventional treatment approach. In addition to existing bone grafts, new alternative bone scaffolds continue to be produced by several manufacturing processes. Various producing methods have been applied to develop porous three-dimensional (3D) scaffolds containing a range of biomaterials that are able to aid and direct bone regeneration. Natural bone structure is a heterogeneous composite material comprising organic components (Type I collagen, lipids, and noncollagenous proteins) and inorganic components (hydroxyapatite, calcium phosphate, etc.). During the fabrication of the scaffold, it would make sense to incorporate a compound of materials to form a composite scaffold that possibly allows for better scaffold biological activity and biomimicry structural design. Therefore, after reviewing the properties of skeletal manufacturing methods, this chapter will investigate some of the crucial composites recently used in bone tissue engineering.

The aim of this study is to evaluate the mechanical and biological performance of cartilage-like constructs produced by 3D printing. During the investigation, poly(ε-caprolactone) (PCL) and polyvinylpyrrolidone (PVP) were used as a matrix polymer and low-molecular-weight chitosan (CS), hyaluronic acid (HA), and alginic acid sodium salt (SA) were integrated separately with the polymer matrix to fabricate the constructs. Thermal, mechanical, morphology, and chemical properties and swelling, degradation, and biocompatibility behaviors were evaluated in detail. With the addition of 3 fillers, the melting temperature of the matrix increased with the addition of fillers, and PCL/3wt.%PVP/1wt.%HA had the highest melting temperature value. Mechanical characterization results demonstrated that the printed PCL/3wt.%PVP/1wt.%CS displayed the highest compressive strength of around 9.51 MPa. The compressive strength difference between the PCL/3wt.%PVP and PCL/3wt.%PVP/1wt.%CS was 5.38 MPa. Biocompatibility properties of the constructs were tested by mitochondrial dehydrogenase activity, and in vitro studies showed that the PCL/3wt.%PVP/1wt.%HA composite construct had more cell viability than the other constructs by making use of the mesenchymal stem cell line.

Parkinson’s disease, the second most common neurodegenerative disease in the world, develops due to decreased dopamine levels in the basal ganglia. Levodopa, a dopamine precursor used in the treatment of Parkinson’s disease, can be used as a drug delivery system. This study presents an approach to the use of 3D-printed levodopa-loaded neural tissue scaffolds produced with polylactic acid (PLA) and chitosan (CS) for the treatment of Parkinson’s disease. Surface morphology and pore sizes were examined by scanning electron microscopy (SEM). Average pore sizes of 100–200 µm were found to be ideal for tissue engineering scaffolds, allowing cell penetration but not drastically altering the mechanical properties. It was observed that the swelling and weight loss behaviors of the scaffolds increased after the addition of CS to the PLA. Levodopa was released from the 3D-printed scaffolds in a controlled manner for 14 days, according to a Fickian diffusion mechanism. Mesenchymal stem cells (hAD-MSCs) derived from human adipose tissue were used in MTT analysis, fluorescence microscopy and SEM studies and confirmed adequate biocompatibility. Overall, the obtained results show that PLA/CS 3D-printed scaffolds have an alternative use for the levodopa delivery system for Parkinson’s disease in neural tissue engineering applications.

Bovine hydroxyapatite (BHA) and BHA blended with clinoptilolite (CLIN) and alumina (Al2O3) coatings were synthesized using pulsed laser deposition (PLD) with a KrF* excimer laser source (lambda = 248 nm, tau(FWHM) <= 25 ns). Physical-chemical characteristics and the potential use of coatings for preventing bacteria adhesion and biofilm formation were investigated. Optimized PLD conditions were selected for coatings with rough morphologies, suitable for good cell adhesion and implant anchorage and good replication of the source target composition. The crystallinity of composite coatings was progressively decreasing with the augment of the Al2O3 and CLIN contents, which in turn can facilitate an efficacious release of active components. Al2O3- and CLIN-containing coatings exhibited high cytocompatibility and specific antibiofilm profiles, preventing the initiation and maturation of bacterial biofilms. Optimum biological activity profiles associated with the use of sustainable and/or inexpensive materials are, in our opinion, of key importance for the future development of performant implant coatings, which should he perfectly compatible with the surrounding tissue while preventing postsurgical endogenous or nosocomial infections.

OBJECTIVE:Development of immunologically smart implants, integrated to biological systems, is a key aim to minimize the inflammatory response of the host to biomaterial implants. METHODS:The aim of this study is to investigate the influence of titanium alloy and stainless steel implants on immunological responses in rats by comparative analysis of nuclear factor kappa B (NF-κB) profiles in the activation of inflammatory signaling pathways and the role of CD4+CD25+Foxp3+. RESULTS:Both Ti alloy and stainless steel alloy group implantation affect Toll-like receptors-4 pathways and CD4+CD25+ regulatory T cells in different ways. CONCLUSIONS:Results show that NF-κB/p65 and NF-κB1/p50 possess potential as a therapeutic target in the prevention of adverse reactions to metal, especially for controlling inflammation after the implantation.

We report on biological-derived hydroxyapatite (HA, of animal bone origin) doped with lithium carbonate (Li-C) and phosphate (Li-P) coatings synthesized by pulsed laser deposition (PLD) onto Ti6Al4V implants, fabricated by the additive manufacturing (AM) technique. After being previously validated by in vitro cytotoxicity tests, the Li-C and Li-P coatings synthesized onto 3D Ti implants were preliminarily investigated in vivo, by insertion into rabbits’ femoral condyles. The in vivo experimental model for testing the extraction force of 3D metallic implants was used for this study. After four and nine weeks of implantation, all structures were mechanically removed from bones, by tensile pull-out tests, and coatings’ surfaces were investigated by scanning electron microscopy. The inferred values of the extraction force corresponding to functionalized 3D implants were compared with controls. The obtained results demonstrated significant and highly significant improvement of functionalized implants’ attachment to bone (p-values ≤0.05 and ≤0.00001), with respect to controls. The correct placement and a good integration of all 3D-printed Ti implants into the surrounding bone was demonstrated by performing computed tomography scans. This is the first report in the dedicated literature on the in vivo assessment of Li-C and Li-P coatings synthesized by PLD onto Ti implants fabricated by the AM technique. Their improved mechanical characteristics, along with a low fabrication cost from natural, sustainable resources, should recommend lithium-doped biological-derived materials as viable substitutes of synthetic HA for the fabrication of a new generation of metallic implant coatings.