A Multidisciplinary Evaluation of Three-Dimensional Polycaprolactone Bioactive Glass Scaffolds for Bone Tissue Engineering Purposes.

In the development of bone graft substitutes, a fundamental step is the use of scaffolds with adequate composition and architecture capable of providing support in regenerative processes both on the tissue scale, where adequate resistance to mechanical stress is required, as well as at the cellular level where compliant chemical-physical and mechanical properties can promote cellular activity. In this study, based on a previous optimization study of this group, the potential of a three-dimensional construct based on polycaprolactone (PCL) and a novel biocompatible Mg- and Sr-containing glass named BGMS10 was explored. Fourier-transform infrared spectroscopy and scanning electron microscopy showed the inclusion of BGMS10 in the scaffold structure. Mesenchymal stem cells cultured on both PCL and PCL-BGMS10 showed similar tendencies in terms of osteogenic differentiation; however, no significant differences were found between the two scaffold types. This circumstance can be explained via X-ray microtomography and atomic force microscopy analyses, which correlated the spatial distribution of the BGMS10 within the bulk with the elastic properties and topography at the cell scale. In conclusion, our study highlights the importance of multidisciplinary approaches to understand the relationship between design parameters, material properties, and cellular response in polymer composites, which is crucial for the development and design of scaffolds for bone regeneration.

Effect of Thermal Treatments and Ion Substitution on Sintering and Crystallization of Bioactive Glasses: A Review.

Bioactive glasses (BGs) are promising materials for bone regeneration due to their ability to bond with living bone tissue. However, thermal stability and mechanical properties of BGs need improvement for better clinical performance. In this paper, we present an overview of the influence of different ions on the sintering and crystallization of BGs. Specifically, this review focuses on the impact of thermal treatments on the crystallization of 45S5 and other significant BG compositions. Potential applications of these thermally treated BGs, such as scaffolds, BG-based composites, and thermally sprayed coatings, are explored. Moreover, the substitution of ions has been investigated as a method to enhance the thermal properties of BGs. Notably, zinc, potassium, and strontium have been studied extensively and have demonstrated promising effects on both the thermal and the mechanical properties of BGs. However, it is important to note that research on ion inclusion in BGs is still in its early stages, and further investigation is necessary to fully comprehend the effects of different ions on sintering and crystallization. Therefore, future studies should focus on optimizing the ion substitution method to improve the thermal, mechanical, and even biological properties of BGs, thereby enhancing their potential for various biomedical applications.

Additive Manufacturing of Polymer/Bioactive Glass Scaffolds for Regenerative Medicine: A Review.

Tissue engineering (TE) is a branch of regenerative medicine with enormous potential to regenerate damaged tissues using synthetic grafts such as scaffolds. Polymers and bioactive glasses (BGs) are popular materials for scaffold production because of their tunable properties and ability to interact with the body for effective tissue regeneration. Due to their composition and amorphous structure, BGs possess a significant affinity with the recipient's tissue. Additive manufacturing (AM), a method that allows the creation of complex shapes and internal structures, is a promising approach for scaffold production. However, despite the promising results obtained so far, several challenges remain in the field of TE. One critical area for improvement is tailoring the mechanical properties of scaffolds to meet specific tissue requirements. In addition, achieving improved cell viability and controlled degradation of scaffolds is necessary to ensure successful tissue regeneration. This review provides a critical summary of the potential and limitations of polymer/BG scaffold production via AM covering extrusion-, lithography-, and laser-based 3D-printing techniques. The review highlights the importance of addressing the current challenges in TE to develop effective and reliable strategies for tissue regeneration.

Surface Analysis of Ti-Alloy Micro-Grooved 12/14 Tapers Assembled to Non-Sleeved and Sleeved Ceramic Heads: A Comparative Study of Retrieved Hip Prostheses.

Ti6Al4V titanium alloy (Ti-alloy) sleeved ceramic heads have become widely used in revision surgery when the hip stem is left in situ. This solution guarantees a new junction between the bore of the ceramic head and the Ti-alloy sleeve, regardless of any possible, slight surface damage to the Ti-alloy taper of the stem. However, this solution introduces an additional Ti-alloy/Ti-alloy interface pairing, which is potentially susceptible to mechanically assisted crevice corrosion. This study evaluated both qualitatively and quantitatively the damage that occurred in vivo on Ti-alloy micro-grooved 12/14 tapers of (i) primary implants with non-sleeved ceramic heads (Group 1), (ii) secondary implants with non-sleeved ceramic heads (Group 2), and (iii) secondary implants with sleeved ceramic heads (Group 3). A total of 45 explants-15 for each group, including short-, medium- and long-neck heads-underwent optical evaluation for surface damage (Goldberg scoring), surface roughness analysis, and SEM/EDX analysis. The Goldberg scores did not reveal different patterns in the tapers' surface damage; surface damage was classified as absent or mild (surface damage score ≤2) in 94%, another 94%, and 92% of the analysed regions for Group 1, Group 2, and Group 3, respectively. Small but significant differences in morphological changes occurred in the tapers of the three groups: reductions no greater than a few percentage points in median values of roughness parameters were found in Group 1 and Group 2, while negligible changes were found in Group 3. SEM/EDX analysis revealed little (i.e., a slight increase in the oxygen content) to undetectable changes in the chemical composition on the Ti-alloy surface independently of the group. These results suggest that the Ti-alloy/Ti-alloy sleeve/taper junction is only mildly susceptible to mechanically assisted crevice corrosion. Assembling a sleeved ceramic head, with variable neck lengths up to a "long-neck", to a Ti-alloy micro-grooved 12/14 taper of a stem left in situ does not seem to increase the risk of revision due to trunnionosis, as long as junction stability (i.e., the proper seating of the sleeved ceramic head on the 12/14 taper) is achieved intraoperatively.

Adhesive Bioinspired Coating for Enhancing Glass-Ceramics Scaffolds Bioactivity.

Bioceramic scaffolds, composed of a biphasic composite containing bioactive glass and hydroxyapatite, were prepared in this work to overcome the intrinsic limits of the two components taken separately (in particular, their specific reactivities and dissolution rates, which should be tunable as a function of the given clinical requirements). To mimic the biological environment and tune the different stages of cellular response, a coating with gelatin and chondroitin sulphate via Layer-by-Layer (LbL) assembly was presented and discussed. The resulting functionalized scaffolds were affected by the coating in terms of microstructure and porosity. In addition, the LbL coating significantly enhanced the seeded cell behaviour, with high adhesion, proliferation and osteogenic activity, as revealed by the alkaline phosphatase activity and overexpression of osteopontin and osteocalcin.

Low-Temperature Sintering of a New Bioactive Glass Enriched with Magnesium Oxide and Strontium Oxide.

The recent research on bioactive glasses (BGs) has mainly moved on two fronts: (1) introducing ions of therapeutic interest in their composition and (2) the development of scaffolds, fibers, coatings and sintered products starting from BGs in powder form. In this case, the main obstacle to overcome is that BGs rapidly crystallize during heat treatments, thus transforming into glass-ceramics with low reactivity, slow ion release and, eventually, poor mechanical properties. Here an innovative bioactive glass (BGMS_LS), capable of responding to the main limitations of commercial BGs, is presented. The new material contains strontium and magnesium, whose therapeutic relevance is well known, and can be sintered at extraordinarily low temperatures without crystallizing, thus keeping all of its biological potential intact.

Bioactive Glasses in Periodontal Regeneration: Existing Strategies and Future Prospects-A Literature Review.

The present review deals with bioactive glasses (BGs), a class of biomaterials renowned for their osteoinductive and osteoconductive capabilities, and thus widely used in tissue engineering, i.e., for the repair and replacement of damaged or missing bone. In particular, the paper deals with applications in periodontal regeneration, with a special focus on in vitro, in vivo and clinical studies. The study reviewed eligible publications, identified on the basis of inclusion/exclusion criteria, over a ranged time of fifteen years (from 1 January 2006 to 31 March 2021). While there are many papers dealing with in vitro tests, only a few have reported in vivo (in animal) research, or even clinical trials. Regardless, BGs seem to be an adequate choice as grafts in periodontal regeneration.

Bioactive Glass Applications: A Literature Review of Human Clinical Trials.

The use of bioactive glasses in dentistry, reconstructive surgery, and in the treatment of infections can be considered broadly beneficial based on the emerging literature about the potential bioactivity and biocompatibility of these materials, particularly with reference to Bioglass® 45S5, BonAlive® and 19-93B3 bioactive glasses. Several investigations have been performed (i) to obtain bioactive glasses in different forms, such as bulk materials, powders, composites, and porous scaffolds and (ii) to investigate their possible applications in the biomedical field. Although in vivo studies in animals provide us with an initial insight into the biological performance of these systems and represent an unavoidable phase to be performed before clinical trials, only clinical studies can demonstrate the behavior of these materials in the complex physiological human environment. This paper aims to carefully review the main published investigations dealing with clinical trials in order to better understand the performance of bioactive glasses, evaluate challenges, and provide an essential source of information for the tailoring of their design in future applications. Finally, the paper highlights the need for further research and for specific studies intended to assess the effect of some specific dissolution products from bioactive glasses, focusing on their osteogenic and angiogenic potential.

Composite Scaffolds for Bone Tissue Regeneration Based on PCL and Mg-Containing Bioactive Glasses.

Polycaprolactone (PCL) is widely used in additive manufacturing for the construction of scaffolds for tissue engineering because of its good bioresorbability, biocompatibility, and processability. Nevertheless, its use is limited by its inadequate mechanical support, slow degradation rate and the lack of bioactivity and ability to induce cell adhesion and, thus, bone tissue regeneration. In this study, we fabricated 3D PCL scaffolds reinforced with a novel Mg-doped bioactive glass (Mg-BG) characterized by good mechanical properties and biological reactivity. An optimization of the printing parameters and scaffold fabrication was performed; furthermore, an extensive microtopography characterization by scanning electron microscopy and atomic force microscopy was carried out. Nano-indentation tests accounted for the mechanical properties of the scaffolds, whereas SBF tests and cytotoxicity tests using human bone-marrow-derived mesenchymal stem cells (BM-MSCs) were performed to evaluate the bioactivity and in vitro viability. Our results showed that a 50/50 wt% of the polymer-to-glass ratio provides scaffolds with a dense and homogeneous distribution of Mg-BG particles at the surface and roughness twice that of pure PCL scaffolds. Compared to pure PCL (hardness H = 35 ± 2 MPa and Young's elastic modulus E = 0.80 ± 0.05 GPa), the 50/50 wt% formulation showed H = 52 ± 11 MPa and E = 2.0 ± 0.2 GPa, hence, it was close to those of trabecular bone. The high level of biocompatibility, bioactivity, and cell adhesion encourages the use of the composite PCL/Mg-BG scaffolds in promoting cell viability and supporting mechanical loading in the host trabecular bone.

A Review of Bioactive Glass/Natural Polymer Composites: State of the Art.

Collagen, gelatin, silk fibroin, hyaluronic acid, chitosan, alginate, and cellulose are biocompatible and non-cytotoxic, being attractive natural polymers for medical devices for both soft and hard tissues. However, such natural polymers have low bioactivity and poor mechanical properties, which limit their applications. To tackle these drawbacks, collagen, gelatin, silk fibroin, hyaluronic acid, chitosan, alginate, and cellulose can be combined with bioactive glass (BG) nanoparticles and microparticles to produce composites. The incorporation of BGs improves the mechanical properties of the final system as well as its bioactivity and regenerative potential. Indeed, several studies have demonstrated that polymer/BG composites may improve angiogenesis, neo-vascularization, cells adhesion, and proliferation. This review presents the state of the art and future perspectives of collagen, gelatin, silk fibroin, hyaluronic acid, chitosan, alginate, and cellulose matrices combined with BG particles to develop composites such as scaffolds, injectable fillers, membranes, hydrogels, and coatings. Emphasis is devoted to the biological potentialities of these hybrid systems, which look rather promising toward a wide spectrum of applications.

A New Generation of Electrospun Fibers Containing Bioactive Glass Particles for Wound Healing.

Chitosan fibers blended with polyethylene oxide (CHIT_PEO) and crosslinked with genipin were fabricated by electrospinning technique. Subsequently, CHIT_PEO bioactive glass composite electrospun mats were fabricated with the aim to achieve flexible structures with adequate mechanical properties and improved biological performance respect to CHIT_PEO fibers, for potential applications in wound healing. Three different compositions of bioactive glasses (BG) were selected and investigated: 45S5 BG, a Sr and Mg containing bioactive glass (BGMS10) and a Zn-containing bioactive glass (BGMS_2Zn). Particulate BGs (particles size < 20 µm) were separately added to the starting CHIT_PEO solution before electrospinning. The two recently developed bioactive glasses (BGMS10 and BGMS_2Zn) showed very promising biological properties in terms of bioactivity and cellular viability; thus, such compositions were added for the first time to CHIT_PEO solution to fabricate composite electrospun mats. The incorporation of bioactive glass particles and their distribution into CHIT_PEO fibers were assessed by SEM and FTIR analyses. Furthermore, CHIT_PEO composite electrospun mats showed improved mechanical properties in terms of Young's Modulus compared to neat CHIT_PEO fibers; on the contrary, the values of tensile strain at break (%) were comparable. Biological performance in terms of cellular viability was investigated by means of WST-8 assay and CHIT_PEO composite electrospun mats showed cytocompatibility and the desired cellular viability.

A Novel Bioactive Glass Containing Therapeutic Ions with Enhanced Biocompatibility.

A novel bioactive glass containing therapeutic ions with enhanced biocompatibility was designed and produced by the classical melt-quenching route. Starting from a very promising composition (Bio_MS), which combined bioactivity and high crystallization temperature, the ratio between some oxides was tailored to obtain a new and more reactive (in terms of dissolution rate) bioactive glass, called BGMSN (composition in mol%: 6.1 Na2O, 31.3 CaO, 5 MgO, 10 SrO, 2.6 P2O5, 45 SiO2). The aim of this work was to produce a bioactive glass with a good biological performance, preserving, at the same time, the high crystallization temperature achieved for Bio_MS; this is strategic in order to avoid undesired crystalline phases during thermal treatments, which can undermine the bioactivity and even the stability of final products. A complete characterization of the novel bioactive glass was performed in terms of thermal, mechanical and biological properties and in vitro bioactivity. The thermal behavior of the bioactive glass was studied by heating microscopy, differential thermal analysis (DTA) and optical dilatometry; BGMSN showed a very high crystallization temperature and a high sinterability parameter, thus being suitable for applications where thermal treatments are required, such as sintered samples, coatings and scaffolds. Mechanical properties were investigated by the micro-indentation technique. The in vitro biological properties were evaluated by means of both direct and indirect cell tests, i.e., neutral red (NR) uptake and MTT assay, using murine long bone osteocyte Y4 (MLO-Y4) cells: the cellular viability of BGMSN was higher compared to cellular viability of 45S5, both in direct and indirect tests. Finally, the in vitro bioactivity test by soaking samples in simulated body fluid (SBF) showed high dissolution rate, with a good rate of formation of hydroxyapatite.

Effects of a Novel Bioactive Glass Composition on Biological Properties of Human Dental Pulp Stem Cells.

Functional reconstruction of bone defects represents a clinical challenge in the regenerative medicine field, which targets tissue repair following traumatic injuries and disease-related bone deficiencies. In this regard, the optimal biomaterial should be safe, biocompatible and tailored in order to promote the activation of host progenitor cells towards bone repair. Bioactive glasses might be suitable biomaterials due to their composition being able to induce the host healing response and, eventually, anti-bacterial properties. In this study we investigated whether and how an innovative bioactive glass composition, called BGMS10, may affect cell adhesion, morphology, proliferation, immunomodulation and osteogenic differentiation of human dental pulp stem cells (hDPSCs). When cultured on BGMS10, hDPSCs maintained their proliferation rate and typical fibroblast-like morphology, showing the expression of stemness markers STRO-1 and c-Kit. Moreover, the expression of FasL, a key molecule in mediating immunomodulation effects of hDPSCs, was maintained. BGMS10 also proved to trigger osteogenic commitment of hDPSCs, as confirmed by the activation of bone-related transcription factors RUNX2 and Osx and the ongoing deposition of extracellular matrix supported by the expression of OPN and OCN. Our findings suggest that BGMS10 not only maintains the typical biological and immunomodulatory properties of hDPSCs but also favors the osteogenic commitment.

Chitosan-Based Bioactive Glass Gauze: Microstructural Properties, In Vitro Bioactivity, and Biological Tests.

Passive commercial gauzes were turned into interactive wound dressings by impregnating them with a chitosan suspension. To further improve healing, and cell adhesion and proliferation, chitosan/bioactive glass wound dressings were produced with the addition of (i) 45S5, (ii) a Sr- and Mg-containing bioactive glass, and (iii) a Zn-containing bioactive glass to the chitosan suspension. SEM and FTIR analyses evidenced positive results in terms of incorporation of bioactive glass particles. Bioactivity was investigated by soaking chitosan-based bioactive glass wound dressings in simulated body fluid (SBF). Cell viability, proliferation, and morphology were investigated using NIH 3T3 (mouse embryonic fibroblast) cells by neutral red (NR) uptake and MTT assays. Furthermore, the wound-healing rate was evaluated by means of the scratch test, using NIH 3T3. The results showed that bioactive glass particles enhance cell adhesion and proliferation, and wound healing compared to pure chitosan. Therefore, chitosan-based bioactive glass wound dressings combine the properties of the organic matrix with the specific biological characteristics of bioactive glasses to achieve chitosan composites suitable for healing devices.

On the in Vitro Biocompatibility Testing of Bioactive Glasses.

In this work, a new 3D cellular model-based on human bone marrow mesenchymal stem cells (BM-MSCs)-was used for the first time to test the 45S5 Bioglass® (45S5). Such a model, previously used to evaluate the biologic performance of two novel bioactive glasses, suggested out the regenerative potential of such materials. In fact, BM-MSCs were able both to adhere and colonize the biomaterials, as well as differentiate toward osteoblasts-even in absence of specific growth factors. Surprisingly, BM-MSCs were not able to colonize 45S5 granules (almost no adhesion and/or colonization was observed), and thus, were not capable of providing any osteogenic differentiation. Therefore, the model seems to suggest that the two novel bioactive glasses have a better biologic performance than 45S5. If this hypothesis is confirmed also by in vivo tests, the 3D model may become a predictive tool for discriminating between different potential bioactive materials by comparatively evaluating them, and preliminarily selecting the best ones in relation to their biocompatibility potential-before proceeding with further experiments in vivo. This approach could favor the reduction of costs and time of pre-clinical and clinical trials.

A new bioactive glass with extremely high crystallization temperature and outstanding biological performance.

In this work, a new bioactive glass was designed, prepared by means of a melt-quenching route and characterized in terms of both thermal properties and biological performance. The main objective was to obtain a novel product with high temperature of crystallization in view of possible thermal treatments, as well as remarkable biological responsiveness. Thermal behavior was investigated by heating microscopy, differential thermal analysis (DTA) and sintering tests. The glass displayed a very high crystallization temperature and the samples remained completely amorphous after sintering. Bioactivity was evaluated by means of Simulated Body Fluid (SBF) assay, which is a widely used method to preliminary investigate samples' reactivity in vitro; the glass showed a strong apatite forming ability. Additionally, in order to exclude cytotoxic effects, biocompatibility was verified according to ISO standard 10993. Finally, the biological potential of the new glass was tested by using an innovative 3D cellular model, that mimicked the potential clinical application of a given biomaterial. Human bone marrow mesenchymal stem cells (BM-MSCs) were employed to study the performance of bioactive glass granules in such 3D cellular model. The results showed that the bioactive glass supported human BM-MSCs adhesion, colonization and bone differentiation. Thus, this new bioactive glass looks particularly promising for orthopedic applications, bone tissue engineering and regenerative medicine, especially when a thermal treatment is necessary for the production of specific devices.

In vitro studies of solution precursor plasma-sprayed copper-doped hydroxyapatite coatings with increasing copper content.

In this present study, in vitro characterization on bioactivity and biocompatibility of the copper-doped hydroxyapatite (Cu-HA) coatings deposited onto Ti6Al4V alloy with increasing copper content obtained using solution precursor plasma spray process were evaluated under simulated body fluid (SBF) and cytotoxicity tests. The growth of flake-like structures was observed at the coatings' surface after 7 days of immersion in SBF. Elemental composition analysis showed a calcium-deficient carbonate-containing apatite for the grown layer. Broadening of the HA peaks were also observed in the X-ray diffraction patterns and infrared absorption spectra of the immersed coatings associated with the possible formation of an amorphous layer at the surface of the coatings. Copper incorporation does not alter the bioactivity of HA but only slowed down for 10 mol% copper content. Cytotoxicity evaluation using MTT assays showed that 3 mol% Cu-HA coating was not toxic under in vitro characterizations.

Comparison of Three Manufacturing Techniques for Sustainable Porous Clay Ceramics.

This study proposes different manufacturing techniques (manual pelletization, powder pressing, and "shell scaffold") to obtain lightweight clay ceramics containing recovery raw materials. The sintering in an electrical furnace (1000 °C, 1 h processing time) was conducted by traditional firing from room temperature, for pressed and shell-scaffold samples, while the flash heating (i.e., samples directly put at 1000 °C) was used only for the pellets. The porous materials (porosity 40-80%), functionalized with nutrients (K and P) in amounts to confer the fertilizer capability, gave suitable results in terms of pH (6.7-8.15) and electrical conductivity (0.29-1.33 mS/cm). Thus, such materials can be considered as feasible lightweight clay ceramics, with a positive effect on the soil. These findings permit us to hypothesize a potential use in green roofs or in agronomic applications.

Human Mesenchymal Stem Cell Combined with a New Strontium-Enriched Bioactive Glass: An ex-vivo Model for Bone Regeneration.

A 3D cellular model that mimics the potential clinical application of a biomaterial is here applied for the first time to a bioactive glass, in order to assess its biological potential. A recently developed bioactive glass (BGMS10), whose composition contained strontium and magnesium, was produced in the form of granules and fully investigated in terms of biocompatibility in vitro. Apart from standard biological characterization (Simulated Body Fluid (SBF) testing and biocompatibility as per ISO10993), human bone marrow mesenchymal stromal/stem cells (BM-MSCs) were used to investigate the performance of the bioactive glass granules in an innovative 3D cellular model. The results showed that BGMS10 supported human BM-MSCs adhesion, colonization, and bone differentiation. Thus, bioactive glass granules seem to drive osteogenic differentiation and thus look particularly promising for orthopedic applications, bone tissue engineering and regenerative medicine.

Advanced Open-Celled Structures from Low-Temperature Sintering of a Crystallization-Resistant Bioactive Glass.

Most materials for bone tissue engineering are in form of highly porous open-celled components (porosity >70%) developed by means of an adequate coupling of formulations and manufacturing technologies. This paper is dedicated to porous components from BGMS10 bioactive glass, originally designed to undergo viscous flow sintering without crystallization, which is generally known to degrade the bioactivity of 45S5 bioglass. The adopted manufacturing technologies were specifically conceived to avoid any contamination and give excellent control on the microstructures by simple operations. More precisely, 'green' components were obtained by digital light processing and direct foaming of glass powders suspended in a photosensitive organic binder or in an aqueous solution, activated with an organic base, respectively. Owing to characteristic quite large sintering window of BGMS10 glass, sintering at 750 °C caused the consolidation of the structures generated at room temperature, without any evidence of viscous collapse.