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Mesoporous bioactive glass nanoparticles (MBGNPs) are attracting significant attention as suitable materials for multifunctional biomedical applications. In this study, cerium was incorporated into MBGNPs using two different approaches. In the first approach, cerium was added to the glass system directly during the synthesis, while in the second approach, cerium was added to the as-synthesized MBGNPs via the template ion-exchange method. The influence of the method of synthesis on the physicochemical properties of nanoparticles was examined by SEM, TEM, XRD, FTIR, and N2 adsorption-desorption analyses. The MBGNPs exhibited spheroidal morphology and disordered mesoporous structure. XRD analysis confirmed the amorphous nature of the nanoparticles. The chemical composition was determined by the acid digestion method using ICP-OES. The influence of the synthesis method on the specific surface area, mesoporosity, and solubility of synthesized nanoparticles in Tris/HCl (pH 7.4) and acetate (pH 4.5) buffer has also been studied. The obtained Ce containing MBGNPs were non-cytotoxic toward ​preosteoblast MC3T3-E1 cells in contact with nanoparticles in a concentration of up to 100 ​μg/mL. The anti-inflammatory effect of Ce containing MBGNPs was tested with lipopolysaccharides (LPS)-induced proinflammatory RAW 264.7 macrophage cells. Ce containing MBGNPs decreased the release of nitric oxide, indicating the anti-inflammatory response of macrophage cells. Ce containing MBGNPs also showed antibacterial activity against S. aureus and E. coli. The mentioned features of the obtained MBGNPs make them useful in a variety of biomedical applications, considering their biocompatibility, anti-inflammatory response, and enhanced antibacterial effect.

Electrospinning is a versatile and straightforward technique to produce nanofibrous mats with different morphologies. In addition, by optimizing the solution, processing, and environmental parameters, three-dimensional (3D) nanofibrous scaffolds can also be created using this method. In this work, the preparation and characterization of bioactive glass (BG) scaffolds based on the SiO 2 -CaO sol-gel system, a biomaterial with a highly reactive surface, is reported. The electrospinning technique was combined with sol-gel methods to obtain nanofibrous 3D cotton wool-like scaffolds. The addition of zinc and copper ions to the silica-calcia system was examined, and the influence of these ions on the material properties and characteristics was investigated by various characterization techniques, from morphological and chemical properties to antibacterial and wound closure capability, cell viability and ion release. Our findings show that the cotton wool-like ion-doped nanofibers are promising for wound healing applications., (Creative Commons Attribution license.)

Mesoporous silica nanoparticles were synthesized using a microemulsion-assisted sol-gel method, and calcium, gallium or a combination of both, were used as dopants. The influence of these metallic ions on the physicochemical properties of the nanoparticles was investigated by scanning and transmission electron microscopy, as well as N 2 adsorption-desorption methods. The presence of calcium had a significant impact on the morphology and textural features of the nanoparticles. The addition of calcium increased the average diameter of the nanoparticles from 80 nm to 150 nm, while decreasing their specific surface area from 972 m 2 /g to 344 m 2 /g. The nanoparticles of all compositions were spheroidal, with a disordered mesoporous structure. An ion release study in cell culture medium demonstrated that gallium was released from the nanoparticles in a sustained manner. In direct contact with concentrations of up to 100 μg/mL of the nanoparticles, gallium-containing nanoparticles did not exhibit cytotoxicity towards pre-osteoblast MC3T3-E1 cells. Moreover, in vitro cell culture tests revealed that the addition of gallium to the nanoparticles enhanced osteogenic activity. Simultaneously, the nanoparticles disrupted the osteoclast differentiation of RAW 264.7 macrophage cells. These findings suggest that gallium-containing nanoparticles possess favorable physicochemical properties and biological characteristics, making them promising candidates for applications in bone tissue regeneration, particularly for unphysiological or pathological conditions such as osteoporosis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Assisted Reproductive Technologies (ARTs) have been validated in human and animal to solve reproductive problems such as infertility, aging, genetic selection/amplification and diseases. The persistent gap in ART biomedical applications lies in recapitulating the early stage of ovarian folliculogenesis, thus providing protocols to drive the large reserve of immature follicles towards the gonadotropin-dependent phase. Tissue engineering is becoming a concrete solution to potentially recapitulate ovarian structure, mostly relying on the use of autologous early follicles on natural or synthetic scaffolds. Based on these premises, the present study has been designed to validate the use of the ovarian bioinspired patterned electrospun fibrous scaffolds fabricated with poly(ε-caprolactone) (PCL) for multiple preantral (PA) follicle development., Methods: PA follicles isolated from lamb ovaries were cultured on PCL scaffold adopting a validated single-follicle protocol (Ctrl) or simulating a multiple-follicle condition by reproducing an artificial ovary engrafted with 5 or 10 PA (AO 5PA and AO 10PA ). The incubations were protracted for 14 and 18 days before assessing scaffold-based microenvironment suitability to assist in vitro folliculogenesis (ivF) and oogenesis at morphological and functional level., Results: The ivF outcomes demonstrated that PCL-scaffolds generate an appropriate biomimetic ovarian microenvironment supporting the transition of multiple PA follicles towards early antral (EA) stage by supporting follicle growth and steroidogenic activation. PCL-multiple bioengineering ivF (AO 10PA ) performed in long term generated, in addition, the greatest percentage of highly specialized gametes by enhancing meiotic competence, large chromatin remodeling and parthenogenetic developmental competence., Conclusions: The study showcased the proof of concept for a next-generation ART use of PCL-patterned scaffold aimed to generate transplantable artificial ovary engrafted with autologous early-stage follicles or to advance ivF technologies holding a 3D bioinspired matrix promoting a physiological long-term multiple PA follicle protocol., (© 2024. The Author(s).)

Introduction: Total ankle arthroplasty (TAR) procedures have become more reliable and incidence is increasing. A growing number of postoperative complications can be expected and should be correctly addressed., Presentation of Case: A 43-year-old woman suffering from severe ankle osteoarthritis underwent TAR (Stryker's Infinity with Prophecy alignment guides, uncemented tibial component and cemented talus component). After a fall, one month after the surgery, she presented with a fracture of the medial malleolus and an anterior periprosthetic fracture of the tibia, with anterior dislocation of the tibial prosthetic component. Fracture fixation and partial revision surgery was planned using the same anterior surgical access. To restore length, rotation and joint articulation, the medial malleolus was first reduced and synthesized with a plate and 6 screws through additional minimally invasive medial ankle incisions. Once the medial malleolus was stabilized, the already mobilized tibial component was removed. Freehand cuts under fluoroscopic guidance on the anterior surface of the tibia were performed. After a final check with a trial component, a Stryker's Inbone II stem, which matched the previously inserted talar component, was implanted and partially cemented to fill the remaining bone gaps., Discussion: Only few case reports of periprosthetic ankle fractures exist, and none of them were similar to ours. There are no published precedents for this revision approach from a stemless to a stemmed tibial implant without changing the talar implant., Conclusion: Partial revision after a complex periprosthetic fracture results in a satisfactory outcome if surgery is performed in specialized centers., Competing Interests: Conflict of interest statement None., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this work, composite electrospun fibers containing innovative bioactive glass nanoparticles were produced and characterized. Poly(ε-caprolactone), benign solvents, and sol-gel B- and Cu-doped bioactive glass powders were used to fabricate fibrous scaffolds. The retention of bioactive glass nanoparticles in the polymer matrix, the electrospinnability of this novel solution and the obtained electrospun composites were extensively characterized. As a result, composite electrospun fibers characterized by biocompatibility, bioactivity, and exhibiting overall properties adequate for both hard and soft tissue engineering applications, have been produced. The addition of these bioactive glass nanoparticles was, indeed, able to impart bioactive properties to the fibers. Cell culture studies show promising results, demonstrating proliferation and growth of cells on the composite fibers. Wettability, degradation rate, and mechanical performance were also tested and are in line with previous results., (© 2023 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC.)

Pasteurella multocida , a zoonotic infectious organism, has most often been described in patients after an animal bite. It can cause a variety of infections ranging from superficial skin infections to more serious systemic infections, such as sepsis and meningitis. P. multocida is a rare but well-recognized cause of prosthetic joint infections. Here, we report the first implant-associated infection caused by drug-resistant (penicillin, ampicillin, amoxicillin/clavulanic acid) P. multocida , which was cured with targeted antimicrobial treatment and debridement, exchange of mobile parts, and retention of the prosthesis. Patients undergoing arthroplasty should be informed of the risks of close contact with pets, especially in light of the worrying phenomena of drug resistance spreading among animals due to the addition of antibiotics in animal feed., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Maritati et al.)

In vitro maturation (IVM) is not a routine assisted reproductive technology (ART) for oocytes collected from early antral (EA) follicles, a large source of potentially available gametes. Despite substantial improvements in IVM in the past decade, the outcomes remain low for EA-derived oocytes due to their reduced developmental competences. To optimize IVM for ovine EA-derived oocytes, a three-dimensional (3D) scaffold-mediated follicle-enclosed oocytes (FEO) system was compared with a validated cumulus-oocyte complex (COC) protocol. Gonadotropin stimulation (eCG and/or hCG) and/or somatic cell coculture (ovarian vs. extraovarian-cell source) were supplied to both systems. The maturation rate and parthenogenetic activation were significantly improved by combining hCG stimulation with ovarian surface epithelium (OSE) cells coculture exclusively on the FEO system. Based on the data, the paracrine factors released specifically from OSE enhanced the hCG-triggering of oocyte maturation mechanisms by acting through the mural compartment (positive effect on FEO and not on COC) by stimulating the EGFR signaling. Overall, the FEO system performed on a developed reproductive scaffold proved feasible and reliable in promoting a synergic cytoplasmatic and nuclear maturation, offering a novel cultural strategy to widen the availability of mature gametes for ART.

Human Adipose-Derived Mesenchymal Stem/Stromal Cells (hAD-MSCs) have great potential for tissue regeneration. Since transplanted hAD-MSCs are likely to be placed in a hypoxic environment, culturing the cells under hypoxic conditions might improve their post-transplantation survival and regenerative performance. The combination of hAD-MSCs and PCL-nHA nanofibers synergically improves the contribution of both components for osteoblast differentiation. In this work, we hypothesized that this biomaterial constitutes a hypoxic environment for hAD-MSCs. We studied the cellular re-arrangement and the subcellular ultrastructure by Transmission Electron Microscopy (TEM) of hAD-MSCs grown into PCL-nHA nanofibers, and we compared them with the same cells grown in two-dimensional cultures, over tissue culture-treated plastic, or glass coverslips. Among the most evident changes, PCL-nHA grown cells showed enlarged mitochondria, and accumulation of glycogen granules, consistent with a hypoxic environment. We observed a 3.5 upregulation (p = 0.0379) of Hypoxia Inducible Factor (HIF)-1A gene expression in PCL-nHA grown cells. This work evidences for the first time intra-cellular changes in three-dimensional compared to two-dimensional cultures, which are adaptive responses of the cells to an environment more closely resembling that of the in vivo niche after transplantation, thus PCL-nHA nanofibers are adequate for hAD-MSCs pre-conditioning., (© 2023 Wiley-VCH GmbH.)

Background: Aliphatic polyesters are widely used for biomedical, pharmaceutical and environmental applications due to their high biodegradability and cost-effective production. Recently, star and hyperbranched polyesters based on glycerol and ω-carboxy fatty diacids have gained considerable interest. Succinic acid and bio-based diacids similar to glycerol are regarded as safe materials according to the US Food and Drug Administration (FDA). Bioactive glass scaffolds utilized in bone tissue engineering are relatively brittle materials. However, their mechanical properties can be improved by using polymer coatings that can further control their degradation rate, tailor their biocompatibility and enhance their performance. The purpose of this study is to explore a new biopolyester poly(glycerol succinate) (PGSuc) reinforced with mesoporous bioactive nanoparticles (MSNs) as a novel coating material to produce hybrid scaffolds for bone tissue engineering., Methods: Bioactive glass scaffolds were coated with neat PGSuc, PGSuc loaded with dexamethasone sodium phosphate (DexSP) and PGSuc loaded with DexSP-laden MSNs. The physicochemical, mechanical and biological properties of the scaffolds were also evaluated., Results: Preliminary data are provided showing that polymer coatings with and without MSNs improved the physicochemical properties of the 1393 bioactive glass scaffolds and increased the ALP activity and alizarin red staining, suggesting osteogenic differentiation potential when cultured with adipose-derived mesenchymal stem cells., Conclusions: PGSuc with incorporated MSNs coated onto 1393 bioactive glass scaffolds could be promising candidates in bone tissue engineering applications.

We report the successful preparation and characterization of chitosan-Zn complex (ChiZn) in the form of films, intended to enhance the biological performance of chitosan by the presence of Zn as antibacterial agent and biologically active ion. The influence of Zn chelation on morphology and structure of chitosan was assessed by scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy and infrared spectroscopy. The biodegradability study of ChiZn showed a sustained release of Zn up to 2 mg/mL. No toxic response was observed toward stromal cell line ST-2 in indirect contact with the ChiZn films. The dissolution product of ChiZn showed improved wound closure (88% closure) compared to the positive control group (70% closure). Moreover, ChiZn exhibited antibacterial activity against S. aureus together with a slight increase (~30%) in the secretion of VEGF and moderate decrease in nitric oxide evolution. Our findings indicate that ChiZn could be used as a safe and effective wound healing agent., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

Current assisted reproduction technologies (ART) are insufficient to cover the slice of the population needing to restore fertility, as well as to amplify the reproductive performance of domestic animals or endangered species. The design of dedicated reproductive scaffolds has opened the possibility to better recapitulate the reproductive 3D ovarian environment, thus potentially innovating in vitro folliculogenesis (ivF) techniques. To this aim, the present research has been designed to compare ovine preantral follicles in vitro culture on poly(epsilon-caprolactone) (PCL)-based electrospun scaffolds designed with different topology (Random vs. Patterned fibers) with a previously validated system. The ivF performances were assessed after 14 days under 3D-oil, Two-Step (7 days in 3D-oil and on scaffold), or One-Step PCL protocols (14 days on PCL-scaffold) by assessing morphological and functional outcomes. The results show that Two- and One-Step PCL ivF protocols, when performed on patterned scaffolds, were both able to support follicle growth, antrum formation, and the upregulation of follicle marker genes leading to a greater oocyte meiotic competence than in the 3D-oil system. In conclusion, the One-Step approach could be proposed as a practical and valid strategy to support a synergic follicle-oocyte in vitro development, providing an innovative tool to enhance the availability of matured gametes on an individual basis for ART purposes.

Electrospun fibers based on biodegradable polyanionic or polycationic biopolymers are highly beneficial for biomedical applications. In this work, electrospun nanofibers made from poly(epsilon caprolactone) (PCL), chitosan (CS) and κ -carrageenan ( κ -C) were successfully fabricated using several mixtures of benign solvents containing formic acid and acetic acid. The addition of κ -C improved the preparation procedure for the production of PCL/CS fibers by electrospinning. Moreover, a polymer mixture was selected to be stored at -20 °C for one month with the purpose to study the properties of the resulting fiber mat. The results indicated that fiber characteristics were not seriously compromised compared to the ones of those fabricated with the original solution, which represents an important reduction in produced waste. Thus, the interactions that occur between positively and negatively charged hydrophilic polysaccharides might induce higher stability to the linear aliphatic polyester in the polymer mixture. All fiber mats were morphologically, physico-chemically and mechanically characterized, showing average fiber diameters in the nano scale. A direct cell viability assay using ST-2 cells demonstrated cell proliferation after seven days of incubation for all prepared fiber mats, confirming their suitability as potential candidates for bone tissue engineering and wound healing applications., (Creative Commons Attribution license.)

Tendon injuries are at the frontier of innovative approaches to public health concerns and sectoral policy objectives. Indeed, these injuries remain difficult to manage due to tendon's poor healing ability ascribable to a hypo-cellularity and low vascularity, leading to the formation of a fibrotic tissue affecting its functionality. Tissue engineering represents a promising solution for the regeneration of damaged tendons with the aim to stimulate tissue regeneration or to produce functional implantable biomaterials. However, any technological advancement must take into consideration the role of the immune system in tissue regeneration and the potential of biomaterial scaffolds to control the immune signaling, creating a pro-regenerative environment. In this context, immunoengineering has emerged as a new discipline, developing innovative strategies for tendon injuries. It aims at designing scaffolds, in combination with engineered bioactive molecules and/or stem cells, able to modulate the interaction between the transplanted biomaterial-scaffold and the host tissue allowing a pro-regenerative immune response, therefore hindering fibrosis occurrence at the injury site and guiding tendon regeneration. Thus, this review is aimed at giving an overview on the role exerted from different tissue engineering actors in leading immunoregeneration by crosstalking with stem and immune cells to generate new paradigms in designing regenerative medicine approaches for tendon injuries.

Background: Yttria-stabilized zirconia nanoparticles can be applied as fillers to improve the mechanical and antibacterial properties of luting cement. The aim of this study was to synthesize yttria-stabilized zirconia nanoparticles by the sol-gel method and to investigate their composition, structure, morphology and biological properties., Methods: Nanopowders of ZrO 2 7 wt% Y 2 O 3 (nY-ZrO) were synthesized by the sol-gel method and were sintered at three different temperatures: 800, 1000 and 1200 °C, and their composition, size and morphology were investigated. The biocompatibility was investigated with human gingival fibroblasts (hGFs), while reactive oxygen species (ROS) production was evaluated through fluorescence analysis., Results: All synthesized materials were composed of tetragonal zirconia, while nanopowders sintered at 800 °C and 1000 °C additionally contained 5 and 20 wt% of the cubic phase. By increasing the calcination temperature, the crystalline size of the nanoparticles increased from 12.1 nm for nY-ZrO800 to 47.2 nm for nY-ZrO1200. Nano-sized particles with good dispersion and low agglomeration were received. Cell culture studies with human gingival fibroblasts verified the nanopowders' biocompatibility and their ROS scavenging activity., Conclusions: the obtained sol-gel derived nanopowders showed suitable properties to be potentially used as nanofillers for dental luting cement.

Electrospun fibers of shape memory triethoxysilane-terminated poly(epsilon-caprolactone) (PCL-TES) loaded with bioactive glasses (BG) are here presented. Unloaded PCL-TES, as well as PCL/BG nanocomposite fibers, are also considered for comparison. It is proposed that hydrolysis and condensation reactions take place between triethoxysilane groups of the polymer and the silanol groups at the BG particle surface, thus generating additional crosslinking points with respect to those present in the PCL-TES system. The as-spun PCL-TES/BG fibers display excellent shape memory properties, in terms of shape fixity and shape recovery ratios, without the need of a thermal crosslinking treatment. BG particles confer in vitro bioactivity to PCL-based nanocomposite fibers and favor the precipitation of hydroxycarbonate apatite on the fiber surface. Preliminary cytocompatibility tests demonstrate that the addition of BG particles to PCL-based polymer does not inhibit ST-2 cell viability. This novel approach of using bioactive glasses not only for their biological properties, but also for the enhancement of shape memory properties of PCL-based polymers, widens the versatility and suitability of the obtained composite fibers for a huge portfolio of biomedical applications., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors.)

Silicone resins, filled with phosphates and other oxide fillers, yield upon firing in air at 1100 °C, a product resembling Biosilicate ® glass-ceramics, one of the most promising systems for tissue engineering applications. The process requires no preliminary synthesis of parent glass, and the polymer route enables the application of direct ink writing (DIW) of silicone-based mixtures, for the manufacturing of reticulated scaffolds at room temperature. The thermal treatment is later applied for the conversion into ceramic scaffolds. The present paper further elucidates the flexibility of the approach. Changes in the reference silicone and firing atmosphere (from air to nitrogen) were studied to obtain functional composite biomaterials featuring a carbon phase embedded in a Biosilicate ® -like matrix. The microstructure was further modified either through a controlled gas release at a low temperature, or by the revision of the adopted additive manufacturing technology (from DIW to digital light processing).

In recent years, mesoporous bioactive glass nanoparticles (MBGNPs) have generated great attention in biomedical applications. In this study, cerium and gallium doped MBGNPs were prepared by microemulsion assisted sol-gel method in the binary SiO 2 -CaO system. MBGNPs with spheroidal and pineal shaped morphology were obtained. Nitrogen sorption analysis elucidated the mesoporous structure of synthesized nanoparticles with high specific surface area. X-ray diffraction analysis confirmed the amorphous nature of the nanoparticles. The chemical compositions of all samples were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES), which revealed that the contents of cerium and gallium could be tailored by adjusting the concentrations of the precursors used for the synthesis. All MBGNPs exhibited in vitro bioactivity when immersed in simulated body fluid, except the particles doped with higher amounts than 1 mol% of cerium. MBGNPs showed antibacterial activity against S. aureus and E. coli without exhibiting cytotoxicity towards MG-63 osteoblast-like cells. Mentioned features of the obtained Ce and Ga-doped MBGNPs make them useful for multifunctional applications such as drug delivery carriers or bioactive fillers for bone tissue engineering applications., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

The aim of this study was to obtain hierarchical scaffolds combining 3D printing and two electrofluidodynamic methods. The multi-layered scaffold is composed by 3D printed struts, electrospun fibers obtained from poly( ϵ -caprolactone) and electrosprayed spheres produced from hydrophobically modified chitosan, namely chitosan grafted with linoleic acid (CHLA). Since CHLA has been used for the first time in the electrospraying (electro dynamic spraying, EDS) process, the formation of spheres needed an optimization process. The EDS process was strongly affected by the solvent mixture composition, concentration of acid used for CHLA dissolution and solution flow rate. By using the optimized electrospraying conditions, uniformly distributed spheres have been obtained, decorating struts and nanofibers. Preliminary biological tests with mouse preosteoblasts (MC3T3-E1) were performed to investigate the effect of the hierarchical scaffold on cell seeding efficacy. Results showed that the hierarchical structure enhances cell seeding efficacy, respect to the 3D printed struts alone, preventing that the cells passed through the struts during the seeding. Moreover, the addition of the electrosprayed nanoparticles does not affect the cell seeding efficiency. The versatility of the proposed structure, with the added value of CHLA nanoparticles decoration could be suitable for several applications in tissue engineering, mainly related to drug delivery systems., (© 2021 IOP Publishing Ltd.)

Wound dressings produced by electrospinning exhibit a fibrous structure close to the one of the extracellular matrix of the skin. In this article, electrospinning was used to fabricate fiber mats based on the well-known biopolymers poly(ɛ-caprolactone) (PCL) and methylcellulose (MC) using benign solvents. The blend fiber mats were cross-linked using Manuka honey and additionally used as a biodegradable platform to deliver bioactive glass particles. It was hypothesized that a dual therapeutic effect can be achieved by combining Manuka honey and bioactive glass. Morphological and chemical examinations confirmed the successful production of submicrometric PCL-MC fiber mats containing Manuka honey and bioactive glass particles. The multifunctional fiber mats exhibited improved wettability and suitable mechanical properties (ultimate tensile strength of 3-5 MPa). By performing dissolution tests using simulated body fluid, the improved bioactivity of the fiber mats by the addition of bioactive glass was confirmed. Additionally, cell biology tests using human dermal fibroblasts and human keratinocytes-like HaCaT cells showed the potential of the fabricated composite fiber mats to be used as wound dressing, specially due to the ability to support wound closure influenced by the presence of bioactive glass. Moreover, based on the results of the antibacterial tests, it is apparent that an optimization of the electrospun fiber mats is required to develop suitable wound dressing for the treatment of infected wounds., (© 2020 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals LLC.)

Although selenium nanoparticles (SeNPs) have gained attention in the scientific community mostly through investigation of their anticancer activity, a great potential of this nanomaterial was recognized recently regarding its antimicrobial activity. The particle form, size, and surface chemistry have been recognized as crucial parameters determining the interaction of nanomaterials with biological entities. Furthermore, considering a narrow boundary between beneficial and toxic effects for selenium per se, it is clear that investigations of biomedical applications of SeNPs are very demanding and must be done with great precautions. The goal of this work is to evaluate the effects of SeNPs surface chemistry and structure on antimicrobial activity against several common bacterial strains, including Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 29212), Bacillus subtilis (ATCC 6633), and Kocuria rhizophila (ATCC 9341), as well as Escherichia coli (ATCC 8739), Salmonella Abony (NCTC 6017), Klebsiella pneumoniae (NCIMB 9111) and Pseudomonas aeruginosa (ATCC 9027), and the standard yeast strain Candida albicans (ATCC 10231). Three types of SeNPs were synthesized by chemical reduction approach using different stabilizers and reducing agents: (i) bovine serum albumin (BSA) + ascorbic acid, (ii) chitosan + ascorbic acid, and (iii) with glucose. A thorough physicochemical characterization of the obtained SeNPs was performed to determine the effects of varying synthesis parameters on their morphology, size, structure, and surface chemistry. All SeNPs were amorphous, with spherical morphology and size in the range 70-300 nm. However, the SeNPs obtained under different synthesis conditions, i.e. by using different stabilizers as well as reducing agents, exhibited different antimicrobial activity as well as cytotoxicity which are crucial for their applications. In this paper, the antimicrobial screening of the selected systems is presented, which was determined by the broth microdilution method, and inhibitory influence on the production of monomicrobial and dual-species biofilm was evaluated. The potential mechanism of action of different systems is proposed. Additionally, the cytotoxicity of SeNPs was examined on the MRC-5 cell line, in the same concentration interval as for antimicrobial testing. It was shown that formulation SeNPs-BSA expressed a significantly lower cytotoxic effect than the other two formulations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Filipović, Ušjak, Milenković, Zheng, Liverani, Boccaccini and Stevanović.)

Mesoporous silica-based nanoparticles (MSNs) are considered promising drug carriers because of their ordered pore structure, which permits high drug loading and release capacity. The dissolution of Si and Ca from MSNs can trigger osteogenic differentiation of stem cells towards extracellular matrix calcification, while Mg and Sr constitute key elements of bone biology and metabolism. The aim of this study was the synthesis and characterization of sol-gel-derived MSNs co-doped with Ca, Mg and Sr. Their physico-chemical properties were investigated by X-ray diffraction (XRD), scanning electron microscopy with energy dispersive X-ray analysis (SEM/EDX), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray fluorescence spectroscopy (XRF), Brunauer Emmett Teller and Brunauer Joyner Halenda (BET/BJH), dynamic light scattering (DLS) and ζ-potential measurements. Moxifloxacin loading and release profiles were assessed with high performance liquid chromatography (HPLC) cell viability on human periodontal ligament fibroblasts and their hemolytic activity in contact with human red blood cells (RBCs) at various concentrations were also investigated. Doped MSNs generally retained their textural characteristics, while different compositions affected particle size, hemolytic activity and moxifloxacin loading/release profiles. All co-doped MSNs revealed the formation of hydroxycarbonate apatite on their surface after immersion in simulated body fluid (SBF) and promoted mitochondrial activity and cell proliferation.

Chitosan fibers blended with polyethylene oxide (CHIT&#95;PEO) and crosslinked with genipin were fabricated by electrospinning technique. Subsequently, CHIT&#95;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&#95;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&#95;2Zn). Particulate BGs (particles size < 20 μm) were separately added to the starting CHIT&#95;PEO solution before electrospinning. The two recently developed bioactive glasses (BGMS10 and BGMS&#95;2Zn) showed very promising biological properties in terms of bioactivity and cellular viability; thus, such compositions were added for the first time to CHIT&#95;PEO solution to fabricate composite electrospun mats. The incorporation of bioactive glass particles and their distribution into CHIT&#95;PEO fibers were assessed by SEM and FTIR analyses. Furthermore, CHIT&#95;PEO composite electrospun mats showed improved mechanical properties in terms of Young's Modulus compared to neat CHIT&#95;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&#95;PEO composite electrospun mats showed cytocompatibility and the desired cellular viability.

Metals are used in several orthopedic applications as fixation elements for the stabilization of fractures or as prostheses. One of the most common orthopedic biomaterials in many developing countries is surgical grade stainless steel (SS). However, its use as permanent implant in orthopedic surgery is conditioned due to its limited corrosion resistance in physiological media, lack of osseointegration, and absence of antibacterial effect. The aim of this work is to generate a degradable coating with antibacterial properties for stainless steel to be used in implants/medical devices. The coating is composed of a biopolymer/silica-gentamicin nanoparticles composite obtained by electrophoretic deposition (EPD) on surgical grade stainless steel plates. The coating surface was characterized by microscopic examination, and in vitro performance was evaluated after immersion in phosphate-buffered saline (PBS) solution, simulated body fluid (SBF), and cell culture medium, to analyze coating degradation, antibiotics release, cell attachment (ST-2 stromal cells), and antibacterial (Escherichia coli and Staphylococcus aureus) properties. EPD coatings were uniform and covered homogeneously the surface of the SS substrate. Also the distribution of silica-gentamicin nanoparticles was homogeneous on the coated area. The degradation of the chitosan-gelatin coatings was evident after 7 days of immersion. The gentamicin release led to excellent antibacterial behavior at 24 h, meanwhile the cell proliferation (at 7 days culture) was not inhibited. The results show that the coating system exhibits promising behavior which could contribute to prevent hospital infections at early implantation times., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Wound dressings able to deliver topically bioactive molecules represent a new generation of wound-regeneration therapies. In this article, foams based on methylcellulose cross-linked with Manuka honey were used as a platform to deliver borate bioactive glass particles doped additionally with copper. Borate bioactive glasses are of great interest in wound-healing applications due to a combination of favorable features, such as angiogenic and antibacterial properties. The multifunctional composite providing the dual effect of the bioactive glass and Manuka honey was produced by freeze-drying, and the resulting foams exhibit suitable morphology characterized by high porosity. Moreover, the performed tests showed improved wettability and mechanical performance with the addition of bioactive glass particles. Dissolution studies using simulated body fluid and cell biology tests using relevant skin cells further proved the excellent bioactivity and positive effects of the foams on cell proliferation and migration. Most interestingly, by the dual release of Manuka honey and ions from the copper-doped bioactive glass, an antibacterial effect against E. coli and S. aureus was achieved. Therefore, the multifunctional foams showed promising outcomes as potential wound dressings for the treatment of infected wounds.

Vascularization is necessary in tissue engineering to keep adequate blood supply in order to maintain the survival and growth of new tissue. The synergy of biologically active ions with multi-target activity may lead to superior angiogenesis promotion in comparison to single-target approaches but it has been rarely investigated. In this study, polycaprolactone (PCL) fiber mats embedded with B and Co co-doped bioactive glass nanoparticles (BCo.BGNs) were fabricated as a tissue regeneration scaffold designed for promoting angiogenesis. BCo.NBGs were successfully prepared with well-defined spherical shape using a sol-gel method. The PCL fiber mats embedding co-doped bioactive glass nanoparticles were fabricated by electrospinning using benign solvents. The Young's moduli of the nanoparticle containing PCL fiber mats were similar to those of the neat fiber mats and suitable for scaffolds utilized in soft tissue repair approaches. The mats also showed non-cytotoxicity to ST-2 cells. PCL fiber mats containing BCo.BGNs with a relatively high content of B and Co promoted the secretion of vascular endothelial growth factor to a greater extent than PCL fiber mats with a relatively low B and Co contents, which demonstrates the potential of dual ion release (B and Co) from bioactive glasses to enhance angiogenesis in soft tissue engineering.

Many elements, such as copper, cobalt (Co), strontium, magnesium and boron (B) have been used for single or multiple doping of bioactive glasses to promote angiogenesis during bone regeneration. However, few works have focused on promoting angiogenesis through stimulating several different signalling pathways which can be called a multi-target approach. In this study, 45S5 bioactive glass (BG) co-doped with B and Co was prepared by conventional melt quenching method. The synergistic effect of the two co-doping elements on angiogenesis promotion was expected. ATR-Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy and inductively coupled plasma-optical emission spectrometry were used to characterize the composition, element distribution and ion release of the samples, respectively. Results showed that the compositions of all samples were consistent with the design compositions and all elements were homogeneously distributed in the bulk glass. Different contents of B and Co led to different release rates of these elements. All samples showed bioactivity after immersion in simulated body fluid, regardless of the amount of doped B and Co. The 1% and 0.1% extracts of all samples did not show any cytotoxicity to MG-63 and ST-2 cells. Compared with single B or Co doping, the B and Co dual doped sample led to a stronger increase of the secretion of vascular endothelial growth factor from ST-2 cells, which supports and confirms the synergistic effect on angiogenesis of B and Co as co-doping elements in 45S5 BG., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

In the present study, we present a comparative analysis among the outputs of porcine follicle isolation using either mechanical technique alone or in combination with enzymes, proposing an optimized protocol useful for all further applications related to follicle in vitro growth and reproductive tissue engineering. The porcine follicles were isolated using mechanical technique alone (hand blender and scalpels) or in combination with collagenase or Liberase Dispase High (DH) at different doses applying different protocols. Finally, the number, morphology, and stage of isolated follicles were compared between the protocols. Moreover, the follicle viability (live/dead assay) and morphology (rhodamine phalloidin and 4',6-diamidino-2-phenylindole staining and scanning electron microscopy analysis) were evaluated after 10 days of culture. We found an optimum protocol for intact follicle isolation using the mechanical technique in combination with enzymes at a concentration of 0.5 mg/mL. However, the number of total isolated follicles and primordial follicles was significantly higher when collagenase was used compared to Liberase DH ( p  < 0.05), while Liberase DH could isolate a significantly higher percentage of preantral follicles. After 10 days of culture, the morphology and health status of follicles were statistically higher when Liberase DH was used in comparison with collagenase. Moreover, on the follicles extracted with Liberase DH, it was possible to observe theca cells covering part of the follicle surface. In conclusion, we demonstrated that the intact primary or secondary follicles could not be obtained using only mechanical methods, which led to the isolation of denuded oocytes and dramatically damaged follicles. We concluded that the collagenase-based follicle isolation could negatively affect the morphology and developmental potential of the follicles. Moreover, the incubation of ovarian cortex tissues with Liberase DH solution is an optimized protocol for porcine ovarian follicle isolation with developmental competence. Impact statement Isolation and in vitro maturation of follicles can pave the way for activities on reproductive tissue engineering (REPROTEN) and developing an artificial ovary. In this regard, the standardization and optimization of the extraction methods are pivotal for the design of experiment of follicle in vitro growth. In the present study, we provided a comparative analysis among the outputs of porcine follicle isolation using either mechanical technique alone or in combination with collagenase or Liberase DH, proposing an optimized protocol useful for all further applications related to follicles' in vitro growth and REPROTEN.

Poly(glycerol-sebacate) (PGS) and poly(epsilon caprolactone) (PCL) have been widely investigated for biomedical applications in combination with the electrospinning process. Among others, one advantage of this blend is its suitability to be processed with benign solvents for electrospinning. In this work, the suitability of PGS/PCL polymers for the fabrication of composite fibers incorporating bioactive glass (BG) particles was investigated. Composite electrospun fibers containing silicate or borosilicate glass particles (13-93 and 13-93BS, respectively) were obtained and characterized. Neat PCL and PCL composite electrospun fibers were used as control to investigate the possible effect of the presence of PGS and the influence of the bioactive glass particles. In fact, with the addition of PGS an increase in the average fiber diameter was observed, while in all the composite fibers, the presence of BG particles induced an increase in the fiber diameter distribution, without changing significantly the average fiber diameter. Results confirmed that the blended fibers are hydrophilic, while the addition of BG particles does not affect fiber wettability. Degradation test and acellular bioactivity test highlight the release of the BG particles from all composite fibers, relevant for all applications related to therapeutic ion release, i.e., wound healing. Because of weak interface between the incorporated BG particles and the polymeric fibers, mechanical properties were not improved in the composite fibers. Promising results were obtained from preliminary biological tests for potential use of the developed mats for soft tissue engineering applications.