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This study aimed to develop and characterize silk fibroin methacrylated/SilkMA electrospun scaffolds associated with metronidazole/MET to control infection in root-end resected periapical lesions while supporting bone regeneration. SilkMA-based formulations (10% w/v) incorporating MET (0-control; 5, 15, or 30% w/w) were electrospun into fibrous scaffolds and photocrosslinked. Scaffolds' morphology, chemical composition, swelling/degradation profiles, mechanical properties, cytocompatibility with alveolar bone-derived mesenchymal stem cells/aBMSCs and stem cells from apical papilla/SCAPs, anti-inflammatory potential, and antibacterial efficacy (direct contact assay against Aggregatibacter actinomycetemcomitans/Aa and Fusobacterium nucleatum/Fn; Aa biofilm model) were assessed. Statistical analysis was conducted using a significance level of 5%. Morphological analysis revealed that MET content influenced fiber diameters post-crosslinking, while the chemical composition analysis confirmed MET integration within the scaffolds. 30%MET-laden scaffolds demonstrated reduced swelling capacity compared to SilkMA/control scaffolds, while complete degradation was observed after 42 days for the formulated scaffolds. Mechanical testing indicated enhanced stiffness and tensile strength in 30%MET-laden scaffolds compared to SilkMA/control (p < 0.05). Cytocompatibility evaluations showed non-cytotoxic effects across all formulations for aBMSCs and SCAPs. Anti-inflammatory assays demonstrated decreased pro-inflammatory cytokine interleukin-6 synthesis by aBMSCs treated with SilkMA + MET30% and Escherichia coli LPS, comparable to negative control (p > 0.05). Antibacterial efficacy assays revealed significant inhibition of Aa and Fn, with 30%MET-laden scaffolds demonstrating biofilm inhibition against Aa (p < 0.05). These findings underscore the potential of SilkMA scaffolds laden with MET as a promising strategy for managing periapical lesions, offering enhanced structural support, antimicrobial properties, and biocompatibility crucial for effective tissue regeneration and infection control after endodontic surgery., Competing Interests: Declarations Conflict of interest The authors declare that they have no conflicts of interest., (© 2024. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Objective: Calcium silicate cements (CSCs) are often used in endodontics despite some limitations related to their physical properties and antibacterial efficacy. This study aimed to develop and demonstrate the viability of a series of CSCs that were produced by sol-gel method and further modified with mesoporous bioactive glass nanoparticles (MBGNs) and collagen, for endodontic therapy., Methods: Calcium silicate (CS) particles and MBGNs were synthesized by the sol-gel method, and their elemental, molecular, and physical microstructure was characterized. Three CSCs were developed by mixing the CS with distilled water (CS+H 2 O), 10 mg/mL collagen solution (CS+colH 2 O), and MBGNs (10 %) (CSmbgn+colH 2 O). The mixing (MT) and setting (ST) times of the CSCs were determined, while the setting reaction was monitored in real-time. Antibacterial efficacy against Enterococcus faecalis (E. faecalis) and regenerative potential on dental pulp stem cells (DPSCs) were also analyzed., Results: The CS+H 2 O displayed a ST comparable to commercial products, while CSmbgn+colH 2 O achieved the longest MT of 68 s and the shortest ST of 8 min. All the experimental CSCs inhibited the growth of E. faecalis. Additionally, compared to the control group, CSCs supported cell proliferation and spreading and mineralized matrix production, regardless of their composition., Significance: Tested CSCs presented potential as candidates for pulp therapy procedures. Future research should investigate the pulp regeneration mechanisms alongside rigorous antibacterial evaluations, preferably with multi-organism biofilms, executed over extended periods., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Assessing the biocompatibility of materials is crucial for ensuring the safety and well-being of patients by preventing undesirable, toxic, immune, or allergic reactions, and ensuring that materials remain functional over time without triggering adverse reactions. To ensure a comprehensive assessment, planning tests that carefully consider the intended application and potential exposure scenarios for selecting relevant assays, cell types, and testing parameters is essential. Moreover, characterizing the composition and properties of biomaterials allows for a more accurate understanding of test outcomes and the identification of factors contributing to cytotoxicity. Precise reporting of methodology and results facilitates research reproducibility and understanding of the findings by the scientific community, regulatory agencies, healthcare providers, and the general public., Aims: This article aims to provide an overview of the key concepts associated with evaluating the biocompatibility of biomaterials while also offering practical guidance on cellular principles, testing methodologies, and biological assays that can support in the planning, execution, and reporting of biocompatibility testing., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The reconstruction of craniomaxillofacial bone defects remains clinically challenging. To date, autogenous grafts are considered the gold standard but present critical drawbacks. These shortcomings have driven recent research on craniomaxillofacial bone reconstruction to focus on synthetic grafts with distinct materials and fabrication techniques. Among the various fabrication methods, additive manufacturing (AM) has shown significant clinical potential. AM technologies build three-dimensional (3D) objects with personalized geometry customizable from a computer-aided design. These layer-by-layer 3D biomaterial structures can support bone formation by guiding cell migration/proliferation, osteogenesis, and angiogenesis. Additionally, these structures can be engineered to degrade concomitantly with the new bone tissue formation, making them ideal as synthetic grafts. This review delves into the key advances of bioceramic grafts/scaffolds obtained by 3D printing for personalized craniomaxillofacial bone reconstruction. In this regard, clinically relevant topics such as ceramic-based biomaterials, graft/scaffold characteristics (macro/micro-features), material extrusion-based 3D printing, and the step-by-step workflow to engineer personalized bioceramic grafts are discussed. Importantly, in vitro models are highlighted in conjunction with a thorough examination of the signaling pathways reported when investigating these bioceramics and their effect on cellular response/behavior. Lastly, we summarize the clinical potential and translation opportunities of personalized bioceramics for craniomaxillofacial bone regeneration., (© 2024. The Author(s).)

Periodontal disease poses significant challenges to the long-term stability of oral health by destroying the supporting structures of teeth. Guided tissue regeneration techniques, particularly barrier membranes, enable local regeneration by providing an isolated, protected compartment for osseous wound healing while excluding epithelial tissue. Here, this study reports on a thermosensitive periodontal membrane (TSPM) technology designed to overcome the mechanical limitations of current membranes through a semi-interpenetrating network of high molecular weight poly(L-lactic acid) (PLLA) and in situ-polymerized mesh of poly(ε-caprolactone)diacrylate (PCL-DA), and poly lactide-co-glycolide diacrylate (PLGA-DA). An optimized composition allows facile reshaping at greater than 52 °C and rigid shape maintenance at physiological temperature. Its unique bilayer morphology is achieved through self-assembly and thermally-induced phase separation, resulting in distinct yet continuous smooth and nanofibrous compartments adequate for epithelial occlusion and regeneration. Incorporating PLGA-DA enhances the membrane's hydrophilicity and degradation properties, facilitating a more rapid and controlled degradation and therapeutic delivery. This study demonstrates its ability to promote local regeneration by serving as a barrier membrane and simultaneously as a scaffolding matrix in a rat orthotopic periodontal defect model. The TSPM outperformed a clinically available material (Epi-Guide) to facilitate robust alveolar bone and periodontal ligament regeneration at 4 and 8 weeks., (© 2024 The Author(s). Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

Barrier membranes (BM) for guided bone regeneration (GBR) aim to support the osteogenic healing process of a defined bony defect by excluding epithelial (gingival) ingrowth and enabling osteoprogenitor and stem cells to proliferate and differentiate into bone tissue. Currently, the most widely used membranes for these approaches are collagen-derived, and there is a discrepancy in defining the optimal collagen membrane in terms of biocompatibility, strength, and degradation rates. Motivated by these clinical observations, we designed a collagen-free membrane based on l-valine- co -l-phenylalanine-poly(ester urea) (PEU) copolymer via electrospinning. Degradation and mechanical properties of these membranes were performed on as-spun and water-aged samples. Alveolar-bone-derived stem cells (AvBMSCs) were seeded on the PEU BM to assess their cell compatibility and osteogenic characteristics, including cell viability, attachment/spreading, proliferation, and mineralized tissue-associated gene expression. In vivo , PEU BMs were subcutaneously implanted in rats to evaluate their potential to cause inflammatory responses and facilitate angiogenesis. Finally, critical-size calvarial defects and a periodontal model were used to assess the regenerative capacity of the electrospun PEU BM compared to clinically available Cytoflex synthetic membranes. PEU BM demonstrated equal biocompatibility to Cytoflex with superior mechanical performance in strength and elasticity. Additionally, after 14 days, PEU BM exhibited a higher expression of BGLAP/osteocalcin and superior in vivo performance-less inflammation and increased CD31 and VWF expression over time. When placed in critical-sized defects in the calvaria of rats, the PEU BM led to robust bone formation with high expression of osteogenesis and angiogenesis markers. Moreover, our membrane enhanced alveolar bone and cementum regeneration in an established periodontal model after 8 weeks. We demonstrate that the PEU BM exhibits favorable clinical properties, including mechanical stability, cytocompatibility, and facilitated bone formation in vitro and in vivo. This highlights its suitability for GBR in periodontal and craniofacial bone defects.

Background: Bibliometric analysis is a critical indicator of the influence and relevance of scientific papers, whilst also highlighting key contributors and gaps in knowledge in a scientific field., Objectives: To update and analyse the 100 most-cited papers in regenerative endodontics from 2019 to 2023., Methods: A search of the most-cited recent papers focusing on regenerative endodontics using journals included in the category, 'Dentistry, Oral Surgery & Medicine', in the Clarivate Web of Science database from 2019 to 2023 was performed. Three researchers conducted the study selection and data extraction. Data extraction included publication title and year, authors, number and mean number of citations, institution, country and continent, study design, journal title, keywords and research topic. Citation counts were also collected in Google Scholar and Scopus databases. Graphical bibliometric networks were created using VOSviewer software., Results: The number of citations of the 100 most-cited articles ranged from 6 to 85. Most were published in 2020 (n = 48), principally in the Journal of Endodontics (47%), followed by International Endodontic Journal (13%), Journal of Dental Research (6%) and Dental Materials (6%). Laboratory study was the most common study design amongst the included papers (n = 47), followed by narrative reviews (n = 17) and observational studies (n = 16). The most frequent first author on the top three most-cited papers was Hacer Aksel, whilst Adham A. Azim (n = 6; 89 citations) contributed most to the top 100 articles. The institution from which most articles originated was the University of Hong Kong (China) (n = 5; 81 citations), whereas the corresponding authors were predominantly from the United States of America (USA) (n = 31; 560 citations). The VOSviewer map of co-authorship demonstrated research collaborative clusters. 'Regenerative endodontics' and 'stem-cells' were the most employed keywords (37 and 36 occurrences respectively)., Discussion: The current study was designed not only to showcase the most influential papers in regenerative endodontics since 2019 but also to provide a better understanding of global research in this area over the last five years., Conclusions: This bibliometric analysis highlighted papers, authors, institutions and keywords in regenerative endodontics. The 100 most-cited papers primarily consisted of laboratory studies published in the USA, focusing on evaluating biomaterials and scaffold design strategies in contact with stem cells. Clinical studies and systematic reviews representing higher levels of scientific evidence are currently not the most influential in the regenerative endodontic field., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Objective: This study examined how range concentrations of Fibroblast Growth Factor-2 (FGF-2) influence the differentiation and activity of human-derived periodontal ligament (hPDLSCs) and alveolar bone-derived stem cells (haBMSCs)., Design: hPDLSCs and haBMSCs were cultured with varying concentrations of FGF-2 (0, 1, 2.5, 5, 10, 20 ng/mL) and monitored for osteogenic differentiation through alkaline phosphatase (ALP) activity and quantification of gene expression (qRT-PCR) for osteogenesis markers. Additionally, alizarin red staining and a hydroxyproline colorimetric assay evaluated and quantified osteogenic matrix mineralization and collagen deposition. Statistical analyses were performed using one-way ANOVA or two-way ANOVA for multiple comparisons between groups., Results: At low FGF-2 concentrations, hPDLSCs differentiated toward an osteogenic lineage, whereas higher concentrations of FGF-2 inhibited osteogenesis and promoted fibroblastic differentiation. The effect of FGF-2 at the lowest concentration tested (1 ng/mL) led to significantly higher ALP activity than osteogenically induced positive controls at early time points and equivalent RUNX2 expression at early and later time points. FGF-2 supplementation of haBMSC cultures was sufficient, at all concentrations, to increase ALP activity at an earlier time point. Mineralization of haBMSC cultures increased significantly within 5-20 ng/mL FGF-2 concentrations under basal growth media conditions (α-minimal essential medium supplemented with 15 % fetal bovine serum and 1 % penicillin/streptomycin)., Conclusions: FGF-2 has a dual capacity in promoting osteogenic and fibroblastic differentiation within hPDLSCs contingent upon the dosage and timing of administration, alongside supporting osteogenic differentiation in haBMSCs. These findings underscore the need for precision growth factors dosing when considering the design of biomaterials for periodontal regeneration., Competing Interests: Declaration of Competing Interest The authors declare that they have no financial or personal interests that compete with the outcomes presented in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objective: To fabricate and characterize an innovative gelatin methacryloyl/GelMA electrospun scaffold containing the citrus flavonoid naringenin/NA with osteogenic and anti-inflammatory properties., Methods: GelMA scaffolds (15 % w/v) containing 0/Control, 5, 10, or 20 % of NA w/w were obtained via electrospinning. The chemical composition, fiber morphology/diameter, swelling/degradation profile, and NA release were investigated. Cytotoxicity, cell proliferation, adhesion and spreading, total protein/TP production, alkaline phosphatase/ALP activity, osteogenic genes expression (OCN, OPN, RUNX2), and mineralized nodules deposition/MND with human alveolar bone-derived mesenchymal stem cells (aBMSCs) seeded on the scaffolds were assessed. Moreover, aBMSCs seeded on the scaffolds and stimulated with tumor necrosis factor-alpha/TNF-α were submitted to collagen, nitric oxide/NO, interleukin/IL-1α, and IL-6 production assessment. Data were analyzed using ANOVA and t-student/post-hoc tests (α = 5 %)., Results: NA-laden scaffolds presented increased fiber diameter, lower swelling capacity, and faster degradation profile over 28 days (p < 0.05). NA release was detected over time. Cell adhesion and spreading, and TP production were similar between GelMA and GelMA+NA5 % scaffolds, while cell proliferation, ALP activity, OCN/OPN/RUNX2 gene expression, and MND were higher for GelMA+NA5 % scaffolds (p < 0.05). Cells seeded on control scaffolds and TNF-α-stimulated presented higher levels of NO, IL-1α/IL-6, and lower levels of collagen (p < 0.05). In contrast, cells seeded on GelMA+NA5 % scaffolds showed downregulation of inflammatory markers and higher collagen synthesis (p < 0.05)., Significance: GelMA+NA5 % scaffold was cytocompatible, stimulated aBMSCs proliferation and differentiation, and downregulated inflammatory mediators' synthesis, suggesting its therapeutic effect as a multi-target bifunctional scaffold with osteogenic and anti-inflammatory properties for bone tissue engineering., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Calcium phosphate (CaP) scaffolds doping with therapeutic ions are one of the focuses of recent bone tissue engineering research. Among the therapeutic ions, strontium stands out for its role in bone remodeling. This work reports a simple method to produce Sr-doped 3D-printed CaP scaffolds, using Sr-doping to induce partial phase transformation from β-tricalcium phosphate (β-TCP) to hydroxyapatite (HA), resulting in a doped biphasic calcium phosphate (BCP) scaffold. Strontium carbonate (SrCO 3 ) was incorporated in the formulation of the 3D-printing ink, studying β-TCP:SrO mass ratios of 100:0, 95:5, and 90:10 (named as β-TCP, β-TCP/5-Sr, and β-TCP/10-Sr, respectively). Adding SrCO 3 in the 3D-printing ink led to a slight increase in viscosity but did not affect its printability, resulting in scaffolds with a high printing fidelity compared to the computational design. Interestingly, Sr was incorporated into the lattice structure of the scaffolds, forming hydroxyapatite (HA). No residual SrO or SrCO 3 were observed in the XRD patterns of any composition, and HA was the majority phase of the β-TCP/10-Sr scaffolds. The addition of Sr increased the compression strength of the scaffolds, with both β-TCP/5-Sr and β-TCP/10-Sr performing better than the β-TCP. Overall, β-TCP/5-Sr presented higher mineralized nodules and mechanical strength, while β-TCP scaffolds presented superior cell viability. The incorporation of SrCO 3 in the ink formulation is a viable method to obtain Sr-BCP scaffolds. Thus, this approach could be explored with other CaP scaffolds aiming to optimize their performance and the addition of alternative therapeutic ions., 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 Elsevier Ltd. All rights reserved.)

Management of skin injuries imposes a substantial financial burden on patients and hospitals, leading to diminished quality of life. Periostin (rhOSF), an extracellular matrix component, regulates cell function, including a proliferative healing phase, representing a key protein to promote wound healing. Despite its proven efficacy in vitro , there is a lack of scaffolds that facilitate the in situ delivery of rhOSF. In addition, there is a need for a scaffold to not only support cell growth, but also to resist the mechanical forces involved in wound healing. In this work, we synthesized rhOSF-loaded mesoporous nanoparticles (MSNs) and incorporated them into a cell-laden gelatin methacryloyl (GelMA) ink that was bioprinted into melt electrowritten poly(ε-caprolactone) (PCL) microfibrous (MF-PCL) meshes to develop mechanically competent constructs. Diffraction light scattering (DLS) analysis showed a narrow nanoparticle size distribution with an average size of 82.7 ± 13.2 nm. The rhOSF-loaded hydrogels showed a steady and controlled release of rhOSF over 16 days at a daily dose of ∼40 ng/mL. Compared with blank MSNs, the incorporation of rhOSF markedly augmented cell proliferation, underscoring its contribution to cellular performance. Our findings suggest a promising approach to address challenges such as prolonged healing, offering a potential solution for developing robust, biocompatible, and cell-laden grafts for burn wound healing applications.

Introduction: Recognizing the necessity of novel disinfection strategies for improved bacterial control to ultimately favor tissue regeneration, this study developed and characterized antibiotics-laden silk fibroin methacrylated (SilkMA) scaffolds for regenerative endodontics., Methods: SilkMA-based solutions (10% w/v) containing clindamycin (CLI) or tinidazole (TIN) (0 - control; 5, 10, or 15% w/w) or the combination of both drugs (BiMix CLI/TIN 10%) were electrospun and photocrosslinked. Morphology and composition were assessed using scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). Additionally, swelling and degradation profiles were also determined. Cytotoxicity was evaluated in stem cells from apical papilla (SCAPs). Antibacterial efficacy was tested using direct and indirect contact assays against Aggregatibacter actinomycetemcomitans/Aa, Actinomyces naeslundii/An, Enterococcus faecalis/Ef, and Fusobacterium nucleatum/Fn. E. faecalis biofilm inhibition on dentin discs was specifically evaluated for BiMix-laden scaffolds. Data were statistically analyzed with a significance level of 5%., Results: SEM revealed that all scaffolds had similar characteristics, including fiber morphology and bead absence. FTIR showed the incorporation of CLI and TIN into the fibers and in BiMix scaffolds. Antibiotic-laden scaffolds exhibited lower swelling capacity than the control and were degraded entirely after 45 days. Scaffolds laden with CLI, TIN, or BiMix throughout all time points did not reduce SCAPs' viability. CLI-laden scaffolds inhibited the growth of Aa, An, and Ef, while TIN-laden scaffolds inhibited Fn growth. BiMix-laden scaffolds significantly inhibited Aa, An, Ef, and Fn in direct contact, and their aliquots inhibited An and Fn through indirect contact, with additional biofilm inhibition against Ef., Conclusions: BiMix-laden SilkMA scaffolds are cytocompatible and exhibit antimicrobial effects against endodontic pathogens, indicating their therapeutic potential as a drug delivery system for regenerative endodontics., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objectives: To synthesize casein enzymatic hydrolysate (CEH)-laden gelatin methacryloyl (GelMA) fibrous scaffolds and evaluate the cytocompatibility and anti-inflammatory effects on dental pulp stem cells (DPSCs)., Materials and Methods: GelMA fibrous scaffolds with 10%, 20%, and 30% CEH (w/w) and without CEH (control) were obtained via electrospinning. Chemo-morphological, degradation, and mechanical analyses were conducted to evaluate the morphology and composition of the fibers, mass loss, and mechanical properties, respectively. Adhesion/spreading and viability of DPSCs seeded on the scaffolds were also assessed. The anti-inflammatory potential on DPSCs was tested after the chronic challenge of cells with lipopolysaccharides (LPS), followed by treatment with extracts obtained after immersing the scaffolds in α-MEM. The synthesis of the pro-inflammatory cytokines IL-6, IL-1α, and TNF-α was measured by ELISA. Data were analyzed by ANOVA/post-hoc tests (α = 5%)., Results: CEH-laden electrospun fibers had a larger diameter than pure GelMA (p ≤ 0.036). GelMA scaffolds laden with 20% and 30% CEH had a greater mass loss. Tensile strength was reduced for the 10% CEH fibers (p = 0.0052), whereas no difference was observed for the 20% and 30% fibers (p ≥ 0.6736) compared to the control. Young's modulus decreased with CEH (p < 0.0001). Elongation at break increased for the 20% and 30% CEH scaffolds (p ≤ 0.0038). Over time, DPSCs viability increased across all groups, indicating cytocompatibility, with CEH-laden scaffolds exhibiting greater cell viability after seven days (p ≤ 0.0166). Also, 10% CEH-GelMA scaffolds decreased the IL-6, IL-1α, and TNF-α synthesis (p ≤ 0.035)., Conclusion: CEH-laden GelMA scaffolds facilitated both adhesion and proliferation of DPSCs, and 10% CEH provided anti-inflammatory potential after chronic LPS challenge., Clinical Relevance: CEH incorporated in GelMA fibrous scaffolds demonstrated the potential to be used as a cytocompatible and anti-inflammatory biomaterial for vital pulp therapy., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study investigated gelatin methacryloyl (GelMA) and polycaprolactone (PCL) blend scaffolds incorporating cerium oxide (CeO) nanoparticles at concentrations of 0%, 5%, and 10% w/w via electrospinning for periodontal tissue engineering. The impact of photocrosslinking on these scaffolds was evaluated by comparing crosslinked (C) and non-crosslinked (NC) versions. Methods included Fourier transform infrared spectroscopy (FTIR) for chemical analysis, scanning electron microscopy (SEM) for fiber morphology/diameters, and assessments of swelling capacity, degradation profile, and biomechanical properties. Biological evaluations with alveolar bone-derived mesenchymal stem cells (aBMSCs) and human gingival fibroblasts (HGFs) encompassed tests for cell viability, mineralized nodule deposition (MND), and collagen production (CP). Statistical analysis was performed using Kruskal-Wallis or ANOVA/post-hoc tests (α = 5%). Results indicate that C scaffolds had larger fiber diameters (~250 nm) compared with NC scaffolds (~150 nm). NC scaffolds exhibited higher swelling capacities than C scaffolds, while both types demonstrated significant mass loss (~50%) after 60 days ( p < 0.05). C scaffolds containing CeO showed increased Young's modulus and tensile strength than NC scaffolds. Cells cultured on C scaffolds with 10% CeO exhibited significantly higher metabolic activity (>400%, p < 0.05) after 7 days among all groups. Furthermore, CeO-containing scaffolds promoted enhanced MND by aBMSCs (>120%, p < 0.05) and increased CP in 5% CeO scaffolds for both variants (>180%, p < 0.05). These findings underscore the promising biomechanical properties, biodegradability, cytocompatibility, and enhanced tissue regenerative potential of CeO-loaded GelMA/PCL scaffolds for periodontal applications.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofibers embedded with borate glasses of 45B5 composition doped with Co 2+ , Cu 2+ , and Zn 2 + (46.1 B₂O₃26.9-X CaO24.4 Na₂O2.6 P₂O₅, X CoO/CuO/ZnO mol % (X = 0-5)) were produced by electrospinning for wound healing applications. Prior to their addition, the glasses exhibited two broad halos typical of a vitreous borate network, which were mainly composed of ring-type metaborate structural units. The particle distribution in the PHBV nanofibers embedded with 45B5 borate bioactive glasses is present in isolated and agglomerated states, being partially coated by a polymeric layer-except for the cobalt-doped glass, which resulted in a successful encapsulation with 100% embedding efficiency. The incorporation of the glasses reduced the PHBV crystallinity degree and its decomposition temperature, as well as its mechanical properties, including Young's modulus, tensile strength, and elongation at break. The neat PHBV fibers and those containing the cobalt-doped glasses demonstrated great cytocompatibility with human keratinocytes (HaCat), as suggested by the high cell viability after 7 days of exposure. Further studies are needed to fully understand the wound healing potential of these fibers, but our results significantly contribute to the area., (© 2024 Wiley Periodicals LLC.)

Apical periodontitis (AP) is a dental-driven condition caused by pathogens and their toxins infecting the inner portion of the tooth (i.e., dental pulp tissue), resulting in inflammation and apical bone resorption affecting 50% of the worldwide population, with more than 15 million root canals performed annually in the United States. Current treatment involves cleaning and decontaminating the infected tissue with chemo-mechanical approaches and materials introduced years ago, such as calcium hydroxide, zinc oxide-eugenol, or even formalin products. Here, we present, for the first time, a nanotherapeutics based on using synthetic high-density lipoprotein (sHDL) as an innovative and safe strategy to manage dental bone inflammation. sHDL application in concentrations ranging from 25 µg to 100 µg/mL decreases nuclear factor Kappa B (NF-κB) activation promoted by an inflammatory stimulus (lipopolysaccharide, LPS). Moreover, sHDL at 500 µg/mL concentration markedly decreases in vitro osteoclastogenesis (P < 0.001), and inhibits IL-1α (P = 0.027), TNF-α (P = 0.004), and IL-6 (P < 0.001) production in an inflammatory state. Notably, sHDL strongly dampens the Toll-Like Receptor signaling pathway facing LPS stimulation, mainly by downregulating at least 3-fold the pro-inflammatory genes, such as Il1b, Il1a, Il6, Ptgs2, and Tnf. In vivo, the lipoprotein nanoparticle applied after NaOCl reduced bone resorption volume to (1.3 ± 0.05) mm 3 and attenuated the inflammatory reaction after treatment to (1 090 ± 184) cells compared to non-treated animals that had (2.9 ± 0.6) mm 3 (P = 0.012 3) and (2 443 ± 931) cells (P = 0.004), thus highlighting its promising clinical potential as an alternative therapeutic for managing dental bone inflammation., (© 2024. The Author(s).)

Gingival recession, a prevalent condition affecting the gum tissues, is characterized by the exposure of tooth root surfaces due to the displacement of the gingival margin. This review explores conventional treatments, highlighting their limitations and the quest for innovative alternatives. Importantly, it emphasizes the critical considerations in gingival tissue engineering leveraging on cells, biomaterials, and signaling factors. Successful tissue-engineered gingival constructs hinge on strategic choices such as cell sources, scaffold design, mechanical properties, and growth factor delivery. Unveiling advancements in recent biofabrication technologies like 3D bioprinting, electrospinning, and microfluidic organ-on-chip systems, this review elucidates their precise control over cell arrangement, biomaterials, and signaling cues. These technologies empower the recapitulation of microphysiological features, enabling the development of gingival constructs that closely emulate the anatomical, physiological, and functional characteristics of native gingival tissues. The review explores diverse engineering strategies aiming at the biofabrication of realistic tissue-engineered gingival grafts. Further, the parallels between the skin and gingival tissues are highlighted, exploring the potential transfer of biofabrication approaches from skin tissue regeneration to gingival tissue engineering. To conclude, the exploration of innovative biofabrication technologies for gingival tissues and inspiration drawn from skin tissue engineering look forward to a transformative era in regenerative dentistry with improved clinical outcomes., (© 2024 The Authors. Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

The field of tissue engineering has witnessed significant advancements in recent years, driven by the pursuit of innovative solutions to address the challenges of bone regeneration. In this study, we developed an electrospun composite scaffold for bone tissue engineering. The composite scaffold is made of a blend of poly(L-lactide-co-ε-caprolactone) (PLCL) and polyethylene glycol (PEG), with the incorporation of calcined and lyophilized silicate-chlorinated bioactive glass (BG) particles. Our investigation involved a comprehensive characterization of the scaffold's physical, chemical, and mechanical properties, alongside an evaluation of its biological efficacy employing alveolar bone-derived mesenchymal stem cells. The incorporation of PEG and BG resulted in elevated swelling ratios, consequently enhancing hydrophilicity. Thermal gravimetric analysis confirmed the efficient incorporation of BG, with the scaffolds demonstrating thermal stability up to 250°C. Mechanical testing revealed enhanced tensile strength and Young's modulus in the presence of BG; however, the elongation at break decreased. Cell viability assays demonstrated improved cytocompatibility, especially in the PLCL/PEG+BG group. Alizarin red staining indicated enhanced osteoinductive potential, and fluorescence analysis confirmed increased cell adhesion in the PLCL/PEG+BG group. Our findings suggest that the PLCL/PEG/BG composite scaffold holds promise as an advanced biomaterial for bone tissue engineering., (© 2024 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals LLC.)

This study aimed to develop gelatin methacryloyl (GelMA)-injectable hydrogels incorporated with 58S bioactive glass/BG-doped with strontium for vital pulp therapy applications. GelMA hydrogels containing 0% (control), 5%, 10%, and 20% BG ( w / v ) were prepared. Their morphological and chemical properties were evaluated by scanning electron microscopy/SEM, energy dispersive spectroscopy/EDS, and Fourier transform infrared spectroscopy/FTIR (n = 3). Their swelling capacity and degradation ratio were also measured (n = 4). Cell viability (n = 8), mineralized matrix formation, cell adhesion, and spreading (n = 6) on DPSCs were evaluated. Data were analyzed using ANOVA/post hoc tests (α = 5%). SEM and EDS characterization confirmed the incorporation of BG particles into the hydrogel matrix, showing GelMA's (C, O) and BG's (Si, Cl, Na, Sr) chemical elements. FTIR revealed the main chemical groups of GelMA and BG, as ~1000 cm -1 corresponds to Si-O and ~1440 cm -1 to C-H. All the formulations were degraded by day 12, with a lower degradation ratio observed for GelMA+BG20%. Increasing the concentration of BG resulted in a lower mass swelling ratio. Biologically, all the groups were compatible with cells ( p > 0.6196), and cell adhesion increased over time, irrespective of BG concentration, indicating great biocompatibility. GelMA+BG5% demonstrated a higher deposition of mineral nodules over 21 days ( p < 0.0001), evidencing the osteogenic potential of hydrogels. GelMA hydrogels incorporated with BG present great cytocompatibility, support cell adhesion, and have a clinically relevant degradation profile and suitable mineralization potential, supporting their therapeutic potential as promising biomaterials for pulp capping.

This study investigated the incorporation of sources of calcium, phosphate, or both into electrospun scaffolds and evaluated their bioactivity on human dental pulp cells (HDPCs). Additionally, scaffolds incorporated with calcium hydroxide (CH) were characterized for degradation, calcium release, and odontogenic differentiation by HDPCs. Polycaprolactone (PCL) was electrospun with or without 0.5% w / v of calcium hydroxide (PCL + CH), nano-hydroxyapatite (PCL + nHA), or β-glycerophosphate (PCL + βGP). SEM/EDS analysis confirmed fibrillar morphology and particle incorporation. HDPCs were cultured on the scaffolds to assess cell viability, adhesion, spreading, and mineralized matrix formation. PCL + CH was also evaluated for gene expression of odontogenic markers (RT-qPCR). Data were submitted to ANOVA and Student's t -test (α = 5%). Added CH increased fiber diameter and interfibrillar spacing, whereas βGP decreased both. PCL + CH and PCL + nHA improved HDPC viability, adhesion, and proliferation. Mineralization was increased eightfold with PCL + CH. Scaffolds containing CH gradually degraded over six months, with calcium release within the first 140 days. CH incorporation upregulated DSPP and DMP1 expression after 7 and 14 days. In conclusion, CH- and nHA-laden PCL fiber scaffolds were cytocompatible and promoted HDPC adhesion, proliferation, and mineralized matrix deposition. PCL + CH scaffolds exhibit a slow degradation profile, providing sustained calcium release and stimulating HDPCs to upregulate odontogenesis marker genes.

Objectives: Customization and the production of patient-specific devices, tailoring the unique anatomy of each patient's jaw and facial structures, are the new frontiers in dentistry and maxillofacial surgery. As a technological advancement, additive manufacturing has been applied to produce customized objects based on 3D computerized models. Therefore, this paper presents advances in additive manufacturing strategies for patient-specific devices in diverse dental specialties., Methods: This paper overviews current 3D printing techniques to fabricate dental and maxillofacial devices. Then, the most recent literature (2018-2023) available in scientific databases reporting advances in 3D-printed patient-specific devices for dental and maxillofacial applications is critically discussed, focusing on the major outcomes, material-related details, and potential clinical advantages., Results: The recent application of 3D-printed customized devices in oral prosthodontics, implantology and maxillofacial surgery, periodontics, orthodontics, and endodontics are presented. Moreover, the potential application of 4D printing as an advanced manufacturing technology and the challenges and future perspectives for additive manufacturing in the dental and maxillofacial area are reported., Significance: Additive manufacturing techniques have been designed to benefit several areas of dentistry, and the technologies, materials, and devices continue to be optimized. Image-based and accurately printed patient-specific devices to replace, repair, and regenerate dental and maxillofacial structures hold significant potential to maximize the standard of care in dentistry., 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 Inc. All rights reserved.)

The quest for finding a suitable scaffold system that supports cell survival and function and, ultimately, the regeneration of the pulp-dentin complex remains challenging. Herein, we hypothesized that dental pulp stem cells (DPSCs) encapsulated in a collagen-based hydrogel with varying stiffness would regenerate functional dental pulp and dentin when concentrically injected into the tooth slices. Collagen hydrogels with concentrations of 3 mg/mL (Col3) and 10 mg/mL (Col10) were prepared, and their stiffness and microstructure were assessed using a rheometer and scanning electron microscopy, respectively. DPSCs were then encapsulated in the hydrogels, and their viability and differentiation capacity toward endothelial and odontogenic lineages were evaluated using live/dead assay and quantitative real-time polymerase chain reaction. For in vivo experiments, DPSC-encapsulated collagen hydrogels with different stiffness, with or without growth factors, were injected into pulp chambers of dentin tooth slices and implanted subcutaneously in severe combined immunodeficient (SCID) mice. Specifically, vascular endothelial growth factor (VEGF [50 ng/mL]) was loaded into Col3 and bone morphogenetic protein (BMP2 [50 ng/mL]) into Col10. Pulp-dentin regeneration was evaluated by histological and immunofluorescence staining. Data were analyzed using 1-way or 2-way analysis of variance accordingly (α = 0.05). Rheology and microscopy data revealed that Col10 had a stiffness of 8,142 Pa with a more condensed and less porous structure, whereas Col3 had a stiffness of 735 Pa with a loose microstructure. Furthermore, both Col3 and Col10 supported DPSCs' survival. Quantitative polymerase chain reaction showed Col3 promoted significantly higher von Willebrand factor (VWF) and CD31 expression after 7 and 14 d under endothelial differentiation conditions ( P < 0.05), whereas Col10 enhanced the expression of dentin sialophosphoprotein (DSPP), alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), and collagen 1 (Col1) after 7, 14, and 21 d of odontogenic differentiation ( P < 0.05). Hematoxylin and eosin and immunofluorescence (CD31 and vWF) staining revealed Col10+Col3+DPSCs+GFs enhanced pulp-dentin tissue regeneration. In conclusion, the collagen-based concentric construct modified by growth factors guided the specific lineage differentiation of DPSCs and promoted pulp-dentin tissue regeneration in vivo., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article. Data AvailabilityAll research data supporting this publication are directly available within this manuscript and supplementary Appendix.

This study aimed to evaluate the effect of dentin hypersensitivity treatments on immediate and long-term shear bond strength (SBS) of composite restorations. Ninety non-carious extracted human molars were cut to expose dentin, which was embedded in acrylic resin, and randomly divided into three groups (n = 30/group) according to surface treatment: 1) no treatment (C and C*; control); 2) silver diamine fluoride with potassium iodide (SDF/KI and SDF/KI*; Riva Star); and 3) nano-hydroxyapatite (nHAp and nHAp*; PrevDent). The specimens were etched through the etch-and-rinse technique, followed by universal adhesive application and resin composite cylinders (2.38 mm in diameter × 3.5 mm high). The SBS was tested immediately (24 h after the restoration) and after thermocycling (*) (5000 cycles, 5 °C to 55 °C) at a 0.5 mm/min crosshead speed using a universal testing machine. A stereomicroscope was used to evaluate the mode of failure, and representative scanning electron microscopy (SEM) images were also acquired. Data normality was verified, and two-way ANOVA and Tukey's post hoc tests were performed for multiple comparisons (α = 0.05). The control group presented the highest SBS (27.10 MPa), while SDF/KI* had the lowest values (6.87 MPa). nHAp-based desensitizer exhibited higher SBS than SDF/KI for both immediate (22.6 MPa) and thermocycled (19.03 MPa) conditions. No intragroup difference was evidenced between immediate and thermocycled samples for any group. Most specimens for the C and nHAp groups presented mixed failure, while the SDF/KI groups presented comparable adhesive and mixed failures. The SBS of adhesive restorations after the application of desensitizing agents is material dependent, where SDF/KI reduces SBS values below the acceptable minimum bond strength, while the nHAp application meets the minimally required bond strength., (© 2023. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Introduction: Citric acid (CA) conditioning may be a promising alternative to ethylenediaminetetraacetic acid (EDTA) in regenerative endodontic procedures, as reported to improve growth factors' release from dentin. This review systematically investigated the effect of CA conditioning on the growth factors release from dentin and cell behavior compared to EDTA conditioning., Methods: Searches were conducted (PubMed/MEDLINE, Scopus, Web of Science, Embase, SciELO, Cochrane Library, and grey literature) until May-2023. Only in vitro studies that evaluated the effects of CA on growth factors' release from dentin and cell behavior outcomes compared to EDTA were included. The studies were critically appraised using a modified Joanna Briggs Institute's checklist. Meta-analysis was unfeasible., Results: Out of the 335 articles screened, nine were included. Among these, three studies used dentin discs/roots from permanent human teeth; the rest combined them with stem cells. 10% CA for 5 or 10 minute was the most used protocol. Meanwhile, EDTA concentrations ranged from 10% to 17%. In eight studies examining the release of growth factors, five reported a significant release of transforming growth factor-β after dentin conditioning with 10% CA compared to 17% EDTA. Regarding cell behavior (6 studies), three studies assessed cell viability. The findings revealed that 10% CA conditioning showed cell viability similar to those of 17% EDTA. Additionally, in two out of three studies, it was observed that 10% CA conditioning did not affect cell morphology. The studies had a low risk of bias., Conclusions: The use of 10% CA to condition dentin for 5-10 minutes resulted in a notable transforming growth factor -β1 release, but its cell responses were similar to those of EDTA., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Pulp capping is a necessary procedure for preserving the vitality and health of the dental pulp, playing a crucial role in preventing the need for root canal treatment or tooth extraction. Here, we developed an electrospun gelatin methacryloyl (GelMA) fibrous scaffold incorporating beta-tricalcium phosphate (TCP) particles for pulp capping. A comprehensive morphological, physical-chemical, and mechanical characterization of the engineered fibrous scaffolds was performed. In vitro bioactivity, cell compatibility, and odontogenic differentiation potential of the scaffolds in dental pulp stem cells (DPSCs) were also evaluated. A pre-clinical in vivo model was used to determine the therapeutic role of the GelMA/TCP scaffolds in promoting hard tissue formation. Morphological, chemical, and thermal analyses confirmed effective TCP incorporation in the GelMA nanofibers. The GelMA+20%TCP nanofibrous scaffold exhibited bead-free morphology and suitable mechanical and degradation properties. In vitro, GelMA+20%TCP scaffolds supported apatite-like formation, improved cell spreading, and increased deposition of mineralization nodules. Gene expression analysis revealed upregulation of ALPL, RUNX2, COL1A1, and DMP1 in the presence of TCP-laden scaffolds. In vivo, analyses showed mild inflammatory reaction upon scaffolds' contact while supporting mineralized tissue formation. Although the levels of Nestin and DMP1 proteins did not exceed those associated with the clinical reference treatment (i.e., mineral trioxide aggregate), the GelMA+20%TCP scaffold exhibited comparable levels, thus suggesting the emergence of differentiated odontoblast-like cells capable of dentin matrix secretion. Our innovative GelMA/TCP scaffold represents a simplified and efficient alternative to conventional pulp-capping biomaterials. STATEMENT OF SIGNIFICANCE: Vital pulp therapy (VPT) aims to preserve dental pulp vitality and avoid root canal treatment. Biomaterials that bolster mineralized tissue regeneration with ease of use are still lacking. We successfully engineered gelatin methacryloyl (GelMA) electrospun scaffolds incorporated with beta-tricalcium phosphate (TCP) for VPT. Notably, electrospun GelMA-based scaffolds containing 20% (w/v) of TCP exhibited favorable mechanical properties and degradation, cytocompatibility, and mineralization potential indicated by apatite-like structures in vitro and mineralized tissue deposition in vivo, although not surpassing those associated with the standard of care. Collectively, our innovative GelMA/TCP scaffold represents a simplified alternative to conventional pulp capping materials such as MTA and Biodentine™ since it is a ready-to-use biomaterial, requires no setting time, and is therapeutically effective., 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 © 2023 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The evolutionary establishment of an internal biological clock is a primordial event tightly associated with a 24-h period. Changes in the circadian rhythm can affect cellular functions, including proliferation, DNA repair and redox state. Even isolated organs, tissues and cells can maintain an autonomous circadian rhythm. These cell-autonomous molecular mechanisms are driven by intracellular clock genes, such as BMAL1. Little is known about the role of core clock genes and epigenetic modifications in the skin. Our focus was to identify BMAL1-driven epigenetic modifications associated with gene transcription by mapping the acetylation landscape of histones in epithelial cells responding to injury. We explored the role of BMAL1 in epidermal wound and tissue regeneration using a loss-of-function approach in vivo. We worked with BMAL1 knockout mice and a contraction-resistance wound healing protocol, determining the histone modifications using specific antibodies to detect the acetylation levels of histones H3 and H4. We found significant differences in the acetylation levels of histones in both homeostatic and injured skin with deregulated BMAL1. The intact skin displayed varied acetylation levels of histones H3 and H4, including hyperacetylation of H3 Lys 9 (H3K9). The most pronounced changes were observed at the repair site, with notable alterations in the acetylation pattern of histone H4. These findings reveal the importance of histone modifications in response to injury and indicate that modulation of BMAL1 and its associated epigenetic events could be therapeutically harnessed to improve skin regeneration., (© 2024 The Authors. Experimental Dermatology published by John Wiley & Sons Ltd.)

Keeping sodium hypochlorite (NaOCl) within the root canal is challenging in regenerative endodontics. In this study, we developed a drug delivery system using a gelatin methacryloyl (GelMA) hydrogel incorporated with aluminosilicate clay nanotubes (HNTs) loaded with NaOCl. Pure GelMA, pure HNTs, and NaOCl-loaded HNTs carrying varying concentrations were assessed for chemo-mechanical properties, degradability, swelling capacity, cytocompatibility, antimicrobial and antibiofilm activities, and in vivo for inflammatory response and degradation. SEM images revealed consistent pore sizes of 70-80 µm for all samples, irrespective of the HNT and NaOCl concentration, while HNT-loaded hydrogels exhibited rougher surfaces. The hydrogel's compressive modulus remained between 100 and 200 kPa, with no significant variations. All hydrogels demonstrated a 6-7-fold mass increase and complete degradation by the seventh day. Despite an initial decrease in cell viability, all groups recovered to 65-80% compared to the control. Regarding antibacterial and antibiofilm properties, 12.5 HNT(Double) showed the highest inhibition zone on agar plates and the most significant reduction in biofilm compared to other groups. In vivo, the 12.5 HNT(Double) group displayed partial degradation after 21 days, with mild localized inflammatory responses but no tissue necrosis. In conclusion, the HNT-NaOCl-loaded GelMA hydrogel retains the disinfectant properties, providing a safer option for endodontic procedures without harmful potential.

Objectives: The goal of this systematic review was to analyze, in randomized controlled clinical trials (RCTs), regenerative techniques used to treat peri -implantitis (PI)., Methods: Three databases (PubMed/Medline, EMBASE, and On-Line Knowledge Library) were accessed, applying the PICO strategy (Population [P], Intervention [I], Comparison [C], and Outcomes [O]), with the following focused questions: (i) "In patients who received regenerative treatments for peri -implantitis (P), is the regenerative surgical treatment (I) clinically effective and predictable compared to non-regenerative (C) to treat PI (O)?"; and (ii) "In patients who received regenerative treatments for peri -implantitis (P), the regenerative approach (I), compared to non-regenerative (C), significantly increase the prognosis and implant survival rate in the mid- and long-term (O)?" The inclusion criteria were RCTs published in English between 2012 and 2022, with at least a one-year follow-up, which applied regenerative techniques to treat peri -implantitis. Cochrane's collaboration tool for assessing the risk of bias was used., Main Results: Nine articles were included with 404 patients (225 females and 179 males; mean age of 60.44 years). One study evaluated patients after 48 months and another after 88 months. The techniques and devices used were: (i) implantoplasty with Er:YAG laser, (ii) blood concentrate (growth factors), and (iii) EMD, with no statistically significant outcome. Two studies considered the use of titanium granules with a significant increase in radiographic bone identification, whereas regenerative techniques with bone graft (autogenous, alloplastic, and xenograft) were the majority chosen, associated or not, with a collagen membrane. Xenograft had better results radiographically when compared to the autogenous bone graft and presented better results for bone level. There was an overall decrease in bleeding on probing, independent of the control or test group, and a reduction in pocket depth in the groups analyzed. Titanium granules, EMD, Er:YAG laser, and CGF had non-significant results; better results were observed when using bone grafts. The RoB showed a low risk in four studies (44.44%), three with moderate (33.33%), and two with high risk (22.23%)., Conclusion: Surgical regenerative treatment was a predictable option in the management of PI and in improving the clinical parameters of peri -implant tissues in the short term, mainly when using porous titanium granules, alloplastic bone grafts, and xenografts., Competing Interests: 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., (© 2023 King Saud University.)

Major advances in the field of periodontal tissue engineering have favored the fabrication of biodegradable membranes with tunable physical and biological properties for guided bone regeneration (GBR). Herein, we engineered innovative nanoscale beta-tricalcium phosphate (β-TCP)-laden gelatin methacryloyl/polycaprolactone (GelMA/PCL-TCP) photocrosslinkable composite fibrous membranes via electrospinning. Chemo-morphological findings showed that the composite microfibers had a uniform porous network and β-TCP particles successfully integrated within the fibers. Compared with pure PCL and GelMA/PCL, GelMA/PCL-TCP membranes led to increased cell attachment, proliferation, mineralization, and osteogenic gene expression in alveolar bone-derived mesenchymal stem cells (aBMSCs). Moreover, our GelMA/PCL-TCP membrane was able to promote robust bone regeneration in rat calvarial critical-size defects, showing remarkable osteogenesis compared to PCL and GelMA/PCL groups. Altogether, the GelMA/PCL-TCP composite fibrous membrane promoted osteogenic differentiation of aBMSCs in vitro and pronounced bone formation in vivo. Our data confirmed that the electrospun GelMA/PCL-TCP composite has a strong potential as a promising membrane for guided bone regeneration.

Objectives: To evaluate the incorporation of halloysite nanotubes (HNTs) loaded with one of two calcium sources (i.e., calcium hydroxide/CaOH 2 or beta-tricalcium phosphate/β-TCP) on the physicochemical and biological properties of an experimental resin-based dual-cured endodontic sealer., Materials and Methods: HNTs were encapsulated with CaOH 2 or β-TCP at 10 wt.%. HNTs containing CaOH 2 or β-TCP were added into the experimental sealers at 50 wt.%. The control sealers were the calcium-free HNT-modified resin-based experimental sealer and AH Plus™, a commercially available endodontic sealer. Degree of conversion, setting time, flow, film thickness, radiopacity, dimensional stability, and calcium ions release were determined. Antibiofilm properties and cytocompatibility of the formulated sealers and commercial control were also evaluated. One and two-way ANOVA analysis followed by Tukey's post hoc test was conducted to evaluate the effect of the independent variable on the evaluated properties., Results: FTIR confirmed the encapsulation of calcium sources into HNTs. Regarding flow and film thickness, the values obtained from these sealers were in accordance with the specifications provided by ISO 6876. For radiopacity, AH Plus™ achieved the highest radiopacity (p<0.05). Among the experimental formulations, all experimental HNT-containing compositions exhibited values below 3 mm Al. The experimental sealers showed greater dimensional changes when compared to the commercial (AH Plus™) control. The release of calcium ions was observed for the HNT&#95;CaOH 2 and HNT&#95;β-TCP sealers without statistical differences. Experimental sealers containing HNT&#95;CaOH 2 and HNT&#95;β-TCP significantly reduced the CFU/mL count and showed cell compatibility., Conclusions: The findings of this study demonstrate that the incorporation of HNT&#95;CaOH 2 or HNT&#95;β-TCP into resin-based experimental sealers promoted antimicrobial effects and gradual calcium release without impairing cytocompatibility or physicochemical properties of the sealers. Still, an adjustment to reach the minimal radiopacity established by ISO 6876 is needed., Clinical Relevance: The experimental resin-based sealers seemed to be an alternative for endodontics. The incorporation of calcium sources exerts promising antimicrobial effects while displaying low cell toxicity., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Currently employed approaches and materials used for vital pulp therapies (VPTs) and regenerative endodontic procedures (REPs) lack the efficacy to predictably achieve successful outcomes due to their inability to achieve adequate disinfection and/or lack of desired immune modulatory effects. Natural polymers and medicinal herbs are biocompatible, biodegradable, and present several therapeutic benefits and immune-modulatory properties; thus, standing out as a clinically viable approach capable of establishing a conducive environment devoid of bacteria and inflammation to support continued root development, dentinal bridge formation, and dental pulp tissue regeneration. However, the low stability and poor mechanical properties of the natural compounds have limited their application as potential biomaterials for endodontic procedures. In this study, Aloe vera (AV), as a natural antimicrobial and anti-inflammatory agent, was incorporated into photocrosslinkable Gelatin methacrylate (GelMA) nanofibers with the purpose of developing a highly biocompatible biomaterial capable of eradicating endodontic infection and modulating inflammation. Stable GelMA/AV nanofibers with optimal properties were obtained at the ratio of (70:30) by electrospinning. In addition to the pronounced antibacterial effect against Enterococcus faecalis, the GelMA/AV (70:30) nanofibers also exhibited a sustained antibacterial activity over 14 days and significant biofilm reduction with minimal cytotoxicity, as well as anti-inflammatory properties and immunomodulatory effects favoring healing. Our results indicate that the novel GelMA/AV (70:30) nanofibers hold great potential as a biomaterial strategy for endodontic infection eradication and enhanced healing., 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 © 2023 Elsevier B.V. All rights reserved.)

The objective of this research was to create and appraise biodegradable polymer-based nanofibers containing distinct concentrations of calcium trimetaphosphate (Ca-TMP) for periodontal tissue engineering. Poly(ester urea) (PEU) (5% w / v ) solutions containing Ca-TMP (15%, 30%, 45% w / w ) were electrospun into fibrous scaffolds. The fibers were evaluated using SEM, EDS, TGA, FTIR, XRD, and mechanical tests. Degradation rate, swelling ratio, and calcium release were also evaluated. Cell/Ca-TMP and cell/scaffold interaction were assessed using stem cells from human exfoliated deciduous teeth (SHEDs) for cell viability, adhesion, and alkaline phosphatase (ALP) activity. Analysis of variance (ANOVA) and post-hoc tests were used (α = 0.05). The PEU and PEU/Ca-TMP-based membranes presented fiber diameters at 469 nm and 414-672 nm, respectively. Chemical characterization attested to the Ca-TMP incorporation into the fibers. Adding Ca-TMP led to higher degradation stability and lower dimensional variation than the pure PEU fibers; however, similar mechanical characteristics were observed. Minimal calcium was released after 21 days of incubation in a lipase-enriched solution. Ca-TMP extracts enhanced cell viability and ALP activity, although no differences were found between the scaffold groups. Overall, Ca-TMP was effectively incorporated into the PEU fibers without compromising the morphological properties but did not promote significant cell function.

Regenerative endodontics represents a paradigm shift in dental pulp therapy for necrotic young permanent teeth. However, there are still challenges associated with attaining maximum root canal disinfection while supporting angiogenesis and preserving resident stem cells viability and differentiation capacity. Here, we developed a hydrogel system by incorporating antibiotic-eluting fiber-based microparticles in gelatin methacryloyl (GelMA) hydrogel to gather antimicrobial and angiogenic properties while prompting minimum cell toxicity. Minocycline (MINO) or clindamycin (CLIN) was introduced into a polymer solution and electrospun into fibers, which were further cryomilled to attain MINO- or CLIN-eluting fibrous microparticles. To obtain hydrogels with multi-therapeutic effects, MINO- or CLIN-eluting microparticles were suspended in GelMA at distinct concentrations. The engineered hydrogels demonstrated antibiotic-dependent swelling and degradability while inhibiting bacterial growth with minimum toxicity in dental-derived stem cells. Notably, compared to MINO, CLIN hydrogels enhanced the formation of capillary-like networks of endothelial cells in vitro and the presence of widespread vascularization with functioning blood vessels in vivo. Our data shed new light onto the clinical potential of antibiotic-eluting gelatin methacryloyl hydrogel as an injectable scaffold with multi-therapeutic effects to promote antimicrobial disinfection and angiogenesis for regenerative endodontics.

Objectives: The current standard for treating irreversibly damaged dental pulp is root canal therapy, which involves complete removal and debridement of the pulp space and filling with an inert biomaterial. A regenerative approach to treating diseased dental pulp may allow for complete healing of the native tooth structure and enhance the long-term outcome of once-necrotic teeth. The aim of this paper is, therefore, to highlight the current state of dental pulp tissue engineering and immunomodulatory biomaterials properties, identifying exciting opportunities for their synergy in developing next-generation biomaterials-driven technologies., Methods: An overview of the inflammatory process focusing on immune responses of the dental pulp, followed by periapical and periodontal tissue inflammation are elaborated. Then, the most recent advances in treating infection-induced inflammatory oral diseases, focusing on biocompatible materials with immunomodulatory properties are discussed. Of note, we highlight some of the most used modifications in biomaterials' surface, or content/drug incorporation focused on immunomodulation based on an extensive literature search over the last decade., Results: We provide the readers with a critical summary of recent advances in immunomodulation related to pulpal, periapical, and periodontal diseases while bringing light to tissue engineering strategies focusing on healing and regenerating multiple tissue types., Significance: Significant advances have been made in developing biomaterials that take advantage of the host's immune system to guide a specific regenerative outcome. Biomaterials that efficiently and predictably modulate cells in the dental pulp complex hold significant clinical promise for improving standards of care compared to endodontic root canal therapy., (Copyright © 2023 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

Periodontitis is a ubiquitous chronic inflammatory, bacteria-triggered oral disease affecting the adult population. If left untreated, periodontitis can lead to severe tissue destruction, eventually resulting in tooth loss. Despite previous efforts in clinically managing the disease, therapeutic strategies are still lacking. Herein, melt electrowriting (MEW) is utilized to develop a compositionally and structurally tailored graded scaffold for regeneration of the periodontal ligament-to-bone interface. The composite scaffolds, consisting of fibers of polycaprolactone (PCL) and fibers of PCL-containing magnesium phosphate (MgP) were fabricated using MEW. To maximize the bond between bone (MgP) and ligament (PCL) regions, we evaluated two different fiber architectures in the interface area. These were a crosshatch pattern at a 0/90° angle and a random pattern. MgP fibrous scaffolds were able to promote in vitro bone formation even in culture media devoid of osteogenic supplements. Mechanical properties after MgP incorporation resulted in an increase of the elastic modulus and yield stress of the scaffolds, and fiber orientation in the interfacial zone affected the interfacial toughness. Composite graded MEW scaffolds enhanced bone fill when they were implanted in an in vivo periodontal fenestration defect model in rats. The presence of an interfacial zone allows coordinated regeneration of multitissues, as indicated by higher expression of bone, ligament, and cementoblastic markers compared to empty defects. Collectively, MEW-fabricated scaffolds having compositionally and structurally tailored zones exhibit a good mimicry of the periodontal complex, with excellent regenerative capacity and great potential as a defect-specific treatment strategy.

Degradation of the collagen fibrils at the dentin-resin interface by the enzymatic activity of matrix metalloproteinases (MMPs) has been known to permit some dental restoration complications, such as microleakage, secondary caries, and, ultimately, restoration failures. This study aimed to evaluate a modified adhesive by adding an MMP inhibitor from green tea extract with and without nanotube encapsulation to sustain the drug release. Epigallocatechin-3-gallate (EGCG) and Halloysite nanotubes (HNTs) were prepared to produce three variant combinations of modified adhesive (EGCG, EGCG-encapsulated HNT, and EGCG-free HNT). The drug loading efficiency and EGCG release over time were evaluated using UV-vis spectrometry. MMP-mediated β-casein (BCN) cleavage rate assays were used to determine the ability of the EGCG in eluates of the adhesive to inhibit MMP-9 activities. For up to 8 weeks, HNT encapsulation reduced release to a statistically significant level. MMP-mediated β-casein cleavage rate assays showed a significant decrease for the EGCG groups compared to the non-EGCG adhesive groups. Furthermore, the use of HNT for EGCG encapsulation to modify a dental adhesive helped slow down the rate of EGCG release without impacting its MMP inhibitory capabilities, which may help to maintain the dentin-resin interface's integrity over the long term after dental restoration placement.

For nearly three decades, tissue engineering strategies have been leveraged to devise effective therapeutics for dental, oral, and craniofacial (DOC) regenerative medicine and treat permanent deformities caused by many debilitating health conditions. In this regard, additive manufacturing (AM) allows the fabrication of personalized scaffolds that have the potential to recapitulate native tissue morphology and biomechanics through the utilization of several 3D printing techniques. Among these, melt electrowriting (MEW) is a versatile direct electrowriting process that permits the development of well-organized fibrous constructs with fiber resolutions ranging from micron to nanoscale. Indeed, MEW offers great prospects for the fabrication of scaffolds mimicking tissue specificity, healthy and pathophysiological microenvironments, personalized multi-scale transitions, and functional interfaces for tissue regeneration in medicine and dentistry. Excitingly, recent work has demonstrated the potential of converging MEW with other AM technologies and/or cell-laden scaffold fabrication (bioprinting) as a favorable route to overcome some of the limitations of MEW for DOC tissue regeneration. In particular, such convergency fabrication strategy has opened great promise in terms of supporting multi-tissue compartmentalization and predetermined cell commitment. In this review, we offer a critical appraisal on the latest advances in MEW and its convergence with other biofabrication technologies for DOC tissue regeneration. We first present the engineering principles of MEW and the most relevant design aspects for transition from flat to more anatomically relevant 3D structures while printing highly-ordered constructs. Secondly, we provide a thorough assessment of contemporary achievements using MEW scaffolds to study and guide soft and hard tissue regeneration, and draw a parallel on how to extrapolate proven concepts for applications in DOC tissue regeneration. Finally, we offer a combined engineering/clinical perspective on the fabrication of hierarchically organized MEW scaffold architectures and the future translational potential of site-specific, single-step scaffold fabrication to address tissue and tissue interfaces in dental, oral, and craniofacial regenerative medicine. STATEMENT OF SIGNIFICANCE: Melt electrowriting (MEW) techniques can further replicate the complexity of native tissues and could be the foundation for novel personalized (defect-specific) and tissue-specific clinical approaches in regenerative dental medicine. This work presents a unique perspective on how MEW has been translated towards the application of highly-ordered personalized multi-scale and functional interfaces for tissue regeneration, targeting the transition from flat to anatomically-relevant three-dimensional structures. Furthermore, we address the value of convergence of biofabrication technologies to overcome the traditional manufacturing limitations provided by multi-tissue complexity. Taken together, this work offers abundant engineering and clinical perspectives on the fabrication of hierarchically MEW architectures aiming towards site-specific implants to address complex tissue damage in regenerative dental medicine., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest or with respect to the authorship and/or publication of this article., (Copyright © 2022 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

This investigation aimed to synthesize poly(D,L-lactide) (PLA)-based fibrous scaffolds containing natural essential oils (i.e., linalool and citral) and determine their antimicrobial properties and cytocompatibility as a clinically viable cell-friendly disinfection strategy for regenerative endodontics. PLA-based fibrous scaffolds were fabricated via electrospinning with different concentrations of linalool and citral. The micromorphology and average diameter of the fibers was investigated through scanning electron microscopy (SEM). The chemical composition of the scaffolds was inferred by Fourier-transform infrared spectroscopy (FTIR). Antimicrobial efficacy against Enterococcus faecalis and Actinomyces naeslundii was also evaluated by agar diffusion and colony-forming units (CFU) assays. The scaffolds' cytocompatibility was determined using dental pulp stem cells (DPSCs). Statistical analyses were performed and the significance level was set at α = 5%. Linalool and citral's incorporation in the PLA fibrous scaffolds was confirmed in the FTIR spectra. SEM images indicate no morphological changes upon inclusion of the essential oils, except the reduced diameter of 40% linalool-laden fibers (p < 0.05). Importantly, significant antimicrobial properties were reported for citral-containing scaffolds for CFU/mL counts (p < 0.05), while only 20% and 40% linalool-laden scaffolds reduced CFU/mL (p < 0.05). Meanwhile, the inhibition halos were verified in a concentration-dependent manner for all monoterpenes-laden scaffolds. Citral- and linalool-laden PLA-based fibrous scaffolds showed acceptable cytocompatibility. The incorporation of natural monoterpenes did not alter the scaffolds' fibrous morphology, promoted antimicrobial action against endodontic pathogens, and preserved DPSCs viability. Linalool- and citral-laden electrospun scaffolds hold promise as naturally derived antimicrobial therapeutics for applications in regenerative endodontics., (© 2022. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Gelatin methacrylate (GelMA) is a biodegradable and biocompatible engineered material with significant promise for its applications in tissue engineering, drug delivery, and 3D bioprinting applications. Gelatin is functionalized with terminal methacrylate groups which allow for its photoinducible crosslinking, and thereby tunable properties. Photocrosslinking of GelMA solution in situ allows for fabrication of hydrogels to fit patient-specific defects. Given its favorable biologic properties, GelMA may be used as a carrier for bioactive substances necessary to induce regenerative phenotypes or augment healing, such as growth factors and biotherapeutics. Gelatin is cleaved by cell-secreted enzymes such that its degradation, and subsequently release of bioactive substances, is well-matched to tissue regeneration processes. GelMA may be mixed with a wide array of additives to enhance and improve the specificity of its biologic activity. Here, we present two protocols for novel fabrications and their uses as clinically relevant drug delivery systems. GelMA hydrogels provides a versatile platform for the development of injectable drug delivery therapeutics for broad applications in regenerative dental medicine., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Objectives: To evaluate the incorporation of repairing capsules containing different monomers and polymerization modulators on the self-healing efficiency of an experimental photopolymerizable resin-based composite., Methods: Self-healing capsules containing different monomers and polymerization modulators were prepared by emulsion polymerization: TC DHEPT (TEGDMA and DHEPT), BTC DHEPT (Bis-GMA, TEGDMA, and DHEPT), and BTC BPO (Bis-GMA, TEGDMA, and BPO). The capsules were analyzed through Fourier transform infrared spectroscopy and scanning electron microscopy. The capsules were added into experimental photopolymerizable resin composites establishing the following groups: ER (Control without capsules), ER+BPO, ER+BPO+TC DHEPT , and ER+BTC BPO +BTC DHEPT . Filtek Z350 resin composite (3 M ESPE) was used as a commercial reference. The materials were tested for degree of conversion (DC), flexural strength (σf), elastic modulus (Ef), fracture toughness (virgin K IC ), self-healing efficiency (healed K IC ), and roughness. For statistical analysis, the significance value was established at an a = 0.05 level., Results: When compared to the control material, the incorporation of repairing capsules did not affect DC, σf, and Ef. Fracture toughness was statistically similar between the experimental groups (p ≤ 0.05). Healed K IC was statistically different between the groups ER+TC DHEP and ER+BTC BPO +BTC DHEPT ; the self-healing efficiency was higher for ER+TC DHEPT . Surface roughness was statistically similar among all groups., Conclusions: The use of self-healing capsules promoted repair of the material. Studies with material aging after the self-healing process are necessary to better demonstrate the effectiveness of this system., Clinical Significance: The self-healing system seemed to be a promising technology to be used in self-repaired restorative materials, which may prevent restoration fractures., Competing Interests: Declaration of Competing Interest The authors report no conflicts of interest in this work., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Fused Filament Fabrication (FFF)-based 3D printing is an efficient technique for developing medical implants, but it is not very useful in developing small yet mechanically robust design-specific fixtures such as dental implants (<15 mm). Specifically, it is challenging to 3D print robust Polyetheretherketone (PEEK) small implants due to PEEK's high melting temperature and melt viscosity. However, in this study, we efficiently utilize high-temperature FFF to develop the first-of-its-kind patient-specific robust PEEK dental implants with high print resolution. Specifically, we explore the effects of critical FFF processing conditions on the mechanical properties of the implants and subsequently determine an optimized set of processing conditions that are essential in developing durable dental implant systems. Our results indicate that the 3D printed dental implants exhibit good fatigue properties and suffice the clinical and industrial requirements for dental implants. Furthermore, we prove that the 3D printed implants exhibit adequate mechanical durability even after simulated (accelerated) aging of 30 years., 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 © 2022 Elsevier Ltd. All rights reserved.)

This study was aimed at engineering photocrosslinkable azithromycin (AZ)-laden gelatin methacryloyl fibers via electrospinning to serve as a localized and biodegradable drug delivery system for endodontic infection control. AZ at three distinct amounts was mixed with solubilized gelatin methacryloyl and the photoinitiator to obtain the following fibers: GelMA+5%AZ, GelMA+10%AZ, and GelMA+15%AZ. Fiber morphology, diameter, AZ incorporation, mechanical properties, degradation profile, and antimicrobial action against Aggregatibacter actinomycetemcomitans and Actinomyces naeslundii were also studied. In vitro compatibility with human-derived dental pulp stem cells and inflammatory response in vivo using a subcutaneous rat model were also determined. A bead-free fibrous microstructure with interconnected pores was observed for all groups. GelMA and GelMA+10%AZ had the highest fiber diameter means. The tensile strength of the GelMA-based fibers was reduced upon AZ addition. A similar pattern was observed for the degradation profile in vitro. GelMA+15%AZ fibers led to the highest bacterial inhibition. The presence of AZ, regardless of the concentration, did not pose significant toxicity. In vivo findings indicated higher blood vessel formation, mild inflammation, and mature and thick well-oriented collagen fibers interweaving with the engineered fibers. Altogether, AZ-laden photocrosslinkable GelMA fibers had adequate mechanical and degradation properties, with 15%AZ displaying significant antimicrobial activity without compromising biocompatibility.