Search Results (28)

This study investigated the effects of acid etching with grape seed extract (GSE)-modified etchants, varying phosphoric acid (PA) concentrations, on endogenous collagenolytic activity of etched dentin, adhesive–dentin (A/D) interfacial formation, and bond strength over time. Three PA concentrations (5%, 10%, and 20%) were combined with 2% GSE (5PA/GSE, 10PA/GSE, and 20PA/GSE) and compared to a control (CT) group using 32% PA gel (3M Universal Scotchbond etchant). Seventy-four caries-free human third molars were sectioned to expose dentin surfaces, which were etched and analyzed. In situ zymography with confocal laser microscopy was used to assess endogenous collagenolytic activity in etched dentin specimens. For A/D interfacial morphology and bond strength, etched dentin was bonded with Adper Single Bond Plus adhesive (3M ESPE) and composite buildup. The interfacial morphology of A/D specimens was evaluated using either Goldner’s trichrome staining under light microscopy after microtomy sectioning or scanning electron microscopy. A/D specimens were stored in either TESCA buffer or collagenase solution and tested immediately (IM) or at multiple time points over one year using the microtensile bond strength (μTBS) test. Data were analyzed by one- or three-way ANOVA followed by Games–Howell or Tukey’s tests (α = 0.05). GSE-modified etchants significantly reduced endogenous collagenolytic activity (p < 0.05). Although GSE-modified etchants resulted in thinner A/D interfaces, the bond strength remained unaffected (p > 0.05). Bond strength stability was prolonged up to one year with 5PA/GSE and 10PA/GSE (p < 0.001), while CT or 20PA/GSE showed significant degradation by 17 weeks (p < 0.01). Storage in the more aggressive collagenase solution did not further reduce the bond strength compared to TESCA buffer (p = 0.966). Acid etching with GSE-modified etchants effectively inhibits endogenous MMP-mediated collagenolytic activity. At 5% and 10% PA, this approach enhances the stability of the A/D bond strength, offering a promising modification for dentin bonding protocols. Full article

Boron-containing compounds (BCCs) have been proposed for the treatment of diabetes and its complications. Recent studies have reported an improvement in the design and development of pharmaceutical formulations (often gels) containing boric acid applied to the foot ulcers of humans diagnosed with diabetes. The proposed mechanisms of action of boric acid include antimicrobial effects, the modulation of inflammation and metabolism, and the induction of cell differentiation. On the other hand, recent studies have suggested that boronic acids are potent antibacterial and antifungal compounds, effective modulators of inflammation, and inducers of vascular regeneration as well as inducers of healing, and they confer attractive properties such as adhesion, interaction, and the formation of complexes in formulations. Moreover, only a handful of studies conducted in animals have suggested the effective role of some BCCs as potent enhancers of wound healing, including their actions on induced and/or infected wounds in animals with disrupted metabolism. Also, it should be mentioned that no strong interactions between boric acid and the boronic acids present in formulations have been described. The developed combination could act as an additive and complementary therapy in the treatment of diabetic ulcers in humans. Further studies are required to support the hypothesis that this combination acts through diverse mechanisms to improve healing while avoiding or limiting a local or disseminated infection. Furthermore, the safety of BCCs used for foot ulcers should be established, as should the role of these formulations as a complementary therapy in current protocols for treating patients with diabetic foot ulcers. Full article

Stimuli-responsive materials, particularly supramolecular hydrogels, exhibit a dynamic adaptability to external factors such as pH and ultrasound. Among these, phenylalanine (Phe)-derived hydrogels are promising due to their biocompatibility, biodegradability, and tunable properties, making them ideal for biomedical applications. This study explores the effects of pH and ultrasound on the gelation properties of N-substituted Phe derivatives, with a primary focus on the role of ultrasound in optimizing the gelation process. A series of N-substituted Phe derivatives were synthesized via reductive amination and hydrolysis. Hydrogel formation was possible with two of these compounds, namely G1 and G2, using the following two methods: heating–cooling (H–C) and heating–ultrasound–cooling (H–US–C). The critical gelation concentration (CGC), gelation kinetics, thermal stability (T_gel), and viscoelastic properties were assessed. Morphological and cytotoxicity analyses were performed to confirm the suitability of these gels for biomedical applications. Both G1 and G2 derivatives demonstrated enhanced gelation under the H–US–C protocol compared to H–C, with notable reductions in CGC (up to 47%) and gelation time (by over 90%). Ultrasound-induced gels led to an improved network density and stability, while maintaining thermal reversibility and mechanical properties comparable to those of hydrogels formed without ultrasound. Cytotoxicity studies confirmed a high biocompatibility, with cell viability rates above 95% across the tested concentrations. Given the similar rheological and morphological properties of the hydrogels regardless of the preparation method, drug release experiments were performed with representative gel samples and demonstrated the efficient encapsulation and controlled release of 5-fluorouracil and methotrexate from the hydrogels, supporting their potential as pH-responsive drug delivery platforms. This study highlights the role of ultrasound as a powerful tool for accelerating and optimizing the gelation process of supramolecular hydrogels, which is particularly relevant for applications requiring rapid gel formation. The developed Phe-based hydrogels also demonstrate promising characteristics as drug delivery systems. Full article

The growing need for sustainable biotechnological solutions to address environmental challenges, such as climate change and resource depletion, has intensified interest in microbial-based production systems. Synthetic biofilms, which mimic natural microbial consortia, offer a promising platform for optimizing complex metabolic processes that can convert renewable feedstocks into valuable chemicals. In this context, understanding and harnessing the interactions between co-immobilized microorganisms are critical for advancing bioprocesses that contribute to circular bioeconomy goals. In this study, we investigated the viability and metabolic activity of Clostridium carboxidivorans and Clostridium kluyveri within a synthetic, dual-layered biofilm composed of agar hydrogel. This setup compartmentalized each bacterial species. Embedding the bacteria in a structured biofilm offers numerous opportunities for bioproduction, but the inability to monitor cell growth or movement within the immobilization matrix limits process insights. To address this, we adapted a fluorescence in situ hybridization (FISH) protocol, enabling precise, species-specific visualization of bacterial distribution and growth within the gel matrix. Batch processes with the dual-layered biofilm in anaerobic flasks, designed with a metabolic advantage for C. kluyveri, revealed distinct growth dynamics. C. kluyveri exhibited significant metabolic activity, forming clusters at low initial cell concentrations and converting ethanol and acetate into 1-butyrate and 1-hexanoate, indicating viability and cell growth. C. carboxidivorans remained evenly distributed without significant growth or product formation, suggesting that while the cells were viable, they were not metabolically active under the experimental conditions. Both bacterial species were confined to their respective compartments throughout the process, with C. kluyveri showing enhanced substrate conversion at higher initial cell densities in the hydrogel. The pH drop throughout the batch experiment likely contributed to incomplete substrate consumption, particularly for C. kluyveri, which thrives within a narrow pH range. These findings highlight synthetic biofilms as a promising platform for optimizing microbial interactions and improving bioprocess efficiency, especially in applications involving complex metabolic exchanges between co-immobilized microorganisms. Further research will focus on applying conditions to support the growth and metabolic activity of C. carboxidivorans to explore spatial dynamics of bacterial migration and cooperative relationships in the synthetic biofilm. Full article

Thirty-two ambient cured alkali-activated engineered composites (AAECs) were developed by incorporating MgO, multi-walled carbon nanotubes (MWCNTs), reduced graphene oxide (rGO), and polyvinyl alcohol (PVA) fiber with a one-part dry mix technique using powder-based activators/reagents. The effects of material variables, namely binary or ternary combination source materials (fly ash C or F and ground granulated blast furnace slag ‘GGBFS’), two types of reagents with varying chemical ratios and dosages of additives (from 0 to 5% MgO and from 0 to 6% MWCNT/rGO), on the physical (slump flow, flow time, flow velocity, and density), hardness (compressive strength from 0 to 180 days and 28-day ultrasonic pulse velocity ‘UPV’), and micro-structural (SEM/EDS, XRD and FTIR) properties were evaluated. All these variables, individually or combined, influenced the properties and microstructural aspects of AAECs. Problems associated with the dispersion and agglomeration of nanomaterials, which could disrupt the microstructure and weaken its mechanical/physical properties, were avoided through the use of defined ultra-sonication with a high-shear mixing protocol. All AAECs achieved a 28-day compressive strength ranging from 26.0 MPa to 48.5 MPa and a slump flow > 800 mm, satisfying the criteria for flowable structural concrete. The addition of 5% MgO and up to 0.3% MWCNT/rGO increased the compressive strength/UPV of AAECs with MgO-MWCNT or rGO combination provided an improved strength at a higher dosage of 0.6%. A linear correlation between compressive strength and UPV was derived. As per SEM/EDS and XRD analyses, besides common C-A-S-H/N-C-A-S-H or C-A-S-H/C-S-H gels, the addition of MgO led to the formation of magnesium-aluminum hydrotalcite (Ht) and M-S-H (demonstrating self-healing potential), while the incorporation of rGO produced zeolites which densified the matrix and increased the compressive strength/UPV of the AAECs. Fourier transform infrared spectrometer (FTIR) analysis also suggested the formation of an aluminosilicate network in the AAECs, indicating a more stable structure. The increased UPV of MWCNT/rGO-incorporated AAECs indicated their better conductivity and ability of self-sensing. The developed AAECs, incorporating carbon-nano materials and MgO additive, have satisfactory properties with self-healing/-sensing potentials. Full article

Organoid technology, as an innovative approach in biomedicine, exhibits promising prospects in disease modeling, pharmaceutical screening, regenerative medicine, and oncology research. However, the use of tumor-derived Matrigel as the primary method for culturing organoids has significantly impeded the clinical translation of organoid technology due to concerns about potential risks, batch-to-batch instability, and high costs. To address these challenges, this study innovatively introduced a photo-crosslinkable hydrogel made from a porcine small intestinal submucosa decellularized matrix (SIS), fish collagen (FC), and methacrylate gelatin (GelMA). The cost-effective hydrogel demonstrated excellent biocompatibility, tunable mechanical properties, rapid gelation properties, and low immunogenicity. Importantly, the proliferation and differentiation capacities of small intestinal organoids cultured in hydrogel were comparable to those in Matrigel, with no significant disparity observed. Furthermore, after one week of transplantation in nude mice, the hydrogel–organoid complex exhibited sustained structural and functional stability while preserving the differentiation characteristics of small intestinal organoids. Our study also demonstrated the effective potential of FC/SIS/GelMA hydrogel in accelerating the repair process of small intestinal defects, reducing the area of scar formation, and promoting the regeneration of both intestinal villi and smooth muscle tissue. In summary, this study presents a novel protocol for culturing small intestinal organoids, offering potential implications for future clinical applications and serving as an experimental foundation for the development of tissue-engineered intestines based on small intestinal organoids. Full article

Kesterite-based semiconductors, particularly copper–zinc–tin–sulfide (CZTS), have garnered considerable attention as potential absorber layers in thin-film solar cells because of their abundance, nontoxicity, and cost-effectiveness. In this study, we explored the synthesis of Ag-alloyed CZTS (ACZTS) materials via the sol–gel method and deposited them on a transparent fluorine-doped tin oxide (FTO) back electrode. A key challenge is the selection and manipulation of metal–salt precursors, with a particular focus on the oxidation states of copper (Cu) and tin (Sn) ions. Two distinct protocols, varying the oxidation states of the Cu and Sn ions, were employed to synthesize the ACZTS materials. The transfer from the solution to the precursor film was analyzed, followed by annealing at different temperatures under a sulfur atmosphere to investigate the behavior and growth of these materials during the final stage of annealing. Our results show that the precursor transformation from solution to film is highly sensitive to the oxidation states of these metal ions, significantly influencing the chemical reactions during sol–gel synthesis and subsequent annealing. Furthermore, the formation pathway of the kesterite phase at elevated temperatures differs between the two protocols. Structural, morphological, and optical properties were characterized via X-ray diffraction (XRD), Raman spectroscopy, and scanning electron microscopy (SEM). Our findings highlight the critical role of the Cu and Sn oxidation states in the formation of high-quality kesterite materials. Additionally, we studied a novel approach for controlling the synthesis and phase evolution of kesterite materials via molecular inks, which could provide new opportunities for enhancing the efficiency of thin-film solar cells. Full article

Objectives: The study addresses the challenge of cross-infection in dentistry, focusing on improving disinfection protocols for dental hydrocolloid gel materials. This research aimed to incorporate chlorhexidine, natural plant extracts, and green-synthesized silver nanoparticles (AgNPs) into dental alginate to enhance safety and hygiene standards in dental practices. Methods: Conventional dental alginate served as the control, with experimental groups including alginate modified with 0.2% chlorhexidine (CHX-group) and alginate incorporating AgNPs synthesized from Syzygium aromaticum (SA) and Zingiber officinale (ZO) extracts (SA + AgNPs and ZO + AgNPs groups). Plant extracts were analyzed via GC/MS to confirm their composition. UV–visible spectroscopy, SEM, and EDX characterized the synthesized AgNPs. Antimicrobial efficacy against Candida albicans, Streptococcus mutans, and methicillin-resistant and methicillin-sensitive Staphylococcus aureus was evaluated using agar well diffusion assays. The dimensional accuracy of alginate impressions was assessed according to ISO 21563:2021 standards. Results: Chemical analysis of SA and ZO extracts identified 60 and 43 active compounds, respectively, supporting their use in AgNP synthesis. UV–visible spectroscopy, SEM, and EDX confirmed the formation of spherical AgNPs using SA and ZO extracts. Modified groups showed inhibitory activity against Streptococcus mutans and methicillin-resistant (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA), in contrast to the unmodified control. Both AgNP-modified groups demonstrated efficacy comparable to the CHX-group against MRSA and MSSA, with SA + AgNP showing superior performance against C. albicans. The dimensional accuracy of all groups was within clinically acceptable ranges as reported in the literature (0.027–0.083 mm). Discussion: CHX, silver nitrate, and green-synthesized AgNPs present promising options for developing self-disinfecting alginate impression materials. Utilizing plant extracts in AgNP synthesis offers a safe, efficient, and synergistic approach between metal ions and phytotherapeutic agents. This approach could potentially enhance disinfection efficacy without compromising material performance, improving dental safety and hygiene. Full article

R-loops, structures that play a crucial role in various biological processes, are integral to gene expression, the maintenance of genome stability, and the formation of epigenomic signatures. When these R-loops are deregulated, they can contribute to the development of serious health conditions, including cancer and neurodegenerative diseases. The detection of R-loops is a complex process that involves several approaches. These include S9.6 antibody- or RNAse H-based immunoprecipitation, non-denaturing bisulfite footprinting, gel electrophoresis, and electron microscopy. Each of these methods offers unique insights into the nature and behavior of R-loops. In our study, we introduce a novel protocol that has been developed based on a single-molecule DNA combing assay. This innovative approach allows for the direct and simultaneous visualization of RNA:DNA hybrids and replication forks, providing a more comprehensive understanding of these structures. Our findings confirm the transcriptional origin of the hybrids, adding to the body of knowledge about their formation. Furthermore, we demonstrate that these hybrids have an inhibitory effect on the progression of replication forks, highlighting their potential impact on DNA replication and cellular function. Full article

The present challenge in dental pulp tissue engineering scaffold materials lies in the development of tissue-specific scaffolds that are conducive to an optimal regenerative microenvironment and capable of accommodating intricate root canal systems. This study utilized porcine dental pulp to derive the decellularized extracellular matrix (dECM) via appropriate decellularization protocols. The resultant dECM was dissolved in an acid pepsin solution to form dECM hydrogels. The analysis encompassed evaluating the microstructure and rheological properties of dECM hydrogels and evaluated their biological properties, including in vitro cell viability, proliferation, migration, tube formation, odontogenic, and neurogenic differentiation. Gelatin methacrylate (GelMA) hydrogel served as the control. Subsequently, hydrogels were injected into treated dentin matrix tubes and transplanted subcutaneously into nude mice to regenerate dental pulp tissue in vivo. The results showed that dECM hydrogels exhibited exceptional injectability and responsiveness to physiological temperature. It supported the survival, odontogenic, and neurogenic differentiation of dental pulp stem cells in a 3D culture setting. Moreover, it exhibited a superior ability to promote cell migration and angiogenesis compared to GelMA hydrogel in vitro. Additionally, the dECM hydrogel demonstrated the capability to regenerate pulp-like tissue with abundant blood vessels and a fully formed odontoblast-like cell layer in vivo. These findings highlight the potential of porcine dental pulp dECM hydrogel as a specialized scaffold material for dental pulp regeneration. Full article

Background: Protein complexes provide valuable biological information, but can be difficult to handle. Therefore, technical advancements designed to improve their manipulation are always useful. Methods: We investigated the opportunity to exploit native agarose gels and the contact blot method for the transfer of native proteins to membranes as means for optimizing the conditions for obtaining stable complexes. As a simple model of protein–protein interactions, an antigen–ligand complex was used in which both proteins were fused to reporters. Results: At each step, it was possible to visualize both the antigen, fused to a fluorescent protein, and the ligand, fused to a monomeric ascorbate peroxidase (APEX) and, as such, a way to tune the protocol. The conditions for the complex formation were adapted by modifying the buffer conditions, the concentration of the proteins and of the cross-linkers. Conclusions: The procedure is rapid, inexpensive, and the several detection opportunities allow for both the monitoring of complex stability and the preservation of the functionality of its components, which is critical for understanding their biomedical implications and supporting drug discovery. The overall protocol represents a handy alternative to gel filtration, uses very standard and ubiquitous equipment, and can be implemented rapidly and without specific training. Full article

Surface chemistry evaluation is crucial in assessing the efficacy of chemical decontamination products for titanium implants. This study aimed to investigate the effectiveness of chemical decontamination solutions in cleaning a contaminated dental implant surface and to evaluate the potential of combining Pluronic gel with hydrogen peroxide (NuBone^®Clean) by evaluating pellicle disruption and re-formation on implant surfaces. In addition, ensuring safety with in vitro and human testing protocols. X-ray Photoelectron Spectroscopy (XPS) was utilised for surface analysis. All the tested gels had some effect on the surface cleanness except for PrefGel^®. Among the tested chemical decontamination candidates, NuBone^®Clean demonstrated effectiveness in providing a cleaner titanium surface. Furthermore, none of the tested chemical agents exhibited cytotoxic effects, and the safety assessment showed no adverse events. The results of this study highlight the significance of conducting comprehensive evaluations, encompassing safety and efficacy, before introducing new chemical agents for dental treatments. The findings suggest that NuBone^®Clean shows potential as a chemical decontamination solution for implant surfaces. However, further investigation through randomised clinical trials is necessary. By adhering to rigorous testing protocols, the development of safe and efficient chemical decontamination strategies can be advanced, benefiting patients and promoting progress in implant dentistry. Full article

Temperature and light responsiveness are widely exploited stimuli to tune the physico-chemical properties of double network hydrogels. In this work, new amphiphilic poly(ether urethane)s bearing photo-sensitive moieties (i.e., thiol, acrylate and norbornene functionalities) were engineered by exploiting the versatility of poly(urethane) chemistry and carbodiimide-mediated green functionalization procedures. Polymers were synthesized according to optimized protocols maximizing photo-sensitive group grafting while preserving their functionality (approx. 1.0 × 10¹⁹, 2.6 × 10¹⁹ and 8.1 × 10¹⁷ thiol, acrylate and norbornene groups/g_polymer), and exploited to prepare thermo- and Vis-light-responsive thiol-ene photo-click hydrogels (18% w/v, 1:1 thiol:ene molar ratio). Green light-induced photo-curing allowed the achievement of a much more developed gel state with improved resistance to deformation (ca. 60% increase in critical deformation, γL). Triethanolamine addition as co-initiator to thiol-acrylate hydrogels improved the photo-click reaction (i.e., achievement of a better-developed gel state). Differently, L-tyrosine addition to thiol-norbornene solutions slightly hindered cross-linking, resulting in less developed gels with worse mechanical performances (~62% γL decrease). In their optimized composition, thiol-norbornene formulations resulted in prevalent elastic behavior at lower frequency compared to thiol-acrylate gels due to the formation of purely bio-orthogonal instead of heterogeneous gel networks. Our findings highlight that exploiting the same thiol-ene photo-click chemistry, a fine tuning of the gel properties is possible by reacting specific functional groups. Full article

Bacterial ghosts (BGS) are empty non-living envelopes produced either genetically or chemically. This study investigated a novel chemical protocol for the production of Neisseria meningitidis ghost vaccine using tween 80 followed by a pH reduction with lactic acid. For our vaccine candidate, both safety and immunogenicity aspects were evaluated. The ghost pellets showed no sign of growth upon cultivation. BGS were visualized by scanning electron microscopy, illustrating the formation of trans-membrane tunnels with maintained cell morphology. Gel electrophoresis showed no distinctive bands of the cytoplasmic proteins and DNA, assuring the formation of ghost cells. In animal model, humoral immune response significantly increased when compared to commercial vaccine (p < 0.01). Moreover, serum bactericidal assay (SBA) recorded 94.67% inhibition compared to 64% only for the commercial vaccine after three vaccination doses. In conclusion, this is the first N. meningitidis ghost vaccine candidate, proven to be effective, economic, and with significant humoral response and efficient SBA values; however, clinical studies should be performed. Full article

Several musculoskeletal conditions are triggered by inflammatory processes that occur along with imbalances between anabolic and catabolic events. Platelet-rich plasma (PRP) is an autologous product derived from peripheral blood with inherent immunomodulatory and anabolic properties. The clinical efficacy of PRP has been evaluated in several musculoskeletal conditions, including osteoarthritis, tendinopathy, and osteonecrosis. When used in combination with hyaluronic acid (HA), a common treatment alternative, the regenerative properties of PRP are significantly enhanced and may provide additional benefits in terms of clinical outcomes. Recently, a new PRP-derived product has been reported in the literature and is being referred to as “plasma gel”. Plasma gels are obtained by polymerizing plasmatic proteins, which form solid thermal aggregates cross-linked with fibrin networks. Plasma gels are considered to be a rich source of growth factors and provide chemotactic, migratory, and proliferative properties. Additionally, clot formation and the associated fibrinolytic reactions play an additional role in tissue repair. There are only a few scientific articles focusing on plasma gels. Historically, they have been utilized in the fields of aesthetics and dentistry. Given that the combination of three products (PRP, HA, and plasma gel) could enhance tissue repair and wound healing, in this technical note, we propose a novel regenerative approach, named “PRP–HA cellular gel matrix” (PRP-GM), in which leukocyte-rich PRP (LR-PRP) is mixed with a plasma gel (obtained by heating the plasma up) and HA in one syringe using a three-way stopcock. The final product contains a fibrin–albumin network entangled with HA’s polymers, in which the cells and biomolecules derived from PRP are attached and released gradually as fibrinolytic reactions and hyaluronic acid degradation occur. The presence of leukocytes, especially monocytes and macrophages, promotes tissue regeneration, as type 2 macrophages (M2) possess an anti-inflammatory feature. In addition, HA promotes the viscosuplementation of the joint and induces an anti-inflammatory response, resulting in pain relief. This unique combination of biological molecules may contribute to the optimization of regenerative protocols suitable for the treatment of degenerative musculoskeletal diseases. Full article