Search Results (282)

There is insufficient evidence regarding the cytotoxicity of restorative 3D-printing resins, used as part of the digital workflow in dentistry. This study presents a novel comparative evaluation of cell viability and adhesion using human Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs), a less commonly used but clinically relevant cell line in dental biomaterials research. The aim of this study was to evaluate the cell viability of WJ-MSCs seeded on 3D-printed resins intended for temporary restorations. Resin discs of three commercial 3D-printing resins (NextDent C&B, Leaf Dental C&B, and UNIZ Temp) and a conventional self-curing acrylic resin (NicTone) were used. WJ-MSCs were cultured on the specimens for 1, 4, and 10 days. Cell viability was assessed using the PrestoBlue assay, Live/Dead immunofluorescence staining, and 7AAD/Annexin V staining. Cell adhesion was evaluated using scanning electron microscopy. Direct exposure to the 3D-printed resins and the self-curing acrylic caused slight reductions in cell viability compared to the control group in both microscopic analyses. 7AAD/Annexin V showed the highest percentage of viable WBCs for the conventional acrylic (34%), followed by UNIZ (35%), NextDent (42%), and Leaf Dental (36%) (ANOVA p < 0.05 Tukey’s post-hoc test p < 0.05). These findings suggest that 3D-printed resins could be considered safe for use in temporary restorations. Full article

To improve implant osseointegration while preventing infection, we developed a strontium (Sr)-doped Ti₃C₂T_x MXene coating on titanium, aiming to synergistically enhance bone integration and antibacterial performance. MXene is a family of two-dimensional transition-metal carbides/nitrides whose abundant surface terminations endow high hydrophilicity and bioactivity. The coating was fabricated via anodic electrophoretic deposition (40 V, 2 min) of Ti₃C₂T_x nanosheets, followed by SrCl₂ immersion to incorporate Sr²⁺. The coating morphology, phase composition, chemistry, hydrophilicity, mechanical stability, and Sr²⁺ release were characterized. In vitro bioactivity was assessed with rat bone marrow mesenchymal stem cells (BMSCs)—with respect to viability, proliferation, migration, alkaline phosphatase (ALP) staining, and Alizarin Red S mineralization—while the antibacterial efficacy was evaluated against Staphylococcus aureus (S. aureus) via live/dead staining, colony-forming-unit enumeration, and AlamarBlue assays. The Sr-doped MXene coating formed a uniform lamellar structure, lowered the water-contact angle to ~69°, and sustained Sr²⁺ release (0.36–1.37 ppm). Compared to undoped MXene, MXene/Sr enhanced BMSC proliferation on day 5, migration by 51%, ALP activity and mineralization by 47%, and reduced S. aureus viability by 49% within 24 h. Greater BMSCs activity accelerates early bone integration, whereas rapid bacterial suppression mitigates peri-implant infection—two critical requirements for implant success. Sr-doped Ti₃C₂T_x MXene thus offers a simple, dual-function surface-engineering strategy for dental and orthopedic implants. Full article

In this comprehensive study, we delve into the intricate binding properties of tannic acid (TA) and examine their dual role in the realm of biomaterial development. While TA’s properties can enhance the functionality and performance of biomaterials, they also raise concerns regarding potential biases in in vitro biocompatibility assessments. We focus on the relevance and constraints of several widely employed cell viability assays, namely the DNA-based PicoGreen assay, the PrestoBlue assay, and the Live/Dead staining technique utilizing fluorescein diacetate (FDA) and propidium iodide (PI). We investigate how these assays perform when applied to TA-coated scaffolds and cell sheets. Through a detailed presentation of our experimental findings, we juxtapose them through a critical review of the existing literature, allowing us to identify and elucidate the limitations these assays face when assessing TA-based biomaterials. In doing so, we aim not only to enhance the understanding of these potential assay biases but also to provide actionable recommendations for accurately evaluating the biocompatibility of TA-modified substances. This dual approach, combining empirical research with literature analysis, offers vital insights for the research community, ensuring that the assessment of TA-coated biomaterials is scientifically sound and reproducible. Full article

UV-cured methacrylated chitosan (MCHIT) hydrogels were achieved in the presence of silanised tellurium-doped silica bioactive glass (BG-Te-Sil) to produce an antimicrobial and osteoconductive scaffold for tissue engineering applications. Methacrylation of chitosan enabled efficient crosslinking, and the curing process was evaluated by means of Fourier-transform infrared spectroscopy (FTIR) and photorheology analyses. Compressive testing on crosslinked hydrogels showed that the silanised, bioactive, doped glass increased the hydrogel’s elastic modulus by up to 200% compared to unreinforced controls. Antibacterial assays against Staphylococcus aureus ATCC 43300 revealed a significant (p < 0.05) reduction in bacterial metabolic activity for hydrogels containing 50 wt% of the Te-doped bioactive glass. In vitro cytocompatibility with human bone-marrow mesenchymal stem cells demonstrated sustained viability and uniform distribution at 72 h (live/dead staining, AlamarBlue). Under H₂O₂-induced oxidative stress, reinforced hydrogels downregulated pro-inflammatory genes (TNF-α, IFN-γ, IL-1β, and PGES-2). These results suggest that the presence of the silanised bioactive glass can significantly enhance mechanical stability, antibacterial properties, and anti-inflammatory responses without affecting cytocompatibility, making these hydrogels promising for tissue engineering applications. Full article

Chronic wounds represent a major clinical challenge due to their prolonged healing process and susceptibility to bacterial colonization, particularly by biofilm-forming bacteria. To address these issues, in this work, silver-treated silk fibroin scaffolds were developed and tested as multifunctional wound dressings, combining antimicrobial and regenerative properties. Silk fibroin, a natural protein derived from Bombyx mori cocoons, is widely recognized for its biocompatibility and suitability for tissue engineering. In this study, porous silk fibroin scaffolds were functionalized with silver nanoparticles through a photo-reduction process and were comprehensively tested for their cytocompatibility and wound healing potential. The excellent antibacterial activity of the silver-treated scaffolds was demonstrated against Escherichia coli and antibiotic-resistant Pseudomonas aeruginosa, as was extensively reported in a previous work. Biological assays were performed using 3T3 fibroblasts cultured on both untreated and silver-treated silk fibroin scaffolds. Biocompatibility assays, such as MTT, Live/Dead, and cytoskeleton analyses, demonstrated biocompatibility in both scaffold types, comparable to the control. Wound healing potential was assessed using in vitro scratch assays, revealing full wound closure (100%) after 24 h in cells cultured with untreated and silver-treated silk fibroin scaffolds, in contrast to 78.5% closure in the control. Notably, silver-treated scaffolds exhibited enhanced fibroblast repopulation within the wound gap, suggesting a synergistic effect of silver and fibroin in promoting tissue regeneration. These findings demonstrate that silver-treated silk fibroin scaffolds possess both anti-microbial and regenerative properties, making them promising candidates for chronic wound management applications. Full article

This study developed a novel water-soluble Cellulose Acetoacetate (CAA)-chitosan (CS) composite hydrogel drug delivery system. In this system, CAA and CS molecules are cross-linked via dynamic enamine bonds, forming a three-dimensional network structure suitable for drug encapsulation and controlled release. The primary objective was to address the challenges associated with the short half-life and significant fluctuations in therapeutic concentration of cytokine drugs, such as interleukin-2 (IL-2). A hydrogel system with a three-dimensional spatial network structure was successfully constructed via dynamic enamine bonds cross-linking between the acetoacetate groups in CAA molecules and the amino groups in CS. This system exhibits the following characteristics: (1) Dynamic covalent bonds impart adjustable mechanical properties to the hydrogel, enabling precise control over gelation time and mechanical performance; (2) A hierarchical pore structure (average pore size of 100–200 μm) provides a three-dimensional confined space for efficient drug encapsulation, achieving an IL-2 encapsulation efficiency of 83.3 ± 3.1%; (3) In vitro release studies demonstrated that the cumulative release of IL-2 within 72 h ranged from 18.4% to 34.7%, indicating sustained-release behavior. Cell viability assays confirmed that the hydrogel maintained the survival rate of L929 cells above 85% (as determined by the CCK-8 method), and live/dead staining revealed no apparent cytotoxicity. Overall, this three-dimensional network hydrogel based on dynamic covalent bonds represents a promising strategy for low-dose, long-lasting cytokine delivery. Full article

The blood–brain barrier (BBB) limits drug delivery to the brain, particularly for large or hydrophilic molecules. Brain microvascular endothelial cells (bEND.3), which form part of the BBB, play a critical role in regulating drug uptake. This study investigates the use of cold atmospheric microplasma (CAM) to enhance membrane permeability and facilitate drug delivery in bEND.3 cells. CAM generates reactive oxygen species (ROS) that modulate membrane properties. We exposed bEND.3 cells to CAM at varying voltages (3, 3.5, 4, and 4.5 kV) and measured drug uptake using the fluorescent drug FD-150, fluorescence intensity, ROS levels, membrane lipid order, and membrane potential. The results showed a significant increase in fluorescence intensity and drug concentration in the plasma-treated cells compared to controls. ROS production, measured by DCFH-DA staining, was higher in the plasma-treated cells, supporting the hypothesis that CAM enhances membrane permeability through ROS-induced changes. Membrane lipid order, assessed using the LipiORDER probe, shifted from the liquid-ordered (Lo) to liquid-disordered (Ld) phase, indicating increased membrane fluidity. Membrane depolarization was detected with DisBAC2(3) dye, showing increased fluorescence in the plasma-treated cells. Cell viability, assessed by trypan blue and LIVE/DEAD™ assays, revealed transient damage at higher voltages (≥4 kV), with recovery after 24 h. These results suggest that CAM enhances drug delivery in bEND.3 cells by modulating membrane properties via ROS production and changes in membrane potential. CAM offers a promising strategy for improving drug delivery to the brain, with potential applications in brain-targeted therapies. Full article

Background/Objectives: Periodontal disease involves chronic immunoinflammatory processes and microbial dysbiosis, making phytochemicals with anti-inflammatory properties potential therapeutic agents. This study aimed to assess the modulatory effects of yellow passion fruit bagasse extract (PFBE) on periodontal cells under microbial condition. Methods: A human periodontal ligament (PDL) cell line was exposed to F. nucleatum ATCC 25586 to simulate a microbial environment in vitro in the presence and absence of PFBE containing three different concentrations (0.25, 0.50, and 1.00 µg/mL) of piceatannol. Pro-inflammatory markers (TNF-α, IL-8, CCL2), the antioxidant enzyme SOD2, and the protease marker MMP-1 were analyzed by real-time PCR. Protein levels were assessed via ELISA and NF-κB nuclear translocation by immunofluorescence. Cell viability was investigated using live/dead and alamarBlue assays, and in vitro wound healing was evaluated by an automated scratch assay. Results: PDL cells exposed to F. nucleatum significantly increased the gene and protein expression of all inflammatory markers. The stimulatory effects of F. nucleatum were significantly reduced when PDL cells were simultaneously exposed to PFBE. F. nucleatum triggered the NF-κB nuclear translocation while PFBE abrogated the F. nucleatum-stimulated NF-κB nuclear translocation at 60 min. Viability assays demonstrated that neither PFBE nor F. nucleatum were toxic or significantly affected PDL cell viability. In vitro wound closure was improved by the addition of PFBE to F. nucleatum. Conclusions: PFBE exhibited anti-inflammatory and anti-proteolytic effects while improving in vitro wound healing, suggesting a potential modulatory role of PFBE in periodontal disease prevention and treatment. Full article

Objective: To investigate the toxicity of cetalkonium chloride (CKC) on primary cultured human corneal epithelial cells (HCECs). Methods: HCECs were subjected to various concentrations (0.03125 × 10⁻⁴ to 2.0 × 10⁻⁴% (w/v)) of CKC for durations ranging from 24 to 72 h. Cell viability was evaluated using the CCK-8 kit along with live and dead cell staining. Intracellular reactive oxygen species (ROS) levels were measured 20 min following CKC exposure. Observations of changes in cell morphology, cytoplasmic actin filaments, and mitochondrial distribution were conducted using immunocytochemistry and MitoTracker assays. Protein expression levels related to cell survival pathways, including mTOR, ERK, Akt, Bcl-xL, and BAX, were examined via Western blot analysis. Results: CKC exhibited dose-dependent toxicity in HCECs. Exposure to CKC concentrations below 0.125 × 10⁻⁴% resulted in no significant decrease in HCEC viability for up to 72 h. Conversely, exposure to CKC at concentrations of 1.0 × 10⁻⁴% or higher led to significantly decreased HCEC viability. Following exposure to higher concentrations of CKC, elevated levels of intracellular ROS and LDH release were observed. This toxicity was further characterized by decreased levels of phosphorylated mTOR, phosphorylated Akt, phosphorylated ERK, and Bcl-xL, as well as an increase in BAX expression. As the CKC concentration increased, HCECs decreased in size, and mitochondria displayed a loss of characteristic punctate staining. Conclusions: Our findings indicated that exposure to CKC caused significant toxicity in HCECs, which varied with concentration and duration of exposure. This toxicity was associated with an increase in ROS, mitochondrial alterations, and a decline in activity of the cell survival pathways. Full article

Proton beam therapy (PBT) is a rapidly advancing modality of hadron therapy. The primary advantage of proton therapy lies in a unique depth-dose distribution characterized by the Bragg peak, which enables a highly targeted irradiation of the area limited to the tumor, while minimizing the impact on healthy tissues. However, a broader clinical adoption of the ion beam therapy is limited by both economic and radiobiological constraints. One of the possible ways to increase the relative biological effectiveness (RBE) of proton therapy involves the use of radiosensitizers. Background/Objectives: In this work, we investigated the efficacy of using colloidal solutions of lanthanum hexaboride (LaB₆) nanoparticles (NPs) coated with polyacrylic acid (PAA) as sensitizers to increase the antitumor biological effectiveness of proton irradiation. This material has not yet been studied extensively so far, despite its promising physical and chemical properties and several reports on its biocompatibility. Methods: LaB₆ NPs were synthesized by femtosecond pulsed laser ablation, functionalized with PAA and characterized. The safety of NPs was evaluated in vitro using a Live/Dead assay on cell cultures: EMT6/P, BT-474, and in vivo in Balb/c mice after intravenous (i.v.) administration. The efficacy of binary proton therapy was evaluated in vitro on cell cultures: EMT6/P, BT-474, and in vivo in the model of human ductal carcinoma of the mammary gland BT-474 in female Nu/j mice after intratumoral (i.t.) administration at a dose of 2.0 mg/mouse and local proton irradiation (fractional exposure of 31 Gy + 15 Gy). The biodistribution of LaB₆-PAA NPs in the animal body was also evaluated. Results: Significant enhancement in cancer cell death following proton beam irradiation was demonstrated in vitro on EMT6/P, BT-474 cell lines. Although the antitumor efficacy observed in vivo was comparatively lower—likely due to the high sensitivity of the BT-474 xenografts—both proton monotherapy and binary treatment were well tolerated. Conclusions: LaB₆-PAA NPs show promise as efficient sensitizers capable of enhancing the biological efficacy of proton therapy, offering a potential path forward for improving therapeutic outcomes. Full article

Cerium oxide nanoparticles (CeO₂ NPs) have gained significant attention in various fields, including biomedicine, semiconductors, cosmetics, and fuel cells, due to their unique physico-chemical properties. Notably, green-synthesized CeO₂ NPs have demonstrated enhanced potential as drug carriers, particularly in biomedical applications such as anti-inflammatory, anticancer, antimicrobial, and anti-oxidant therapies. This study aimed to investigate the anticancer effects of cerium oxide nanoparticles synthesized using turmeric rhizomes on human lung cancer cells. The cytotoxicity and proliferation inhibition of these nanoparticles were assessed using MTT and Live/Dead assays, revealing a dose-dependent reduction in cell viability. Additionally, reactive oxygen species (ROS) generation was quantified through ROS assays, confirming oxidative stress induction as a key mechanism of cytotoxicity. Cell proliferation analysis further demonstrated that increasing concentrations of CeO₂ NPs significantly reduced the multiplication of healthy lung cancer cells. These findings highlight the potential of turmeric-derived CeO₂ NPs as a promising therapeutic agent for lung cancer treatment, warranting further exploration of their mechanism of action and in vivo efficacy. Full article

Cell culture in two dimensions has been the main instrument in cellular and molecular biology. But there are limitations to two-dimensional culture when it comes to tissue engineering and in vivo reproduction. Tissue engineering technology enabled the creation of biomedical scaffolds, which are mostly utilized to biofabricate different artificial human organs. Tissue architecture that encourage cell proliferation can be produced using direct bioprinting technology. The development of bioinks for 3D bioprinting is consistently seen as a problem in the domains of biofabrication and tissue engineering. This study aimed to determine if Fibroblasts and Keratinocytes could grow on hydrogel scaffolds as efficiently as they can in the culture plates. Melanocytes were co-cultured, and the production of melanin was assessed in a two- and three-dimensional culture system. Scaffolds were fabricated using 8% alginate and 6% gelatin and 3D-printed using a cell link printer. FTIR was used to determine the precise composition of the gels. SEM analysis was performed for the cells present in gel and the topology of the cells. In addition, 8% alginate and 6% alginate gel scaffolds were analyzed for swelling and degradation over time in the cell growth medium and PBS. Furthermore, a gene expression study of cell cultures on scaffolds was performed through qPCR. A live/dead assay was performed to determine cell viability for cells grown on scaffolds for 7, 14, and 21 days. Most of the cells were shown to be viable, similar to the control cells grown on a plate. The findings from the SEM showed that cells were grown on the gel surface, remained viable even after 21 days, and displayed circular cells stacked three-dimensionally on the gel surface in the 3D scaffold. The MTT assay was performed to check the viability of cells cultured on a 3D-printed scaffold for 1, 5, and 15 days. We observed about 40% viable cells after 15 days, as shown by the MTT assay. Furthermore, a co-culture study with Melanocyte showed an increased production of melanin in a 3D culture as compared to a 2D culture. Our findings suggest that an alginate and gelatin polymer can be used as a cellular matrix for epithelial cell culture. Further, in vivo and ex vivo experiments are needed to validate the results for future applications in tissue engineering for wound healing and other tissue engineering domains. Full article

Thin films of ReN were synthesized by DC sputtering at different nitrogen pressures (120, 140, 160, and 180 mTorr) on silicon and Ti6Al4V substrates. The coatings were evaluated for their microstructural and mechanical properties. Additionally, the biocompatibility and electrochemical properties of the films were studied using Hanks’ lactate solution at 37 °C. X-ray diffraction (XRD) confirmed the formation of cubic ReN with higher nitrogen content. The optimized nitrogen pressure (180 mTorr) allowed the complete formation of the cubic phase of ReN. Regarding electrochemical behavior, ReN coatings significantly improve corrosion resistance, reducing the corrosion rate as nitrogen content increases, reaching 0.0145 µm/year at 180 mTorr. Regarding mechanical properties, the deposited ReN films presented an optimal combination of hardness and elastic modulus for the highest nitrogen contents. Cell viability was assessed by comparing uncoated and coated samples using a live/dead staining assay, demonstrating the biocompatibility of the coatings. To complement this study, scanning electron microscopy (SEM) was used to analyze the protein–coating interaction and cell morphology on the surface of the samples. Full article

(1) Autologous chondrocyte implantation (ACI) is a prominent method for treating cartilage damage, but periosteal patches can cause chondrocyte leakage. This study evaluates the potential of a decellularized membrane derived from the cell-produced extracellular matrix of 1-day-old porcine cartilage (pcECM-DM) to act as a substitute for periosteal patches. (2) The interaction between young rabbit chondrocyte cells and pcECM-DM was assessed through cytotoxicity, differentiation, cell viability, cell migration, and adhesive ability. Rabbit chondrocyte cells, cultivated until passage two, were seeded onto a 6 mm diameter membrane. Assessments included DAPI-PKH26 staining, histological staining, live/dead assay, WST-1 assay, and proteomics analysis. (3) Results: DAPI-PKH26 staining showed successful adhesion and the uniform distribution of cells on the membrane. Safranin-O and H&E staining confirmed that the membrane supports chondrocyte adhesion and extracellular matrix production with high cell density and typical chondrocyte morphology. The live/dead assay demonstrated a high proportion of viable cells at 24 and 48 h, with increased cell proliferation over time. The WST-1 assay showed a significant increase in OD450 values, confirming cell proliferation and biocompatibility. Proteomic analysis revealed the significant enrichment of genes associated with extracellular matrix organization, cell adhesion, and cartilage development. (4) Conclusions: This novel biomaterial holds the potential to enhance cartilage regeneration and offer a viable alternative to periosteal patches. Full article

Pseudomonas syringae pv. actinidiae (Psa) is responsible for causing kiwifruit canker disease. The detection of Psa is commonly carried out using normal PCR and culture-based isolation. However, normal PCR does not differentiate between live and dead cells, potentially resulting in the incorrect estimation of the amount of infectious substance in a sample. Such an incorrect estimation could result in unnecessary phytosanitary strategies and control measures. This study attempts to establish a specific assay for detecting only live Psa bacterial cells. To achieve this, a pair of strain-specific primers designed from HopZ3 effector were used, and the traditional PCR method was assessed using a nucleic acid-binding dye (propidium monoazide—PMA), establishing a PMA–PCR system and conditions for detecting live Psa in this study. Sensitivity tests showed a detection limit of 10 cfu/mL and 1 pg/μL. This method was also tested in diseased kiwifruit tissues and can be seen as a rapid and dependable replacement to PCR methods for detecting only those infective kiwifruit materials with viable Psa. Full article