Search Results (3,417)

Background/Objectives: Endodontic sealers can interact with periapical tissues through extrusion, yet the molecular mechanisms underlying their biological effects remain poorly defined. This study investigated how commonly used sealers influence mitogen-activated protein kinase (MAPK) signaling, cell viability, and osteogenic-associated responses in MC3T3-E1 pre-osteoblasts. Methods: Four commercial sealers, Calcium-silicate-based Bioceramic Sealer (EndoSequence^® BC Sealer, BC), Zinc oxide eugenol sealer (Kerr Pulp Canal Sealer, ZOE), Sealapex™, and AH26^®, were applied as standardized pellets, allowed to set, and cultured with MC3T3-E1 cells. Calcium deposition was assessed by Alizarin Red S (ARS) staining, and MAPK activation was evaluated by Western blotting. Due to excessive solubility (Sealapex™) or poor cell survival (AH26^®), mechanistic analyses were performed only for BC and ZOE. Osteogenic-associated gene expression was measured by qRT-PCR, and the functional role of MAPK signaling was assessed using ERK, JNK, and p38 inhibitors. Results: BC and Sealapex™ produced robust ARS staining, while ZOE and AH26^® produced minimal mineral-associated staining. Both BC and ZOE activated ERK, JNK, and p38, with ZOE inducing higher phosphorylation. However, BC maintained greater cell viability and increased Runx2 and Osx expression, whereas ZOE impaired early cell attachment and viability. MAPK inhibition in BC-treated cultures reduced osteogenic-associated gene expression and ARS staining, indicating MAPK involvement in BC-mediated responses. Conclusions: BC and ZOE elicit distinct MAPK activation patterns and cellular responses. Under the conditions tested, BC promoted a more favorable osteogenic-associated response, whereas ZOE compromised early cell viability. These mechanistic insights may help explain clinical differences in periapical tissue responses to sealer extrusion. Full article

Background/Objectives: The formation of a soft tissue seal through mucosal integration around dental implants is critical for potentially achieving long-term peri-implant health and clinical success. Understanding how different implant and abutment surfaces interact with individual layers of the oral mucosa remains limited. This study aimed to compare the differential attachment of tissue-engineered oral epithelium, connective tissue, and full-thickness human oral mucosa to various implant and abutment materials and surface topographies. Methods: Sand-blasted, large-grit, acid-etched (TiZr-SLA), machined TiZr (TiZr-M), machined zirconia (ZrO₂-M), polished zirconia (ZrO₂-P), and machined PEEK rods, along with commercially available titanium and ZrO₂ healing abutments, were inserted into 3D oral mucosal models following a 4-mm punch biopsy. Inflammation was induced using Escherichia coli lipopolysaccharide. Analyses included histology, PrestoBlue viability assay, scanning electron microscopy, and ELISA quantification of cytokines IL-1β, IL-6, and IL-8. Results: Epithelial attachment was greater on TiZr-SLA, ZrO₂-P, and PEEK-M (p < 0.05 and p < 0.01) surfaces compared with TiZr-M and ZrO₂-M. TiZr-SLA exhibited the highest connective tissue attachment (p < 0.05). Commercial titanium and ZrO₂ healing abutments demonstrated the highest post-pull PrestoBlue viability and overall soft tissue attachment. SEM confirmed cell retention on all implant surfaces. Elevated IL-1β levels were detected in models exposed to ZrO₂-M and PEEK-M, whereas IL-6 and IL-8 levels were not influenced by any material or surface topography. Conclusions: In vitro epithelial and connective tissue responses are influenced by implant material, surface topography, and design. Rough TiZr-SLA surfaces promote superior connective tissue attachment, while smooth commercial abutments support optimal overall soft tissue integration. These findings highlight the importance of surface engineering in preclinical optimization of peri-implant soft tissue attachment. Full article

Primary hepatocyte cultures serve as an ex vivo model of liver physiology. This study aims to employ poly(vinyl alcohol) (PVA) nanofiber membranes (NMs) to establish a three-dimensional (3D) culture system that supports the long-term functionality of primary hepatocytes. Primary hepatocytes were monocultured on a PVA NM or indirectly cocultured with NIH3T3 fibroblasts on a distinct polycaprolactone (PCL) NM layer. Monocultured and cocultured hepatocytes maintained prolonged survival without supplemental growth factors. Cocultured hepatocytes formed larger aggregates composed of cell clusters attached to untreated nanofibers than monocultured cells. However, most primary hepatocytes cultured on NaOH-treated PVA NM and Arg–Gly–Asp (RGD) peptide-blended PVA (RGD-PVA) NM, under monoculture and coculture conditions, formed non-aggregated cells in a single-cell layer. In a bioinert assay, unstimulated dendritic cells were activated on untreated but not NaOH-treated PVA NM. CYP3A4 activity was higher in cocultured cells on RGD-PVA NM with fibroblasts than in monocultured cells on PVA and RGD-PVA NM. Functional hepatocyte cultures were successfully maintained in a 3D single-cell layer on RGD-PVA NM, along with fibroblasts in a layer-by-layer coculture, for a prolonged period. The prolonged culture of hepatocytes in a 3D single-cell layer may facilitate further drug discovery, toxicity studies, and translational liver research. Full article

End-stage liver disease caused by advanced fibrosis and cirrhosis remains a major global burden, yet its treatment is limited by donor organ shortages. Bioengineered liver scaffolds offer a promising alternative, but their efficacy is often limited by thrombosis, insufficient vascularization, and poor graft integration due to inadequate endothelialization. To overcome these challenges, we employed LXW7 αvβ3 integrin targeting peptide with high endothelial cell specificity and low platelet affinity to enhance re-endothelialization and hemocompatibility of decellularized liver scaffold (DLS) and thereby improve hepatic integration and function. LXW7 was covalently conjugated to the decellularized rat liver scaffold via EDC/NHS-mediated carbodiimide coupling and subsequently reseeded with human umbilical vein endothelial cells (HUVECs) and cultured in a perfusion bioreactor to promote endothelialization. LXW7 immobilization significantly improved HUVECs attachment and proliferation, achieving approximately 81% vascular coverage, while sustaining the endothelial function. Ex vivo blood perfusion showed minimal thrombus formation and markedly reduced platelet adhesion, demonstrating enhanced hemocompatibility. Following confirmation of endothelialization, scaffolds were recellularized with hepatocellular carcinoma (HepG2) cells and HUVECs. LXW7 modified scaffolds promote organized hepatocyte distribution, sustained albumin expression, and increased urea secretion. In vivo implantation of LXW7-DLS into the omentum of mice promoted robust host endothelial recruitment and enhanced neovascularization, highlighting the scaffold’s excellent biocompatibility and good integration with surrounding tissues. Moreover, in vivo implantation of LXW7 recellularized scaffolds into a thioacetamide-induced fibrotic mouse liver resulted in reduced collagen deposition and lowered serum ALT/AST levels, demonstrating hepatic regeneration and extracellular matrix remodeling. Overall, our results showed that LXW7-modified DLS promotes stable endothelialization, improves hemocompatibility, and enhances hepatic function, underscoring its translational potential for the development of vascularized transplantable liver grafts. Full article

This study aimed to establish a model for investigating X chromosome inactivation using transgenic mouse strains expressing green fluorescent protein (GFP). The D4/XGFP-Tg (XGFP) strain carries the GFP transgene on the X chromosome; therefore, due to random X chromosome inactivation, female offspring from crosses between XGFP males and CD-1 females exhibit mosaic GFP expression. In contrast, the B5/EGFP-Tg (EGFP) strain harbours autosomal integration of the same reporter construct, resulting in uniform GFP expression in progenies. Analysis of CD-1 × XGFP attached blastocysts revealed strong GFP expression in giant trophoblast cells and primordial germ cells (PGCs) at E6.5, demonstrating paternal X-chromosome reactivation. In 14.5-day-old CD-1 × XGFP female embryos and CD-1 × EGFP embryos, intense CAG promoter-driven GFP signals were detected in the brain, heart, gonads, somites, and limbs. In line with random X-chromosome inactivation, only 56% of embryonic fibroblast cells, derived from CD-1 × XGFP female embryos, exhibited GFP expression. These findings validate that CD-1 × XGFP mice represent a valuable in vivo model for studying X chromosome inactivation during early embryonic development and PGC specification. Furthermore, CD-1 × XGFP embryonic fibroblasts represent a valuable in vitro model for investigating the molecular mechanisms governing X-chromosome activation and inactivation. Full article

Stilbenoids are plant-derived chemical compounds that are classified as phytoalexins; recent focus has been drawn, especially on astringin, piceid, piceatannol, pterostilbene, pinosylvin, and resveratrol. These substances have been extensively studied for a variety of beneficial properties, including their effects on pathogenic microorganisms, parasites, and viruses. In their antifungal capacity, they are effective against Aspergillus spp., Botrytis spp., Candida spp., Trichophyton spp., and other fungi; tested stilbenoids have exhibited fungicidal and fungistatic effects, and inhibition of biofilm formation. Against parasites, they are effective against Echinococcus spp., Leishmania spp., Schistosoma spp., Trypanosoma spp., Toxoplasma spp., among others. Relevant action mechanisms include a reduction in parasitic enzymatic activity and inhibition of proliferation. They are also effective against different DNA and RNA viruses; the relevant mechanisms comprise reduction in viral replication, inhibition of viral genome expression, and viral attachment to cells. The toxicity of stilbenoids has been reviewed in recent papers, and, in most cases, the effective concentrations applied are well below the toxicity limit. Full article

Daphnis nerii can severely damage pine forests worldwide. Daphnis nerii cypovirus-23 (DnCPV-23) is an important viral pathogen for controlling D. nerii. However, the mechanism underlying DnCPV-23 cell entry has not been elucidated. In this study, we determined that VP3 mediates the binding of DnCPV-23 to host brush border membrane vesicles. Far-Western blotting and mass spectrometry results revealed that a Manduca sexta alpha-glucosidase (MsAGL) can interact with VP3. The interaction between MsAGL and VP3 was verified by co-immunoprecipitation and glutathione S-transferase pull-down assays. Notably, MsAGL influenced DnCPV-23 entry into host cells, including attachment and the subsequent internalization of the virus. Furthermore, MsAGL inhibited DnCPV-23 infections of M. sexta cells and D. nerii larvae. In summary, we confirmed that VP3 of DnCPV-23 mediates cell entry, while also identifying MsAGL as an entry factor for DnCPV-23. The study findings provide useful insights relevant for further elucidating the cell entry mechanisms of cypoviruses. Full article

Background/Objectives: Drug development and delivery remain critical areas of research for addressing modern bioanalytical challenges. Understanding drug biodistribution, stability, and metabolism within biological systems is essential for optimising therapeutic efficacy. This study focuses on synthesising and characterising a novel fluorescent conjugate derived from commercially available rapid-acting insulin glulisine (Apidra^®) and fluorescein isothiocyanate (FITC). The objective was to produce a mono-labelled FITC-insulin glulisine conjugate without employing complex protective group strategies or multi-step processes. Methods: The conjugation was optimised by varying molar ratios (1:1 to 3:1) and reaction times (18–24 h) at pH 7. Results: The desired B1 mono-labelled conjugate was successfully achieved at a 2:1 molar ratio, pH 7, and 18 h reaction time. MALDI-TOF mass spectrometry confirmed the molecular weight and conjugation site, with fragmentation analysis identifying FITC attachment at phenylalanine (B1) on the β-chain (m/z = 537.11). Western blots performed on C2C12 skeletal cell lysates stimulated with the FITC–insulin glulisine conjugate showed Akt and IRS-1 activity similar to that of cells treated with native commercial insulin glulisine. Confocal imaging also demonstrated translocation of GLUT4 in FITC–insulin glulisine conjugate-treated C2C12 cells similar to that of commercial native insulin glulisine. Octanol-water partitioning studies assessed the physicochemical properties of the conjugate. Conclusions: This approach demonstrates an efficient method for fluorescent labelling of insulin analogues, enabling future applications in imaging, biodistribution studies, and pharmacokinetic profiling. Full article

Background: The aim of this study was to comprehensively investigate the potential degenerative effects of periodontitis severity on retinal and choroidal structures in patients with different types of diabetic retinopathy (DR). Materials and Methods: The study’s Clinical Trials Registration Number is NCT07137013. A total of 100 participants (56 females and 44 males), each group consisting of 20 individuals, were allocated into five groups: systemically healthy controls (G1), diabetic patients without DR (G2: DM+ DR−), non-proliferative DR without diabetic macular edema (G3: NPDR DME−), non-proliferative DR with diabetic macular edema (G4: NPDR DME+), and proliferative DR (G5: PDR). Ocular examinations were performed using optical coherence tomography (OCT) and OCT angiography (OCTA). Retinal layer thicknesses, choroid-sclera interface (CSI), ganglion cell layer (GCL), retinal nerve fiber layer (RNFL), and peripapillary CSI were assessed by OCT, whereas superficial and deep retinal vessel densities and the foveal avascular zone (FAZ) were evaluated by OCTA. Clinical periodontal status was assessed using plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment loss (CAL). Results: In the G3 and G5 groups, the presence of stage III–IV periodontitis was associated with a marked increase in retinal layer thickness. GCL + Inner Plexiform Layer (GCL+) thickness was significantly reduced in individuals with stage III–IV periodontitis in almost all regions of the G5 group, except for the 3 mm nasal and inferior areas. Peripapillary CSI values showed a significant decrease with increasing periodontitis severity. RNFL thickness was significantly reduced in individuals with stage III–IV periodontitis, particularly in the G5 group. OCTA analyses demonstrated significant reductions in superficial and deep retinal vessel densities in several regions in the presence of stage III–IV periodontitis. Moreover, FAZ areas were significantly enlarged in individuals with stage III–IV periodontitis in the G2 and G5 groups. Conclusions: Periodontal inflammation, particularly in advanced periodontitis (stage III–IV), induces degenerative changes in the retinal microvasculature and neural tissues. Increasing periodontitis severity may represent a potential provoking factor in the pathogenesis of DR. Full article

Surgical-site infections (SSIs) represent a significant healthcare burden, often complicating wound healing and recovery. To overcome the limitations of systemic antibiotic administration, such as toxicity and poor localization, this study aimed to develop a bioactive dressing utilizing bacterial nanocellulose (BNC) impregnated with gentamicin-loaded chitosan nanoparticles (GNP). Chitosan nanoparticles were synthesized via ionic gelation with sodium tripolyphosphate (TPP) and optimized using a one-factor-at-a-time (OFAT) approach to control particle size, polydispersity index (PDI) and zeta potential. The optimized nanoparticles were impregnated into BNC disks, and the resulting composite was characterized using FTIR and Raman spectroscopy, XRD and SEM. Antimicrobial efficacy was evaluated against Klebsiella pneumoniae, while biocompatibility was assessed using MTT assays and cell-adhesion studies on human fibroblasts. The optimization process yielded stable, monodisperse nanoparticles with a mean size of 80.07 nm and a PDI of 0.192. SEM imaging confirmed the successful integration of nanoparticles into the BNC nanofibrillar network without compromising the membrane’s structural integrity. The BNC-GNP composite demonstrated significant antimicrobial activity against K. pneumoniae, comparable to free gentamicin solution. Furthermore, in vitro studies revealed good biocompatibility, with cell viability exceeding 70% and sustained fibroblast adhesion, although cell-attachment density decreased with higher nanoparticle concentrations. The developed BNC dressing containing gentamicin-loaded chitosan nanoparticles presents a promising multifunctional biomaterial. It effectively combines local infection control with a biocompatible environment suitable for tissue regeneration, offering a novel approach for the postoperative treatment of surgical-site infections. Full article

A tissue-engineered heart valve is a fully functional tissue facilitated through the cultivation of autologous cells on appropriate scaffolds. Scaffold’s surface charge and wettability are the main factors that significantly affect cell adhesion, which is known to be favourable on hydrophilic surfaces. Moreover, biocompatible scaffolds that induce minimal immunogenic response are also essential for successful tissue engineering (TE). However, commonly used biocompatible polymers with preferable bulk properties lack desirable surface properties. For example, poly-ε-caprolactone (PCL), which is widely used as a scaffold in TE, is known for its satisfying structural and mechanical properties, but due to its surface characteristics, cell attachment and, consequently, cell growth on this polymer are limited. In this study, we investigated the possible effect of H₂-N₂ plasma treatment on the surface wettability of electrospun PCL nanofibres to see the feasibility of improvement in cell adhesion and proliferation. Our results showed an increase in the hydrophilicity of the 650 nm PCL specimens after plasma treatment, which was followed by a significant enhancement in cell attachment without altering PCL mechanical properties. Plasma surface modification is a promising approach that can be used to improve hiMSCs growth without altering the desired bulk properties and fibre morphology of 650 nm PCL specimens. Full article

Embryo implantation presents a cell biological paradox: contact formation between the trophoblast of the blastocyst and the epithelial lining of the endometrium contradicts typical epithelial cell behaviour, as does the subsequent invasion needed for placenta formation in most species (including the human). Explaining this conundrum became a challenge for investigation since its recognition about 40 years ago and it receives increasing interest because implantation failure appears to be a major cause for low success in assisted reproduction. The present article reviews the main findings that have directed attention of researchers on epithelial cell polarity and on the theoretical concept of epithelial–mesenchymal transition (EMT). Apart from trophoblast attachment competence, a special focus is on endometrial receptivity. Comparison with epithelial fusion processes (EFPs) in development and with tumour cell invasion has been and is still considered helpful in order to take advantage of the progress made in those fields. Concerning the mechanisms involved, it must be emphasized that trophoblast and uterine luminal epithelium (ULE) do not undergo a complete switch to a mesenchymal programme (do not undergo a complete EMT) but make use of partial changes in the epithelial programme. The large number of data accumulated recently should allow us to now make progress in identifying what these partial programme changes are exactly and how they are regulated; also, they may offer chances for obtaining deeper insights into the regulation of implantation. Full article

Vitamin D₃ is a fat-soluble steroid essential for bone metabolism, immune modulation, and inflammation control, all critical for periodontal health. Its active form, 1,25-dihydroxyvitamin D₃, binds to the vitamin D receptor (VDR) in periodontal cells, including periodontal ligament stromal cells, fibroblasts, osteoblasts, and macrophages, enhancing osteogenesis, antimicrobial defenses, and anti-inflammatory responses. Clinical and experimental evidence demonstrates that adequate systemic vitamin D₃ levels and local activation in gingival tissues improve outcomes of nonsurgical and surgical periodontal therapies, reducing probing pocket depth (PPD), clinical attachment loss (CAL), and gingival inflammation. Dose-dependent supplementation shows greater clinical efficacy, and emerging evidence supports potential topical applications. This review integrates molecular mechanisms with clinical findings, highlighting the therapeutic potential of vitamin D₃ in periodontal disease management. Full article

Background: Hyaluronic acid (HA) and β-tricalcium phosphate (β-TCP) are widely used biomaterials in periodontal and alveolar regeneration; however, their complementary biological roles across soft- and hard-tissue healing have not been jointly assessed in a single review. Objective: to systematically evaluate clinical and translational evidence regarding the adjunctive use of HA in periodontal therapy and the regenerative performance of β-TCP in alveolar bone reconstruction. Methods: A systematic search was conducted across PubMed/MEDLINE, Scopus, Web of Science Core Collection, and Embase for studies published between 1 January 2015 and 1 October 2025. Randomized and non-randomized clinical studies evaluating HA as an adjunct to periodontal therapy and β-TCP in ridge preservation or augmentation were included. In vitro studies were considered when providing mechanistic insight relevant to clinical outcomes. Screening, data extraction, and qualitative synthesis were performed according to PRISMA 2020 guidelines. Results: Database searching identified 312 records. After removal of duplicates, 241 records were screened, of which 179 were excluded. Sixty-two full-text articles were assessed for eligibility, and twenty studies met the inclusion criteria (twelve clinical; eight in vitro). Across non-surgical periodontal therapy trials, adjunctive HA demonstrated modest but consistent additional improvements in probing depth reduction (~0.8–1.5 mm) and clinical attachment gain (~0.5–1.2 mm) compared with mechanical therapy alone, particularly in deeper defects and systemically compromised patients. Clinical studies on β-TCP reported predictable dimensional bone preservation and stable implant feasibility, supported by histologic evidence of scaffold-guided new bone formation. In vitro findings indicated that HA modulates biofilm-induced inflammation and supports fibroblast and epithelial cell function, whereas β-TCP promotes osteoblast activity and controlled osteoclast-mediated remodeling. Conclusions: HA and β-TCP demonstrate complementary regenerative roles, with HA primarily enhancing soft-tissue resolution and inflammatory modulation and β-TCP providing osteoconductive structural support for bone regeneration. Current evidence supports their selective integration in personalized regenerative approaches; however, standardized outcome reporting and longer-term trials are required to establish the clinical value of sequential or combined application. Full article

Non-absorbable sutures provide a site for bacterial attachment and increase the risk of surgical site infections. An alternative prevention of infections requires plant-extract coatings on sutures. The objectives of this study were to develop P. betle leaf extract-coated non-absorbable sutures and to investigate their activities on staphylococci. P. betle leaves were extracted and analyzed for the phytochemicals. P. betle extract was coated on sutures, including polyester and polypropylene. The stability of hydroxychavicol on coated sutures was evaluated. Four treatments were designed, including (1) uncoated, (2) antibiotic/extract-free-coated, (3) extract-coated, and (4) gentamicin-coated sutures. Each treatment was tested for antibacterial, antibiofilm, and anti-adhesion activities on Staphylococcus aureus and Staphylococcus pseudintermedius. In addition, the cytotoxicity of extract-coated sutures was tested. Analysis of the extract identified hydroxychavicol (40.07%) as the primary phytochemical. Stability tests indicated higher hydroxychavicol on Day 1 of extract-coated polyester compared to polypropylene, and the levels decreased on the subsequent days (p < 0.05). Antibacterial activity of extract-coated polyester showed antibacterial effects during the experiment period (5.16 ± 2.35 mm), while polypropylene showed no effectiveness. Additionally, biofilm inhibition was found to be 36.63 ± 27.08% and 37.34 ± 26.98% in tested staphylococci for extract-coated polyester and polypropylene, respectively. Anti-adhesion showed that the extract-coated sutures had a higher ability to decrease tested bacteria attachment (56.25–60.42% living cell reduction). The cytotoxicity study revealed that extract-coated sutures of ≤1.5 mg/1.5 cm had a 99% survival rate. The findings indicate that the coated sutures showed antibacterial, antibiofilm, and anti-adhesion effects against staphylococci causing canine skin infections and might lead to alternative surgical use in veterinary medicine. Full article