Search Results (620)

Metformin is a promising small molecule for dentin regeneration, but an effective local delivery system for pulp applications has been underexplored. This study encapsulated metformin in poly(lactic-co-glycolic acid) (PLGA) microspheres and incorporated them into calcium phosphate–chitosan cement (CPCC) as a direct pulp-capping material (DPC). Metformin-PLGA microspheres were prepared by double emulsion and mixed with CPCC at a concentration of 0% to 20% by weight. Microsphere morphology, encapsulation efficiency, chemical composition, and physico-mechanical properties were characterized, and compatibility with human dental pulp stem cells (hDPSCs) was evaluated by live/dead assay and SEM. The microspheres were spherical (5.43 ± 0.17 µm) with (51 ± 3.69%) encapsulation efficiency, and FTIR confirmed metformin incorporation. The 15% Met-PLGA-CPCC group showed flexural strength (15.22 ± 1.98 MPa), elastic modulus (4.60 ± 0.73 GPa), and work of fracture (104.96 ± 12.48 J/m²) comparable to or higher than CPCC and MTA, while all Met-PLGA-CPCC groups had shorter setting times ranging from 18 min to 27 min than CPCC (39.15 ± 2.10 min) and MTA (123 ± 4.2 min). Metformin release increased proportionally with Met-PLGA content. hDPSCs exhibited good attachment and high viability on all materials over the evaluated period. In conclusion, Met-PLGA-CPCC provides fast-setting and favorable physico-mechanical properties, sustained metformin delivery, and excellent hDPSC compatibility. These properties support its potential as a bioactive direct pulp-capping material and as a versatile platform for regenerative applications. Full article

Horses’ hypsodont (high-crowned) teeth face permanent dental wear. This is compensated for by a continuous eruption, which requires a high adaptability of odontoblasts; otherwise, the dental pulp would be exposed. Here, we report on how equine odontoblasts respond to the challenge of maintaining a high production rate of dentin. We analyzed CD90, a marker of odontoblastic differentiation, and nestin, a marker of mature odontoblasts, in equine pulpal tissue via immunofluorescence. For comparison, we examined the hypselodont (ever-growing) incisors and brachydont (short-crowned) molars of rats. Immunofluorescence and Western blot analysis of pulpal tissue revealed a higher content of CD90-positive cells in hypsodont equine teeth than in brachydont and hypselodont rat teeth. The odontoblastic layer of hypsodont teeth was positive for CD90 (marker for differentiating odontoblasts), which was not the case for brachydont and hypselodont rat teeth. Most samples of hypsodont teeth were negative for nestin, whereas in hypselodont and brachydont teeth, odontoblasts were positive for nestin (marker for mature odontoblasts). Our findings suggest that there is a constant replacement of odontoblasts in the equine dentition, enabling a continuous high production rate of dentin. These results contradict the idea of lifelong vital, postmitotic and productive odontoblasts. Full article

Hydraulic calcium silicate cements (HCSCs) are contemporary materials in vital pulp therapy (VPT) and regenerative endodontic therapy (RET) due to their favorable effects on pulpal and periodontal cells, including cell differentiation and hard tissue formation. Recent studies also indicated the involvement of several S100A proteins in inflammatory, differentiation, and mineralization processes of the pulp. The aim of the present study was to investigate the effects of HCSCs on S100A gene expression in human dental pulp stem cells (hDPSCs). Human DPSCs were isolated and characterized by multi-lineage stem-cell markers and differentiation protocols. In stimulation experiments hDPSCs were exposed to ProRoot^®MTA, Biodentine^®, IL-1β, and dexamethasone. Cell viability was determined by XTT assay. IL-6 and IL-8 mRNA expression was measured to analyze proinflammatory response. In addition, odontogenic differentiation and biomineralization assays were conducted (DSPP- and ALP-mRNA expression, ALP activity, and Alizarin Red staining). Differential expression of 13 S100A genes was examined using qPCR. Low concentrations of HCSCs enhanced the proliferation of hDPSCs, whereas higher concentrations exhibited cytotoxic effects. HCSCs induced a pro-inflammatory response and led to odontogenic differentiation and biomineralization. This was accompanied by significant alterations in the expression levels of various S100A genes. ProRoot^®MTA and Biodentine^® significantly affect the expression of several S100A genes in hDPSCs, supporting their role in inflammation, differentiation, and mineralization. These findings indicate a link between the effects of HCSCs on human pulp cells during VPT or RET and S100A proteins. Full article

Mesenchymal stem cells (MSCs) are multipotent adult cells that are highly valued for their immunomodulatory potential and intrinsic ability to home to inflamed sites. This study specifically utilized human dental pulp stem cells (hDPSCs), a unique MSC subtype derived from the neural crest, due to their reported superior anti-inflammatory capacity. To rigorously test their efficacy, we employed the AIRmax murine model, which exhibits a genetically determined high-inflammatory phenotype. Acute inflammation was induced by subcutaneous injection of the polyacrylamide suspension Biogel P-100. Two hours post-induction, AIRmax mice were treated intravenously with hDPSCs. Our results demonstrate that hDPSC treatment produced significant anti-inflammatory effects evident at 24 h. The treated group showed a pronounced reduction in leukocyte migration and decreased protein extravasation in the inflammatory exudate. Crucially, hDPSCs also modulated molecular mediators, significantly decreasing the pro-inflammatory cytokine TNF-alpha and reactive oxygen species (ROS) production. Furthermore, while hDPSCs efficiently and rapidly homed to the inflammation site within 2 h, their maximal therapeutic benefits only manifested after 24 h. This suggests that their robust capacity to modulate acute inflammatory responses relies not only on rapid migration but also on a paracrine “hit-and-run” mechanism that suppresses cellular infiltration and oxidative stress over time. This study reinforces the potential of hDPSCs as a powerful, multi-target therapeutic agent for inflammatory conditions, supporting further investigation into their precise mechanisms and clinical application. Full article

Extracellular vesicle (EV)-based therapies have emerged as promising, cell-free approaches for dental tissue regeneration. This narrative review integrates mechanistic insights, therapeutic efficacy data, and safety and delivery considerations from in vitro and in vivo studies to elucidate the molecular mechanisms by which EVs, particularly those from dental pulp stem cells (DPSCs) and mesenchymal stem cells (MSCs), drive regenerative processes via key signalling axes (PI3K/Akt, MAPK, BMP/Smad, and Hedgehog). Preclinical studies demonstrate that unmodified and engineered EVs enhance odontogenic differentiation, angiogenesis, bone repair, and immunomodulation in models of pulp regeneration, alveolar bone defects, osteonecrosis, and periodontitis. Isolation and purification methodologies were also evaluated, comparing ultracentrifugation, size-exclusion chromatography, and density-cushion approaches, and discussing how protocol variations affect EV purity, dosing metrics, and functional reproducibility. Early-phase clinical evaluations report only low-grade transient adverse events, underscoring a generally favourable safety profile. Despite these encouraging results, significant challenges remain: heterogeneity in EV cargo composition, lack of standardised potency assays, and incomplete long-term safety data. The review highlights the urgent need for rigorous, harmonised regulatory frameworks and robust quality control measures to ensure that EV-based modalities can be translated into safe, effective, and reproducible therapies in regenerative dentistry. Full article

Background/Objectives: We investigated whether palmitic acid (PA) induced the expression of inflammatory cytokines and was involved in pyroptosis in a human dental pulp cell line. Methods: Human dental pulp cells cultured in Minimum Essential Medium Alpha (αMEM) were treated with 1 µg/mL LPS and/or PA (100, 300 and 500 µM). As a control, αMEM was added in the culture medium. The WST-1 assay was performed to assess cell proliferation, and morphological changes in cells were examined. RNA expression of IL-1β, IL-6, TNF-α, caspase-4 and gasdermin d were detected by quantitative RT-PCR (qPCR). Results: The WST-1 assay showed that cell viability decreased by 36% at 300 µM and 47% at 500 µM PA compared to the control (p < 0.05). Cell morphology revealed slight shrinkage in 100, 300 and 500 µM PA groups. RNA expression of IL-1β and IL-6 in the PA groups was significantly higher than that in the control groups (p < 0.05), while RNA expression of TNF-α in the PA group was the same as that of control group. The mRNA expression of caspase-4 and gasdermin d in PA groups was significantly higher than that in control group (p < 0.05). Likewise, the concentration of IL-1β and IL-6 was significantly higher in both LPS and PA groups than that in the LPS or PA groups (p < 0.05). Conclusions: The results of this study suggest that PA induces the expression of inflammatory cytokines and is involved in pyroptosis in a human dental pulp cell line. Full article

Bone defects in the maxillofacial region severely impair patient function and esthetics. Free autologous bone grafting remains the gold-standard treatment; however, surgical intervention at donor sites limits clinical applicability. Treatment using artificial materials also presents challenges, including insufficient bone regeneration and poor biocompatibility. Bio three-dimensional (3D) printing, which enables the fabrication of scaffold-free 3D constructs from cellular spheroids has emerged as a promising regenerative approach. This study investigated the osteogenic potential of scaffold-free constructs composed of human dental pulp stem cell (DPSC) spheroids in a rat mandibular defect model. DPSCs isolated from extracted human teeth were used to generate spheroids, which were assembled into 3D constructs using a Bio 3D printer. The spheroids exhibited higher mRNA expression of stem cells and early osteogenic markers than monolayer cultures. The constructs were transplanted into mandibular defects of immunodeficient rats, and bone regeneration was assessed eight weeks post-transplantation. Radiographic and micro-Computed Tomography analyses revealed significantly greater bone volume and mineral density in the 3D construct group. Histological and immunohistochemical examinations confirmed newly formed bone containing osteogenic cells derived from the transplanted DPSCs. These findings indicate that Bio 3D-printed, scaffold-free DPSC constructs promote mandibular bone regeneration and may provide a novel strategy for maxillofacial reconstruction. Full article

Autophagy is a well-preserved biological mechanism that is essential for sustaining homeostasis by degradation and recycling damaged organelles, misfolded proteins, and other cytoplasmic detritus. Cannabinoid signaling has emerged as a prospective regulator of diverse cellular functions, including immunological modulation, oxidative stress response, apoptosis, and autophagy. Dysregulation of autophagy contributes to pathogenesis and treatment resistance of several oral diseases, including oral squamous cell carcinoma (OSCC), periodontitis, and gingival inflammation. This review delineates the molecular crosstalk between cannabinoid receptor type I (CB1) and type II (CB2) activation and autophagic pathways across oral tissues. Cannabinoids, including cannabidiol (CBD) and tetrahydrocannabinol (THC), modulate key regulators like mTOR, AMPK, and Beclin-1, thereby influencing autophagic flux, inflammation, and apoptosis. Experimental studies indicate that cannabinoids inhibit the PI3K/AKT/mTOR pathway, promote reactive oxygen species (ROS)-induced autophagy, and modulate cytokine secretion, mechanisms that underline their dual anti-inflammatory and anti-cancer capabilities. In addition, cannabinoid-induced autophagy has been shown to enhance stem cell survival and differentiation, offering promise for dental pulp regeneration. Despite these promising prospects, several challenges remain, including receptor selectivity, dose-dependent variability, limited oral bioavailability, and ongoing regulatory constraints. A deeper understanding of the context-dependent regulation of autophagy by cannabinoid signaling could pave the way for innovative therapeutic interventions in dentistry. Tailored cannabinoid-based formulations, engineered for receptor specificity, tissue selectivity, and optimized delivery, hold significant potential to revolutionize oral healthcare by modulating autophagy-related molecular pathways involved in disease resolution and tissue regeneration. Full article

Dental pain often arises from the compromised integrity of the tooth pulp due to dental injury or caries. The dentin–pulp complex has long been considered to be central to the unique biology of dental pain. Most trigeminal ganglion afferents projecting into tooth pulp are myelinated neurons, which lose their myelination at the site of peripheral dentin innervation. The pulpal afferents likely combine multiple internal and external stimuli to mediate nociception and maintain pulp homeostasis. Transient receptor potential (TRP) ion channels in neurons and odontoblasts, along with mechanosensitive ion channels such as Piezo, form a key molecular hub for pulpal nociception by sensing thermal, chemical, and hydrodynamic stimuli. Among these, TRP vanilloid 1 (TRPV1) mediates nociception and the release of calcitonin-gene-related peptides (CGRPs), while TRP canonical 5 (TRPC5) mediates cold pain. TRP melastatin 8 (TRPM8) mediates the transduction of hyperosmotic stimuli. Pulpitis elevates endogenous TRPV1 and TRPA1 agonists, while inflammatory mediators sensitize TRP channels, amplifying pain. CGRP recruits immune cells and promotes bacterial clearance and reparative dentinogenesis, yet the roles of TRP channels in these processes remain unclear. Future studies should use advanced multi-omics and in vivo or organotypic models in animal and human teeth to define TRP channel contributions to pain, immune responses, and regeneration. Understanding neuronal and non-neuronal TRP channel interactions and their integration with other ion channels may enable novel analgesic and regenerative strategies in dentistry. Full article

The intra- and intercellular signaling molecule nitric oxide (NO) is produced in endothelial cells by the activity of endothelial NO synthase (eNOS). Upon formation, NO diffuses into the underlying vascular smooth muscle cells, where it activates NO-sensitive guanylyl cyclase (NO-GC) resulting in the production of cyclic guanosine 3′,5′-monophosphate (cGMP) from guanosine 5′-triphosphate (GTP). Inducing vasodilatation, inhibiting platelet aggregation and leukocyte adhesion, and inhibiting the proliferation and migration of vascular smooth muscle cells, the NO-cGMP signaling leads to a number of anti-inflammatory processes. Inflammation-dependent elevated concentrations of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in blood vessels of inflamed dental pulp induce an uncoupling of eNOS and oxidized NO-GC, leading to a disruption of NO-cGMP signaling. Endothelial dysfunction in inflamed dental pulp alters cell–cell and cell–matrix interactions, reducing the regenerative and reparative potential of the dentin–pulp complex in response to carious lesions. In the therapeutic management of caries, it is essential to consider the presence of endothelial dysfunction in the inflamed dental pulp. The utilization of NO-GC stimulators and activators in indirect and direct pulp capping materials may enhance the regeneration and repair potential of inflamed dental pulp. Full article

Direct pulp capping using mineral trioxide aggregate (MTA) is commonly used to preserve dental pulp tissue, but the molecular mechanisms underlying reparative dentin formation during this procedure and the restoration of dental pulp homeostasis remain unclear. To elucidate these mechanisms, we investigated the expression and function of intercellular adhesion molecule 2 (ICAM2) in dental pulp cells and tissue. ICAM2 expression in human dental pulp cells (HDPCs) was confirmed by the gene and protein expression analysis. ICAM2 expression during reparative dentin formation after direct pulp capping was investigated using a rat direct pulp capping model. The effect of ICAM2 on odontoblast-like differentiation of HDPCs was assessed using siRNA and magnetic cell sorting (MACS). The gene and protein expression analysis showed that ICAM2 is expressed in approximately 10% of HDPCs. Immunofluorescence staining of rat mandibular bone sections showed that ICAM2 is expressed in dental pulp tissue. During reparative dentin formation, ICAM2 expression significantly increased to nearly three-fold higher than pretreatment levels on the 3 days after direct pulp capping and then returned to normal levels. ICAM2 knockdown by siRNA enhanced odontoblast-like differentiation of HDPCs. In contrast, culture supernatant from ICAM2-positive HDPCs separated by MACS inhibited odontoblast-like differentiation of HDPCs. These findings suggest that ICAM2 may regulate reparative dentin formation in dental pulp tissue. Full article

Objectives: Bone defects, resulting from many causes, represent a challenge in maxillofacial and orthopedic surgery. Regenerative medicine offers promising strategies by introducing exogenous materials to modify the tissue environment and modulate the body’s natural healing mechanisms. Dental pulp stem cells (DPSCs) are considered an effective source for tissue repair. Small molecules such as caffeic acid phenethyl ester (CAPE), although having promising effects in promoting bone regeneration, are characterized by low chemical stability, which impairs their clinical application. This study aimed to investigate the bone regenerative capability of four CAPE derivatives, recently synthesized in our laboratory and selected based on previous studies. Methods: DPSCs were induced to osteogenic differentiation in the presence of these compounds (0–5 μM), and cell viability, matrix deposition, alkaline phosphatase activity, and osteogenic marker gene expression were evaluated. In addition, bone biomaterials composed of a chitosan/agarose matrix reinforced with nanohydroxyapatite and enriched with these CAPE derivatives were fabricated and assessed for cytotoxicity and cell adhesion. Results: Two of the tested compounds effectively enhanced DPSC differentiation toward the osteogenic lineage. The fabricated bone biomaterials showed no cytotoxicity and supported cell adhesion. Furthermore, these compounds demonstrated stability under various conditions, confirming their suitability for incorporation into bone biomaterials. Conclusions: The tested CAPE derivatives exhibit promising osteoinductive properties and stability, offering a valid alternative to traditional therapeutic strategies in regenerative medicine. Full article

The use of chitosan nanoparticles (CH-NPs) loaded with antibiotics as irrigants in endodontics offers a unique combination, demonstrating effective antibacterial activity and low toxicity. Therefore, the aim of this study was to develop chitosan nanoparticles loaded with metronidazole, ciprofloxacin, and minocycline for use as endodontic irrigants to improve antibacterial activity against Enterococcus faecalis and to evaluate their cytotoxicity in human dental pulp stem cells (hDPSCs). Methods: The CH-NPs loaded with antibiotics were synthesized and analyzed using Fourier Transform Infrared Spectroscopy (FTIR) and Ultraviolet-visible spectroscopy (UV-Vis). Agar diffusion and microdilution assays were performed to determine the minimum inhibitory concentration (MIC), and a cytotoxicity assay was conducted to establish the median cytotoxic dose. Results: Peaks corresponding to the functional groups of the chitosan-antibiotic complex’s chemical structure were identified. A one-way ANOVA (p ≤ 0.05) with Tukey’s post hoc test was used to analyze the antibacterial effect. CH-NPs-ciprofloxacin showed the greatest antibacterial activity against E. faecalis in both agar diffusion and microdilution assays. CH-NPs-metronidazole demonstrated lower cytotoxicity against hDPSCs. CH-NPs-minocycline showed superior antibacterial effects compared to plain chitosan in microdilution assays, although they exhibited greater cytotoxicity. Conclusions: The ongoing search for an irrigating solution with effective antibacterial properties and low cytotoxicity could transform traditional techniques. However, this field is still developing and underexplored. It is essential to reevaluate decisions about irrigation solutions, as evidence on the use of chitosan nanoparticles with antibiotics is limited. This study provides valuable data for endodontics and is a crucial step for future research. Full article

Preserving dental pulp vitality is a key goal in minimally invasive dentistry. Conventional materials such as calcium hydroxide and mineral trioxide aggregate (MTA) are effective but limited in bioactivity and mechanical strength. This systematic review evaluated the biological efficacy of chitosan-based nanoparticles and biomaterials for pulp capping and regeneration. Following PRISMA 2020 guidelines, electronic searches were conducted across five databases up to April 2025. Controlled in vitro and animal studies using chitosan-based nanoparticles, hydrogels, or composite scaffolds were included. Risk of bias was assessed using SYRCLE (animal) and ToxRTool (in vitro), and certainty of evidence was rated via the GRADE-Preclinical framework. Due to methodological heterogeneity, data were synthesized using direction-of-effect coding and visualized through Albatross and heatmap plots. Sixteen studies met the criteria, consistently demonstrating enhanced cell viability, mineralization, and upregulation of odontogenic and angiogenic markers (BMP-2, TGF-β₁, VEGF, DSPP) compared with MTA or calcium hydroxide. Animal models confirmed improved angiogenesis, reparative dentin formation, and pulp vitality preservation. Despite uniformly positive biological outcomes, the overall certainty was rated Low to Very Low owing to small samples and unclear randomization. Chitosan-based biomaterials show promising regenerative potential, warranting well-designed preclinical and clinical studies for translational validation. Full article

The BMP2/7 heterodimer is known as a stronger inducer of osteogenic differentiation than BMP2 or BMP7 homodimers. Our aim was to establish BMP2/7-expressing human dental pulp stem cells (DPSCs) to evaluate tReceived: 23 October 2025; Revised: 29 November 2025; Accepted: 3 December 2025; Published: 3 December 2025 he osteogenic potential of the genetically modified cells. Lentiviral transduction was used to introduce the Tet-ON-regulated transgene-containing vector to the cells. Endogenous heterodimers were detected at the mRNA and protein levels using RNA-seq, qPCR, and Western blot, while secreted heterodimers were detected using ELISA assays. Osteogenic differentiation was monitored by measuring alkaline phosphatase (ALP) activity, mineralization, and the expression levels of RUNX2 and ALPL genes. Our results showed that ALP activity did not change in the transduced DPSCs; however, increased mineralization was detected, which correlates with the results obtained by RNA sequencing. Based on our results, BMP2/7-expressing DPSCs could be used in the treatment of bone defects, where heterodimers may have not only autocrine but also paracrine effects. Full article