Classification of Caries Based on CBCT: A Deep Learning Network Interpretability Study.

This study aimed to create a caries classification scheme based on cone-beam computed tomography (CBCT) and develop two deep learning models to improve caries classification accuracy. A total of 2713 axial slices were obtained from CBCT images of 204 carious teeth. Both classification models were trained and tested using the same pretrained classification networks on the dataset, including ResNet50_vd, MobileNetV3_large_ssld, and ResNet50_vd_ssld. The first model was used directly to classify the original images (direct classification model). The second model incorporated a presegmentation step for interpretation (interpretable classification model). Performance evaluation metrics including accuracy, precision, recall, and F1 score were calculated. The Local Interpretable Model-agnostic Explanations (LIME) method was employed to elucidate the decision-making process of the two models. In addition, a minimum distance between caries and pulp was introduced for determining the treatment strategies for type II carious teeth. The direct model that utilized the ResNet50_vd_ssld network achieved top accuracy, precision, recall, and F1 score of 0.700, 0.786, 0.606, and 0.616, respectively. Conversely, the interpretable model consistently yielded metrics surpassing 0.917, irrespective of the network employed. The LIME algorithm confirmed the interpretability of the classification models by identifying key image features for caries classification. Evaluation of treatment strategies for type II carious teeth revealed a significant negative correlation (p < 0.01) with the minimum distance. These results demonstrated that the CBCT-based caries classification scheme and the two classification models appeared to be acceptable tools for the diagnosis and categorization of dental caries.

Nanoparticles in tumor microenvironment remodeling and cancer immunotherapy.

Cancer immunotherapy and vaccine development have significantly improved the fight against cancers. Despite these advancements, challenges remain, particularly in the clinical delivery of immunomodulatory compounds. The tumor microenvironment (TME), comprising macrophages, fibroblasts, and immune cells, plays a crucial role in immune response modulation. Nanoparticles, engineered to reshape the TME, have shown promising results in enhancing immunotherapy by facilitating targeted delivery and immune modulation. These nanoparticles can suppress fibroblast activation, promote M1 macrophage polarization, aid dendritic cell maturation, and encourage T cell infiltration. Biomimetic nanoparticles further enhance immunotherapy by increasing the internalization of immunomodulatory agents in immune cells such as dendritic cells. Moreover, exosomes, whether naturally secreted by cells in the body or bioengineered, have been explored to regulate the TME and immune-related cells to affect cancer immunotherapy. Stimuli-responsive nanocarriers, activated by pH, redox, and light conditions, exhibit the potential to accelerate immunotherapy. The co-application of nanoparticles with immune checkpoint inhibitors is an emerging strategy to boost anti-tumor immunity. With their ability to induce long-term immunity, nanoarchitectures are promising structures in vaccine development. This review underscores the critical role of nanoparticles in overcoming current challenges and driving the advancement of cancer immunotherapy and TME modification.

Design and evaluation of an MMP-9-responsive hydrogel for vital pulp therapy.

OBJECTIVE: To design and evaluate a matrix metalloproteinase 9 (MMP-9)-responsive hydrogel for vital pulp therapy. METHODS: A peptide linker with optimized sensitivity toward MMP-9 was crosslinked with 4-arm poly (ethylene glycol)-norbornene (PEG-NB) by thiol-norbornene photo-polymerization. This resulted in the formation of a hydrogel network in which the peptide IDR-1002 was incorporated. Hydrogel characterization and gelation kinetics were examined with Fourier-transform infrared spectroscopy, scanning electron microscopy, rheological testing, and swelling evaluation. Hydrogel degradation was examined through multiple exposure to pre-activated MMP-9, to simulate flare-ups of dental pulp inflammation. The IDR-1002 released from degraded hydrogels was measured with high-performance liquid chromatography. Effect of IDR-1002 released from hydrogels on one-week-old multispecies oral biofilms was evaluated using confocal laser scanning microscopy. RESULTS: MMP-9-responsive, injectable, and photo-crosslinkable hydrogels were successfully synthesized. When hydrogel degradation and release of IDR-1002 were examined with exposure to pre-activated MMP-9, IDR-1002 release was significantly correlated with elevated levels of MMP-9 (p < 0.05). The effectiveness of IDR-1002 in killing bacteria in multispecies oral biofilms was significantly enhanced when the hydrogels were immersed in 10 nM or 20 nM pre-activated MMP-9, compared to immersion in phosphate-buffered saline (p < 0.05). CONCLUSIONS: The MMP-9-responsive hydrogel is a promising candidate for on-demand delivery of bioactive agent in vital pulp therapy. CLINICAL SIGNIFICANCE: MMP-9 is one of the most important diagnostic and prognostic biomarkers for pulpitis. An MMP-9-responsive hydrogel has potential to be used as an in-situ on-demand release system for the diagnosis and treatment of dental pulp inflammation.

Bacteria-mediated resistance of neutrophil extracellular traps to enzymatic degradation drives the formation of dental calculi.

Dental calculi can cause gingival bleeding and periodontitis, yet the mechanism underlying the formation of such mineral build-ups, and in particular the role of the local microenvironment, are unclear. Here we show that the formation of dental calculi involves bacteria in local mature biofilms converting the DNA in neutrophil extracellular traps (NETs) from being degradable by the enzyme DNase I to being degradation resistant, promoting the nucleation and growth of apatite. DNase I inhibited NET-induced mineralization in vitro and ex vivo, yet plasma DNases were ineffective at inhibiting ectopic mineralization in the oral cavity in rodents. The topical application of the DNA-intercalating agent chloroquine in rodents fed with a dental calculogenic diet reverted NET DNA to its degradable form, inhibiting the formation of calculi. Our findings may motivate therapeutic strategies for the reduction of the prevalence of the deposition of bacteria-driven calculi in the oral cavity.

Efficacy of pulpotomy in managing irreversible pulpitis in mature permanent teeth: A systematic review and meta-analysis.

OBJECTIVES: This paper evaluated the success rates of pulpotomy, compared its efficacy with non-surgical root canal treatment (NSRCT), evaluated different pulpotomy techniques, and analyzed the effectiveness of contemporary bioactive materials in managing irreversible pulpitis in mature permanent teeth. DATA SOURCES: A comprehensive literature search was conducted across multiple databases including PubMed, Web of Science, Scopus, and the Cochrane Library. Search was conducted from the inception of each database to the present, adhering to PRISMA 2020 guidelines. STUDY SELECTION: Studies were selected through a multi-step screening process, focusing on adult populations, randomized controlled trials, and single-arm trials. DATA: Fifteen randomized controlled trials and eight single-arm trials were included. For a follow-up period of more than 24 months, pooled clinical success rate of pulpotomy was 92.9 % (95 %CI;82.1-99.0 %), whereas pooled radiographic success rate was 78.5 % (95 %CI;66.7-88.4 %). Meta-analyses showed that there was no significant difference in success rates between pulpotomy and NSRCT, between full and partial pulpotomy techniques, or between Mineral Trioxide Aggregate pulpotomy and Calcium Enriched Mixture pulpotomy. The results indicated comparable efficacy across these variables. CONCLUSIONS: The study highlights the potential of less invasive treatments. Pulpotomy may be a viable alternative to NSRCT for managing irreversible pulpitis in mature permanent teeth. Limitations such as the low quality of some single-arm trials and the high risk of bias in some randomized controlled trials highlight the need for further research to standardize methodologies and broaden literature inclusion for a more comprehensive understanding of the efficacy of pulpotomy, considering the high success rates reported. Clinical Significance This quantitative systematic review recognizes the potential of full or partial pulpotomy as a viable treatment alternative to root canal therapy for managing irreversible pulpitis in mature permanent teeth. Future studies should aim for standardized protocols to validate these findings and improve patient treatment outcomes.

Interorgan communication in neurogenic heterotopic ossification: the role of brain-derived extracellular vesicles.

Brain-derived extracellular vesicles participate in interorgan communication after traumatic brain injury by transporting pathogens to initiate secondary injury. Inflammasome-related proteins encapsulated in brain-derived extracellular vesicles can cross the blood‒brain barrier to reach distal tissues. These proteins initiate inflammatory dysfunction, such as neurogenic heterotopic ossification. This recurrent condition is highly debilitating to patients because of its relatively unknown pathogenesis and the lack of effective prophylactic intervention strategies. Accordingly, a rat model of neurogenic heterotopic ossification induced by combined traumatic brain injury and achillotenotomy was developed to address these two issues. Histological examination of the injured tendon revealed the coexistence of ectopic calcification and fibroblast pyroptosis. The relationships among brain-derived extracellular vesicles, fibroblast pyroptosis and ectopic calcification were further investigated in vitro and in vivo. Intravenous injection of the pyroptosis inhibitor Ac-YVAD-cmk reversed the development of neurogenic heterotopic ossification in vivo. The present work highlights the role of brain-derived extracellular vesicles in the pathogenesis of neurogenic heterotopic ossification and offers a potential strategy for preventing neurogenic heterotopic ossification after traumatic brain injury. Brain-derived extracellular vesicles (BEVs) are released after traumatic brain injury. These BEVs contain pathogens and participate in interorgan communication to initiate secondary injury in distal tissues. After achillotenotomy, the phagocytosis of BEVs by fibroblasts induces pyroptosis, which is a highly inflammatory form of lytic programmed cell death, in the injured tendon. Fibroblast pyroptosis leads to an increase in calcium and phosphorus concentrations and creates a microenvironment that promotes osteogenesis. Intravenous injection of the pyroptosis inhibitor Ac-YVAD-cmk suppressed fibroblast pyroptosis and effectively prevented the onset of heterotopic ossification after neuronal injury. The use of a pyroptosis inhibitor represents a potential strategy for the treatment of neurogenic heterotopic ossification.

A Versatile Chitosan-Based Hydrogel Accelerates Infected Wound Healing via Bacterial Elimination, Antioxidation, Immunoregulation, and Angiogenesis.

Drug-resistant bacterial infection of cutaneous wounds causes great harm to the human body. These infections are characterized by a microenvironment with recalcitrant bacterial infections, persistent oxidative stress, imbalance of immune regulation, and suboptimal angiogenesis. Treatment strategies available to date are incapable of handling the healing dynamics of infected wounds. A Schiff base and borate ester cross-linked hydrogel, based on phenylboronic acid-grafted chitosan (CS-PBA), dibenzaldehyde-grafted poly(ethylene glycol), and tannic acid (TA), is fabricated in the present study. Customized phenylboronic acid-modified zinc oxide nanoparticles (ZnO) are embedded in the hydrogel prior to gelation. The CPP@ZnO-P-TA hydrogel effectively eliminates methicillin-resistant Staphylococcus aureus (MRSA) due to the pH-responsive release of Zn2+ and TA. Killing is achieved via membrane damage, adenosine triphosphate reduction, leakage of intracellular components, and hydrolysis of bacterial o-nitrophenyl-ß-d-galactopyranoside. The CPP@ZnO-P-TA hydrogel is capable of scavenging reactive oxygen and nitrogen species, alleviating oxidative stress, and stimulating M2 polarization of macrophages. The released Zn2+ and TA also induce neovascularization via the PI3K/Akt pathway. The CPP@ZnO-P-TA hydrogel improves tissue regeneration in vivo by alleviating inflammatory responses, stimulating angiogenesis, and facilitating collagen deposition. These findings suggest that this versatile hydrogel possesses therapeutic potential for the treatment of MRSA-infected cutaneous wounds.

The dynamics of bacterial proliferation, viability, and extracellular polymeric substances in oral biofilm development.

OBJECTIVES: This study investigated the relationship between bacterial growth, viability, and extracellular polymeric substances (EPS) formation in biofilms, particularly regarding resistance development. It also examined the impact of chemical factors on the EPS matrix and bacterial proliferation in oral biofilms. METHODS: Three multi-species oral biofilms were incubated in anaerobic conditions. Three strains of Enterococcus faecalis were incubated in aerobic conditions. The incubation periods ranged from 0 h to 7 days for short-term biofilms, and from 3 to 90 days for long-term biofilms. Fluorescent labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) and flow cytometry were used to track EPS and bacterial growth. Confocal laser scanning microscopy (CLSM) assessed bacterial viability and EPS structure. Biofilms aged 7, 14, and 21 days were treated with 2 % chlorhexidine (CHX) and 1 % sodium hypochlorite (NaOCl) to evaluate their effects on EPS and bacterial proliferation. RESULTS: Short-term biofilms showed rapid bacterial proliferation and a gradual increase in EPS, maintaining stable viability. In the first two weeks, a significant rise in CFSE indicated growing maturity. From 14 to 90 days, EPS and CFSE levels stabilized. Following treatment, CHX significantly reduced bacterial proliferation, while NaOCl decreased EPS volume. CONCLUSIONS: Biofilm development involves a balance between bacterial proliferation and EPS production. The complexity of this process poses challenges in treating biofilm-associated infections, requiring strategies tailored to the biofilm's developmental stage. CLINICAL SIGNIFICANCE: For effective root canal treatment, it is imperative to focus on reducing bacterial proliferation during the early stages of oral infections. In contrast, strategies aimed at minimizing EPS production could be more beneficial for long-term management of these conditions.

Effect of Nisin-based pretreatment solution on dentin bond strength, antibacterial property, and MMP activity of the adhesive interface.

OBJECTIVE: To evaluate the effect of a Nisin-based dentin pretreatment solution on microtensile bond strength, antibacterial activity, and matrix metalloproteinase (MMP) activity of the adhesive interface. MATERIALS AND METHODS: 100 human molars were sectioned to expose dentin. The teeth were assigned to five groups (n = 20), according to the dentin pretreatment: 0.5%, 1.0%, or 1.5% Nisin; 0.12% chlorhexidine (positive control), and no solution (negative control), and divided into 2 subgroups: no aging, and thermomechanical aging. Specimens were etched with 37% H3PO4 for 15 s and submitted to the dentin pretreatment. Then, they were bonded with an adhesive (Adper Single Bond 2) and a resin composite for microtensile bond strength (µTBS) evaluation. Antibacterial activity against Streptococcus mutans was qualitatively examined using an agar diffusion test. Anti-MMP activity within hybrid layers was examined using in-situ zymography. Data were analyzed with two-factor ANOVA and post-hoc Tukey's test (α = 0.050). RESULTS: For µTBS, significant differences were identified for the factors "solutions" (p = 0.002), "aging" (p = 0.017), and interaction of the two factors (p = 0.002). In the absence of aging, higher µTBS was observed for the group 0.5% Nisin. In the presence of aging, all groups showed similar µTBS values. All Nisin concentrations were effective in inhibiting the growth of S. mutans. Endogenous MMP activity was more significantly inhibited using 0.5% and 1.0% Nisin (p < 0.050). CONCLUSION: 0.5% and 1.0% Nisin solutions do not adversely affect resin-dentin bond strength and exhibit a potential bactericidal effect against S. mutans. Both concentrations effectively reduce endogenous gelatinolytic activity within the hybrid layer. CLINICAL RELEVANCE: The use of 0.5% and 1.0% Nisin solutions for dentin pretreatment potentially contributes to preserving the adhesive interface, increasing the longevity of composite restorations.

Biomimetic Self-Maturation Mineralization System for Enamel Repair.

Enamel repair is crucial for restoring tooth function and halting dental caries. However, contemporary research often overlooks the retention of organic residues within the repair layer, which hinders the growth of dense crystals and compromises the properties of the repaired enamel. During the maturation of natural enamel, the organic matrix undergoes enzymatic processing to facilitate further crystal growth, resulting in a highly mineralized tissue. Inspired by this process, a biomimetic self-maturation mineralization system is developed, comprising ribonucleic acid-stabilized amorphous calcium phosphate (RNA-ACP) and ribonuclease (RNase). The RNA-ACP induces initial mineralization in the form of epitaxial crystal growth, while the RNase present in saliva automatically triggers a biomimetic self-maturation process. The mechanistic study further indicates that RNA degradation prompts conformational rearrangement of the RNA-ACP, effectively excluding the organic matter introduced earlier. This exclusion process promotes lateral crystal growth, resulting in the generation of denser enamel-like apatite crystals that are devoid of organic residues. This strategy of eliminating organic residues from enamel crystals enhances the mechanical and physiochemical properties of the repaired enamel. The present study introduces a conceptual biomimetic mineralization strategy for effective enamel repair in clinical practice and offers potential insights into the mechanisms of biomineral formation.

Lysosomal destabilization: A missing link between pathological calcification and osteoarthritis.

Calcification of cartilage by hydroxyapatite is a hallmark of osteoarthritis and its deposition strongly correlates with the severity of osteoarthritis. However, no effective strategies are available to date on the prevention of hydroxyapatite deposition within the osteoarthritic cartilage and its role in the pathogenesis of this degenerative condition is still controversial. Therefore, the present work aims at uncovering the pathogenic mechanism of intra-cartilaginous hydroxyapatite in osteoarthritis and developing feasible strategies to counter its detrimental effects. With the use of in vitro and in vivo models of osteoarthritis, hydroxyapatite crystallites deposited in the cartilage are found to be phagocytized by resident chondrocytes and processed by the lysosomes of those cells. This results in lysosomal membrane permeabilization (LMP) and release of cathepsin B (CTSB) into the cytosol. The cytosolic CTSB, in turn, activates NOD-like receptor protein-3 (NLRP3) inflammasomes and subsequently instigates chondrocyte pyroptosis. Inhibition of LMP and CTSB in vivo are effective in managing the progression of osteoarthritis. The present work provides a conceptual therapeutic solution for the prevention of osteoarthritis via alleviation of lysosomal destabilization.

A glycol chitosan derivative with extrafibrillar demineralization potential for self-etch dentin bonding.

OBJECTIVES: Extrafibrillar demineralization is an etching technique that removes only minerals from around the collagen fibrils for resin infiltration. The intrafibrillar minerals are left intact to avoid their replacement by water that is hard for adhesive resin monomers to displace. The present work reported the synthesis of a water-soluble methacryloyloxy glycol chitosan-EDTA conjugate (GCE-MA) and evaluated its potential as an extrafibrillar demineralization agent for self-etch dentin bonding. METHODS: Glycol chitosan-EDTA was functionalized with a methacryloyloxy functionality. Conjugation was confirmed using Fourier transform-infrared spectroscopy. The GCE-MA was used to prepare experimental self-etch primers. Extrafibrillar demineralization of the primers was evaluated with scaning electron microscopy and transmission electron microscopy. The feasibility of this new self-etch bonding approach was evaluated using microtensile bond strength testing and inhibition of dentin gelatinolytic activity. The antibacterial activity and cytotoxicity of GCE-MA were also analyzed. RESULTS: Conjugation of EDTA and the methacryloyloxy functionality to glycol chitosan was successful. The functionalized conjugate was capable of extrafibrillar demineralization of mineralized collagen fibrils. Tensile bond strength of the experimental self-etch primer to dentin was comparable to that of phosphoric acid-etched dentin and the commercial self-etch primer Clearfil SE Bond 2. The GCE-MA also inhibited soluble rhMMP-9. In-situ zymography detected minimal fluorescence in hybrid layers conditioned with the experimental primer. The GCE-MA was noncytotoxic and possessed antibacterial activities against planktonic bacteria. SIGNIFICANCE: Synthesis of GCE-MA brought into fruition a self-etch conditioner that selectively demineralizes the extrafibrillar mineral component of dentin. A self-etch primer prepared with GCE-MA achieved bond strengths comparable to commercial reference adhesive systems.

Bacteria debridement efficacy of two sonic root canal irrigant activation systems.

OBJECTIVE: To evaluate the bacteria debridement efficacy of two generations of sonic root canal irrigant activation systems: EndoActivator (Dentsply Sirona), the first generation, and SmartLite Pro EndoActivator, the second generation. METHODS: Instrumented, autoclaved, single-rooted human premolars were inoculated with Enterococcus faecalis (ATCC-29212) for 21 days. The bacteria biofilm-containing teeth were randomly divided into 5 groups (N=8): Group 1: Syringe-side-vented needle (S-N) delivery of saline for 1 min; Group 2: S-N delivery of 2% NaOCl for 1 min; Group 3: S-N delivery of 2% NaOCl for 5 min; Group 4: EndoActivator activation of 2% NaOCl for 1 min; Group 5: SmartLite Pro EndoActivator activation of 2% NaOCl for 1 min. The teeth were evaluated for bacterial reduction using CFU counts, and the percentages of dead bacteria within the dentinal tubules using confocal laser scanning microscopy. RESULTS: Activation of NaOCl with EndoActivator or SmartLite Pro EndoActivator significantly reduced the overall intracanal bacterial load, compared with S-N irrigant delivery (P<0.05), with no significant difference between the two agitation devices (P>0.05). Nevertheless, S-N delivery of 2% NaOCl for 5 min produced better bacteria debridement than either sonic agitation system. Different degrees of bacteria kill were identified in the coronal-middle portions and apical portion of the canal space. CONCLUSION: Delivery time of NaOCl affects the efficacy of bacteria disinfection. Activation for 1 min with the EndoActivator or SmartLite Pro EndoActivator demonstrated comparable canal wall biofilm and intracanal bacteria reduction efficacy when 2% NaOCl was used as irrigant for disinfecting E. faecalis in single-rooted teeth. CLINICAL SIGNIFICANCE: Although the sonic root canal irrigant activation devices investigated do not completely eliminate live bacteria biofilms from the canal space, they help reduce bacteria load during irrigant activation.