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This study aimed to synthesize a novel elastomeric ligature with dimethylaminohexadecyl methacrylate (DMAHDM) grafted, providing a new strategy for improving the issue of enamel demineralization during fixed orthodontics. DMAHDM was incorporated into elastomeric ligatures at different mass fractions using ultraviolet photochemical grafting. The antibacterial properties were evaluated and the optimal DMAHDM amount was determined based on cytotoxicity assays. Moreover, tests were conducted to evaluate the in vivo changes in the mechanical properties of the elastomeric ligatures. To assess the actual in vivo effectiveness in preventing enamel demineralization, a rat demineralization model was established, with analyses focusing on changes in surface microstructure, elemental composition, and nanomechanical properties. Elastomeric ligatures with 2% DMAHDM showed excellent biocompatibility and the best antibacterial properties, reducing lactic acid production by 65.3% and biofilm bacteria by 50.0% within 24 h, without significant mechanical property differences from the control group (p > 0.05). Most importantly, they effectively prevented enamel demineralization in vivo, enhancing elastic modulus by 73.2% and hardness by 204.8%. Elastomeric ligatures incorporating DMAHDM have shown great potential for application in preventing enamel demineralization, providing a new strategy to solve this issue during fixed orthodontics.
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Esmalte Dental , Elastómeros , Desmineralización Dental , Desmineralización Dental/prevención & control , Animales , Elastómeros/química , Ratas , Esmalte Dental/efectos de los fármacos , Antibacterianos/farmacología , Antibacterianos/química , Metacrilatos/química , Metacrilatos/farmacología , Aparatos Ortodóncicos , Biopelículas/efectos de los fármacos , MasculinoRESUMEN
OBJECTIVES: To synthesize a novel antibacterial orthodontic elastomeric ligature incorporating dimethylaminohexadecyl methacrylate (DMAHDM) for the first time to prevent enamel demineralization during orthodontic therapy. METHODS: Various mass fractions of DMAHDM (ranging from 0 % to 20 %) were grafted onto commercial elastomeric ligatures using an ultraviolet photochemical grafting method and were characterized. The optimal DMAHDM concentration was determined based on biocompatibility and mechanical properties, and the antibacterial efficacy was evaluated in a whole-plaque biofilm model. TaqMan real-time polymerase chain reaction and fluorescence in situ hybridization were used to assess the microbial regulatory ability of the multispecies biofilms. Furthermore, an in vitro tooth demineralization model was established to explore its preventive effects on enamel demineralization. Statistical analysis involved a one-way analysis of variance and LSD post hoc tests at a significance level of 0.05. RESULTS: The elastomeric ligature containing 2 % mass fraction of DMAHDM exhibited excellent mechanical properties, favorable biocompatibility, and the most effective antibacterial ability against microorganisms, which decreased by almost two logarithms (P < 0.05). It significantly reduced the proportion of Streptococcus mutans in the multispecies plaque biofilm by 25 % at 72 h, leading to an enhanced biofilm microenvironment. Moreover, the novel elastomeric ligature demonstrated an obvious preventive effect on enamel demineralization, with an elastic modulus 30 % higher and hardness 62 % higher than those of the control group within 3 months (P < 0.05). SIGNIFICANCE: The integration of DMAHDM with an elastomeric ligature holds significant promise for regulating biofilms and preventing enamel demineralization in orthodontic applications.
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OBJECTIVE: Streptococcus mutans (S. mutans) is a major contributor to dental caries, with its ability to synthesize extracellular polysaccharides (EPS) and biofilms. The gcrR gene is a regulator of EPS synthesis and biofilm formation. The objectives of this study were to investigate a novel strategy of combining gcrR gene over-expression with dimethylaminohexadecyl methacrylate (DMAHDM), and to determine their in vivo efficacy in reducing caries in rats for the first time. METHODS: Two types of S. mutans were tested: Parent S. mutans; and gcrR gene over-expressed S. mutans (gcrR OE S. mutans). Bacterial minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were measured with DMAHDM and chlorhexidine (CHX). Biofilm biomass, polysaccharide, lactic acid production, live/dead staining, colony-forming units (CFUs), and metabolic activity (MTT) were evaluated. A Sprague-Dawley rat model was used with parent S. mutans and gcrR OE S. mutans colonization to determine caries-inhibition in vivo. RESULTS: Drug-susceptibility of gcrR OE S. mutans to DMAHDM or CHX was 2-fold higher than that of parent S. mutans. DMAHDM reduced biofilm CFU by 3-4 logs. Importantly, the combined gcrR OE S. mutans+ DMAHDM dual strategy reduced biofilm CFU by 5 logs. In the rat model, the parent S. mutans group had a higher cariogenicity in dentinal (Dm) and extensive dentinal (Dx) regions. The DMAHDM + gcrR OE group reduced the Dm and Dx caries to only 20 % and 0 %, those of parent S. mutans + PBS control group (p < 0.05). The total caries severity of gcrR OE + DMAHDM group was decreased to 51 % that of parent S. mutans control (p < 0.05). SIGNIFICANCE: The strategy of combining S. mutans gcrR over-expression with antibacterial monomer reducing biofilm acids by 97 %, and reduced in vivo total caries in rats by 48 %. The gcrR over-expression + DMAHDM strategy is promising for a wide range of dental applications to inhibit caries and protect tooth structures.
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OBJECTIVE: The article reviewed novel orthodontic devices and materials with bioactive capacities in recent years and elaborated on their properties, aiming to provide guidance and reference for future scientific research and clinical applications. DATA, SOURCES AND STUDY SELECTION: Researches on remineralization, protein repellent, antimicrobial activity and multifunctional novel bioactive orthodontic devices and materials were included. The search of articles was carried out in Web of Science, PubMed, Medline and Scopus. CONCLUSIONS: The new generation of orthodontic devices and materials with bioactive capacities has broad application prospects. However, most of the current studies are limited to in vitro studies and cannot explore the true effects of various bioactive devices and materials applied in oral environments. More research, especially in vivo researches, is needed to assist in clinical application. CLINICAL SIGNIFICANCE: Enamel demineralization (ED) is a common complication in orthodontic treatments. Prolonged ED can lead to dental caries, impacting both the aesthetics and health of teeth. It is of great significance to develop antibacterial orthodontic devices and materials that can inhibit bacterial accumulation and prevent ED. However, materials with only preventive effect may fall short of addressing actual needs. Hence, the development of novel bioactive orthodontic materials with remineralizing abilities is imperative. The article reviewed the recent advancements in bioactive orthodontic devices and materials, offering guidance and serving as a reference for future scientific research and clinical applications.
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Caries Dental , Soportes Ortodóncicos , Desmineralización Dental , Humanos , Caries Dental/prevención & control , Estética Dental , Esmalte Dental , Desmineralización Dental/prevención & controlRESUMEN
PURPOSE: To characterize the microbiome composition in peri-implant pocket of peri-implantitis and peri-implant sulcus controls using 16S rRNA gene sequencing. MATERIALS AND METHODS: In this controlled clinical cross-sectional study, 23 subjects with control implants (n = 14) and diseased implants (peri-implantitis, n = 21) were included. The peri-implant pocket/sulcus was sampled and used to extract DNA and amplify the 16S rRNA gene using universal primers targeting the V3-V4 regions. The resulting 16S PCR amplicons were sequenced on Illumina MiSeq, and the sequences were processed using DADA2 and the Human Oral Microbiome Database (HOMD) as references. Alpha and Beta diversity, as well as core microbiome and differential abundance analyses, were performed using the MicrobiomeAnalyst workflow. RESULTS: There were no significant differences in microbial diversity between control implants and implants with peri-implantitis (Shannon p = 0.82). Overall bacterial community structure assessed through beta diversity analysis was also not significantly different between the two groups (p = 0.18). However, high levels of Gram-negative bacteria were detected in peri-implant pockets compared to the control sulcus. Abundant species in peri-implantitis were Capnocytophaga leadbetteri, Treponema maltophilum, Peptostreptococcus, Neisseria, P. gingivalis, and Porphyromonas endodontali, Lactococcus lactis and Filifactor alocis (p < 0.05). Gram-positive bacteria such as Streptococcus salivaris, Prevotella melaninogenica, L. wadei, and Actinomyces spp. serve were more abundant in peri-implant control sulcus. CONCLUSIONS: Peri-implant sulcus in control implants harbors predominantly Gram-positive bacteria, whereas pockets of implants with peri-implantitis harbor predominantly Gram-negative bacteria.
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Implantes Dentales , Microbiota , Periimplantitis , Humanos , Periimplantitis/microbiología , Implantes Dentales/efectos adversos , ARN Ribosómico 16S/genética , Estudios Transversales , Microbiota/genéticaRESUMEN
PURPOSE: There are different methods to transfer occlusal vertical dimension (OVD) when fabricating complete dentures, but it is not clear which method yields the most accurate transfer. This study investigated the accuracy of transferring the OVD of the maxillary/mandibular relationship when designing and fabricating digital dentures by evaluating two commonly used workflows: duplicate dentures (DDs) and occlusion rims (ORs). MATERIALS AND METHODS: Mounted edentulous casts were used to simulate a completely edentulous patient. These casts were used in two workflows: (1) DD workflow where complete dentures were fabricated and relined (n = 15), and (2) OR where final impressions and ORs were constructed (n = 15). Relined dentures and ORs with fiduciary markers were then scanned and virtually articulated, then exported as STL files. A scan of the articulated casts (simulated patients) with fiduciary markers embedded was also exported as an STL file and used as a control. STL files for each workflow were superimposed over the control and analyzed in 3D inspection software (Geomagic Control X, 3D Systems, NC) for deviations (mm) at three different locations. Average 3D deviations were compared among the groups using a two-way ANOVA (α-0.05). RESULTS: The average deviation for OR was significantly larger than DD at all locations of measurement [F = 46.00, p < 0.0001]. The majority of the measurements (98.9%) had a positive deviation value, indicating increased OVD. The overall deviation for the OR group from the control was 0.72 ± 0.13 mm versus 0.45 ± 0.23 mm for the DD group. There was no difference in deviation between the points of measurement and no interaction between the location and method [F = 0.02, p < 0.98]. CONCLUSIONS: The DD workflow exhibited less deviation in the transfer of OVD than the OR workflow. However, the deviation was small and may not be clinically significant.
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Diseño de Dentadura , Boca Edéntula , Humanos , Dimensión Vertical , Diseño de Dentadura/métodos , Dentadura Completa , Diseño Asistido por ComputadoraRESUMEN
OBJECTIVES: Dental caries is caused by acids from biofilms. pH-sensitive nanoparticle carriers could achieve improved targeted effectiveness. The objectives of this study were to develop novel mesoporous silica nanoparticles carrying nanosilver and chlorhexidine (nMS-nAg-Chx), and investigate the inhibition of biofilms as well as the modulation of biofilm to suppress acidogenic and promote benign species for the first time. METHODS: nMS-nAg was synthesized via a modified sol-gel method. Carboxylate group functionalized nMS-nAg (COOH-nMS-nAg) was prepared and Chx was added via electrostatic interaction. Minimal inhibitory concentration (MIC), inhibition zone, and growth curves were evaluated. Streptococcus mutans (S. mutans), Streptococcus gordonii (S. gordonii), and Streptococcus sanguinis (S. sanguinis) formed multispecies biofilms. Metabolic activity, biofilm lactic acid, exopolysaccharides (EPS), and TaqMan real-time polymerase chain reaction (RT-PCR) were tested. Biofilm structures and biomass were observed by scanning electron microscopy (SEM) and live/dead bacteria staining. RESULTS: nMS-nAg-Chx possessed pH-responsive properties, where Chx release increased at lower pH. nMS-nAg-Chx showed good biocompatibility. nMS-nAg-Chx exhibited a strong antibacterial function, reducing biofilm metabolic activity and lactic acid as compared to control (p < 0.05, n = 6). Moreso, biofilm biomass was dramatically suppressed in nMS-nAg-Chx groups. In control group, there was an increasing trend of S. mutans proportion in the multispecies biofilm, with S. mutans reaching 89.1% at 72 h. In sharp contrast, in nMS-nAg-Chx group of 25 µg/mL, the ratio of S. mutans dropped to 43.7% and the proportion of S. gordonii and S. sanguinis increased from 19.8% and 10.9 to 69.8% and 56.3%, correspondingly. CONCLUSION: pH-sensitive nMS-nAg-Chx had potent antibacterial effects and modulated biofilm toward a non-cariogenic tendency, decreasing the cariogenic species nearly halved and increasing the benign species approximately twofold. nMS-nAg-Chx is promising for applications in mouth rinse and endodontic irrigants, and as fillers in resins to prevent caries.
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Caries Dental , Nanopartículas , Plata , Humanos , Clorhexidina/farmacología , Clorhexidina/química , Caries Dental/microbiología , Dióxido de Silicio/farmacología , Dióxido de Silicio/química , Antibacterianos/farmacología , Antibacterianos/química , Streptococcus mutans , Nanopartículas/química , Ácido Láctico/análisis , Biopelículas , Concentración de Iones de HidrógenoRESUMEN
OBJECTIVE: Implant-related infections from the adhesion and proliferation of dental plaque are a major challenge for dental implants. The objectives of this study were to: (1) develop novel antibacterial titanium (Ti) healing abutment; (2) investigate the inhibition of implant infection-related pathogenic bacteria and saliva-derived biofilm, and evaluate the biocompatibility of the new material for the first time. METHODS: Dimethylaminohexadecyl methacrylate (DMAHDM) and hydroxyapatite (HAP) were polymerized via polydopamine (PDA) on Ti. Staphylococcus aureus (S. aureus), Streptococcus sanguinis (S. sanguinis) and human saliva-derived biofilms were tested. After 4 weeks of DMAHDM release, the antibacterial efficacy of the DMAHDM remaining on Ti surface and the DMADHM in medium was tested. Biocompatibility was determined using human gingival fibroblasts (HGFs) and periodontal ligament stem cells (PDLSCs). RESULTS: The DMAHDM-loaded coating filled into the nano-voids in Ti surfaces. The modified Ti showed potent antibacterial activity, reducing the CFU of S. aureus, S. sanguinis and saliva-derived biofilms by 8, 7 and 4 log, respectively (P < 0.05). After 4 weeks of release, the modified Ti was still able to reduce S. aureus and S. sanguinis biofilm CFU by 1-3 log (P < 0.05). This provided strong antibacterial function for more than 4 weeks, which were the high-risk period for implant infections. The new material showed excellent biocompatibility when compared to control (P > 0.05). CONCLUSION: Novel DMAHDM-loaded Ti healing abutment had strong antibacterial effects, reducing biofilm CFUs by orders of magnitude, and lasting for over four weeks to cover the high-risk period for implant infections. The novel antibacterial Ti is promising to combat implant-related infections in dental, craniofacial and orthopedic applications.
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Implantes Dentales , Metilaminas , Titanio , Humanos , Titanio/farmacología , Staphylococcus aureus , Antibacterianos/farmacología , Metacrilatos/farmacología , BiopelículasRESUMEN
Serotonergic neurons in the rostral ventral medulla (RVM) contribute to bidirectional control of pain through modulation of spinal and trigeminal nociceptive networks. Deficits in this pathway are believed to contribute to pathologic pain states, but whether changes in serotonergic mechanisms are pro- or antinociceptive is debated. We used a combination of optogenetics and fiber photometry to examine these mechanisms more closely. We find that optogenetic activation of RVM serotonergic afferents in the spinal cord of naive mice produces mechanical hypersensitivity and conditioned place aversion (CPA). Neuropathic pain, produced by chronic constriction injury of the infraorbital nerve (CCI-ION), evoked a tonic increase in serotonin (5HT) concentrations within the spinal trigeminal nucleus caudalis (SpVc), measured with liquid chromatography-tandem mass spectroscopy (LC-MS/MS). By contract, CCI-ION had no effect on the phasic serotonin transients in SpVc, evoked by noxious pinch, and measured with fiber photometry of a serotonin sensor. These findings suggest that serotonin release in the spinal cord is pronociceptive and that an increase in sustained serotonin signaling, rather than phasic or event driven increases, potentiate nociception in models of chronic pain.
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Neuralgia , Serotonina , Ratones , Animales , Serotonina/metabolismo , Hiperalgesia/metabolismo , Cromatografía Liquida , Espectrometría de Masas en Tándem , Asta Dorsal de la Médula Espinal , Médula Espinal/metabolismo , Neuralgia/metabolismoRESUMEN
Recurrent caries remain a persistent concern, often linked to microleakage and a lack of bioactivity in contemporary dental composites. Our study aims to address this issue by developing a low-shrinkage-stress nanocomposite with antibiofilm and remineralization capabilities, thus countering the progression of recurrent caries. In the present study, we formulated low-shrinkage-stress nanocomposites by combining triethylene glycol divinylbenzyl ether and urethane dimethacrylate, incorporating dimethylaminododecyl methacrylate (DMADDM), along with nanoparticles of calcium fluoride (nCaF2) and nanoparticles of amorphous calcium phosphate (NACP). The biofilm viability, biofilm metabolic activity, lactic acid production, and ion release were evaluated. The novel formulations containing 3% DMADDM exhibited a potent antibiofilm activity, exhibiting a 4-log reduction in the human salivary biofilm CFUs compared to controls (p < 0.001). Additionally, significant reductions were observed in biofilm biomass and lactic acid (p < 0.05). By integrating both 10% NACP and 10% nCaF2 into one formulation, efficient ion release was achieved, yielding concentrations of 3.02 ± 0.21 mmol/L for Ca, 0.5 ± 0.05 mmol/L for P, and 0.37 ± 0.01 mmol/L for F ions. The innovative mixture of DMADDM, NACP, and nCaF2 displayed strong antibiofilm effects on salivary biofilm while concomitantly releasing a significant amount of remineralizing ions. This nanocomposite is a promising dental material with antibiofilm and remineralization capacities, with the potential to reduce polymerization-related microleakage and recurrent caries.
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OBJECTIVES: Composites are commonly used for tooth restorations, but recurrent caries often lead to restoration failures due to polymerization shrinkage-stress-induced marginal leakage. The aims of this research were to: (1) develop novel low-shrinkage-stress (L.S.S.) nanocomposites containing dimethylaminododecyl methacrylate (DMADDM) with nanoparticles of calcium fluoride (nCaF2) or amorphous calcium phosphate (NACP) for remineralization; (2) investigate antibacterial and cytocompatibility properties. METHODS: Nanocomposites were made by mixing triethylene glycol divinylbenzyl ether with urethane dimethacrylate containing 3% DMADDM, 20% nCaF2, and 20% NACP. Flexural strength, elastic modulus, antibacterial properties against Streptococcus mutans biofilms, and cytotoxicity against human gingival fibroblasts and dental pulp stem cells were tested. RESULTS: Nanocomposites with DMADDM and nCaF2 or NACP had flexural strengths matching commercial composite control without bioactivity. The new nanocomposite provided potent antibacterial properties, reducing biofilm CFU by 6 logs, and reducing lactic acid synthesis and metabolic function of biofilms by 90%, compared to controls (p < 0.05). The new nanocomposites produced excellent cell viability matching commercial control (p > 0.05). CONCLUSIONS: Bioactive L.S.S. antibacterial nanocomposites with nCaF2 and NACP had excellent bioactivity without compromising mechanical and cytocompatible properties. The new nanocomposites are promising for a wide range of dental restorations by improving marginal integrity by reducing shrinkage stress, defending tooth structures, and minimizing cariogenic biofilms.
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BACKGROUND: Vascularization plays an important role in dental and craniofacial regenerations. Human periodontal ligament stem cells (hPDLSCs) are a promising cell source and, when co-cultured with human umbilical vein endothelial cells (hUVECs), could promote vascularization. The objectives of this study were to develop a novel prevascularized hPDLSC-hUVEC-calcium phosphate construct, and investigate the osteogenic and angiogenic efficacy of this construct with human platelet lysate (hPL) in cranial defects in rats for the first time. METHODS: hPDLSCs and hUVECs were co-cultured on calcium phosphate cement (CPC) scaffolds with hPL. Cell proliferation, angiogenic gene expression, angiogenesis, alkaline phosphatase activity, and cell-synthesized minerals were determined. Bone and vascular regenerations were investigated in rat critical-sized cranial defects in vivo. RESULTS: hPDLSC-hUVEC-CPC-hPL group had 2-fold greater angiogenic expressions and cell-synthesized mineral synthesis than hPDLSC-hUVEC-CPC group (p < 0.05). Microcapillary-like structures were formed on scaffolds in vitro. hPDLSC-hUVEC-CPC-hPL group had more vessels than hPDLSC-hUVEC-CPC group (p < 0.05). In cranial defects in rats, hPDLSC-hUVEC-CPC-hPL group regenerated new bone amount that was 2.1 folds and 4.0 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). New blood vessel density of hPDLSC-hUVEC-CPC-hPL group was 2 folds and 7.9 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). CONCLUSION: The hPL pre-culture method is promising to enhance bone regeneration via prevascularized CPC. Novel hPDLSC-hUVEC-CPC-hPL prevascularized construct increased new bone formation and blood vessel density by 4-8 folds over CPC control. CLINICAL SIGNIFICANCE: Novel hPDLSC-hUVEC-hPL-CPC prevascularized construct greatly increased bone and vascular regeneration in vivo and hence is promising for a wide range of craniofacial applications.
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Ligamento Periodontal , Andamios del Tejido , Humanos , Animales , Ratas , Ratas Desnudas , Andamios del Tejido/química , Células Madre , Osteogénesis , Regeneración Ósea , Células Endoteliales de la Vena Umbilical Humana , Fosfatos de Calcio/farmacología , Fosfatos de Calcio/química , Cráneo/cirugía , Diferenciación Celular , Células CultivadasRESUMEN
OBJECTIVE: We leveraged an artificial intelligence deep-learning convolutional neural network (DL CNN) to detect calcified carotid artery atheromas (CCAAs) on cone beam computed tomography (CBCT) images. STUDY DESIGN: We obtained 137 full-volume CBCT scans with previously diagnosed CCAAs. The DL model was trained on 170 single axial CBCT slices, 90 with extracranial CCAAs and 80 with intracranial CCAAs. A board-certified oral and maxillofacial radiologist confirmed the presence of each CCAA. Transfer learning through a U-Net-based CNN architecture was utilized. Data allocation was 60% training, 10% validation, and 30% testing. We determined the accuracy of the DL model in detecting CCAA by calculating the mean training and validation accuracy and the area under the receiver operating characteristic curve (AUC). We reserved 5 randomly selected unseen full CBCT volumes for final testing. RESULTS: The mean training and validation accuracy of the model in detecting extracranial CCAAs was 92% and 82%, respectively, and the AUC was 0.84 with 1.0 sensitivity and 0.69 specificity. The mean training and validation accuracy in detecting intracranial CCAAs was 61% and 70%, respectively, and the AUC was 0.5 with 0.93 sensitivity and 0.08 specificity. Testing of full-volume scans yielded an AUC of 0.72 and 0.55 for extracranial and intracranial CCAAs, respectively. CONCLUSION: Our DL model showed excellent discrimination in detecting extracranial CCAAs on axial CBCT images and acceptable discrimination on full-volumes but poor discrimination in detecting intracranial CCAAs, for which further research is required.
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Serotonergic neurons in the rostral ventral medulla (RVM) contribute to bidirectional control of pain through modulation of spinal and trigeminal nociceptive networks. Deficits in this pathway are believed to contribute to pathological pain states, but whether changes in serotonergic mechanisms are pro or anti-nociceptive are debated. We used a combination of optogenetics and fiber photometry to examine these mechanisms more closely. We find that optogenetic activation of RVM serotonergic afferents in the spinal cord of naïve mice produces mechanical hypersensitivity and conditioned place aversion. Neuropathic pain, produced by chronic constriction injury of the infraorbital nerve (CCI-ION), evoked a tonic increase in serotonin concentrations within the spinal trigeminal nucleus caudalis (SpVc), measured with liquid chromatography-tandem mass spectroscopy (LC-MS/MS). By contract, CCI-ION had no effect on the phasic serotonin transients in SpVc, evoked by noxious pinch, and measured with fiber photometry of a serotonin sensor. These findings suggest that serotonin release in the spinal cord is pronociceptive and that an increase is sustained serotonin signaling, rather than phasic or event driven increases, potentiate nociception in models of chronic pain.
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The parabrachial nuclear complex (PBN) is a nexus for aversion and for the sensory and affective components of pain perception. We have previously shown that during chronic pain PBN neurons in anesthetized rodents have amplified activity. We report a method to record from PBN neurons of behaving, head-restrained mice while applying reproducible noxious stimuli. We find that both spontaneous and evoked activity are higher in awake animals compared with urethane anesthetized mice. Fiber photometry of calcium responses from calcitonin-gene-related peptide-expressing PBN neurons demonstrates that these neurons respond to noxious stimuli. In both males and females with neuropathic or inflammatory pain, responses of PBN neurons remain amplified for at least 5 weeks, in parallel with increased pain metrics. We also show that PBN neurons can be rapidly conditioned to respond to innocuous stimuli after pairing with noxious stimuli. Finally, we demonstrate that changes in PBN neuronal activity are correlated with changes in arousal, measured as changes in pupil area.SIGNIFICANCE STATEMENT The parabrachial complex is a nexus of aversion, including pain. We report a method to record from parabrachial nucleus neurons of behaving mice while applying reproducible noxious stimuli. This allowed us to track parabrachial activity over time in animals with neuropathic or inflammatory pain. It also allowed us to show that the activity of these neurons correlates with arousal states and that these neurons can be conditioned to respond to innocuous stimuli.
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Dolor Crónico , Núcleos Parabraquiales , Masculino , Femenino , Ratones , Animales , Núcleos Parabraquiales/fisiología , Nocicepción , Vigilia , Péptido Relacionado con Gen de Calcitonina/metabolismoRESUMEN
The parabrachial complex (PB) is critically involved in aversive processes, and chronic pain is associated with amplified activity of PB neurons in rodent models of neuropathic pain. Here, we demonstrate that catecholaminergic input from the caudal nucleus of the solitary tract (cNTScat), a stress responsive region that integrates interoceptive and exteroceptive signals, causes amplification of PB activity and their sensory afferents. We used a virally mediated expression of a norepinephrine (NE) sensor, NE2h, fiber photometry, and extracellular recordings in anesthetized mice to show that noxious mechanical and thermal stimuli activate cNTS neurons. These stimuli also produce prolonged NE transients in PB that far outlast the noxious stimuli. Similar NE transients can be evoked by focal electrical stimulation of cNTS, a region that contains the noradrenergic A2 cell group that projects densely on PB. In vitro, optical stimulation of cNTScat terminals depolarized PB neurons and caused a prolonged increase the frequency of excitatory synaptic activity. A dual opsin approach showed that sensory afferents from the caudal spinal trigeminal nucleus are potentiated by cNTScat terminal activation. This potentiation was coupled with a decrease in the paired pulse ratio (PPR), consistent with an cNTScat-mediated increase in the probability of release at SpVc synapses. Together, these data suggest that A2 neurons of the cNTS generate long lasting NE transients in PB which increase excitability and potentiate responses of PB neurons to sensory inputs. These reveal a mechanism through which stressors from multiple modalities may potentiate the aversiveness of nociceptive stimuli.
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Dolor Crónico , Núcleo Solitario , Ratones , Animales , Neuronas/fisiología , Nervio Vago , NorepinefrinaRESUMEN
The parabrachial nuclear complex (PBN) is a nexus for aversion, and for the sensory and affective components of pain perception. We have previously shown that, during chronic pain, PBN neurons in anesthetized rodents have amplified activity. We report a method to record from PBN neurons of behaving, head-restrained mice, while applying reproducible noxious stimuli. We find that both spontaneous and evoked activity are higher in awake animals, compared to urethane anesthetized mice. Fiber photometry of calcium responses from CGRP-expressing PBN neurons demonstrates that these neurons respond to nociceptive stimuli. In both males and females with neuropathic or inflammatory pain, responses of PBN neurons remain amplified for at least 5 weeks, in parallel with increased pain metrics. We also show that PBN neurons can be rapidly conditioned to respond to innocuous stimuli, after pairing with nociceptive stimuli. Finally, we demonstrate that changes in PBN neuronal activity are correlated with changes in arousal, measured as changes in pupil diameter. Significance Statement: The parabrachial complex is a nexus of aversion, including pain. We report a method to record from parabrachial nucleus neurons of behaving mice, while applying reproducible noxious stimuli. This allowed, for the first time, tracking the activity of these neurons over time in animals with neuropathic or inflammatory pain. It also allowed us to show that the activity of these neurons correlates with arousal states, and that these neurons can be conditioned to respond to innocuous stimuli.
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STATEMENT OF PROBLEM: Delamination of veneering ceramic is one of the most common challenges relating to veneered zirconia restorations. Additive manufacturing (AM) is a fast-expanding technology that has gained widespread acceptance in dentistry and is increasingly being used to produce dental restorations. However, information about bonding of porcelain to AM zirconia is lacking. PURPOSE: The purpose of this in vitro study was to investigate the shear bond strength (SBS) of porcelain to milled and additively manufactured zirconia, and the effect of surface treatment on bond strength. MATERIAL AND METHODS: A Ø12×5-mm disk was designed virtually to fabricate all specimens, which were divided into 2 groups according to the manufacturing technique: additively manufactured or milled zirconia. The effect of airborne-particle abrasion and a zirconia liner before porcelain application was investigated in both groups. Veneering porcelain was fired into an alumina ring mold on the zirconia surface. SBS was measured by using a universal testing machine at a crosshead speed of 1 mm/min before and after aging (n=10). SBS data were analyzed with 3-way ANOVA (α=.05) RESULTS: A significant difference was found between milled and AM zirconia. The SBS of porcelain to milled zirconia was significantly higher (1.38 MPa) than to AM zirconia (0.68 MPa) (P<.001). The surface treatment of zirconia had no significant effect on porcelain SBS in either group (P=.254), whereas thermocycling significantly reduced the SBS of porcelain to zirconia in both milled and AM groups (P=.001). CONCLUSIONS: Porcelain bonding to milled zirconia was better than to AM zirconia. Pretreating the zirconia substrate before porcelain application did not improve the porcelain bond.
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PURPOSE: To characterize the microbiome composition within dental implants of peri-implantitis subjects and healthy controls using 16S rRNA gene sequencing. MATERIALS AND METHODS: Twenty-three subjects with healthy (n = 11 implants) and diseased (peri-implantitis, n = 21) implants were included in this controlled clinical cross-sectional study. Samples were obtained from internal surfaces of dental implants using sterile paper points for microbiological analysis. DNA was extracted, and the16S rRNA gene was amplified using universal primers targeting the V3-V4 regions. The resulting 16S polymerize chain reaction amplicons were sequenced on Illumina MiSeq, and the sequences were processed using DADA2 and the Human Oral Microbiome Database (HOMD) as references. Alpha and Beta diversity, as well as core microbiome and differential abundance analyses were then performed using the MicrobiomeAnalyst workflow. RESULTS: A significant increase in microbial diversity was observed in the internal implant surface of healthy implants compared with the internal surfaces of peri-implantitis (Shannon p = 0.02). Bacterial community structure was significantly different among groups (p = 0.012). High levels of Gram-positive bacteria were detected inside implants with peri-implantitis compared to healthy implants, especially Enterococci. CONCLUSIONS: There is a shift in bacterial diversity inside implants with peri-implantitis from the healthy control. The microbial colonization within that space might contribute to the etiology of peri-implant disease.