Neurotoxic Microglial Activation via IFNγ-Induced Nrf2 Reduction Exacerbating Alzheimer's Disease.

Microglial neuroinflammation appears to be neuroprotective in the early pathological stage, yet neurotoxic, which often precedes neurodegeneration in Alzheimer's disease (AD). However, it remains unclear how the microglial activities transit to the neurotoxic state during AD progression, due to complex neuron-glia interactions. Here, the mechanism of detrimental microgliosis in AD by employing 3D human AD mini-brains, brain tissues of AD patients, and 5XFAD mice is explored. In the human and animal AD models, amyloid-beta (Aß)-overexpressing neurons and reactive astrocytes produce interferon-gamma (IFNγ) and excessive oxidative stress. IFNγ results in the downregulation of mitogen-activated protein kinase (MAPK) and the upregulation of Kelch-like ECH-associated Protein 1 (Keap1) in microglia, which inactivate nuclear factor erythroid-2-related factor 2 (Nrf2) and sensitize microglia to the oxidative stress and induces a proinflammatory microglia via nuclear factor kappa B (NFκB)-axis. The proinflammatory microglia in turn produce neurotoxic nitric oxide and proinflammatory mediators exacerbating synaptic impairment, phosphorylated-tau accumulation, and discernable neuronal loss. Interestingly, recovering Nrf2 in the microglia prevents the activation of proinflammatory microglia and significantly blocks the tauopathy in AD minibrains. Taken together, it is envisioned that IFNγ-driven Nrf2 downregulation in microglia as a key target to ameliorate AD pathology.

Botulinum Neurotoxin Induces Neurotoxic Microglia Mediated by Exogenous Inflammatory Responses.

Botulinum neurotoxin serotype A (BoNT/A) is widely used in therapeutics and cosmetics. The effects of multi-dosed BoNT/A treatment are well documented on the peripheral nervous system (PNS), but much less is known on the central nervous system (CNS). Here, the mechanism of multi-dosed BoNT/A leading to CNS neurodegeneration is explored by using the 3D human neuron-glia model. BoNT/A treatment reduces acetylcholine, triggers astrocytic transforming growth factor beta, and upregulates C1q, C3, and C5 expression, inducing microglial proinflammation. The disintegration of the neuronal microtubules is escorted by microglial nitric oxide, interleukin 1ß, tumor necrosis factor α, and interleukin 8. The microglial proinflammation eventually causes synaptic impairment, phosphorylated tau (pTau) aggregation, and the loss of the BoNT/A-treated neurons. Taking a more holistic approach, the model will allow to assess therapeutics for the CNS neurodegeneration under the prolonged use of BoNT/A.

Environmental aluminum oxide inducing neurodegeneration in human neurovascular unit with immunity.

Aluminum oxide nanoparticle (AlNP), a ubiquitous neurotoxin highly enriched in air pollution, is often produced as an inevitable byproduct in the manufacturing of industrial products such as cosmetics and metal materials. Meanwhile, ALNP has emerged as a significant public health concern due to its potential association with neurological diseases. However, the studies about the neurotoxic effects of AlNP are limited, partially due to the lack of physiologically relevant human neurovascular unit with innate immunity (hNVUI). Here, we employed our AlNP-treated hNVUI model to investigate the underlying mechanism of AlNP-driven neurodegeneration. First, we validated the penetration of AlNP across a blood-brain barrier (BBB) compartment and found AlNP-derived endothelial cellular senescence through the p16 and p53/p21 pathways. Our study showed that BBB-penetrating AlNP promoted reactive astrocytes, which produced a significant level of reactive oxygen species (ROS). The astrocytic neurotoxic factors caused neuronal damage, including the synaptic impairment, the accumulation of phosphoric-tau proteins, and even neuronal death. Our study suggests that AlNP could be a potential environmental risk factor of neurological disorders mediated by neuroinflammation.

Laminin-Augmented Decellularized Extracellular Matrix Ameliorating Neural Differentiation and Neuroinflammation in Human Mini-Brains.

Non-neural extracellular matrix (ECM) has limited application in humanized physiological neural modeling due to insufficient brain-specificity and safety concerns. Although brain-derived ECM contains enriched neural components, certain essential components are partially lost during the decellularization process, necessitating augmentation. Here, it is demonstrated that the laminin-augmented porcine brain-decellularized ECM (P-BdECM) is xenogeneic factor-depleted as well as favorable for the regulation of human neurons, astrocytes, and microglia. P-BdECM composition is comparable to human BdECM regarding brain-specificity through the matrisome and gene ontology-biological process analysis. As augmenting strategy, laminin 111 supplement promotes neural function by synergic effect with laminin 521 in P-BdECM. Annexin A1(ANXA1) and Peroxiredoxin(PRDX) in P-BdECM stabilized microglial and astrocytic behavior under normal while promoting active neuroinflammation in response to neuropathological factors. Further, supplementation of the brain-specific molecule to non-neural matrix also ameliorated glial cell inflammation as in P-BdECM. In conclusion, P-BdECM-augmentation strategy can be used to recapitulate humanized pathophysiological cerebral environments for neurological study.

Comparison of the accuracy of a cone beam computed tomography-based virtual mounting technique with that of the conventional mounting technique using a facebow.

STATEMENT OF PROBLEM: The introduction of digital technology in dentistry has resulted in a shift from conventional methods to digital techniques. However, mounting a digitized dental cast on a virtual articulator is challenging. Several techniques have been suggested to resolve this problem, but in the absence of a standardized method, digitized dental casts are often mounted arbitrarily on a virtual articulator. PURPOSE: The purpose of this clinical study was to compare the accuracy of a novel virtual facebow transfer (VM) technique based on cone beam computed tomography (CBCT) with that of the conventional mounting (CM) technique using a facebow. MATERIAL AND METHODS: Five repeated mountings were performed with each technique for 15 participants. In the CM group, dental casts were mounted using a facebow record and scanned for transmission to the virtual dental space. In the VM group, digital dental casts were mounted on the standard tessellation language file of a reference articulator by reconstructing a file of the participant's skull from CBCT data. In this group, a virtual facebow, prepared by scanning the articulator and facebow complex, was used. After the CM and VM casts had been aligned, the coordinates of target points set on the maxillary right central incisor, maxillary right first molar, and maxillary left first molar were determined, and the mean ±standard deviation distance between the target points was calculated to compare the precision of the techniques. Additionally, vectors of the target point on the maxillary right central incisor were compared to analyze the spatial difference between the techniques. Finally, the occlusal plane angle was calculated. For the correlation analysis of repeated measured data, a 1-way repeated measures analysis of variance (ANOVA) was first performed. The Kolmogorov-Smirnov test was performed to determine normality, and a paired t test and the Wilcoxon signed rank test were performed for normally and nonnormally distributed variables, respectively (α=.05). RESULTS: The mean distance between target points was significantly greater in the CM group (4.72 ±1.45 to 5.17 ±1.54 mm) than in the VM group (2.14 ±0.58 to 2.35 ±0.60 mm) (P<.05). The standard deviation between target points was significantly greater in the CM group (1.60 ±0.64 to 2.30 ±0.87 mm) than in the VM group (0.74 ±0.23 to 1.12 ±0.45 mm) (P<.05). The maxillary right central incisor was located more anteriorly in the VM group than in the CM (100%, P<.05) group. The occlusal plane angle was significantly steeper in the CM group than in the VM group (8.14 degrees versus 2.13 degrees, P<.05). CONCLUSIONS: The VM technique was more precise than the CM technique. VM casts were positioned ahead of CM casts. Further, the occlusal plane angle tended to be steeper with the CM technique than with the VM technique.

Three-dimensional human neural culture on a chip recapitulating neuroinflammation and neurodegeneration.

Neuroinflammation has either beneficial or detrimental effects, depending on risk factors and neuron-glia interactions in neurological disorders. However, studying neuroinflammation has been challenging due to the complexity of cell-cell interactions and lack of physio-pathologically relevant neuroinflammatory models. Here, we describe our three-dimensional microfluidic multicellular human neural culture model, referred to as a 'brain-on-a-chip' (BoC). This elucidates neuron-glia interactions in a controlled manner and recapitulates pathological signatures of the major neurological disorders: dementia, brain tumor and brain edema. This platform includes a chemotaxis module offering a week-long, stable chemo-gradient compared with the few hours in other chemotaxis models. Additionally, compared with conventional brain models cultured with mixed phenotypes of microglia, our BoC can separate the disease-associated microglia out of heterogeneous population and allow selective neuro-glial engagement in three dimensions. This provides benefits of interpreting the neuro-glia interactions while revealing that the prominent activation of innate immune cells is the risk factor leading to synaptic impairment and neuronal loss, validated in our BoC models of disorders. This protocol describes how to fabricate and implement our human BoC, manipulate in real time and perform end-point analyses. It takes 2 d to set up the device and cell preparations, 1-9 weeks to develop brain models under disease conditions and 2-3 d to carry out analyses. This protocol requires at least 1 month training for researchers with basic molecular biology techniques. Taken together, our human BoCs serve as reliable and valuable platforms to investigate pathological mechanisms involving neuroinflammation and to assess therapeutic strategies modulating neuroinflammation in neurological disorders.

Astrocytic scar restricting glioblastoma via glutamate-MAO-B activity in glioblastoma-microglia assembloid.

BACKGROUND: Glial scar formation is a reactive glial response confining injured regions in a central nervous system. However, it remains challenging to identify key factors formulating glial scar in response to glioblastoma (GBM) due to complex glia-GBM crosstalk. METHODS: Here, we constructed an astrocytic scar enclosing GBM in a human assembloid and a mouse xenograft model. GBM spheroids were preformed and then co-cultured with microglia and astrocytes in 3D Matrigel. For the xenograft model, U87-MG cells were subcutaneously injected to the Balb/C nude female mice. RESULTS: Additional glutamate was released from GBM-microglia assembloid by 3.2-folds compared to GBM alone. The glutamate upregulated astrocytic monoamine oxidase-B (MAO-B) activity and chondroitin sulfate proteoglycans (CSPGs) deposition, forming the astrocytic scar and restricting GBM growth. Attenuating scar formation by the glutamate-MAO-B inhibition increased drug penetration into GBM assembloid, while reducing GBM confinement. CONCLUSIONS: Taken together, our study suggests that astrocytic scar could be a critical modulator in GBM therapeutics.

Retentive strength of cement-retained implant-supported fixed dental prostheses according to different cement types and cementation protocols: An in vitro study.

PURPOSE: To develop the most compatible cementation protocol for ensuring minimal residual cement and optimal retention of cement-retained implant-supported fixed dental prostheses. MATERIALS AND METHODS: Thirty custom implant abutments and zirconia crowns with bilateral wings were prepared. Three cement types were used for cementation: non-eugenol resin cement (Premier Implant Cement; Group IC), dual-polymerizing self-adhesive resin cement (SmartCem 2; Group SC), and zinc oxide eugenol cement (Temp-Bond; Group TB) (n=30 per group). Three cementation methods were applied for each cement type and the samples were divided into subgroups: 1) cement was injected using a graduated syringe (IC-N, SC-N, and TB-N); 2) a cementation jig made with a silicone impression material and temporary resin material was used (IC-CJ, SC-CJ, and TB-CJ); 3) three dimensionally (3D) printed abutments were used as replicas for cementation (IC-3DP, SC-3DP, and TB-3DP). The amount of cement injected, surface area of the residual cement, and retentive strength were measured. Kruskal-Wallis and post-hoc Mann-Whitney tests were used for statistical analyses. RESULTS: Excess cement was not observed when cementation jig or 3D-printed replicas were used. For IC and SC subgroups, non-use of these auxiliary tools resulted in significantly higher amounts of injected cement. The retentive strength differed significantly among the IC subgroups, but not among the SC subgroups. The retentive strength of subgroups TB-N and TB-CJ was significantly higher than that of subgroup TB-3DP. CONCLUSIONS: To prolong the main purpose of each cement type, a cementation jig or 3D-printed replica is highly recommended regardless of the cement type. Int J Prosthodont 2023. doi: 10.11607/ijp.8344.

Effect of cement space settings on the marginal and internal fit of 3D-printed definitive resin crowns.

STATEMENT OF PROBLEM: The cement gap setting affects the marginal and internal fits depending on the crown material and manufacturing method (subtractive or additive manufacturing). However, information on the effects of cement space settings in the computer-aided design (CAD) software program, which is used to aid the manufacturing with 3-dimensional (3D) printing-type resin material, is lacking, and recommendations for optimal marginal and internal fit are needed. PURPOSE: The purpose of this in vitro study was to evaluate how cement gap settings affect the marginal and internal fit of a 3D-printed definitive resin crown. MATERIAL AND METHODS: After scanning a prepared typodont left maxillary first molar, a crown was designed with cement spaces of 35, 50, 70, and 100 µm by using a CAD software program. A total of 14 specimens per group were 3D printed from definitive 3D-printing resin. By using the replica technique, the intaglio surface of the crown was duplicated, and the duplicated specimen was sectioned in the buccolingual and mesiodistal directions. Statistical analyses were performed using the Kruskal-Wallis and the Mann-Whitney post hoc tests (α=.05). RESULTS: Although the median values of the marginal gaps were within the clinically acceptable limit (<120 µm) for all the groups, the smallest marginal gaps were obtained with the 70-µm setting. For the axial gaps, there was no observed difference in the 35-, 50-, and 70-µm groups, and the 100-µm group showed the largest gap. The smallest axio-occlusal and occlusal gaps were obtained with the 70-µm setting. CONCLUSIONS: Based on the findings of this in vitro study, a 70-µm cement gap setting is recommended for optimal marginal and internal fit of 3D-printed resin crowns.

Effect of airborne particle abrasion treatment of two types of 3D-printing resin materials for permanent restoration materials on flexural strength.

OBJECTIVES: This study aimed to assess the effects of airborne-particle abrasion (APA) on the flexural strength of two types of 3D-printing resins for permanent restoration. METHODS: Two types of 3D printing resins (urethane dimethacrylate oligomer; UDMA, ethoxylated bisphenol-A dimethacrylate; BEMA) constituting different components were printed. The specimen surfaces were subjected to APA using 50 and 110 µm alumina particles under different pressures. The three-point flexural strength was measured for each surface treatment group, and a Weibull analysis was performed. Surface characteristics were analyzed via surface roughness measurements and scanning electron microscopy. Dynamic mechanical analysis and nano-indentation measurements were limited to the control group. RESULTS: The three-point flexural strength according to the surface treatment was significantly lower in the UDMA group for large particle sizes and at high pressures; the BEMA group demonstrated low flexural strength for large particle sizes regardless of the pressure. After thermocycling, the flexural strengths of UDMA and BEMA significantly decreased in the group subjected to surface treatment. The Weibull modulus and characteristic strength of UDMA were higher than those of BEMA under different APA and thermocycling conditions. As the abrasion pressure and particle size increased, a porous surface formed, and the surface roughness increased. Compared with BEMA, UDMA featured a lower strain, greater strain recovery, and a negligible increase in modulus according to strain. SIGNIFICANCE: Thus, surface roughness increased with the sandblasting particle size and pressure of the 3D-printing resin. Hence, a suitable surface treatment method to improve adhesion can be determined by considering physical property changes.

Effect of Adhesion Conditions on the Shear Bond Strength of 3D Printing Resins after Thermocycling Used for Definitive Prosthesis.

Three-dimensional (3D) printing polymers such as urethane dimethacrylate (UDMA) and ethoxylated bisphenol A dimethacrylate (Bis-EMA) are typically used in definitive prosthesis and require surface treatments before bonding. However, surface treatment and adhesion conditions often affect long-term use. Herein, polymers were divided into Groups 1 and 2 for the UDMA and Bis-EMA components, respectively. The shear bond strength (SBS) between two types of 3D printing resins and resin cements was measured using Rely X Ultimate Cement and Rely X U200, according to adhesion conditions such as single bond universal (SBU) and airborne-particle abrasion (APA) treatments. Thermocycling was performed to evaluate the long-term stability. Sample surface changes were observed using a scanning electron microscope and surface roughness measuring instrument. The effect of interaction between the resin material and adhesion conditions on the SBS was analyzed via a two-way analysis of variance. The optimal adhesion condition for Group 1 was achieved when U200 was used after APA and SBU, whereas Group 2 was not significantly affected by the adhesion conditions. After thermocycling, the SBS significantly decreased in Group 1 without APA treatment and in the entire Group 2. Additionally, porosity, along with increased roughness, was observed on both material surfaces after APA.

Human mini-brains for reconstituting central nervous system disorders.

Neurological disorders in the central nervous system (CNS) are progressive and irreversible diseases leading to devastating impacts on patients' life as they cause cognitive impairment, dementia, and even loss of essential body functions. The development of effective medicines curing CNS disorders is, however, one of the most ambitious challenges due to the extremely complex functions and structures of the human brain. In this regard, there are unmet needs to develop simplified but physiopathologically-relevant brain models. Recent advances in the microfluidic techniques allow multicellular culture forming miniaturized 3D human brains by aligning parts of brain regions with specific cells serving suitable functions. In this review, we overview designs and strategies of microfluidics-based human mini-brains for reconstituting CNS disorders, particularly Alzheimer's disease (AD), Parkinson's disease (PD), traumatic brain injury (TBI), vascular dementia (VD), and environmental risk factor-driven dementia (ERFD). Afterward, the applications of the mini-brains in the area of medical science are introduced in terms of the clarification of pathogenic mechanisms and identification of promising biomarkers. We also present expanded model systems ranging from the CNS to CNS-connecting organ axes to study the entry pathways of pathological risk factors into the brain. Lastly, the advantages and potential challenges of current model systems are addressed with future perspectives.

Engineered Human Intervertebral Disc Model Inducing Degenerative Microglial Proinflammation.

Degeneration of the intervertebral disc (IVD) is a major contributor to low back pain (LBP). IVD degeneration is characterized by abnormal production of inflammatory cytokines secreted by IVD cells. Although the underlying molecular mechanisms of LBP have not been elucidated, increasing evidence suggests that LBP is associated particularly with microglia in IVD tissues and the peridiscal space, aggravating the cascade of degenerative events. In this study, we implemented our microfluidic chemotaxis platform to investigate microglial inflammation in response to our reconstituted degenerative IVD models. The IVD models were constructed by stimulating human nucleus pulposus (NP) cells with interleukin-1ß and producing interleukin-6 (129.93 folds), interleukin-8 (18.31 folds), C-C motif chemokine ligand-2 (CCL-2) (6.12 folds), and CCL-5 (5.68 folds). We measured microglial chemotaxis (p < 0.05) toward the conditioned media of the IVD models. In addition, we observed considerable activation of neurodegenerative and deactivation of protective microglia via upregulated expression of CD11b (p < 0.001) and down-regulation of CD206 protein (p < 0.001) by soluble factors from IVD models. This, in turn, enhances the inflammatory milieu in IVD tissues, causing matrix degradation and cellular damage. Our findings indicate that degenerative IVD may induce degenerative microglial proinflammation, leading to LBP development.

Comparative evaluation of digitally fabricated complete dentures versus conventional complete dentures: A randomized, single-blinded, cross-over clinical trial.

STATEMENT OF PROBLEM: Clinical trials comparing outcomes associated with digital complete dentures (CDs) fabricated from intraoral scan data with those of CDs fabricated by using the conventional workflow are lacking. PURPOSE: The purpose of this randomized clinical trial was to evaluate the clinical performance of and patient satisfaction associated with digitally versus conventionally fabricated CDs. MATERIAL AND METHODS: Eight participants requiring CDs were enrolled in this study. Two sets of CDs were fabricated for each participant. One set was fabricated by using a digital workflow, which involved digital scanning with an intraoral scanner, whereas the other set was made by using the conventional workflow. The participants were given 1 set of CDs for 1 month and another set for the next month. The order of placing CDs was randomly selected for each participant. The internal adaptation, masticatory force, and masticatory efficiency of the CDs in each group were evaluated for objective analysis. Additionally, a questionnaire was provided to the participants, and the responses were evaluated for subjective satisfaction analysis. All parameters were analyzed by using t tests (α=.05). RESULTS: The internal adaptation did not statistically significantly differ between the conventional and digital CDs with regard to the maxillary arches (P=.406) and mandibular arches (P=.412). The average masticatory force (P=.051) and maximum masticatory force (P=.110) likewise did not statistically significantly differ between the 2 types of CDs. Masticatory efficiency, expressed via the mixing ability index, was statistically better for conventional CDs than the digital CDs (P=.009). No statistically significant differences were observed between the 2 types of CDs in terms of overall patient satisfaction as assessed by using the study questionnaire (P=.172 for maxillary CD and P=.161 for mandibular CD). However, the conventional CDs were statistically significantly better than the digital CDs with regard to subjective satisfaction with pronunciation ability (P=.006). CONCLUSIONS: The digital CDs were inferior to the conventional CDs in terms of masticatory efficiency and pronunciation. However, internal adaptation and overall patient satisfaction were comparable between conventional and digital CDs. This finding suggests that intraoral scanning and additively manufactured CDs may be suitable for edentulous patients, at least for interim use.

TREM2 regulates purinergic receptor-mediated calcium signaling and motility in human iPSC-derived microglia.

The membrane protein TREM2 (Triggering Receptor Expressed on Myeloid cells 2) regulates key microglial functions including phagocytosis and chemotaxis. Loss-of-function variants of TREM2 are associated with increased risk of Alzheimer's disease (AD). Because abnormalities in Ca2+ signaling have been observed in several AD models, we investigated TREM2 regulation of Ca2+ signaling in human induced pluripotent stem cell-derived microglia (iPSC-microglia) with genetic deletion of TREM2. We found that iPSC-microglia lacking TREM2 (TREM2 KO) show exaggerated Ca2+ signals in response to purinergic agonists, such as ADP, that shape microglial injury responses. This ADP hypersensitivity, driven by increased expression of P2Y12 and P2Y13 receptors, results in greater release of Ca2+ from the endoplasmic reticulum stores, which triggers sustained Ca2+ influx through Orai channels and alters cell motility in TREM2 KO microglia. Using iPSC-microglia expressing the genetically encoded Ca2+ probe, Salsa6f, we found that cytosolic Ca2+ tunes motility to a greater extent in TREM2 KO microglia. Despite showing greater overall displacement, TREM2 KO microglia exhibit reduced directional chemotaxis along ADP gradients. Accordingly, the chemotactic defect in TREM2 KO microglia was rescued by reducing cytosolic Ca2+ using a P2Y12 receptor antagonist. Our results show that loss of TREM2 confers a defect in microglial Ca2+ response to purinergic signals, suggesting a window of Ca2+ signaling for optimal microglial motility.

An Air Particulate Pollutant Induces Neuroinflammation and Neurodegeneration in Human Brain Models.

Fine particulate matter (PM2.5), a major component among air pollutants, highlights as a global health concern. Several epidemiological studies show the correlation between chronical PM2.5 exposure and incidents of neurological disorders including Alzheimer's disease. However, the mechanisms have not been well understood, partly due to the lack of model systems that reflect the physiologically relevant innate immunity in human brains. Here, PM2.5-polluted human brain models (PMBs) are created in a 3D microfluidic platform reconstituting key aspects of human brain immunity under the PM2.5 exposure. PM2.5 penetration across a blood-brain barrier (BBB) model and accumulation in the brain tissue side of the model are first validated. Second, the PMB model shows that the BBB-penetrating PM2.5 initiates astrogliosis, resulting in slight neuronal loss and microglial infiltration. Third, it is demonstrated that the infiltrating microglia obtain M1 phenotype induced by interleukin-1ß and interferon-γ from neurons and reactive astrocytes under the PM2.5 exposure. Finally, it is observed that additional proinflammatory mediators and nitric oxide released from the M1 microglia exacerbate neuronal damages, such as synaptic impairment, phosphoric tau accumulation, and neuronal death. This study suggests that PM2.5 can be a potential environmental risk factor for dementia mediated by the detrimental neuroinflammation.

Oral Pathogenic Bacteria-Inducing Neurodegenerative Microgliosis in Human Neural Cell Platform.

Porphyromonas gingivalis is a gram-negative bacterium found in the human oral cavity and is responsible for the development of chronic periodontitis as well as neurological diseases, including Alzheimer's disease (AD). Given the significance of the roles of P. gingivalis in AD pathogenesis, it is critical to understand the underlying mechanisms of P. gingivalis-driven neuroinflammation and their contribution to neurodegeneration. Herein, we hypothesize that P. gingivalis produces secondary metabolites that may cause neurodegeneration through direct or indirect pathways mediated by microglia. To test our hypothesis, we treated human neural cells with bacterial conditioned media on our brain platforms and assessed microgliosis, astrogliosis and neurodegeneration. We found that bacteria-mediated microgliosis induced the production of nitric oxide, which causes neurodegeneration assessed with high pTau level. Our study demonstrated the elevation of detrimental protein mediators, CD86 and iNOS and the production of several pro-inflammatory markers from stimulated microglia. Through inhibition of LPS and succinate dehydrogenase in a bacterial conditioned medium, we showed a decrease in neurodegenerative microgliosis. In addition, we demonstrated the bidirectional effect of microgliosis and astrogliosis on each other exacerbating neurodegeneration. Overall, our study suggests that the mouth-brain axis may contribute to the pathogenesis of AD.

Influence of shoulder coverage difference of abutment on stress distribution and screw stability in tissue-level internal connection implants: A finite element analysis and in vitro study.

STATEMENT OF PROBLEM: Tissue-level internal connection implants are widely used, but the difference in abutment screw stability because of the shoulder coverage formed by the contact between the shoulder of the implant collar and the abutment remains unclear. PURPOSE: The purpose of this finite element analysis (FEA) and in vitro study was to investigate stress distribution and abutment screw stability as per the difference in shoulder coverage of the abutment in tissue-level internal connection implants. MATERIAL AND METHODS: Abutments were designed in 3 groups as per the shoulder coverage of the implant collar, yielding complete coverage (complete group), half coverage (half group), no coverage (no group) groups. In the FEA, a tightening torque of 30.0 Ncm was applied to the abutment screw, a force of 250 N was applied to the crown at a 30-degree angle, and the von Mises stresses and the stress distribution patterns were evaluated. In the in vitro study, the groups were tested (n=12). A total of 200 000 cyclic loads were applied at 250 N, 14 Hz, and at a 30-degree angle. Removal torque values and scanning electron microscopy (SEM) images were assessed. Removal torque values were analyzed by ANOVA and paired t tests. RESULTS: The maximum von Mises stress of the abutment screw was the lowest in the complete group, slightly higher in the half group, and highest in the no group. High stresses were concentrated in 1 location in the implant abutment connection area of the no group. The removal torque values after loading were significantly lower in the no group than in the complete group (P=.047). The SEM images revealed concentrated structural loss and wear in 1 location of the no group. CONCLUSIONS: FEA and in vitro studies confirmed that the shoulder coverage of the abutment in the tissue-level internal connection implant helped improve screw stability. Cyclic loading reduced the removal torque of the abutment screw.

Tumor Targeted Delivery of an Anti-Cancer Therapeutic: An In Vitro and In Vivo Evaluation.

The limited effectiveness of current therapeutics against malignant brain gliomas has led to an urgent need for development of new formulations against these tumors. Chelator Dp44mT (di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone) presents a promising candidate to defeat gliomas due to its exceptional anti-tumor activity and its unique ability to overcome multidrug resistance. The goal of this study is to develop a targeted nano-carrier for Dp44mT delivery to glioma tumors and to assess its therapeutic efficacy in vitro and in vivo. Dp44mT is loaded into poly(ethylene glycol) (PEG)ylated poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) decorated with glioma-targeting ligand Interlukin 13 (IL13). IL13-conjugation enhanced the NP uptake by glioma cells and also improved their transport across an in vitro blood-brain-barrier (BBB) model. This targeted formulation showed an outstanding toxicity towards glioma cell lines and patient-derived stem cells in vitro, with IC50 values less than 125 nM, and caused no significant death in healthy brain microvascular endothelial cells. In vivo, when tested on a xenograft mouse model, IL13-conjugated Dp44mT-NPs reduced the glioma tumor growth by ≈62% while their untargeted counterparts reduced the tumor growth by only ≈16%. Notably, this formulation does not cause any significant weight loss or kidney/liver toxicity in mice, demonstrating its great therapeutic potential.