Effects of dentin bonding agents and silanization on bond strength between 3D printed resin and composite resin.

This study aimed to evaluate the effects of dentin bonding agents and silanization on the bond strength between 3D printed resin and composite resin and compare it with a conventional composite resin. 3D printed resin cylinders (PCB) and composite resin substrates (Z250) were prepared and divided into eight subgroups based on the bonding agents used (n=12). The shear bond strength was measured using a universal testing machine, and the failure modes were evaluated. The bond strength was found to vary significantly among the bonding agents and substrate types. Silane application did not significantly improve the bond strength. Among the bonding agents, the universal adhesives exhibited the highest bond strengths for both substrates. Compared to PCB, Z250 demonstrated stronger bonds and exhibited more cohesive failures. Further research is needed to optimize the surface treatments and resin formulations for enhanced bond strength and durability between 3D printed and composite resins.

Isolation of Murine Adipose-Derived Stromal/Stem Cells for Adipogenic and Osteogenic Differentiation or Flow Cytometry-Based Analysis.

Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Adipose tissue stromal vascular cells or primary adipocytes derived from murine adipose tissue and grown in culture are essential tools for studying the mechanisms underlying adipocyte development and function. Herein, we describe methods for the isolation, expansion, and long-term storage of murine adipose-derived stromal/stem cells, along with protocols for inducing adipogenesis to white or beige adipocytes in this cell population and osteogenic differentiation. Isolation of the adipose stromal vascular fraction cells for flow cytometric analysis is also described.

Translucency and Strength of Lithium Disilicate for Computer-Aided Design and Manufacturing at Different Thermal Temperatures and Thicknesses: An In Vitro Study.

To manufacture dental restorations composed of lithium disilicate (LD) through the computer-aided design and manufacturing (CAD/CAM) process, thermal refinement is an essential process that can affect the optical and mechanical properties of ceramics. In this study, we aimed to evaluate the translucency and flexural strength of lithium disilicate glass-ceramic for CAD/CAM using different thermal refinement schedules and thicknesses by measuring the total transmission of light through the specimen and calculating the peak load of the specimen until fracture in a piston-on-three-ball test, respectively. The results showed that a lower translucency was exhibited in thicker specimens, and the flexural strength decreased in the order of 1.0, 0.5, and 2.0 mm (p < 0.05). The lithium disilicates thermally refined at a heat of 820 degrees were shown to have the highest biaxial flexural strength (p < 0.05). These findings suggest that it is possible to adjust transparency and strength according to the clinical situation by choosing an appropriate thickness and thermal refinement process.

Bacillus-Derived Manganese Superoxide Dismutase Relieves Ocular-Surface Inflammation and Damage by Reducing Oxidative Stress and Apoptosis in Dry Eye.

Purpose: We hypothesized that antioxidative enzymes supplementation could be a treatment option for dry eye. We investigated the efficacy of oral administration of Bacillus-derived superoxide dismutase (Bd-SOD) in a murine experimental dry eye (EDE). Methods: In part I, mice were randomly assigned to normal control, EDE, and mice groups that were treated with oral Bd-SOD after induction of EDE (EDE + Bd-SOD group; four mice in each group). Expression of SOD2, a major antioxidant enzyme with manganese as a cofactor, was assessed by immunofluorescence staining. In part II, mice were divided into seven groups (six mice in each group): normal control, EDE, vehicle-treated, topical 0.05% cyclosporin A (CsA)-treated, and oral Bd-SOD-treated (2.5, 5.0, and 10.0 mg/kg Bd-SOD) groups. Tear volume, tear-film break-up time (TBUT), and corneal fluorescein-staining scores (CFS) were measured at zero, five, and 10 days after treatment. Ten days after treatment, 2',7'-dichlorodihydrofluorescein diacetate for reactive oxygen species (ROS), enzyme-linked immunosorbent for malondialdehyde, and TUNEL assays for corneal apoptosis, flow cytometry inflammatory T cells, and histological assessment were performed. Results: Compared to the normal control group in part I, the EDE group showed significantly decreased SOD2 expression by immunofluorescence staining. However, the EDE + Bd-SOD group recovered similar to the normal control group. In part II, ROS, malondialdehyde, and corneal apoptosis were decreased in CsA and all Bd-SOD-treated groups. Corneal and conjunctival inflammatory T cells decreased, and conjunctival goblet cell density increased in CsA-treated and Bd-SOD-treated groups. Compared to the CsA-treated group, the 2.5 mg/kg Bd-SOD-treated group showed increased TBUT and decreased inflammatory T cells, and the 5.0 mg/kg Bd-SOD-treated group showed decreased CFS and increased conjunctival goblet cells. Conclusions: Oral Bd-SOD administration might increase autogenous SOD2 expression in ocular surface tissue in EDE and could be developed as a complementary treatment for DE in the future.

Evaluation of Implant Stability According to Implant Placement Site and Duration in Elderly Patients: A Prospective Multi-Center Cohort Study.

The aim of this prospective study is to investigate implant stability and the reliability of different measuring devices according to implant placement site and duration in patients aged over 65 years. The study evaluated 60 implants (diameter: 3.5/4.0/4.5/5.0 mm and length: 8.5/10.0/11.5 mm) in 60 patients aged ≥ 65 years. The implant placement sites were divided into six evenly distributed sections (n = 10), i.e., maxillary right-posterior, A; maxillary anterior, B; maxillary left-posterior, C; mandibular right-posterior, D; mandibular anterior, E; mandibular left-posterior, F. Participants visited the hospital six times: implant surgery, 1V; stitch removal, 2V; 1-month follow-up, 3V; 2-month follow-up, 4V; before final restoration delivery, 5V; and after final restoration delivery, 6V. The implant stability was evaluated with the Osstell Mentor (ISQ), Periotest M (PTV), and Anycheck (IST). The mean values of ISQ, PTV, and IST were analyzed (α = 0.05). ISQ, PTV, and IST results of 4V and 5V were significantly higher than those of 1V (p < 0.05). The lowest ISQ results occurred in the E location at 4V and 5V (p < 0.05). In all mandibular locations, IST results of 6V were significantly higher than those of 1V, 2V, 3V, and 4V (p < 0.05). ISQ results were negatively correlated with PTV and positively correlated with IST, and PTV was negatively correlated with IST. By considering various factors affecting the stability of the implant, it is necessary to determine the appropriate implant load application time. This could help increase the implant success rate in elderly patients. And as a diagnostic device for implant stability and the evaluation of osseointegration in elderly patients, Anycheck was also able to prove its relative reliability compared to Osstell ISQ Mentor and Periotest M.

Reliability of implant stability measuring devices depending on various clinical conditions: an in vitro study.

PURPOSE: The aim of this study was to evaluate the reliability of implant stability measuring devices depending on the location of the implant and the position of the patient. MATERIALS AND METHODS: Six implants were installed in different dentate sextants of six artificial bone models. Implant stability was measured in three conditions of the bone model (without mounting on a phantom head, mounted on a phantom head in supine position, and mounted on a phantom head in upright position). A resonance frequency analysis device (Osstell) and two damping capacity analysis devices (Periotest and Anycheck) were used to measure implant stability. The values measured outside the phantom head were treated as controls, and the values inside the phantom head were compared using an independent t-test. RESULTS: Osstell showed different results in two of the six divisions in both the supine and upright positions compared to outside of the mouth (P < .05). Periotest showed different results in all six parts in the supine position and in five parts in the upright position compared to outside of the mouth (P < .05). While Anycheck showed different results in five areas in the supine position compared to outside of the mouth, it showed different results in only one area in the upright position (P < .05). CONCLUSION: In the difficult implant position for the operator to access, the implant stability measuring devices show less reliability. The accessibility of implant is greatly affected in the order of Osstell, Anycheck, and Periotest.

Characteristics of the Salivary Microbiota in Periodontal Diseases and Potential Roles of Individual Bacterial Species To Predict the Severity of Periodontal Disease.

The purposes of this study were to examine the compositional changes in the salivary microbiota according to the severity of periodontal disease and to verify whether the distribution of specific bacterial species in saliva can distinguish the severity of disease. Saliva samples were collected from 8 periodontally healthy controls, 16 patients with gingivitis, 19 patients with moderate periodontitis, and 29 patients with severe periodontitis. The V3 and V4 regions of the 16S rRNA gene in the samples were sequenced, and the levels of 9 bacterial species showing significant differences among the groups by sequencing analysis were identified using quantitative real-time PCR (qPCR). The predictive performance of each bacterial species in distinguishing the severity of disease was evaluated using a receiver operating characteristic curve. Twenty-nine species, including Porphyromonas gingivalis, increased as the severity of disease increased, whereas 6 species, including Rothia denticola, decreased. The relative abundances of P. gingivalis, Tannerella forsythia, Filifactor alocis, and Prevotella intermedia determined by qPCR were significantly different among the groups. The three bacterial species P. gingivalis, T. forsythia, and F. alocis were positively correlated with the sum of the full-mouth probing depth and were moderately accurate at distinguishing the severity of periodontal disease. In conclusion, the salivary microbiota showed gradual compositional changes according to the severity of periodontitis, and the levels of P. gingivalis, T. forsythia, and F. alocis in mouth rinse saliva had the ability to distinguish the severity of periodontal disease. IMPORTANCE Periodontal disease is one of the most widespread medical conditions and the leading cause of tooth loss, imposing high economic costs and an increasing burden worldwide as life expectancy increases. Changes in the subgingival bacterial community during the progression of periodontal disease can affect the entire oral ecosystem, and bacteria in saliva can reflect the degree of bacterial imbalance in the oral cavity. This study explored whether the specific bacterial species in saliva can distinguish the severity of periodontal disease by analyzing the salivary microbiota and suggested P. gingivalis, T. forsythia, and F. alocis as biomarkers for distinguishing the severity of periodontal disease in saliva.

Recent insights into the molecular mechanisms of simultaneous fatty acid oxidation and synthesis in brown adipocytes.

Brown adipocytes is a specialized fat cell that dissipates nutrient-derived chemical energy in the form of heat, instead of ATP synthesis. This unique feature provides a marked capacity for brown adipocyte mitochondria to oxidize substrates independent of ADP availability. Upon cold exposure, brown adipocytes preferentially oxidize free fatty acids (FFA) liberated from triacylglycerol (TAG) in lipid droplets to support thermogenesis. In addition, brown adipocytes take up large amounts of circulating glucose, concurrently increasing glycolysis and de novo FA synthesis from glucose. Given that FA oxidation and glucose-derived FA synthesis are two antagonistic mitochondrial processes in the same cell, it has long been questioned how brown adipocytes run FA oxidation and FA synthesis simultaneously. In this review, I summarize mechanisms regulating mitochondrial substrate selection and describe recent findings of two distinct populations of brown adipocyte mitochondria with different substrate preferences. I further discuss how these mechanisms may permit a concurrent increase in glycolysis, FA synthesis, and FA oxidation in brown adipocytes.

Biochemical responses and phytoremediation potential of Azolla imbricata (Roxb.) Nakai in water and nutrient media exposed to waste metal cutting fluid along with temperature and humidity stress.

Phytoremediation of metals from water (WM) and nutrient (NM) media exposed to waste metal cutting fluid (WMCF) along with temperature (T) and humidity (H) stress was tested using Azolla imbricata (Roxb.) Nakai. In the absence of WMCF, biomass was higher in NM than in WM during all tests. Surprisingly, opposite results were noted in the presence of WMCF, with growth failing at exposure to > 0.1% and > 0.5% in NM and WM, respectively. Further, correlation analysis of the growth data following WM exposure revealed that biomass was affected positively by T and negatively by H and metal accumulation. Simultaneously, metal accumulation was affected negatively by T and positively by H. The average accumulations of Al, Cd, Cr, Fe, Pb, and Zn across all T/H tests were 540, 282, 71, 1645, 2494 and 1110 mg·kg-1, respectively. The observed bioconcentration factor indicated that A. imbricata acts as a hyperaccumulator or accumulator of Zn (>10) and as either accumulator (>1) or excluder (<1) of the other metals. Overall, the phytoremediation performance of A. imbricata in multi-metal-contaminated WMCF was high in WM under all environmental conditions. Therefore, the use of WM is an economically feasible approach for the removal of metals from WMCF.

Implant stability changes over time following implant placement in elderly patients: A prospective study.

OBJECTIVES: This study aimed to evaluate the effect of aging on the changes in implant stability over time following implant placement. MATERIALS AND METHODS: A total of 104 patients in four age ranges (group 1: <60 years, group 2: 61-70 years, group 3: 71-80 years, and group 4: >80 years) were included. Bone-level tapered implants were placed without implementing any bone augmentation procedure. The final torque value displayed on the implant engine during implant insertion was recorded. Cone-beam computed tomography (CBCT) was performed immediately after surgery to analyze the bone quality around the implant. Implant stability was measured immediately after surgery and 2, 4, and 8 weeks after surgery. RESULTS: In the CBCT image, higher grayscale values were observed in the order of group 1, group 2, and groups 3/4, with statistical significance (p < .05). There was no significant difference in the insertion torque values between age groups (p ≥ .05). Groups 1 and 2 showed lower implant stability values after 2 and 4 weeks compared to immediately and 8 weeks after surgery (p < .05); however, groups 3 and 4 showed no significant difference between the results measured at different timepoints (p ≥ .05). CONCLUSIONS: Implant treatment in elderly patients is successful showing a settled implant stability over time following implant placement when the implant is appropriately engaged in the alveolar bone in the absence of bone augmentation.

Assessment of Esophageal Reconstruction via Bioreactor Cultivation of a Synthetic Scaffold in a Canine Model.

OBJECTIVES: Using tissue-engineered materials for esophageal reconstruction is a technically challenging task in animals that requires bioreactor training to enhance cellular reactivity. There have been many attempts at esophageal tissue engineering, but the success rate has been limited due to difficulty in initial epithelialization in the special environment of peristalsis. The purpose of this study was to evaluate the potential of an artificial esophagus that can enhance the regeneration of esophageal mucosa and muscle through the optimal combination of a double-layered polymeric scaffold and a custom-designed mesenchymal stem cell-based bioreactor system in a canine model. METHODS: We fabricated a novel double-layered scaffold as a tissue-engineered esophagus using an electrospinning technique. Prior to transplantation, human-derived mesenchymal stem cells were seeded into the lumen of the scaffold, and bioreactor cultivation was performed to enhance cellular reactivity. After 3 days of cultivation using the bioreactor system, tissue-engineered artificial esophagus was transplanted into a partial esophageal defect (5×3 cm-long resection) in a canine model. RESULTS: Scanning electron microscopy (SEM) showed that the electrospun fibers in a tubular scaffold were randomly and circumferentially located toward the inner and outer surfaces. Complete recovery of the esophageal mucosa was confirmed by endoscopic analysis and SEM. Esophagogastroduodenoscopy and computed tomography also showed that there were no signs of leakage or stricture and that there was a normal lumen with complete epithelialization. Significant regeneration of the mucosal layer was observed by keratin-5 immunostaining. Alpha-smooth muscle actin immunostaining showed significantly greater esophageal muscle regeneration at 12 months than at 6 months. CONCLUSION: Custom-designed bioreactor cultured electrospun polyurethane scaffolds can be a promising approach for esophageal tissue engineering.

Clinical Validation of Dental Implant Stability by Newly Designed Damping Capacity Assessment Device during the Healing Period.

Background and Objectives: To evaluate the stability of a dental implant and the effectiveness of a newly designed damping capacity assessment device by improving the number of blows and strength evaluated by a prospective clinical study. Materials and Method: The stability of dental implants was measured in 50 implants in a total of 38 patients. Measurements were performed using Anycheck and Periotest M devices, twice in total, divided into buccal and lingual directions. In addition, measurements were performed on the day of surgery, two weeks, one month, two months, and three months after surgery for a total of five times. After the standardization of the measured values, the differences and changes over time for each device were observed. Result: No difference in standardized values between the two devices was observed at any time point. In both devices, stability decreased at two weeks postoperatively but gradually increased thereafter. No differences were observed in the values according to the measurement direction. Conclusions: The damping capacity of Anycheck was similar to that of Periotest M. After a slight decrease in stability two weeks after implant placement, implant stability increased over time.

Melatonin Type 2 Receptor Activation Regulates Blue Light Exposure-Induced Mouse Corneal Epithelial Damage by Modulating Impaired Autophagy and Apoptosis.

The MT1/2 receptors, members of the melatonin receptor, belong to G protein-coupled receptors and mainly regulate circadian rhythms and sleep in the brain. Previous studies have shown that in many other cells and tissues, such as HEK293T cells and the retina, MT1/2 receptors can be involved in mitochondrial homeostasis, antioxidant, and anti-inflammatory responses. In our study, we aimed to investigate the effects of blue light (BL) exposure on the expression of melatonin and its receptors in the mouse cornea and to evaluate their functional role in corneal epithelial damage. After exposing 8-week-old C57BL/6 mice to BL at 25 and 100 J/cm2 twice a day for 14 days, a significant increase in the expression of 4-HNE and MT2 was observed in the cornea. MT2 antagonist-treated mice exposed to BL showed an increased expression of p62 and decreased expression of BAX and cleaved caspase 3 compared with mice exposed only to BL. In addition, MT2 antagonist-treated mice showed more enhanced MDA and corneal damage. In conclusion, BL exposure can induce MT2 expression in the mouse cornea. MT2 activation can modulate impaired autophagy and apoptosis by increasing the expression of BAX, an apoptosis activator, thereby regulating the progression of corneal epithelial damage induced by BL exposure.

Relationship Between Nursing Students' Awareness of Disaster, Preparedness for Disaster, Willingness to Participate in Disaster Response, and Disaster Nursing Competency.

OBJECTIVE: This study was conducted to provide basic data for preparing a disaster nursing education program. It examined the degree of nursing students' disaster awareness, disaster preparedness, willingness to participate in disaster response, and disaster nursing competency, aiming to determine the relationship between these attributes. METHODS: This was a descriptive research study. The participants were 163 nursing students. The data collected from the participants were analyzed via descriptive statistics and Pearson's correlation coefficients. RESULTS: Disaster awareness showed a positive correlation with a willingness to participate in a disaster response. Further, disaster preparedness and willingness to participate in a disaster response showed a positive correlation with disaster nursing capacity. Disaster awareness did not show a significant correlation with disaster preparedness and disaster nursing competency. Last, disaster preparedness did not show a significant correlation with willingness to participate in a disaster response. CONCLUSIONS: It is necessary to improve nursing students' disaster awareness, disaster preparation, disaster response participation willingness, and disaster nursing competency. It is imperative to develop disaster nursing education programs to strengthen students' capabilities in a comprehensive manner.

Evaluation of microleakage and push-out bond strength of various composite resins for sealing the screw-access channel in implant-supported restorations.

STATEMENT OF PROBLEM: Microleakage and loss of the composite resin sealing the screw-access channel are frequent complications of screw-retained implant-supported prostheses. How the screw-access channel should be best restored to reduce such complications is unclear. PURPOSE: The purpose of this in vitro study was to evaluate the microleakage and bond strength of 3 types of composite resins (flowable, packable, and bulk-fill) with or without a bonding agent treatment to seal the screw-access channel of 2 types of restorative materials (zirconia and Co-Cr alloy) with or without thermocycling. MATERIAL AND METHODS: In total, 240 yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) specimens (IPS e.max ZirCAD) and 240 Co-Cr alloy (Vera PDS) specimens were prepared with a Ø3×3-mm cylindrical cavity at the center to simulate the screw-access channel. Three types of composite resins (flowable, packable, and bulk-fill resin) (Filtek Z350 XT Flowable Restorative, Filtek Z350 XT Universal Restorative, and Filtek One Bulk Fill Restorative) were applied to restore the access channel of the zirconia and Co-Cr specimens with or without a bonding agent (Single Bond Universal Adhesive). Microleakage and push-out bond strength were determined and compared by dividing the specimens into experimental groups with or without thermocycling (1000 times with 30 seconds at 5 ±2 °C and 55 ±2 °C). The results were analyzed by using a 1-way ANOVA and 4-way ANOVA. Adjustment for multiple comparisons was made with the Tukey Honestly Significant Difference (HSD) test. RESULTS: The specimens subjected to thermocycling showed a lower bond strength (P<.001) and higher microleakage (P<.001) than specimens stored in a constant-temperature water bath. Specimens treated with bonding agents showed a higher bond strength (P<.001) and lower microleakage (P<.001) than specimens not treated with a bonding agent. Higher bond strengths were observed in the order of bulk-fill resin, packable resin, and flowable resin (P<.001). Packable resin showed higher microleakage than flowable resin and bulk-fill resin (P<.05). No significant difference in microleakage was found between the flowable resin and bulk-fill resin (P>.05). CONCLUSIONS: Higher bond strengths were observed in the order of bulk-fill resin, packable resin, and flowable resin. Less microleakage was observed in the flowable resin and bulk-fill resin than in the packable composite resin. Bonding agent treatment was effective in increasing bond strength and decreasing microleakage. Zirconia and Co-Cr showed a bond strength similar to that of composite resins, but zirconia showed higher microleakage than Co-Cr. Restoring the screw-access channel with the bulk-fill resin should increase bond strength and reduce microleakage.

Analysis of Strategies to Increase User Retention of Fitness Mobile Apps during and after the COVID-19 Pandemic.

The COVID-19 pandemic has changed the fitness-related field. More people started working out at home, and the use of fitness mobile apps that can measure the amount of exercise through a scientific method has increased compared to before the COVID-19 pandemic. This phenomenon is likely to continue even after the COVID-19 pandemic, and therefore this study aimed to investigate the importance of and satisfaction with a fitness app's functions according to consumers while using the fitness mobile app. Through this study, we intended to provide data for creating an environment where users can use fitness mobile apps consistently. A total of 420 questionnaires were distributed through Google Survey for about 3 months, from 13 September to 20 November 2020, and a total of 399 complete questionnaires were analyzed in this study. Regarding the data processing methods, frequency analysis, exploratory factor analysis, reliability analysis, descriptive statistical analysis, and IPA were used. The results are as follows. First, the first quadrant of the IPA matrix indicated the high importance of and satisfaction with the fitness mobile app, and included five attributes: cost-effectiveness, easy-to-understand information, ease of use and application, privacy protection, and compatibility with other devices. Second, the second quadrant of the matrix indicated relatively low satisfaction in association to high importance and included five attributes: accurate exercise information provision, design efficiency, daily exercise amount setting, convenient icons and interface, and provision of images and videos in appropriate proportions. Third, the third quadrant of the matrix, indicating low importance and low satisfaction, included five attributes: not sharing personal information, overall design composition and color, customer service, reliable security level, and providing information on goal achievement after exercising. Fourth, in the quadrant of the matrix, indicating low importance and high satisfaction, five attributes were included: exercise notification function, continuous service provision, step count and heart rate information, individual exercise recommendation, and individual body type analysis information.

Guided Bone Regeneration with a Nitric-Oxide Releasing Polymer Inducing Angiogenesis and Osteogenesis in Critical-Sized Bone Defects.

Synthetic scaffolds, as bone grafts, provide a favorable environment for the repair and growth of new bone tissue at defect sites. However, the lack of angio- and osteo-induction limits the usefulness of artificial scaffolds for bone regeneration. Nitric oxide (NO) performs essential roles in healing processes, such as regulating inflammation and addressing incomplete revascularization. In this study, a polymer capable of controlled NO release is developed to promote the osteogenic capacity in artificial scaffolds. The biological efficiency of the NO compound is assessed by its effect on pre-osteoblasts and macrophages in vitro and the extent of vascularization and bone formation in the calvaria defect model in vivo. The compound does not inhibit cell adhesion or proliferation. NO treatment significantly increases both alkaline phosphatase activity and mineralization in pre-osteoblasts. Macrophages treated with NO secrete high levels of anti-inflammatory factors and adopt the pro-regenerative phenotype. In the critical-sized defect model, the collagen scaffold containing the NO compound enhances neovascularization and bone formation. The developed NO-releasing system promotes osteogenesis and regeneration of damaged bone tissue. As the multiple functions of NO involve macrophage modulation and angiogenesis, such release systems may be valuable for guiding bone regeneration in critical-sized defects.

Injection laryngoplasty of human adipose-derived stem cell spheroids with hyaluronic acid-based hydrogel improves the morphological and functional characteristics of geriatric larynx.

AIM: As the geriatric population increased, the need of treatment for laryngeal atrophy and dysfunction increased. This study was performed to evaluate the effects of injection of human adipose-derived stem cell (hASC) spheroid-loaded catechol-conjugated hyaluronic acid (HA-CA) hydrogel on therapeutic rejuvenation of the geriatric larynx. METHODS: Stem cell spheroids with hyaluronic acid-based hydrogel were injected into the laryngeal muscles of 18-month-old Sprague-Dawley rats. The effects of hASC spheroids were examined in the following four groups: SHAM, injected with PBS; GEL, injected with HA-CA hydrogel; MONO, injected with single hASCs in HA-CA hydrogel; and SP, injected with hASCs spheroids in HA-CA hydrogel. The rejuvenation efficacy in geriatric laryngeal muscle tissues at 12 weeks postinjection was evaluated and compared by histology, immunofluorescence staining, and functionality analysis. RESULTS: Total myofiber cross-sectional area and myofiber number/density, evaluated by detection of myosin heavy chain with antibodies against laminin and fast myosin heavy chain, were significantly higher in the SP group than in the other groups. The lamina propria of the larynx was evaluated by alcian blue staining, which showed that the HA was increased significantly in the SP group compared to the other groups. In functional analysis, the glottal gap area was significantly reduced in the SP group compared to the other groups. The phase difference in the vocal fold during vibration was also smaller in the SP group than in the other groups, but the difference did not reach statistical significance. CONCLUSION: Injection of hASC spheroids with hyaluronic acid-based hydrogel improves the morphological and functional characteristics of geriatric larynx.

Polarization of human gingival fibroblasts by Th1-, Th2-, Th17-, and Treg-derived cytokines.

BACKGROUND AND OBJECTIVE: The purpose of this study was to evaluate whether gingival fibroblasts (GFs) can be differently activated and polarized into distinct functional subtypes by T-helper (Th) cytokines. METHODS: Gingival fibroblasts were stimulated with interferon (IFN)-γ, interleukin (IL)-4, IL-17, and transforming growth factor (TGF)-ß, representative cytokines of Th1, Th2, Th17, and regulatory T cells, respectively, and the gene expression profiles were analyzed by microarray. Differentially expressed genes (DEGs) in GFs stimulated by 4 cytokines were screened, and a gene ontology (GO) analysis of the DEGs was conducted. To confirm the reliability of the microarray results, the DEGs that showed the largest differences compared with non-stimulated GFs were further analyzed by RT-PCR. To evaluate the effect of polarization on GFs responses to lipopolysaccharide (LPS), GFs stimulated by 4 cytokines were further stimulated with Escherichia coli LPS and mRNA levels of several genes were analyzed using RT-PCR. RESULTS: Differentially expressed genes by 4 Th cytokines were enriched in different GO terms, and the patterns of gene expression on GFs were shown functionally different. GFs stimulated with IFN-γ (GF(IFN-γ)) up-regulated the expression of chemokines (chemokine (C-X-C motif) ligand (CXCL)9, -10, -11, chemokine (C-C motif) ligand (CCL)8), molecules involved in antigen presentation, complement component 3 (C3), and other immune response-related molecules, whereas they down-regulated the expression of several types of collagen, extracellular matrix (ECM) components, and DNA replication and nuclear protein-related molecules. By contrast, GF(IL-4) up-regulated the expression of ECM components, cell adhesion molecules, and tissue development-related molecules and down-regulated the expression of chemokines (CXCL10 and CXCL8) and adaptive immune response-related molecules. GF(IL-17) up-regulated the expression of chemokines and other molecules for neutrophil infiltration and activation, the pro-inflammatory cytokine IL-6, and C3. GF(TGF-ß) up-regulated the expression of cell growth-related molecules, ECM components, several types of collagen, and cell adhesion molecules and down-regulated the expression of molecules related to complement activation and bacterial recognition. GFs stimulated by 4 cytokines responded differently to LPS. CONCLUSION: These results show that Th cytokines can polarize GFs into cells with functionally distinct features: immune-activating but tissue-destructive GF(IFN-γ), tissue-reparative, and immune-inhibiting GF(IL-4), highly pro-inflammatory GF(IL-17), and potent tissue-reparative GF(TGF-ß).

Protein Kinase SGK2 Is Induced by the ß3 Adrenergic Receptor-cAMP-PKA-PGC-1α/NT-PGC-1α Axis but Dispensable for Brown/Beige Adipose Tissue Thermogenesis.

Brown and beige adipocytes are specialized to dissipate energy as heat. Sgk2, encoding a serine/threonine kinase, has been identified as a brown and beige adipocyte-specific gene in rodents and humans; however, its function in brown/beige adipocytes remains unraveled. Here, we examined the regulation and role of Sgk2 in brown/beige adipose tissue thermogenesis. We found that transcriptional coactivators PGC-1α and NT-PGC-1α activated by the ß3 adrenergic receptor-cAMP-PKA pathway are recruited to the Sgk2 promoter, triggering Sgk2 transcription in response to cold. SGK2 elevation was closely associated with increased serine/threonine phosphorylation of proteins carrying the consensus RxRxxS/T phosphorylation site. However, despite cold-dependent activation of SGK2, mice lacking Sgk2 exhibited normal cold tolerance at 4°C. In addition, Sgk2+/+ and Sgk2-/- mice induced comparable increases in energy expenditure during pharmacological activation of brown and beige adipose tissue with a ß3AR agonist. In vitro loss- and gain-of-function studies further demonstrated that Sgk2 ablation or activation does not alter thermogenic gene expression and mitochondrial respiration in brown adipocytes. Collectively, our results reveal a new signaling component SGK2, although dispensable for cold-induced thermogenesis that adds an additional layer of complexity to the ß3AR signaling network in brown/beige adipose tissue.