Inhibition of nucleotide biosynthesis disrupts lipid accumulation and adipogenesis.

Energy balance and nutrient availability are key determinants of cellular decisions to remain quiescent, proliferate, or differentiate into a mature cell. After assessing its environmental state, the cell must rewire its metabolism to support distinct cellular outcomes. Mechanistically, how metabolites regulate cell fate decisions is poorly understood. We used adipogenesis as our model system to ascertain the role of metabolism in differentiation. We isolated adipose tissue stromal vascular fraction cells and profiled metabolites before and after adipogenic differentiation to identify metabolic signatures associated with these distinct cellular states. We found that differentiation alters nucleotide accumulation. Furthermore, inhibition of nucleotide biosynthesis prevented lipid storage within adipocytes and downregulated the expression of lipogenic factors. In contrast to proliferating cells, in which mechanistic target of rapamycin complex 1 is activated by purine accumulation, mechanistic target of rapamycin complex 1 signaling was unaffected by purine levels in differentiating adipocytes. Rather, our data indicated that purines regulate transcriptional activators of adipogenesis, peroxisome proliferator-activated receptor γ and CCAAT/enhancer-binding protein α, to promote differentiation. Although de novo nucleotide biosynthesis has mainly been studied in proliferation, our study points to its requirement in adipocyte differentiation.

Bi-directional and multi-modal effects of dexamphetamine on spatial binding windows in healthy individuals.

OBJECTIVES: Stimuli that are separated by a short window of space or time, known as spatial and temporal binding windows (SBW/TBWs), may be perceived as separate. Widened TBWs are evidenced in schizophrenia, although it is unclear if the SBW is similarly affected. The current study aimed to assess if dexamphetamine (DEX) may increase SBWs in a multimodal visuo-tactile illusion, potentially validating usefulness as an experimental model for multimodal visuo-tactile hallucinations in schizophrenia, and to examine a possible association between altered binding windows (BWs) and working memory (WM) suggested by previous research. METHODS: A placebo-controlled, double-blinded, and counter-balanced crossover design was employed. Permuted block randomisation was used for drug order. Healthy participants received DEX (0.45 mg/kg, PO, b.i.d.) or placebo (glucose powder) in capsules. The Rubber Hand Illusion (RHI) and Wechsler Adult Intelligence Scale Spatial Span was employed to determine whether DEX would alter SBWs and WM, respectively. Schizotypy was assessed with a variety of psychological scales. RESULTS: Most participants did not experience the RHI even under normal circumstances. Bi-directional and multimodal effects of DEX on individual SBWs and schizotypy were observed, but not on WM. CONCLUSIONS: Bidirectional multimodal effects of DEX on the RHI and SBWs were observed in individuals, although not associated with alterations in WM.

Dexamphetamine increased speech and visual unimodal illusions in healthy participants without affecting temporal binding window.

OBJECTIVE: Stimuli received beyond a very short timeframe, known as temporal binding windows (TBWs), are perceived as separate events. In previous audio-visual multisensory integration (McGurk effect) studies, widening of TBWs has been observed in people with schizophrenia. The present study aimed to determine if dexamphetamine could increase TBWs in unimodal auditory and unimodal visual illusions that may have some validity as experimental models for auditory and visual hallucinations in psychotic disorders. METHODS: A double-blind, placebo-controlled, counter-balanced crossover design with permuted block randomisation for drug order was followed. Dexamphetamine (0.45 mg/kg, PO, q.d.) was administered to healthy participants. Phantom word illusion (speech illusion) and visual-induced flash illusion/VIFI (visual illusion) tests were measured to determine if TBWs were altered as a function of delay between stimuli presentations. Word emotional content for phantom word illusions was also analysed. RESULTS: Dexamphetamine significantly increased the total number of phantom words/speech illusions (p < 0.01) for pooled 220-1100 ms ISIs in kernel density estimation and the number of positive valence words heard (beta = 2.20, 95% CI [1.86, 2.55], t = 12.46, p < 0.001) with a large effect size (std. beta = 1.05, 95% CI [0.89, 1.22]) relative to placebo without affecting the TBWs. For the VIFI test, kernel density estimation for pooled 0-801 ms ISIs showed a significant difference (p < 0.01) in the data distributions of number of target flash (es) perceived by participants after receiving dexamphetamine as compared with placebo. CONCLUSIONS: Overall, healthy participants who were administered dexamphetamine (0.45 mg/kg, PO, q.d.) experienced increases in auditory and visual illusions in both phantom word illusion and VIFI tests without affecting their TBWs.

Evaluation of different tightening protocols on angled screw channel abutments after cyclic loading: An in vitro study.

STATEMENT OF PROBLEM: Screw loosening remains one of the most common complications associated with implant-supported restorations. The available literature is scarce regarding the most efficient method of tightening definitive abutment screws for angled screw channel (ASC) abutments. PURPOSE: The purpose of this in vitro study was to identify the optimal screw tightening protocol for ASC abutments by evaluating the influence of different protocols on reverse tightening values (RTVs). MATERIAL AND METHODS: Fifty implants were randomly assigned to 5 groups (n=10). Implants were mounted and received a cover screw and impression coping to simulate the clinical scenario. Each group was allocated to one of the following protocols: tightened once (1×), tightened twice (2×), tightened twice at a 10-minute interval (2× [10 min]), tightened, countertightened, then tightened again (TCT) and tightened, countertightened, tightened, countertightened, then tightened (TCTCT). A monolithic zirconia central incisor implant crown was fabricated by using computer-aided design and computer-aided manufacturing (CAD-CAM) and cemented onto the ASC abutment. The crown-abutment assemblies were secured onto the implants with the Gold-Tite abutment screw by using their assigned tightening protocols. Each screw was tightened to the recommended value of 20 Ncm with a calibrated digital torque measuring device. Specimens were subjected to 250 000 cycles of cyclic loading simulating 3 months of function with a universal testing machine. RTV was measured for each specimen. RESULTS: The mean RTV for all the groups ranged from 13.5 Ncm to 15.9 Ncm. The highest mean RTV was found with the (1×) protocol. However, the difference was not statistically significant among any of the groups (P>.05). CONCLUSIONS: Multiple retightening and time intervals between tightening events of these ASC abutment screws did not have a significant influence on RTV under simulated function of 3 months. The one time (1×) tightening protocol can be recommended as a suitable method for ASC abutments, thus avoiding unnecessary retightening.

Evaluating the effects of splinting implant scan bodies intraorally on the trueness of complete arch digital scans: A clinical study.

STATEMENT OF PROBLEM: A predictable protocol for accurately scanning implants in a complete edentulous arch has not been established. PURPOSE: The purpose of this clinical study was to investigate the effect of splinting implant scan bodies intraorally on the accuracy and scan time for digital scans of edentulous arches. MATERIAL AND METHODS: This single center, nonrandomized, clinical trial included a total of 19 arches. Definitive casts with scan bodies were fabricated and scanned with a laboratory scanner as the reference (control) scan. Each participant received 2 intraoral scans, the first with unsplinted scan bodies and the second with resin-splinted scan bodies. The scan time was also recorded for each scan. To compare the accuracy of the scans, the standard tessellation language (STL) files of the 2 scans were superimposed on the control scan, and positional and angular deviations were analyzed by using a 3-dimensional (3D) metrology software program. The Mann-Whitney U test was used to compare the distance and angular deviations between the splinted group and the unsplinted group with the control. The ANOVA test was conducted to examine the effect of the scan technique on trueness (distance deviation and angular deviation) and scan time (α=.05 for all tests). RESULTS: Statistically significant differences were found in the overall 3D positional and angular deviations of the unsplinted and splinted digital scans when compared with the reference scans (P<.05). No statistically significant differences in overall 3-dimensional positional deviations (P=.644) and angular deviations (P=.665) were found between the splinted and unsplinted experimental groups. A faster scan time was found with the splinted group in the maxillary arch. CONCLUSIONS: Conventional complete arch implant impressions were more accurate than digital complete arch implant scans. Splinting implant scan bodies did not significantly affect the trueness of complete arch digital scans, but splinting appeared to reduce the scan time. However, fabricating the splint was not considered in the time measurement.

Digital workflow for duplication of gold onlay anatomy to monolithic zirconia onlay for a complete denture: A technique paper.

Digital replication of an existing cast gold onlay anatomy to a more esthetic material for a new complete denture is challenging. A technique is presented that uses digital technology to fabricate a new maxillary complete denture with monolithic zirconia onlays that duplicate the anatomy of an existing complete denture with cast gold onlays.

Identifying marginal adaptation discrepancies of lithium disilicate crowns using seven different vertical X-ray angulations.

PURPOSE: To study the effect of different vertical angulations on the ability to radiographically assess vertical marginal discrepancies of lithium disilicate crowns. MATERIALS AND METHODS: Twenty-one lithium disilicate crowns were fabricated for three different prepared natural teeth: incisor, canine, and premolar. Vertical marginal discrepancies ranging from 0 to 300 µm were intentionally created. The seated crowns were radiographed using seven different vertical angulations, totaling 147 images. Thirty experienced evaluators scored each image for marginal discrepancy, and values were statistically analyzed. RESULTS: Significant differences in the ability to accurately assess marginal discrepancies from radiographs were observed for the study factors of angulation, tooth type, and degree of marginal discrepancy (p < 0.001). CONCLUSIONS: The radiographic interpretation of the marginal discrepancies of lithium disilicate crowns is significantly affected by the dimension of the marginal discrepancy. Specifically on premolar crowns, it is significantly affected by different vertical angulations of the X-ray beam. When evaluating marginal discrepancy on lithium disilicate crowns radiographically, vertical beam angulation within ±10° to the cemento-enamel junctionCEJ plane is recommended.

Effect of finishing and polishing techniques on the fit accuracy and dimensions of conventional and CAD-CAM removable partial denture frameworks.

PURPOSE: The purpose of this study was to compare the effects of finishing and polishing techniques on the fit accuracy, metal loss, and surface roughness of conventional versus CAD-CAM removable partial denture (RPD) frameworks. MATERIALS AND METHODS: A 3D-printed maxillary Kennedy class III modification I model served as the master cast. Forty impressions (20 conventional and 20 digital) were divided into four groups: lost-wax technique (Group I: LWT), CAD-printed (Group II: CAD-RP), CAD-printed from a stone cast (Group III: CAD-RPS), and lost-wax technique from resin-printed models (Group IV: LWTR). Various finishing and polishing techniques were applied, followed by digital scanning for fit accuracy assessment using surface matching software. Metal thickness loss and surface roughness were evaluated pre- and post-finishing and polishing. The Kruskal-Wallis test followed by the Scheffe post-hoc test were conducted to evaluate the fit accuracy between groups (α = 0.05). RESULTS: Color mapping revealed significant differences (p < 0.001) between conventionally casted RPD frameworks and 3D-printed groups post-finishing and polishing. The most significant gap was observed with the guide plates from printed RPD frameworks. The D-Lyte technique resulted in less metal loss compared to the conventional finishing and polishing technique (p < 0.001). CONCLUSION: Within the limitations of this study, conventionally processed RPD groups exhibited better overall fit accuracy post-finishing and polishing. Both conventional cast and 3D-printed RPD frameworks showed clinically acceptable fit accuracy. The D-Lyte technique presented less metal loss and smoother surfaces compared to other groups, suggesting it as a viable alternative.

Dexamphetamine widens temporal and spatial binding windows in healthy participants.

BACKGROUND: The pathophysiology of psychosis is complex, but a better understanding of stimulus binding windows (BWs) could help to improve our knowledge base. Previous studies have shown that dopamine release is associated with psychosis and widened BWs. We can probe BW mechanisms using drugs of specific interest to psychosis. Therefore, we were interested in understanding how manipulation of the dopamine or catecholamine systems affect psychosis and BWs. We aimed to investigate the effect of dexamphetamine, as a dopamine-releasing stimulant, on the BWs in a unimodal illusion: the tactile funneling illusion (TFI). METHODS: We conducted a randomized, double-blind, counterbalanced placebo-controlled crossover study to investigate funnelling and errors of localization. We administered dexamphetamine (0.45 mg/kg) to 46 participants. We manipulated 5 spatial (5-1 cm) and 3 temporal (0, 500 and 750 ms) conditions in the TFI. RESULTS: We found that dexamphetamine increased funnelling illusion (p = 0.009) and increased the error of localization in a delay-dependent manner (p = 0.03). We also found that dexamphetamine significantly increased the error of localization at 500 ms temporal separation and 4 cm spatial separation (p interaction = 0.009; p 500ms|4cm v. baseline = 0.01). LIMITATIONS: Although amphetamine-induced models of psychosis are a useful approach to understanding the physiology of psychosis related to dopamine hyperactivity, dexamphetamine is equally effective at releasing noradrenaline and dopamine, and, therefore, we were unable to tease apart the effects of the 2 systems on BWs in our study. CONCLUSION: We found that dexamphetamine increases illusory perception on the unimodal TFI in healthy participants, which suggests that dopamine or other catecholamines have a role in increasing tactile spatial and temporal BWs.

Dexamphetamine influences funneling illusion based on psychometric score.

OBJECTIVES: Our team previously showed that like the experience of the rubber hand illusion (RHI) in people with schizophrenia and their offspring¸ dexamphetamine administration to healthy volunteers increases the stimulus binding windows (BWs) in RHI. It is not clear if similar expansions of BWs are present for unimodal illusions. Studies have also shown that subjective or objective effects of amphetamine would be linked to between-person variations in personality measures. Therefore, we aimed to examine the effect of dexamphetamine (DEX), a dopamine-releasing stimulant, on illusory perception using unimodal sensory stimuli (Tactile Funneling Illusion [TFI]) across both temporal and spatial variables. We further examined the relationship between changes in psychometric scores and changes in illusion perception induced by dexamphetamine. METHODS: Healthy subjects (N = 20) participated in a randomized, double-blind, counter-balanced, placebo-controlled, cross-over study. The effects of dexamphetamine (0.45 mg/kg, PO, q.d.) on funneling and error of spatial localization (EL) were examined using TFI. Psychotomimetic effects were assessed using a battery of psychological measures. RESULTS: Dexamphetamine did not significantly increased the funneling illusion (p = 0.88) or EL (p = 0.5), relative to placebo. However, the degree of change in psychometric scores following dexamphetamine positively correlated with changes in funneling (ρ = 0.48, p = 0.03, n = 20), mainly at 0 ms delay condition (ρ = 0.6, p = 0.004, n = 20). CONCLUSION: Unlike multimodal illusions, alteration of BWs does not occur for unimodal illusions after administration of a dopamine-releasing agent. However, our findings indicate that moderate release of dopamine, through its psychotomimetic effect, indirectly influences unimodal illusion.

Effect of implant abutment screw materials and tightening protocols on reverse tightening values: An in vitro study.

STATEMENT OF PROBLEM: Implant abutment screw loosening is the most common prosthetic complication of implant-supported single crowns. However, few studies have objectively evaluated the effectiveness of different tightening protocols on reverse tightening values (RTVs). PURPOSE: The purpose of this in vitro study was to determine the optimal tightening protocol for implant abutment screws with different screw materials. MATERIAL AND METHODS: Sixty implants from 2 implant systems (Keystone and Nobel Biocare) with different definitive screw materials were selected. One group used diamond-like carbon (DLC) coated screws (DLC Group), and the other used titanium nitride (TiN) screws (TiN Group). Each group consisted of 30 implants. The implants in each group were distributed randomly into 3 subgroups (n=10). The implants from both manufacturers were mounted in resin blocks by following a clinical component connection protocol: a cover screw was placed, then an impression coping, and finally an original manufacturer prefabricated abutment. The abutment screws were tightened to the manufacturer's recommended tightening value using 3 different protocols: tighten the screw once (1T); tighten the abutment screw to the recommended tightening value, wait 10 minutes, and then retighten (2T); and tighten the abutment screw to the recommended tightening value, countertighten, tighten, countertighten, and then tighten (3TC). RTVs were measured after 3 hours. The Shapiro-Wilk test was performed to test for normal distribution of the data. The Kruskal-Wallis test was applied to each system's group that was not normally distributed (P<.05). Where differences existed, a post hoc analysis using the Dwass-Steel-Critchlow-Flinger (DSCF) pairwise comparisons test was conducted. RESULTS: No significant differences were found among the 3 different tightening groups in the TiN group (P>.05). However, significant differences were found among the 3 different tightening protocols in the DLC group (P<.05). CONCLUSIONS: Abutment screw systems from different manufacturers behave differently with respect to how they are tightened. For the TiN screw group, statistically similar RTVs were found for the 3 tightening protocols. The most efficient tightening protocol for the DLC-coated screw was the 3TC-DLC.

Digital beading for a CAD-CAM framework for an implant-supported overdenture: A dental technique.

The analog technique of making bead lines for a metal superstructure framework for the maxillary implant-supported overdenture is created by scribing shallow grooves on the surface of a definitive gypsum cast. This report describes a digital technique that uses computer-aided design and computer-aided manufacturing technology to make dental bead lines on an intraoral impression without using the gypsum cast.

Complications Associated with Metal Resin Fixed Complete Dentures Based on Implant Distribution.

PURPOSE: To compare prevalence and type of complications associated with metal resin fixed complete dentures based on implant distribution. MATERIALS AND METHODS: This retrospective study included data collected for a period of 12 years for patients treated with maxillary and mandibular implant-supported fixed complete dentures at Loma Linda University School of Dentistry. In total, 223 patient's charts were reviewed which included 100 maxillary and 123 mandibular fixed complete dentures. Implant distribution and prevalence of complications associated with each implant-supported fixed complete denture were documented. Tooth delamination, the most common complication associated with fixed complete dentures, was compared between 2 implant distribution groups in each arch. Covariates including age, sex, opposing arch, cantilever occluding units, and number of implants were evaluated to determine their association with tooth delamination. Data and hypotheses were statistically analyzed using descriptive statistics along with logistic regression model. All tests of hypotheses were considered statistically significant at an alpha level of 0.05. RESULTS: In the maxillary arch, some effect of the prevalence of tooth delamination was seen for the group that had implants placed posterior to canine eminence but it was not statistically significant. Denture tooth delamination had the highest prevalence among complications irrespective of implant distribution. Significant denture tooth delamination was seen for patients with opposing fixed complete dentures in comparison to patients with removable opposing arch prostheses. CONCLUSIONS: Implant distribution is not a significant factor related to metal resin fixed complete dentures. Opposing fixed complete dentures have a significantly higher association with denture tooth delamination.

Alloys Used in Different Temporomandibular Joint Reconstruction Replacement Prostheses Exhibit Variable Microstructures and Electrochemical Properties.

PURPOSE: Metallic temporomandibular joint replacement (TMJR) systems vary depending on design, material composition, and manufacturing methods such as casting, forging, and additive manufacturing. Therefore, the purpose of this study was to measure the association between manufacturing process of TMJR systems in terms of microstructure and electrochemical properties. MATERIALS AND METHODS: The sample was composed of new or surgically retrieved TMJ replacement devices of either titanium alloy (Ti6Al4V) or cobalt-chromium-molybdenum (CoCrMo) alloy from 8 different manufacturers. The primary predictor variable was alloy type, according to its manufacturing process (wrought, cast, additively manufactured [AM]). The primary outcome variables were 1) microstructure (grain size, aspect ratio, and phase content) and 2) corrosion potential and current, polarization resistance, and capacitance. Differences between alloy groups were determined by t tests, Kruskal-Wallis, and Mann-Whitney tests. RESULTS: We demonstrated that the TMJR CoCrMo and Ti6Al4V alloy microstructures can vary broadly within American Society for Testing and Materials specifications, where the components made of Ti6Al4V had 3 types of microstructures (equiaxial, bimodal, and martensitic) out of 10 samples, and the components made of CoCrMo had 2 types of microstructure (equiaxial and dendritic) out of 16 samples. Some CoCrMo alloys exhibited preferential corrosion sites, while wrought Ti6Al4V alloys trended toward a superior corrosion behavior (corrosion rate: 2 × 10-9 A/cm2, polarization resistance: 5,000,000 kΩcm2, and capacitance: 10 µSsa/cm2) compared with AM alloys (39 × 10-9 A/cm2, 1676 kΩcm2, 36 µSsa/cm2, respectively), where 4 samples of each group were tested and repeated 5 times. Among four AM devices, two exhibited a significantly inferior corrosion behavior. CONCLUSIONS: Although AM is an exciting emerging new technology that allows manufacturing of custom-made TMJR, their corrosion behavior is still inferior in comparison to that of traditional wrought alloys. Preventing corrosion is crucial because it can cause surface defects that may lead to implant fracture.

Evaluating the effect of the protector cap for castable implant abutments on reverse tightening values.

STATEMENT OF PROBLEM: Screw loosening is the most common mechanical complication with implant prostheses. How the alteration of implant-to-abutment connection surfaces that occurs during laboratory procedures affects screw loosening is unclear. PURPOSE: The purpose of this in vitro study was to compare the reverse tightening value (RTV) differences between custom castable abutments before casting, after casting in a conventional manner, and after casting with custom protector caps and pegs. MATERIAL AND METHODS: Thirty implants with a standard-diameter conical connection (NobelReplace Conical Connection 4.3×13 mm; Nobel Biocare AG) and 30 premachined 4.3-mm GoldAdapt abutments (GoldAdapt; Nobel Biocare AG) were selected for this study. Specimens were divided into 3 groups (n=10): the uncast custom castable abutment group (UCCA) in which abutments were new and not cast; the unprotected custom castable abutment group (UPCCA) in which abutments were cast and devested with airborne-particle abrasion; and the protected custom castable abutment group (PCCA) in which abutments were cast by using protector caps and pegs made by milling zirconia and then devested with airborne-particle abrasion. All abutments in each group were tightened to 35 Ncm with a calibrated digital tightening device. After 10 minutes, all screws were retightened to 35 Ncm. At 3 hours, each screw was loosened, and the value at which the initial loosening occurred was documented as the RTV. The results were statistically analyzed with 1-way ANOVA to explore differences, and post hoc tests with Tukey adjustment were used for multiple comparisons. RESULTS: Among the tested groups, the mean RTV ranged from 19.89 Ncm to 27.19 Ncm: UCCA 27.19 Ncm, UPCCA 19.89 Ncm, and PCCA 24.24 Ncm. A significant difference was found among the tested groups (P<.05). CONCLUSIONS: Casting procedures, especially devestment with airborne-particle abrasion, affected implant-abutment connections and the seat site of the screw. Protecting the implant connection site and the seat site of the abutment screw with protector caps and pegs prevented a significant loss of the RTV.

Scanning accuracy with splinted and unsplinted implant scan bodies for the edentulous arch at implant level: an in vitro study.

Accuracy of completely edentulous arch scanning with implant scan bodies has not been completely validated for intraoral scanners. For desktop laboratory scanners validations were found in the literature. The aim of this in vitro study was to compare the dimensional accuracy of scanning with splinted and unsplinted scan bodies on a completely edentulous maxillary arch with 6 implants. A maxillary edentulous master cast with 6 implants was used as control for all implant level impressions. 6 implants were digitally planned and placed at different angulation of 0, 0  17° and 30°. A contact coordinate measuring machine (CMM) was used to generate baseline linear measurements of the master cast. Four test groups included: 2 intraoral scanners Trios (3shape, Copenhagen, Denmark) and True Definition (3M ESPE, St Paul, MN), one industrial scanner Atos (Core optical 3D scanner) and one conventional impression group scanned with laboratory scanner Dental Wings 7 Series. Each scan recording was made with splinted and unsplinted scan bodies. Digital datasets of all measurements were compared with the CMM baseline values using PolyWorks® InnovMetric Software in order to assess the linear and angular deviations and determine the accuracy of complete arch digital impression. Factorial ANOVA showed significant effect with splinting, scanner type, inter-implant distance and implant angulation for linear deviations. Splinted scan bodies revealed elevated linear and angular deviation values for all scanners with significantly highest deviations for True Definition scanner. Significant correlation was found between inter-implant distance and linear deviation (r =0.45, P =<0.001) with increased linear deviations among all scanners, regardless of splinting. ( P <0.05). Significant effect on accuracy was seen for angular deviation with splinting, scanner type and implant angulation; significant difference was found between 0° and 30° implant angulation ( P =0.035) with more deviation with 30° implant angulation. The accuracy of the complete arch implant digital impression using splinted scan bodies was significantly reduced for measurements that crossed the arch midline. The digital impression technique using a broad splint design to connect scan bodies was not found to improve the scan accuracy for intraoral scanners.

Dynamic microfluidic bioreactor-Hip simulator (DMBH) system for implant toxicity monitoring.

The generation of degradation products (DPs) like ions and organo-metallic particles from corroding metallic implants is an important healthcare concern. These DPs generate local and systemic toxicity. The impact on local toxicity is well documented, however, little is known about systemic toxicity. This is mainly due to the limited scope of the current microtiter plate-based (static) toxicity assay techniques. These methods do not mimic the systemic (dynamic) conditions. In this study, it is hypothesized that DPs incubated with cells in static conditions might provide improper systemic toxicity results, as there is no movement mimicking the blood circulation around cells. This study reports the development of a three-chambered prototype microfluidic system connected to the operational hip implant simulator to test the cellular response induced by the DPs. This setup is called a dynamic microfluidic bioreactor-hip simulator system. We hypothesize that a dynamic microfluidic system will provide a realistic toxicology response induced by DPs than a static cell culture plate. To prove the hypothesis, Neuro2a (N2a) cells were used as representative cells to study systemic neurotoxicity by the implant DPs. The microfluidic bioreactor system was validated by comparing the cell toxicity against the traditional static system and using COMSOL modeling for media flow with DPs. The hip implant simulator used in this study was a state-of-the-art sliding hip simulator developed in our lab. The results suggested that static toxicity was significantly more compared to dynamic microfluidic-based toxicity. The newly developed DMBH system tested for in situ systemic toxicity on N2a cells and demonstrated very minimum toxicity level (5.23%) compared to static systems (31.16%). Thus, the new DMBH system is an efficient tool for in situ implant metal systemic toxicity testing.

Evolution of apetaly in the cosmopolitan genus Stellaria.

PREMISE: Apetaly is widespread across distantly related lineages of flowering plants and is associated with abiotic (or self-) pollination. It is particularly prevalent in the carnation family, and the cosmopolitan genus Stellaria contains many lineages that are hypothesized to have lost petals from showy petalous ancestors. But the pollination biology of apetalous species of Stellaria remains unclear. METHODS: Using a substantial species-level sampling (~92% of known taxonomic diversity), we describe the pattern of petal evolution within Stellaria using ancestral character state reconstructions. To help shed light on the reproductive biology of apetalous Stellaria, we conducted a field experiment at an alpine tundra site in the southern Rocky Mountains to test whether an apetalous species (S. irrigua) exhibits higher levels of selfing than a sympatric, showy petalous congener (S. longipes). RESULTS: Analyses indicated that the ancestor of Stellaria was likely showy petalous and that repeated, parallel reductions of petals occurred in clades across much of the world, with uncommon reversal back to showy petals. Field experiments supported high rates of selfing in the apetalous species and high rates of outcrossing in the petalous species. CONCLUSIONS: Petal loss is rampant across major clades of Stellaria and is potentially linked with self-pollination worldwide. Self-pollination occurs within the buds in S. irrigua, and high propensities for this and other forms of selfing known in many other taxa of arctic-alpine habitats may reflect erratic availability of pollinators.

Implant-derived CoCrMo alloy nanoparticle disrupts DNA replication dynamics in neuronal cells.

The complexity of cobalt-chromium-molybdenum (CoCrMo) nanoparticles generated from the hip modular taper interfaces resulted in inconclusive outcomes on the level of toxicity in orthopedic patients. We used a hip simulator to generate physiologically relevant CoCrMo degradation products (DPs) to demonstrate the variation in the level of toxicity in neurons in comparison to processed degradation products (PDPs). The study outcomes indicate that DP induces a higher level of DNA damage in the form of double- and single-stranded DNA breaks and alkaline labile DNA adducts versus PDPs. The scientific advancements of this study are the following: (i) how DPs mimic more closely to the implant debris from hip implants in terms of bioactivity, (ii) how hip implant debris causes local and systemic issues, and (iii) methods to augment the biologic impact of implant debris. We discovered that DP is bioactive compared with PDP, and this should be considered in the toxicity evaluation related to implants. • The physicochemical characteristics of the CoCrMo is a major factor to consider for implant-related cytotoxicity or genotoxicity experimental design. • Elevated levels of intracellular ROS induced by the physiologically relevant wear particle are detrimental to the neuronal cells. • The DP can induce variation in DNA replication dynamics compared to PDP.

Are Damage Modes Related to Microstructure and Material Loss in Severely Damaged CoCrMo Femoral Heads?

BACKGROUND: Fretting and corrosion in metal-on-polyethylene total hip arthoplasty (THA) modular junctions can cause adverse tissue reactions that are responsible for 2% to 5% of revision surgeries. Damage within cobalt-chromium-molybdenum (CoCrMo) alloy femoral heads can progress chemically and mechanically, leading to damage modes such as column damage, imprinting, and uniform fretting damage. At present, it is unclear which of these damage modes are most detrimental and how they may be linked to implant alloy metallurgy. The alloy microstructure exhibits microstructural features such as grain boundaries, hard phases, and segregation bands, which may enable different damage modes, higher material loss, and the potential risk of adverse local tissue reactions. QUESTIONS/PURPOSES: In this study, we asked: (1) How prevalent is chemically dominated column damage compared with mechanically dominated damage modes in severely damaged metal-on-polyethylene THA femoral heads made from wrought CoCrMo alloy? (2) Is material loss greater in femoral heads that underwent column damage? (3) Do material loss and the presence of column damage depend on alloy microstructure as characterized by grain size, hard phase content, and/or banding? METHODS: Surgically retrieved wrought CoCrMo modular femoral heads removed between June 2004 and June 2019 were scored using a modified version of the Goldberg visually based scoring system. Of the total 1002 heads retrieved over this period, 19% (190 of 1002) were identified as severely damaged, exhibiting large areas of fretting scars, black debris, pits, and/or etch marks. Of these, 43% (81 of 190) were excluded for metal-on-metal articulations, alternate designs (such as bipolar, dual-mobility, hemiarthroplasty, metal adaptor sleeves), or previous sectioning of the implant for past studies. One sample was excluded retroactively as metallurgical analysis revealed that it was made of cast alloy, yielding a total of 108 for further analysis. Information on patient age (57 ± 11 years) and sex (56% [61 of 108] were males), reason for removal, implant time in situ (99 ± 78 months), implant manufacturer, head size, and the CoCrMo or titanium-based stem alloy pairing were collected. Damage modes and volumetric material loss within the head tapers were identified using an optical coordinate measuring machine. Samples were categorized by damage mode groups by column damage, imprinting, a combination of column damage and imprinting, or uniform fretting. Metallurgical samples were processed to identify microstructural characteristics of grain size, hard phase content, and banding. Nonparametric Mann-Whitney U and Kruskal-Wallis statistical tests were used to examine volumetric material loss compared with damage mode and microstructural features, and linear regression was performed to correlate patient- and manufacturer-specific factors with volumetric material loss. RESULTS: Chemically driven column damage was seen in 48% (52 of 108) of femoral heads, with 34% (37 of 108) exhibiting a combination of column damage and imprinting, 12% (13 of 108) of heads displaying column damage and uniform fretting, and 2% (2 of 108) exhibiting such widespread column damage that potentially underlying mechanical damage modes could not be verified. Implants with column damage showed greater material loss than those with mechanically driven damage alone, with median (range) values of 1.2 mm3 (0.2 to 11.7) versus 0.6 mm3 (0 to 20.7; p = 0.03). Median (range) volume loss across all femoral heads was 0.9 mm3 (0 to 20.7). Time in situ, contact area, patient age, sex, head size, manufacturer, and stem alloy type were not associated with volumetric material loss. Banding of the alloy microstructure, with a median (range) material loss of 1.1 mm3 (0 to 20.7), was associated with five times higher material loss compared with those with a homogeneous microstructure, which had a volume loss of 0.2 mm3 (0 to 4.1; p = 0.02). Hard phase content and grain size showed no correlation with material loss. CONCLUSION: Chemically dominated column damage was a clear indicator of greater volume loss in this study sample of 108 severely damaged heads. Volumetric material loss strongly depended on banding (microstructural segregations) within the alloy. Banding of the wrought CoCrMo microstructure should be avoided during the manufacturing process to reduce volumetric material loss and the release of corrosion products to the periprosthetic tissue. CLINICAL RELEVANCE: Approximately 30% of THAs rely on wrought CoCrMo femoral heads. Most femoral heads in this study exhibited a banded microstructure that was associated with larger material loss and the occurrence of chemically dominated column damage. This study suggests that elimination of banding from the alloy could substantially reduce the release of implant debris in vivo, which could potentially also reduce the risk of adverse local tissue reactions to implant debris.