Evaluation of two printing techniques for maxillary removable partial denture frameworks.

STATEMENT OF PROBLEM: Three-dimensional (3D) printing technology has gained popularity in producing removable partial denture (RPD) frameworks, including direct 3D printing of the metal framework and framework printing using castable resin, subsequently cast and processed. However, whether the technology is sufficiently accurate and precise to supersede traditional methods is unclear. PURPOSE: The purpose of this in vitro study was to determine the accuracy and precision of 2 different methods in the fabrication of RPD frameworks, including 3D printing by selective laser melting (SLM) and digital light processing (DLP). MATERIAL AND METHODS: Maxillary casts were digitized to design RPD frameworks. Thereby, 8 frameworks were produced for each group. The SLM group underwent a thermal finishing process after printing. In the DLP group, castable resin was printed but not cast. All frameworks were scanned to generate digital files, which were then compared with the original design using a metrology software program and manual measurements. Statistical analysis was executed using the t-test for independent specimens (α=.05) and by comparing heatmaps of the overlaid meshes. RESULTS: The analysis of the frameworks indicated minor deviations across all specimens. Regarding accuracy, there were no significant differences between the groups (P=.986). The SLM frameworks demonstrated greater precision, with absolute deviation values of 0.13 mm compared with 0.17 mm in the DLP group (P<.001). CONCLUSIONS: The findings underscored a high consistency between the 2 printing techniques, demonstrating a sufficiently advanced production process to yield predictable results. While the accuracy of both techniques was at a comparably high level and did not differ significantly, the SLM technique delivered RPDs with higher precision.

A randomized clinical trial of phycogenic materials for sinus grafting with hydroxyapatite versus biphasic calcium phosphate: 2 years clinical outcomes.

OBJECTIVES: To assess in a prospective randomized trial two phycogenic bone substitutes-biphasic calcium phosphate (BCP) versus almost pure hydroxyapatite (HA)-for their volume stability and clinical implications after sinus floor elevation (SFE). MATERIALS AND METHODS: Twenty patients requiring lateral-window SFE 6 months prior to implant surgery were randomized to a BCP or HA group. As primary outcome, the grafts were analyzed for volume stability, using four cone-beam computed tomography scans obtained immediately/6/12/24 months after SFE. Secondary outcomes were implant survivval, success, periotest values, oral-health-related quality of life (OHIP-G14), and pain (VAS). RESULTS: Kolmogorov-Smirnov goodness-of-fit test revealed normal distribution of samples (p = .200). At 6/12/24 months, the augmented volumes decreased to 96/92/90% (HA) or 99/96/96% (BCP). Volume changes were significantly a factor of time (p < .001; generalized linear model with repeated measures) and reached significantly lower values in HA group (p = .018). Significant intergroup difference in volume losses was notable at 24 months (p = .021; t-test for independent samples). Periotest values decreased from -3/-4.1 (HA/BCP) after implant placement to -6.3/-4.5 (HA/BCP) after 6 months. OHIP scores diverged at 2 months (HA: 9.5; BCP: 5.2) and largely resolved by 24 months (HA: 1.3; BCP: 1.9). VAS scores were comparable, 2.2 at 1 week after SFE being their highest mean level. CONCLUSIONS: After 2 years, both groups experienced no biological or technical complications, demonstrating a consistent healing trajectory without notable symptoms. Although no significant differences were observed in implant stability and survival, BCP demonstrated higher volume stability than HA.

Consequences of somatic mutations of GIRK1 detected in primary malign tumors on expression and function of G-protein activated, inwardly rectifying, K+ channels.

A search in the GDC Data Portal revealed 304 documented somatic mutations of the KCNJ3 gene in primary tumors (out of 10.202 cases). Most affected tumor types were carcinomas from uterus, skin and lung, while breast cancer exerted the lowest number of somatic mutations. We focused our research on 15 missense mutations within the region between TM1 and TM2, comprising the pore helix and ion selectivity signature. Expression was measured by confocal laser scan microscopy of eGFP tagged GIRK1 subunits, expressed with and without GIRK4 in oocytes of Xenopus laevis. GIRK ion currents were activated via coexpressed m2Rs and measured by the Two Electrode Voltage Clamp technique. Magnitude of the total GIRK current, as well as the fraction of current inducible by the agonist, were measured. Ion selectivity was gauged by assessment of the PNa+/PK+ ratio, calculated by the GIRK current reversal potential in extracellular media at different Na+ and K+ concentrations. None of the tested mutations was able to form functional GIRK1 homooligomeric ion channels. One of the mutations, G145A, which locates directly to the ion selectivity signature, exerted an increased PNa+/PK+ ratio. Generally, the missense mutations studied can be categorized into three groups: (i) normal/reduced expression accompanied by reduced/absent function (S132Y, F136L, E139K, G145A, R149Q, R149P, G178D, S185Y, Q186R), (ii) normal/increased expression as well as increased function (E140M, A142T, M184I) and (iii) miniscule expression but increased function relative to expression levels (I151N, G158S). We conclude, that gain of function mutations, identical or similar to categories (ii) and (iii), may potentially be involved in genesis and progression of malignancies in tissues that exert a high rate of occurrence of somatic mutations of KCNJ3.

Dynamic Control of Mitochondrial Ca2+ Levels as a Survival Strategy of Cancer Cells.

Cancer cells have increased energy requirements due to their enhanced proliferation activity. This energy demand is, among others, met by mitochondrial ATP production. Since the second messenger Ca2+ maintains the activity of Krebs cycle dehydrogenases that fuel mitochondrial respiration, proper mitochondrial Ca2+ uptake is crucial for a cancer cell survival. However, a mitochondrial Ca2+ overload induces mitochondrial dysfunction and, ultimately, apoptotic cell death. Because of the vital importance of balancing mitochondrial Ca2+ levels, a highly sophisticated machinery of multiple proteins manages mitochondrial Ca2+ homeostasis. Notably, mitochondria sequester Ca2+ preferentially at the interaction sites between mitochondria and the endoplasmic reticulum (ER), the largest internal Ca2+ store, thus, pointing to mitochondrial-associated membranes (MAMs) as crucial hubs between cancer prosperity and cell death. To investigate potential regulatory mechanisms of the mitochondrial Ca2+ uptake routes in cancer cells, we modulated mitochondria-ER tethering and the expression of UCP2 and analyzed mitochondrial Ca2+ homeostasis under the various conditions. Hence, the expression of contributors to mitochondrial Ca2+ regulation machinery was quantified by qRT-PCR. We further used data from The Cancer Genome Atlas (TCGA) to correlate these in vitro findings with expression patterns in human breast invasive cancer and human prostate adenocarcinoma. ER-mitochondrial linkage was found to support a mitochondrial Ca2+ uptake route dependent on uncoupling protein 2 (UCP2) in cancer cells. Notably, combined overexpression of Rab32, a protein kinase A-anchoring protein fostering the ER-mitochondrial tethering, and UCP2 caused a significant drop in cancer cells' viability. Artificially enhanced ER-mitochondrial tethering further initiated a sudden decline in the expression of UCP2, probably as an adaptive response to avoid mitochondrial Ca2+ overload. Besides, TCGA analysis revealed an inverse expression correlation between proteins stabilizing mitochondrial-ER linkage and UCP2 in tissues of human breast invasive cancer and prostate adenocarcinoma. Based on these results, we assume that cancer cells successfully manage mitochondrial Ca2+ uptake to stimulate Ca2+-dependent mitochondrial metabolism while avoiding Ca2+-triggered cell death by fine-tuning ER-mitochondrial tethering and the expression of UCP2 in an inversed manner. Disruption of this equilibrium yields cancer cell death and may serve as a treatment strategy to specifically kill cancer cells.

Phycogenic bone substitutes for sinus floor augmentation: Histomorphometric comparison of hydroxyapatite and biphasic calcium phosphate in a randomised clinical pilot study.

Aims: While numerous materials are available for sinus floor elevation, plant-based alternatives still hold promise of overcoming concerns about allogeneic or xenogeneic materials. Thus, the present authors designed a randomised clinical trial to histologically compare an almost pure hydroxyapatite (HA) to a biphasic calcium phosphate comprising 80% ß-tricalcium phosphate (ß-TCP) and 20% hydroxyapatite (ß-TCP/HA), all of phycogenic origin. Materials and methods: Twenty patients scheduled for lateral window sinus floor elevation were randomised to either an HA or a ß-TCP/HA group. Biopsy specimens were taken 3 months after sinus floor elevation and during implant surgery after 6 months. One ground section per biopsy specimen (N = 40) was stained, scanned and histomorphometrically analysed for new bone, old bone, soft tissue, graft, bone infiltration of graft, bone-to-graft contact and penetration depth. Results: At 6 months, more new bone was seen in the ß-TCP/HA group (P = 0.011), whereas more residual graft was present and in more extensive contact with new bone in the HA group. More pronounced alterations, and smaller particle sizes, of graft surrounded and infiltrated by bone were seen in the ß-TCP/HA group. The less extensive bone-to-graft contact in the ß-TCP/ HA group reflected a more advanced state of resorption, while infiltration of residual graft material by bone was also increased in this group. Conclusions: Proper healing was seen in both groups, with the graft materials guiding the formation of new bone, which grew especially well through the particles of the highly osteoconductive and resorptive ß-TCP/HA material. HA was very stable, without significant resorption, but was extensively in contact with new bone after 6 months.

Accuracy assessment of 3D-printed tooth replicas.

AIM: The production of individual tooth replicas has two applications in dental practice: tooth autotransplantations and dental root analogue implants. These applications require a particularly high degree of precision. The purpose of this study was to establish and evaluate a method for fabricating individual 3D-printed tooth replicas. MATERIALS AND METHODS: 10 patients requiring extraction of a wisdom tooth and a preoperative cone beam computed tomography (CBCT) scan were included; exclusion criteria were intraoperative fragmentation or fracture of the tooth. 3D Slicer 4.6.2 was used for tooth segmentation and model generation based on CBCT data. The tooth replicas were manufactured by selective laser melting (SLM). The extracted teeth and 3D-printed replicas were scanned and tested for surface deviations in CloudCompare 2.8.1. RESULTS: The mean absolute surface deviation between the 3D-printed teeth and the corresponding extracted teeth ranged from 0.13 to 0.25 mm, with standard deviations of 0.10 to 0.21 mm; 95% of the measured surface points deviated less than 0.474 mm; the surface area was reduced by -6.0% and the volume by -3.4%. The root mean square was 0.238 mm and the mean maximum absolute surface deviation was 0.927 mm. The SLM technique showed a high precision with a mean absolute deviation of 0.045 mm and a standard deviation of 0.04 mm. CONCLUSION: 3D-printed tooth replicas with a very high accuracy could be produced based on CBCT data. The described method is suitable for manufacturing tooth replicas for use in tooth autotransplantations or for fabricating root analogue implants.

Influence of Extraoral Apicoectomy on Revascularization of an Autotransplanted Tooth: A Case Report.

Autotransplantation is considered a standard procedure with a poorer prognosis for mature than for immature teeth. In this case report, the root tip of an almost fully developed tooth was resected during autotransplantation to fit into a deficient recipient site. As a positive side effect, the apical foramen was enlarged, with potential improvement of the probability of revascularization. Clinical and radiologic follow-up examinations up to 18 months indicated revascularization and uneventful periodontal regeneration and thus successful autotransplantation. This case supports the hypothesis that intraoperative apicoectomy can improve the prognosis for revascularization of mature autotransplanted teeth. Further prospective controlled clinical studies are needed to confirm this innovative surgical approach.

Targeting Mitochondria to Counteract Age-Related Cellular Dysfunction.

Senescence is related to the loss of cellular homeostasis and functions, which leads to a progressive decline in physiological ability and to aging-associated diseases. Since mitochondria are essential to energy supply, cell differentiation, cell cycle control, intracellular signaling and Ca2+ sequestration, fine-tuning mitochondrial activity appropriately, is a tightrope walk during aging. For instance, the mitochondrial oxidative phosphorylation (OXPHOS) ensures a supply of adenosine triphosphate (ATP), but is also the main source of potentially harmful levels of reactive oxygen species (ROS). Moreover, mitochondrial function is strongly linked to mitochondrial Ca2+ homeostasis and mitochondrial shape, which undergo various alterations during aging. Since mitochondria play such a critical role in an organism's process of aging, they also offer promising targets for manipulation of senescent cellular functions. Accordingly, interventions delaying the onset of age-associated disorders involve the manipulation of mitochondrial function, including caloric restriction (CR) or exercise, as well as drugs, such as metformin, aspirin, and polyphenols. In this review, we discuss mitochondria's role in and impact on cellular aging and their potential to serve as a target for therapeutic interventions against age-related cellular dysfunction.

UCP2 and PRMT1 are key prognostic markers for lung carcinoma patients.

Cancer cells have developed unique strategies to meet their high energy demand. Therefore, they have established a setting of Ca2+-triggered high mitochondrial activity. But mitochondrial Ca2+ uptake has to be strictly controlled to avoid mitochondrial Ca2+ overload that would cause apoptotic cell death. Methylation by protein arginine methyl transferase 1 (PRMT1) desensitizes the mitochondrial Ca2+ uptake machinery and reduces mitochondrial Ca2+ accumulation in cancer cells. In case of PRMT1-driven methylation, proper mitochondrial Ca2+ uptake is reestablished by increased activity of uncoupling protein 2 (UCP2), pointing to an importance of these proteins for cancer cell survival and activity. Accordingly, in this study we investigated the impact of UCP2 and PRMT1 on the fate of human lung cancer cells (A549, Calu-3 and H1299) as well as on patients suffering from lung carcinoma. We show that combined overexpression of UCP2 and PRMT1 significantly enhances viability, proliferation as well as mitochondrial respiration. In line with these findings, the overall survival probability of lung carcinoma patients with high mRNA expression levels of UCP2 and PRMT1 is strongly reduced. Furthermore, analysis via The Cancer Genome Atlas (TCGA) reveals upregulation of both proteins, UCP2 and PRMT1, as common feature of various cancer types. These findings suggest that proper mitochondrial Ca2+ uptake is essential for devastating tumor growth, and highlight the importance of a tightly controlled mitochondrial Ca2+ uptake to ensure proper ATP biosynthesis while avoiding dangerous mitochondrial Ca2+ overload. By that, the study unveils proteins of the mitochondrial Ca2+ uptake as potential targets for cancer treatment.

Dosis Facit Sanitatem-Concentration-Dependent Effects of Resveratrol on Mitochondria.

The naturally occurring polyphenol, resveratrol (RSV), is known for a broad range of actions. These include a positive impact on lifespan and health, but also pro-apoptotic anti-cancer properties. Interestingly, cell culture experiments have revealed a strong impact of RSV on mitochondrial function. The compound was demonstrated to affect mitochondrial respiration, structure and mass of mitochondria as well as mitochondrial membrane potential and, ultimately, mitochondria-associated cell death pathways. Notably, the mitochondrial effects of RSV show a very strict and remarkable concentration dependency: At low concentrations, RSV (<50 µM) fosters cellular antioxidant defense mechanisms, activates AMP-activated protein kinase (AMPK)- and sirtuin 1 (SIRT1)-linked pathways and enhances mitochondrial network formation. These mechanisms crucially contribute to the cytoprotective effects of RSV against toxins and disease-related damage, in vitro and in vivo. However, at higher concentrations, RSV (>50 µM) triggers changes in (sub-)cellular Ca2+ homeostasis, disruption of mitochondrial membrane potential and activation of caspases selectively yielding apoptotic cancer cell death, in vitro and in vivo. In this review, we discuss the promising therapeutic potential of RSV, which is most probably related to the compound's concentration-dependent manipulation of mitochondrial function and structure.

KCNJ3 is a new independent prognostic marker for estrogen receptor positive breast cancer patients.

Numerous studies showed abnormal expression of ion channels in different cancer types. Amongst these, the potassium channel gene KCNJ3 (encoding for GIRK1 proteins) has been reported to be upregulated in tumors of patients with breast cancer and to correlate with positive lymph node status. We aimed to study KCNJ3 levels in different breast cancer subtypes using gene expression data from the TCGA, to validate our findings using RNA in situ hybridization in a validation cohort (GEO ID GSE17705), and to study the prognostic value of KCNJ3using survival analysis. In a total of > 1000 breast cancer patients of two independent data sets we showed a) that KCNJ3 expression is upregulated in tumor tissue compared to corresponding normal tissue (p < 0.001), b) that KCNJ3 expression is associated with estrogen receptor (ER) positive tumors (p < 0.001), but that KCNJ3 expression is variable within this group, and c) that ER positive patients with high KCNJ3 levels have worse overall (p < 0.05) and disease free survival probabilities (p < 0.01), whereby KCNJ3 is an independent prognostic factor (p <0.05). In conclusion, our data suggest that patients with ER positive breast cancer might be stratified into high risk and low risk groups based on the KCNJ3 levels in the tumor.

Piezo1 forms mechanosensitive ion channels in the human MCF-7 breast cancer cell line.

Mechanical interaction between cells - specifically distortion of tensional homeostasis-emerged as an important aspect of breast cancer genesis and progression. We investigated the biophysical characteristics of mechanosensitive ion channels (MSCs) in the malignant MCF-7 breast cancer cell line. MSCs turned out to be the most abundant ion channel species and could be activated by negative pressure at the outer side of the cell membrane in a saturable manner. Assessing single channel conductance (GΛ) for different monovalent cations revealed an increase in the succession: Li(+) < Na(+) < K(+) ≈Rb(+) ≈ Cs(+). Divalent cations permeated also with the order: Ca(2+) < Ba(2+). Comparison of biophysical properties enabled us to identify MSCs in MCF-7 as ion channels formed by the Piezo1 protein. Using patch clamp technique no functional MSCs were observed in the benign MCF-10A mammary epithelial cell line. Blocking of MSCs by GsMTx-4 resulted in decreased motility of MCF-7, but not of MCF-10A cells, underscoring a possible role of Piezo1 in invasion and metastatic propagation. The role of Piezo1 in biology and progression of breast cancer is further substantiated by markedly reduced overall survival in patients with increased Piezo1 mRNA levels in the primary tumor.