Targeted knock-in of NCF1 cDNA into the NCF2 locus leads to myeloid phenotypic correction of p47 phox -deficient chronic granulomatous disease.

p47 phox -deficient chronic granulomatous disease (p47-CGD) is a primary immunodeficiency caused by mutations in the neutrophil cytosolic factor 1 (NCF1) gene, resulting in defective NADPH oxidase function in phagocytes. Due to its complex genomic context, the NCF1 locus is not suited for safe gene editing with current genome editing technologies. Therefore, we developed a targeted NCF1 coding sequence knock-in by CRISPR-Cas9 ribonucleoprotein and viral vector template delivery, to restore p47 phox expression under the control of the endogenous NCF2 locus. NCF2 encodes for p67 phox , an NADPH oxidase subunit that closely interacts with p47 phox and is predominantly expressed in myeloid cells. This approach restored p47 phox expression and NADPH oxidase function in p47-CGD patient hematopoietic stem and progenitor cells (HSPCs) and in p47 phox -deficient mouse HSPCs, with the transgene expression following a myeloid differentiation pattern. Adeno-associated viral vectors performed favorably over integration-deficient lentiviral vectors for template delivery, with fewer off-target integrations and higher correction efficacy in HSPCs. Such myeloid-directed gene editing is promising for clinical CGD gene therapy, as it leads to the co-expression of p47 phox and p67 phox , ensuring spatiotemporal and near-physiological transgene expression in myeloid cells.

Effect of the Different Dietary Supplements on the Average Surface Roughness and Color Stability of Direct Restorative Materials Used in Pediatric Dentistry.

Increased surface roughness and discoloration of the direct restorative materials used in pediatric patients affect the longevity of restorations and impair children's oral health. Many factors can alter these properties. One of these factors is the intake of dietary supplements. It is crucial to predict the properties of restorative materials when exposed to dietary supplements to maintain the dental care of children. Thus, this study aimed to investigate the effect of various syrup-formed dietary supplements on the average surface roughness and color stability of current restorative materials used in pediatric dentistry. Seven different restorative materials (conventional glass ionomer [Fuji IX GP], resin-modified glass ionomer, [Fuji II LC], zirconia-reinforced glass ionomer [Zirconomer Improved], polyacid-modified composite resin [Dyract®XTRA], bulk-fill glass hybrid restorative [Equia Forte HT Fill], conventional resin composite [Charisma Smart], and resin composite with reactive glass fillers [Cention N]) were tested. The specimens prepared from each type of restorative material were divided into five subgroups according to dietary supplements (Sambucol Kids, Resverol, Imunol, Umca, and Microfer). These specimens were immersed daily in supplement solution over a period of 28 days. Surface roughness and color difference measurements were performed at baseline and at the 7th and 28th days. The color difference and Ra values showed that there was an interaction among the type of restorative material, type of dietary supplement, and immersion time factors (p < 0.05). Whereas lower Ra values were found in the composite resin group, the highest Ra values were found in the conventional glass ionomer group. All supplements caused increasing color difference values, and Resverol and Umca showed higher discoloration values above the clinically acceptable threshold. The intake of dietary supplement type, the immersion time of the dietary supplement, and the restorative material type affected the surface roughness and color stability of the tested direct restorative materials. All of the experimental groups showed higher Ra values than clinically acceptable surface roughness values (0.2 µm). The color difference values also increased with the immersion time.

Effects of 3D-Printing Technology and Cement Type on the Fracture Resistance of Permanent Resin Crowns for Primary Teeth.

PURPOSE: To evaluate the fracture resistance of permanent resin crowns for primary teeth produced using two different 3D-printing technologies (digital light processing [DLP] and stereolithography [SLA]) and cemented with various luting cements (glass ionomer, resin-modified glass ionomer, and self-adhesive resin cement), whether thermally aged or not. MATERIALS AND METHODS: A typodont primary mandibular second molar tooth was prepared and scanned, and a restoration design was created with web-based artificial intelligence (AI) dental software. A total of 96 crowns were prepared, and 12 experimental groups were generated according to the cement type, 3Dprinting technology (DLP or SLA), and thermal aging. Fracture resistance values and failure types of the specimens were noted. The results were statistically analyzed with three-way ANOVA and Tukey HSD tests (α = .05). RESULTS: The results of the three-way ANOVA showed that there was an interaction among the factors (3D-printing technology, cement type, and thermal aging) (P = .003). Thermal aging significantly decreased the fracture resistance values in all experimental groups. DLP-printed crowns showed higher fracture resistance values than SLA-printed crowns. Cement type also affected the fracture resistance, with glass ionomer cement showing the lowest values after aging. Resin-modified glass ionomer and resin cements were more preferable for 3D-printed crowns. CONCLUSIONS: The type of cement and the 3D-printing technology significantly influenced the fracture resistance of 3D-printed permanent resin crowns for primary teeth, and it was decided that these crowns would be able to withstand masticatory forces in children.

Effects of design software program and manufacturing method on the marginal and internal adaptation of esthetic crowns for primary teeth: A microcomputed tomography evaluation.

STATEMENT OF PROBLEM: The adaptation of digitally produced crowns is affected by the design software program and manufacturing method. The effect of artificial intelligence (AI) software program design on the adaptation of the crowns is unclear and comparative evaluations should be documented. PURPOSE: The purpose of this study was to assess the marginal and internal gaps, the absolute marginal discrepancies, and the 3-dimensional (3D) discrepancy volumes of the resin-based milled and 3D printed crowns for primary teeth designed with computer-aided design (CAD) and AI software programs by using microcomputed tomography (µCT). MATERIAL AND METHODS: A total of 40 resin-based esthetic crowns were produced for a prepared typodont tooth (right mandibular primary second molar) according to the design software program (CAD and AI) and manufacturing method (milling and 3D printing) (n=10). Four experimental groups were generated as CAD-milled, CAD-3D printed, AI-milled, and AI-3D printed. The marginal, axial, and occlusal gap values, the absolute marginal discrepancies, and the 3D discrepancy volumes of the specimens were measured by using µCT. The data were analyzed by using 2-way ANOVA and the Tukey HSD tests (α=.05). RESULTS: The lowest value for the marginal gap (54 ±43 µm) was observed in the CAD-milled group and the marginal gap value of the AI-3D printed group was significantly lower than the AI-milled group (P<.05). The lowest value for the axial gap (63 ±7 µm) was observed in the AI-3D printed group, and the highest value (145 ±58 µm) was observed in the CAD-milled group; the result for the occlusal gap value was opposite. The highest absolute marginal discrepancy value was observed in the CAD-milled group. The 3D discrepancy volumes increased in the order of the CAD-3D printed, AI-milled, CAD-milled, and AI-3D printed groups. CONCLUSIONS: The marginal and internal gap values of the resin-based crowns were affected by the design software program and manufacturing method; however, tested groups showed clinically acceptable gap values.