Decellularized kidney capsule as a three-dimensional scaffold for tissue regeneration.

Tissue regeneration is thought to have considerable promise with the use of scaffolds designed for tissue engineering. Although polymer-based scaffolds for tissue engineering have been used extensively and developed quickly, their ability to mimic the in-vivo milieu, overcome immunogenicity, and have comparable mechanical or biochemical properties has limited their capability for repair. Fortunately, there is a compelling method to get around these challenges thanks to the development of extracellular matrix (ECM) scaffolds made from decellularized tissues. We used ECM decellularized sheep kidney capsule tissue in our research. Using detergents such as Triton-X100 and sodium dodecyl sulfate (SDS), these scaffolds were decellularized. DNA content, histology, mechanical properties analysis, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), biocompatibility, hemocompatibility and scanning electron microscope (SEM) imaging were measured. The results showed that the three-dimensional (3D) structure of the ECM remained largely intact. The scaffolds mentioned above had several hydrophilic properties. The best biocompatibility and blood compatibility properties were reported in the SDS method of 0.5%. The best decellularization scaffold was introduced with 0.5% SDS. Therefore, it can be proposed as a scaffold that has ECM like natural tissue, for tissue engineering applications.

Age and sex related change in tooth enamel thickness of maxillary incisors measured by cone beam computed tomography.

BACKGROUND: To measure adequate enamel thickness of maxillary incisors in planning enamel reduction for a porcelain laminate veneer restoration in relation to chronological age and sex by using cone beam computed tomography (CBCT) in an Iraqi subpopulation. METHODS: From 81 CBCT images, 324 maxillary incisors were examined. Enamel thickness was measured at both mesial and distal regions of the tooth in three different levels: cervical, middle, and incisal (occlusal) 1/3 at a sagittal section. Measurements were made for the following tooth areas using CBCT: facial enamel thickness at 1, 3, and 5 mm from the cementoenamel junction (CEJ), palatal enamel thickness at 5 mm from the CEJ (5 mm P), facial and palatal enamel thickness at the incisal edge (IFP), mid incisal enamel thickness (IET), and the incisal edge enamel-pulp distance (IEPD). Relationships of enamel thickness with age and sex were evaluated using Independent t-test, Mann-Whitney U-test and the Pearson correlation coefficient, a simple linear regression analysis used for statistical analysis. RESULTS: Significant differences (P < 0.05) were found in terms of an inverse association between enamel thickness and chronological age at all measurements above the CEJ and the regression model for the mid-incisal enamel thickness was (R2 of 0.4). In contrast, there was an increase in IFP, palatal, and IPED enamel thickness with age. Also, significant differences were found in enamel thickness between males and females, the enamel being thicker in females in relation to facial enamel thickness, enamel palatal thickness above CEJ and IET, while for IEPD, the enamel thickness was greater in males compared to females. CONCLUSION: The measurements for enamel thickness outcome variables in relation to chronological age revealed significant differences for each measured distance and there were statistically significant differences in enamel thickness between males and females at all measurements except at IFP. These results demonstrate that CBCT can be used for noninvasive, accurate measurements of enamel thickness in both sex.

A novel population of local pericyte precursor cells in tumor stroma that require Notch signaling for differentiation.

Pericytes are perivascular support cells, the origin of which in tumor tissue is not clear. Recently, we identified a Tie1(+) precursor cell that differentiates into vascular smooth muscle, in a Notch-dependent manner. To understand the involvement of Notch in the ontogeny of tumor pericytes we used a novel flow immunophenotyping strategy to define CD146(+)/CD45(-)/CD31(-/lo) pericytes in the tumor stroma. This strategy combined with ex vivo co-culture experiments identified a novel pericyte progenitor cell population defined as Sca1(hi)/CD146(-)/CD45(-)/CD31(-). The differentiation of these progenitor cells was stimulated by co-culture with endothelial cells. Overexpression of the Notch ligand Jagged1 in endothelial cells further stimulated the differentiation of Sca1(hi)/CD146(-)/CD45(-)/CD31(-) cells into pericytes, while inhibition of Notch signaling with a γ-secretase inhibitor reduced this differentiation. However, Notch inhibition specifically in Tie1-expressing cells did not change the abundance of pericytes in tumors, suggesting that the pericyte precursor is distinct from the vascular smooth muscle cell precursor. Transplant experiments showed that the bone marrow contributes minimally to tumor pericytes. Immunophenotyping revealed that Sca1(hi)/CD146(-)/CD45(-)/CD31(-) cells have greater potential to differentiate into pericytes and have increased expression of classic mesenchymal stem cell markers (CD13, CD44, Nt5e and Thy-1) compared to Sca1(-/lo)/CD146(-)/CD45(-)/CD31(-) cells. Our results suggest that a local Sca1(hi)/CD146(-)/CD45(-)/CD31(-) pericyte progenitor resides in the tumor microenvironment and requires Notch signaling for differentiation into mature pericytes.

Protocol for performing consecutive bone marrow transplants in mice to study the role of marrow niche in supporting hematopoiesis.

Hematopoietic stem and progenitor cells depend on bone marrow (BM) stromal cells for survival. Here, we present a protocol for performing three consecutive BM transplants in mice to study the role of BM niche in supporting hematopoiesis. We describe steps for transplanting cells to condition the marrow of the recipient mice and transplanting wild-type cells to examine the effect of the conditioned marrow in supporting hematopoiesis. We then detail procedures for transplanting into wild-type recipients to measure bone marrow chimerism. For complete details on the use and execution of this protocol, please refer to Gopal et al. (2022).1.

Evaluation of video teaching on resident competency for all-ceramic crown tooth preparation.

PURPOSE: This study aimed to analyze the impact of live video instruction using digital evaluation of residents' performance in full ceramic crown preparation. METHODS: Digital evaluation using CEREC computer-aided design/computer-assisted manufacturing (CAD/CAM) 5.1.3 software was conducted of preparation on a typodont for all-ceramic crowns of mandibular first molars (MFMs) with a radial shoulder finish line, carried out by 30 residents. Each participant prepared two MFMs: group A prepared the right side without live video instruction, and group B prepared the left side after instruction. All prepared teeth were scanned by Dentsply Sirona chairside CAD/CAM system with Omnicom to assess the inter-occlusal space, undercut, the finish line of the preparation, and surface texture. The data were analyzed using Pearson Chi-square, Wilcoxon signed-rank test, and paired t-test. P-values less than 0.05 were considered to be statistically significant in all tests. RESULTS: Based on the Pearson Chi-square test, there were significant differences between the two groups in terms of inter-occlusal space on the buccal and lingual surface of the prepared tooth, in the presence of rough surfaces before and after the preparation, and difference in the type of finish line. The Wilcoxon signed-rank test revealed a significant difference in convergence angle buccolingually and the remaining height of the prepared teeth before and after the video instruction. CONCLUSIONS: The use of educational live video instruction can be helpful to residents in learning the principles of tooth preparation.

TIRAP drives myelosuppression through an Ifnγ-Hmgb1 axis that disrupts the endothelial niche in mice.

Inflammation is associated with bone marrow failure syndromes, but how specific molecules impact the bone marrow microenvironment is not well elucidated. We report a novel role for the miR-145 target, Toll/interleukin-1 receptor domain containing adaptor protein (TIRAP), in driving bone marrow failure. We show that TIRAP is overexpressed in various types of myelodysplastic syndromes (MDS) and suppresses all three major hematopoietic lineages. TIRAP expression promotes up-regulation of Ifnγ, leading to myelosuppression through Ifnγ-Ifnγr-mediated release of the alarmin, Hmgb1, which disrupts the bone marrow endothelial niche. Deletion of Ifnγ blocks Hmgb1 release and is sufficient to reverse the endothelial defect and restore myelopoiesis. Contrary to current dogma, TIRAP-activated Ifnγ-driven bone marrow suppression is independent of T cell function or pyroptosis. In the absence of Ifnγ, TIRAP drives myeloproliferation, implicating Ifnγ in suppressing the transformation of MDS to acute leukemia. These findings reveal novel, noncanonical roles of TIRAP, Hmgb1, and Ifnγ in the bone marrow microenvironment and provide insight into the pathophysiology of preleukemic syndromes.

Mechanical and thermal stress evaluation of PEEK prefabricated post with different head design in endodontically treated tooth: 3D-finite element analysis.

An endodontic post is required to retain and support the core restoration in case of insufficient remaining coronal dentin after root canal therapy. This study analyzed the biomechanical and thermal behavior of PEEK prefabricated post after choosing the head design that produces the least amount of stress on the core and remaining tooth structure. These results were compared with the most common commercially available prefabricated post, which is titanium and glass fiber post. Thus a CBCT scanning of a maxillary central incisor with its supporting structure was used to construct a 3D solid model of an endodontically treated teeth for finite element analysis (FEA). The restored tooth with the spherical head design of PEEK prefabricated post yielded a more benign stress distribution and repairable failure mode on the crown, luting cement, core, and dentin under both mechanical and thermal loads, followed by glass fiber post and titanium post respectively.

miR-143/145 differentially regulate hematopoietic stem and progenitor activity through suppression of canonical TGFß signaling.

Expression of miR-143 and miR-145 is reduced in hematopoietic stem/progenitor cells (HSPCs) of myelodysplastic syndrome patients with a deletion in the long arm of chromosome 5. Here we show that mice lacking miR-143/145 have impaired HSPC activity with depletion of functional hematopoietic stem cells (HSCs), but activation of progenitor cells (HPCs). We identify components of the transforming growth factor ß (TGFß) pathway as key targets of miR-143/145. Enforced expression of the TGFß adaptor protein and miR-145 target, Disabled-2 (DAB2), recapitulates the HSC defect seen in miR-143/145-/- mice. Despite reduced HSC activity, older miR-143/145-/- and DAB2-expressing mice show elevated leukocyte counts associated with increased HPC activity. A subset of mice develop a serially transplantable myeloid malignancy, associated with expansion of HPC. Thus, miR-143/145 play a cell context-dependent role in HSPC function through regulation of TGFß/DAB2 activation, and loss of these miRNAs creates a preleukemic state.