Local application of gingiva-derived mesenchymal stem cells on experimental periodontitis in rats.

BACKGROUND: Stem cell-based approaches in regenerative periodontal therapy have been used in different experimental models. In this study, the effect of local application of gingival mesenchymal stem cells (GMSC) in fibroin/chitosan oligosaccharide lactate hydrogel (F/COS) on periodontal regeneration was evaluated using experimental periodontitis model in rats. METHODS: Mesenchymal stem cells were isolated from the gingiva of rats and characterized. Viability tests and confocal imaging of GMSC in hydrogels were performed. Healthy control without periodontitis (Health; H; n=10), control with periodontitis but no application (Periodontitis; P; n=10), only hydrogel application (F/COS; n=10), and GMSC+F/COS (n=10) four groups were formed for in vivo studies. Experimental periodontitis was created with silk sutures around the maxillary second molars. GMSC labeled with green fluorescent protein (GFP) (250,000 cells/50 µL) in F/COS were applied to the defect. Animals were sacrificed at 2nd and 8th weeks and maxillae of the animals were evaluated by micro-computed tomography (micro-CT) and histologically. The presence of GFP-labeled GMSC was confirmed at the end of 8 weeks. RESULTS: Micro-CT analysis showed statistically significant new bone formation in the F/COS+GMSC treated group compared with the P group at the end of 8 weeks (p < 0.05). New bone formation was also observed in the F/COS group, but the statistical analysis revealed that this difference was not significant when compared with the P group (p > 0.05). Long junctional epithelium formation was less in the F/COS+GMSC group compared with the P group. Periodontal ligament and connective tissue were well-organized in F/COS+GMSC group. CONCLUSION: The results showed that local GMSC application in hydrogel contributed to the formation of new periodontal ligament and alveolar bone in rats with experimental periodontitis. Since gingiva is easly accessible tissue, it is promising for autologous cell-based treatments in clinical applications.

The Value of Micro-CT in the Diagnosis of Lung Carcinoma: A Radio-Histopathological Perspective.

Micro-computed tomography (micro-CT) is a relatively new imaging modality and the three-dimensional (3D) images obtained via micro-CT allow researchers to collect both quantitative and qualitative information on various types of samples. Micro-CT could potentially be used to examine human diseases and several studies have been published on this topic in the last decade. In this study, the potential uses of micro-CT in understanding and evaluating lung carcinoma and the relevant studies conducted on lung and other tumors are summarized. Currently, the resolution of benchtop laboratory micro-CT units has not reached the levels that can be obtained with light microscopy, and it is not possible to detect the histopathological features (e.g., tumor type, adenocarcinoma pattern, spread through air spaces) required for lung cancer management. However, its ability to provide 3D images in any plane of section, without disturbing the integrity of the specimen, suggests that it can be used as an auxiliary technique, especially in surgical margin examination, the evaluation of tumor invasion in the entire specimen, and calculation of primary and metastatic tumor volume. Along with future developments in micro-CT technology, it can be expected that the image resolution will gradually improve, the examination time will decrease, and the relevant software will be more user friendly. As a result of these developments, micro-CT may enter pathology laboratories as an auxiliary method in the pathological evaluation of lung tumors. However, the safety, performance, and cost effectiveness of micro-CT in the areas of possible clinical application should be investigated. If micro-CT passes all these tests, it may lead to the convergence of radiology and pathology applications performed independently in separate units today, and the birth of a new type of diagnostician who has equal knowledge of the histological and radiological features of tumors.

Multifunctional Silk Fibroin/Carbon Nanofiber Scaffolds for In Vitro Cardiomyogenic Differentiation of Induced Pluripotent Stem Cells and Energy Harvesting from Simulated Cardiac Motion.

In this proof-of-concept study, cardiomyogenic differentiation of induced pluripotent stem cells (iPSCs) is combined with energy harvesting from simulated cardiac motion in vitro. To achieve this, silk fibroin (SF)-based porous scaffolds are designed to mimic the mechanical and physical properties of cardiac tissue and used as triboelectric nanogenerator (TENG) electrodes. The load-carrying mechanism, ß-sheet content, degradation characteristics, and iPSC interactions of the scaffolds are observed to be interrelated and regulated by their pore architecture. The SF scaffolds with a pore size of 379 ± 34 µm, a porosity of 79 ± 1%, and a pore interconnectivity of 67 ± 1% upregulated the expression of cardiac-specific gene markers TNNT2 and NKX2.5 from iPSCs. Incorporating carbon nanofibers (CNFs) enhances the elastic modulus of the scaffolds to 45 ± 3 kPa and results in an electrical conductivity of 0.021 ± 0.006 S/cm. The SF and SF/CNF scaffolds are used as conjugate TENG electrodes and generate a maximum power output of 0.37 × 10-3 mW/m2, with an open-circuit voltage and a short circuit current of 0.46 V and 4.5 nA, respectively, under simulated cardiac motion. A novel approach is demonstrated for fabricating scaffold-based cardiac patches that can serve as tissue scaffolds and simultaneously allow energy harvesting.

The assessment of internal adaptation and fracture resistance of glass ionomer and resin-based restorative materials applied after different caries removal techniques in primary teeth: an in-vitro study.

Background: The aim of this study was to evaluate the 3-dimensional (3D) internal adaptation (IA) and fracture resistance (FR) of compomer and glass ionomers applied after conventional caries removal to sound dentin (CCRSD) and selective caries removal to firm dentin (SCRFD) in in-vitro. Methods: Thirty extracted primary molars were randomly assigned to three main groups (n = 10) as glass hybrid restorative (GHR) (Equia Forte® HT), conventional glass ionomer (CGIR) (Voco Ionofil Molar) and compomer (Dyract XP). Each group was randomly divided into two subgroups according to caries removal technique as CCRSD (n = 5) and SCRFD (n = 5). The restoration procedures were completed after caries removal (CCRSD or SCRFD) in all samples. Then, specimens were subjected to IA and FR tests. Data were analyzed with Student's t, one-way ANOVA, and Kruskal Wallis-H tests. The correlation between IA and FR results was analyzed with a Pearson test. The statistical significance level was considered as 5%. Results: While CCRSD showed superior IA results than SCRFD for all restorative materials (p < 0.05), no statistical difference was found between CCRSD and SCRFD in FR assessment (p > 0.05). In CCRSD, compomer showed superior results for IA and FR than glass ionomers (p < 0.05). In SCRFD, it was found no significant difference between the restoratives for IA (p > 0.05). However, compomer showed superior FR results than glass ionomers (p < 0.05). There was moderate negative correlation between internal voids and FR without statistically significant difference (r = -0.333, p = 0.072). Conclusions: Despite the advantages of SCRFD, it was found to be less superior than CCRSD in IA assessment. Therefore, when SCRFD is preferred, a peripheral seal should be provided for ideal restorative treatment. On the other hand, compomer mostly showed superior results compared to others.

Fracture Resistance of Esthetic Prefabricated and Custom-Made Crowns for Primary Molars After Artificial Aging.

Purpose: The purpose of this in vitro study was to compare the fracture resistance and survival of various esthetic crowns for primary molars after artificial aging via chewing simulation. Methods: A typodont tooth (mandibular primary second molar) was prepared to receive five different types of crowns as follows (n equals 10): prefabricated fiberglass (PF); CAD/CAM zirconia (CZ); CAD/CAM resin-ceramic (CR); composite- strip (CS); and prefabricated zirconia (PZ) as control. All specimens were subjected to 750,000 cycles of thermomechanical loading to artificially simulate three years of clinical service. None of the crowns from the CS group survived artificial aging. Surviving crowns were evaluated via micro-CT considering microcrack formation, and a load-to-fracture test was applied. Data were analyzed using one-way analysis of variance followed by the Duncan test. Results: Group PZ (557.4±170.1 N) and CR (669.6±255.2) were found to have comparable results (P >0.05), which were lower than the other groups (P<0.05). Group CZ resulted in the highest mean load-to-fracture value (1126.2±180.6; P <0.05). At the end of three years of artificial aging, microcracks were observed for only CS and PF groups. Conclusions: These in vitro data suggest that all tested crowns, except CS crowns, survived three years of artificial aging; however, CAD/CAM zirconia crowns may provide longer service, as they showed the highest fracture resistance with no microcrack formation.

Micro-computed tomographic evaluation of the effects of pre-heating and sonic delivery on the internal void formation of bulk-fill composites.

The aim of this study was to compare the effects of conventional, sonic or pre-heating insertion techniques on internal void formation of bulk-fill composites with micro-computed tomography. Standardized cylindrical cavities were prepared in 160 human third molars. Four groups received different paste-like bulk-fill composites: SonicFill 2 (SF2); VisCalor Bulk (VCB); Filtek One Bulk-fill restorative (FBF); Tetric EvoCeram Bulk Fill (TEB); and a conventional posterior composite, Clearfil Majesty Posterior (CMP). A hybrid CAD/CAM block was selected as a control (n=10). Composite restorations were built according to each resin composite type and insertion technique (n=10). Micro-CT was used to assess internal void rates. Data was analyzed with two-way ANOVA and Tukey's multiple comparisons test (α=0.05). CAD/CAM blocks were free of voids. For each composite, the highest void rates were observed for the sonic delivery method (p<0.05) except for SF2. SF2 was not affected by insertion techniques (p>0.05). Other composites showed the lowest void rates with pre-heating technique.

A two-compartment bone tumor model to investigate interactions between healthy and tumor cells.

We produced a novel three-dimensional (3D) bone tumor model (BTM) to study the interactions between healthy and tumor cells in a tumor microenvironment, the migration tendency of the tumor cells, and the efficacy of an anticancer drug, Doxorubicin, on the cancer cells. The model consisted of two compartments: (a) a healthy bone tissue mimic, made of poly(lactic acid-co-glycolic acid) (PLGA)/beta-tricalcium phosphate (ß-TCP) sponge seeded with human fetal osteoblastic cells (hFOB) and human umbilical vein endothelial cells (HUVECs), and (b) a tumor mimic, made of lyophilized collagen sponge seeded with human osteosarcoma cells (Saos-2). The tumor mimic component was placed into a central cavity created in the healthy bone mimic and together they constituted the complete 3D bone tumor model (3D-BTM). The porosities of both sponges were higher than 85% and the diameters of the pores were 199 ± 52 µm for the PLGA/TCP and 50-150 µm for the collagen scaffolds. The compression Young's modulus of the PLGA/TCP and the collagen sponges were determined to be 4.76 MPa and 140 kPa, respectively. Cell proliferation, morphology, calcium phosphate forming capacity and alkaline phosphatase production were studied separately on both the healthy and tumor mimics. All cells demonstrated cellular extensions and spread well in porous scaffolds indicating good cell-material interactions. Confocal microscopy analysis showed direct contact between the cells present in different parts of the 3D-BTM. Migration of HUVECs from the healthy bone mimic to the tumor compartment was confirmed by the increase in the levels of angiogenic factors vascular endothelial growth factor, basic fibroblast growth factor, and interleukin 8 in the tumor component. Doxorubicin (2.7 µg.ml-1) administered to the 3D-BTM caused a seven-fold decrease in the cell number after 24 h of interaction with the anticancer drug. Caspase-3 enzyme activity assay results demonstrated apoptosis of the osteosarcoma cells. This novel 3D-BTM has a high potential for use in studying the metastatic capabilities of cancer cells, and in determining the effective drug types and combinations for personalized treatments.

Evaluation of Threshold Values for Root Canal Filling Voids in Micro-CT and Nano-CT Images.

While several materials and techniques have been used to assess the quality of root canal fillings in micro-CT images, the lack of standardization in scanning protocols has produced conflicting results. Hence, the aim of this study was to determine a cutoff voxel size value for the assessment of root canal filling voids in micro-CT and nano-CT images. Twenty freshly extracted mandibular central incisors were used. Root canals were prepared with nickel titanium files to an ISO size 40/0.06 taper and then filled with a single cone (40/0.06 taper) and AH Plus sealer. The teeth were scanned with different voxel sizes with either micro-CT (5.2, 8.1, 11.2, and 16.73 µm) or nano-CT (1.5 and 5.0 µm) equipment. Images were reconstructed and analyzed with the NRecon and CTAn software. Void proportion and void volume were calculated for each tooth in the apical, middle, and coronal thirds of the root canal. Kruskal-Wallis and post hoc Mann-Whitney U tests were performed with a significance level of 5%. In micro-CT images, significantly different results were detected among the tested voxel sizes for void proportion and void volume, whereas no such differences were found in nano-CT images (p > 0.05). Micro-CT images showed higher void numbers over the entire root length, with statistically significant differences between the voxel size of 16.73 µm and the other sizes (p < 0.05). The values of the different nano-CT voxel sizes did not significantly differ from those of the micro-CT (5.2, 8.1, and 11.2 µm), except for the voxel size of 16.73 µm (p < 0.05). All tested voxel sizes enabled the detection of root canal filling voids except for the voxel size of 16.73 µm. Bearing in mind the limitations of this study, it seems that a voxel size of 11.2 µm can be used as a reliable cutoff value for the assessment of root canal filling voids in micro-CT imaging.

Nuclear targeting peptide-modified, DOX-loaded, PHBV nanoparticles enhance drug efficacy by targeting to Saos-2 cell nuclear membranes.

The aim of this study was to target nano sized (266 ± 25 nm diameter) poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) particles carrying Doxorubicin (DOX), an anticancer agent, to human osteosarcoma cells (Saos-2). A nuclear targeting molecule (Nuclear Localization Signal, NLS), a 17 a.a. peptide, was attached onto the doxorubicin loaded nanoparticles. NLS conjugated nanoparticles surrounded the cell nuclei, but did not penetrate them. Free doxorubicin and doxorubicin loaded nanoparticles entered the cytoplasm and were evenly distributed within the cytoplasm. The localization of the NLS-targeted particles around the nuclear membrane caused a significantly higher decrease in the cancer cell numbers due to apoptosis or necrosis than the untargeted and free doxorubicin formulations showing the importance of targeting the nanoparticles to the nuclear membrane in the treatment of cancer.

3D printed poly(ε-caprolactone) scaffolds modified with hydroxyapatite and poly(propylene fumarate) and their effects on the healing of rabbit femur defects.

A large variety of approaches have been used to treat large and irregular shaped bone defects with less than optimal success due to material or design issues. In recent years patient specific constructs prepared by additive manufacturing provided a solution to the need for shaping implants to fit irregular defects in the surgery theater. In this study, cylindrical disks of poly(ε-caprolactone) (PCL) were printed by fused deposition modeling and modified with nanohydroxyapatite (HAp) and poly(propylene fumarate) (PPF) to create a mechanically strong implant with well-defined pore size and porosity, controllable surface hydrophilicity (with PPF) and osteoconductivity (with HAp). Cytotoxicity, irritation and inflammation tests demonstrated that the scaffolds were biocompatible. PCL/HAp and PCL/HAp/PPF scaffolds were implanted in the femurs of rabbits with and without seeding with rabbit Bone Marrow Stem Cells (BMSC) and examined after 4 and 8 weeks with micro-CT, mechanically and histologically. BMSC seeded PCL/HAp/PPF scaffolds showed improved tissue regeneration as determined by bone mineral density and micro-CT. Compressive and tension stiffness values (394 and 463 N mm-1) were significantly higher than those of the healthy rabbit femur (316 and 392 N mm-1, respectively) after 8 weeks of implantation. These 3D implants have great potential for patient-specific bone defect treatments.

Micro-CT and nano-CT analysis of filling quality of three different endodontic sealers.

OBJECTIVES: To investigate voids in different root canal sealers using micro-CT and nano-CT, and to explore the feasibility of using nano-CT for quantitative analysis of sealer filling quality. METHODS: 30 extracted mandibular central incisors were randomly assigned into three groups according to the applied root canal sealers (Total BC Sealer, Sure Seal Root, AH Plus) by the single cone technique. Subsequently, micro-CT and nano-CT were performed to analyse the incidence rate of voids, void fraction, void volume and their distribution in each sample. RESULTS: Micro-CT evaluation showed no significant difference among sealers for the incidence rate of voids or void fraction in the whole filling materials (p > 0.05), whereas a significant difference was found between AH Plus and the other two sealers using nano-CT (p < 0.05). All three sealers presented less void volume in the apical third; however, higher void volumes were observed in the apical and coronal thirds in AH Plus using micro-CT (p < 0.05), while nano-CT results displayed higher void volume in AH Plus among all the sealers and regions (p < 0.05). CONCLUSIONS: Bioactive sealers showed higher root filling rate, lower incidence rate of voids, void fraction and void volume than AH Plus under nano-CT analysis, when round root canals were treated by the single cone technique. The disparate results suggest that the higher resolution of nano-CT have a greater ability of distinguishing internal porosity, and therefore suggesting the potential use of nano-CT in quantitative analysis of filling quality of sealers.

Advanced cell therapies with and without scaffolds.

Tissue engineering and regenerative medicine aim to produce tissue substitutes to restore lost functions of tissues and organs. This includes cell therapies, induction of tissue/organ regeneration by biologically active molecules, or transplantation of in vitro grown tissues. This review article discusses advanced cell therapies that make use of scaffolds and scaffold-free approaches. The first part of this article covers the basic characteristics of scaffolds, including characteristics of scaffold material, fabrication and surface functionalization, and their applications in the construction of hard (bone and cartilage) and soft (nerve, skin, blood vessel, heart muscle) tissue substitutes. In addition, cell sources as well as bioreactive agents, such as growth factors, that guide cell functions are presented. The second part in turn, examines scaffold-free applications, with a focus on the recently discovered cell sheet engineering. This article serves as a good reference for all applications of advanced cell therapies and as well as advantages and limitations of scaffold-based and scaffold-free strategies.