The interaction between dental pulp derived mesenchymal stem cells (DP-MSCs) and three different tantalum nanotopographies with and without a fibronectin coating is examined: sputter-coated tantalum surfaces with low surface roughness <0.2 nm, hut-nanostructured surfaces with a height of 2.9 +/- 0.6 nm and a width of 35 +/- 8 nm, and dome structures with a height of 13 +/- 2 nm and a width of 52 +/- 14 nm. Using ellipsometry, the adsorption and the availability of fibronectin cell-binding domains on the tantalum surfaces were examined, as well as cellular attachment, proliferation, and vinculin focal adhesion spot assembly on the respective surfaces. The results showed the highest fibronectin mass uptake on the hut structures, with a slightly higher availability of cell-binding domains and the most pronounced formation of vinculin focal adhesion spots as compared to the other surfaces. The proliferation of DP-MSCs was found to be significantly higher on dome and hut surfaces coated with fibronectin compared to the uncoated flat tantalum surfaces. Consequently, the results presented in this study indicate that fibronectin-coated nanotopographies with a vertical dimension of less than 5 nm influence cell adhesion. This rather interesting behavior is argued to originate from the more available fibronectin cell-binding domains observed on the hut structures.
Aim To assess the diagnostic accuracy of Cone Beam Computed Tomography (CBCT) to diagnose apical periodontitis (AP) using histopathology of ex vivo human jaws as the reference standard. Methodology Based on periapical radiographs of jaw specimens from human bodies donated for science, a sample of 223 teeth with 340 roots including all tooth groups, and different disease and treatment statuses was selected. Cone Beam Computed Tomography was performed using Cranex® 3Dx (Soredex Oy, Tuusula, Finland), small field‐of‐view (5 × 5 cm), and isotropic resolution 0.085 mm. Three observers assessed the presence of AP using a probability index. Histopathological examination of the periapical area was used as a reference standard to calculate estimates of diagnostic accuracy. Results For non‐root filled teeth all estimates of diagnostic accuracy; sensitivity (SENS), specificity (SPEC), positive predictive value (PPV) and negative predictive value (NPV) were high. All estimates were lower for root filled teeth. When mild AP was classified as ‘AP’, SENS, SPEC and NPV were significantly lower in root filled roots (P < 0.001 in all cases). The same tendency was seen when mild AP was classified as ‘No AP’ but here only the difference in SPEC was significant (P < 0.001). Conclusion The diagnostic accuracy of CBCT used for diagnosis of AP is dependent on the treatment status of the tooth. For non‐root filled teeth the diagnostic accuracy of CBCT is high and almost all cases of AP can be diagnosed correctly with only a very small risk of over‐diagnosis. All diagnostic accuracy parameters were lower for root filled roots, hence the diagnosis of AP on root filled roots using CBCT was less accurate.
Introduction: The osteogenic differentiation of bone marrow-derived mesenchymal stromal cells (BMSCs) was compared with that of dental pulp-derived stromal cells (DPSCs) in vitro and in a pig calvaria critical-size bone defect model. Methods: BMSCs and DPSCs were extracted from the tibia bone marrow and the molar teeth of each pig, respectively. BMSCs and DPSCs were cultured in monolayer and on a three-dimensional (3D) polycaprolactone (PCL) – hyaluronic acid – tricalcium phosphate (HT-PCL) scaffold. Population doubling (PD), alkaline phosphatase (ALP) activity, and calcium deposition were measured in monolayer. In the 3D culture ALP activity, DNA content, and calcium deposition were evaluated. Six non-penetrating critical-size defects were made in each calvarium of 14 pigs. Three paired sub-studies were conducted: (1) empty defects vs. HT-PCL scaffolds; (2) PCL scaffolds vs. HT-PCL scaffolds; and (3) autologous BMSCs on HT-PCL scaffolds vs. autologous DPSCs on HT-PCL scaffolds. The observation time was five weeks. Bone volume fractions (BV/TV) were assessed with micro-computed tomography (μCT) and histomorphometry. Results and discussion: The results from the in vitro study revealed a higher ALP activity and calcium deposition of the DPSC cultures compared with BMSC cultures. Significantly more bone was present in the HT-PCL group than in both the pure PCL scaffold group and the empty defect group in vivo. DPSCs generated more bone than BMSCs when seeded on HT-PCL. In conclusion, DPSCs exhibited a higher osteogenic potential compared with BMSCs both in vitro and in vivo, making it a potential cell source for future bone tissue engineering.
In this study, we sought to assess the osteogenic potential of human dental pulp stem cells (DPSCs) on three different polycaprolactone (PCL) scaffolds. The backbone structure of the scaffolds was manufactured by fused deposition modeling (PCL scaffold). The composition and morphology was functionalized in two of the scaffolds. The first underwent thermal induced phase separation of PCL infused into the pores of the PCL scaffold. This procedure resulted in a highly variable micro- and nanostructured porous (NSP), interconnected, and isotropic tubular morphology (NSP-PCL scaffold). The second scaffold type was functionalized by dip-coating the PCL scaffold with a mixture of hyaluronic acid and β-TCP (HT-PCL scaffold). The scaffolds were cylindrical and measured 5 mm in height and 10 mm in diameter. They were seeded with 1×10(6) human DPSCs, a cell type known to express bone-related markers, differentiate into osteoblasts-like cells, and to produce a mineralized bone-like extracellular matrix. DPSCs were phenotypically characterized by flow cytometry for CD90(+), CD73(+), CD105(+), and CD14(-). DNA, ALP, and Ca(2+) assays and real-time quantitative polymerase chain reaction for genes involved in osteogenic differentiation were analyzed on day 1, 7, 14, and 21. Cell viability and distribution were assessed on day 1, 7, 14, and 21 by fluorescent-, scanning electron-, and confocal microscopy. The results revealed that the DPSCs expressed relevant gene expression consistent with osteogenic differentiation. The NSP-PCL and HT-PCL scaffolds promoted osteogenic differentiation and Ca(2+) deposition after 21 days of cultivation. Different gene expressions associated with mature osteoblasts were upregulated in these two scaffold types, suggesting that the methods in which the scaffolds promote osteogenic differentiation, depends on functionalization approaches. However, only the HT-PCL scaffold was also able to support cell proliferation and cell migration resulting in even cell dispersion throughout the scaffold. In conclusion, DPSCs could be a possible alternate cell source for bone tissue engineering. The HT-PCL scaffold showed promising results in terms of promoting cell migration and osteogenic differentiation, which warrants future in vivo studies.
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