Comparison of the Migration Potential through Microperforated Membranes of CD146+ GMSC Population versus Heterogeneous GMSC Population

Background. Guided tissue regeneration (GTR) is a powerful modality for periodontal regeneration, but it blocks the periosteum and gingival stem cells (GMSCs), from supporting periodontal wound by the nutrients, growth factors, and regenerative cells. The microperforated membrane considered a rewarding solution for this major drawback; GMSCs can migrate through a GTR microperforated membrane toward a chemoattractant, with the blocking of other unfavorable epithelial cells and fibroblasts. In the absence of a sole marker for MSC, a homogeneous population of GMSC is difficult to isolate; using CD146 as confirmatory markers for MSC identification, testing the behaviour of such homogeneous population in migration dynamics was the question to answer in this study. Materials and Methods. GMSCs from healthy crown lengthening tissue was isolated ( n = 3 ), its stem cell nature was confirmed, CD146 and CD271 markers were confirmatory markers to confirm homogenous stem cell population, and magnetic sorting was used to isolate GMSC with CD146 markers. A homogenous CD146 population was compared to heterogeneous GMSCs of origin; the population doubling time and MTT test of the two populations were compared. Migration dynamics were examined in a transwell migration chamber through 8 μm perforated polycarbonic acid membrane, and 0.4 μm and 3 μm perforated collagen-coated polytetrafluoroethylene membrane (PTFE) and 10% fetal bovine serum (FBS) were the chemoattractants used in the lower compartment to induce cell migration, were incubated in a humidified environment for 24 hours, then migrated the cell in the lower compartment examined by a light and electron microscope. Results. GMSCs fulfilled all the minimal criteria of stem cells and showed low signal 10% for CD146 on average and extremely low signal 2% for CD271 on average. Magnetic sorting optimized the signal of CD146 marker to 55%. GMSC CD146 population showed nonstatistically significant shorter population doubling time. CD146 homogeneous population migrated cell numbers were statistically significant compared to the heterogeneous population, through 0.4 μm and 3 μm perforated collagen membrane and 8 μm perforated polycarbonate membrane. Scanning electron microscopy proved the migration of the cells. Conclusions. A subset of the isolated GMSC showed a CD146 marker, which is considered a dependable confirmatory marker for the stem cells. In terms of GMSC migration through the microperforated membrane, a homogeneous CD146 population migrates more statistically significant than a heterogeneous GMSC population.

Trans-basement membrane migration of eosinophils induced by LPS-stimulated neutrophils from human peripheral bloodin vitro

In the airways of severe asthmatics, an increase of neutrophils and eosinophils is often observed despite high-dose corticosteroid therapy. We previously reported that interleukin-8-stimulated neutrophils induced trans-basement membrane migration (TBM) of eosinophils, suggesting the link between neutrophils and eosinophils. Concentrations of lipopolysaccharide (LPS) in the airway increase in severe asthma. As neutrophils express Toll-like receptor (TLR)4 and can release chemoattractants for eosinophils, we investigated whether LPS-stimulated neutrophils modify eosinophil TBM.Neutrophils and eosinophils were isolated from peripheral blood of healthy volunteers and severe asthmatics. Eosinophil TBM was examined using a modified Boyden's chamber technique. Eosinophils were added to the upper compartment, and neutrophils and LPS were added to the lower compartment. Migrated eosinophils were measured by eosinophil peroxidase assays.LPS-stimulated neutrophils induced eosinophil TBM (about 10-fold increase), although LPS or neutrophils alone did not. A leukotriene B4receptor antagonist, a platelet-activating factor receptor antagonist or an anti-TLR4 antibody decreased eosinophil TBM enhanced by LPS-stimulated neutrophils by almost half. Neutrophils from severe asthmatics induced eosinophil TBM and lower concentrations of LPS augmented neutrophil-induced eosinophil TBM.These results suggest that the combination of neutrophils and LPS leads eosinophils to accumulate in the airways, possibly involved the pathogenesis of severe asthma.

CFD ANALYSIS OF FLOW THROUGH HORIZONTAL CEILING OPENING DUE TO BUOYANCY AND PRESSURE DIFFERENCE

The buoyancy driven exchange flow through the large openings in horizontal partitions occurs in many practical situations such as in enclosed regions with a ceiling opening and a heat source such as fire. The density difference between two compartments arises partly due to difference in composition and partly from the difference in temperature. A heavier fluid located on the top of a lighter fluid and separated by a horizontal vent constitutes a gravitationally unstable system. Horizontal vents produce flow, which are unstable with irregular oscillatory behavior. However, when lower compartment is slightly pressurized the flow becomes stable and unidirectional. A numerical study has been performed to characterize the bi-directional flow and transition to unidirectional flow through a horizontal vent in an enclosure, due to differences in pressure and density across the vent. Fresh and salt water has been considered as working fluids to create density difference across the vent with a pressure field imposed in the lower compartment. The pressure in the lower region was increased to find the critical pressure corresponding to transition to unidirectional from bi-directional flow. Unsteady, 2D axisymmetric, incompressible Navier–Stokes equation along with species, turbulence and continuity equation have been solved with finite volume method using the in-house computational fluid dynamics (CFD) code. Several cases were examined to compute the critical pressure for various density differences for low opening aspect ratio. The code has been validated with reported experiments and used to simulate various other practical cases occurred during fire induced flow through such openings.

Application of Zone Models for Under-Ventilated Compartment Fires

A model is presented that explains the mass loss rate in a compartment as a function of fuel type and scale. The effect of ventilation is included in the model by the inclusion of the ambient oxygen concentration in the lower layer that results due to vent mixing. The model is executed in BRI2002, a zone model, capable of computing species and thermal conditions in the upper and lower compartment gas layers. Computations show good agreement with small-scale compartment data for heptane pools. The results can accurately portray extinction, oscillations in burning, reduction in the flaming area, and quasi-steady behaviors.