Flow-dependent shear stress affects the biological properties of pericyte-like cells isolated from human dental pulp

Abstract

Background Human dental pulp stem cells represent a mesenchymal stem cell niche localized in the perivascular area of dental pulp and are characterized by low immunogenicity and immunomodulatory/anti-inflammatory properties. Pericytes, mural cells surrounding the endothelium of small vessels, regulate numerous functions including vessel growth, stabilization and permeability. It is well established that pericytes have a tight cross talk with endothelial cells in neoangiogenesis and vessel stabilization, which are regulated by different factors, i.e., microenvironment and flow-dependent shear stress. The aim of this study was to evaluate the effects of a pulsatile unidirectional flow in the presence or not of an inflammatory microenvironment on the biological properties of pericyte-like cells isolated from human dental pulp (hDPSCs). Methods Human DPSCs were cultured under both static and dynamic conditions with or without pre-activated peripheral blood mononuclear cells (PBMCs). Pulsatile unidirectional flow shear stress was generated by using a specific peristaltic pump. The angiogenic potential and inflammatory properties of hDPSCs were evaluated through reverse phase protein microarrays (RPPA), confocal immunofluorescence and western blot analyses. Results Our data showed that hDPSCs expressed the typical endothelial markers, which were up-regulated after endothelial induction, and were able to form tube-like structures. RPPA analyses revealed that these properties were modulated when a pulsatile unidirectional flow shear stress was applied to hDPSCs. Stem cells also revealed a downregulation of the immune-modulatory molecule PD-L1, in parallel with an up-regulation of the pro-inflammatory molecule NF-kB. Immune-modulatory properties of hDPSCs were also reduced after culture under flow-dependent shear stress and exposure to an inflammatory microenvironment. This evidence was strengthened by the detection of up-regulated levels of expression of pro-inflammatory cytokines in PBMCs. Conclusions In conclusion, the application of a pulsatile unidirectional flow shear stress induced a modulation of immunomodulatory/inflammatory properties of dental pulp pericyte-like cells.

Zirconia Hybrid Dental Implants Influence the Biological Properties of Neural Crest-Derived Mesenchymal Stromal Cells

“…This study was carried out using commercial human dental pulp stem cells (hDPSCs; CTIbiotech, Meyzieu-Lyon, France) cultured according to previously established protocols [ 32 ]. After cell expansion, a MACS ® separation kit (Miltenyi Biotec S.r.l, Bologna, Italy) was used to obtain a homogenous hDPSCs population through an immune selection against the surface antigens STRO-1 and c-Kit [ 32 ].…”

“…Peripheral blood mononuclear cells (PBMCs) were isolated from the fresh whole blood of healthy adult volunteers (n = 5) by using Histopaque ® (Sigma Aldrich, St. Louis, MO, USA) according to the manufacturer’s instructions and preactivated by adding anti-CD3 and the costimulatory anti-CD28 monoclonal antibodies (BD Biosciences, Franklin Lakes, NJ, USA) to the culture medium [ 32 ]. PBMCs, resting or preactivated, according to each experimental evaluation, were then seeded and cultured in 6-well plates (3.5 × 10 6 cell/well) in the presence of both SLA titanium and CERID disks in order to investigate the biological effects of both implants on PBMCs.…”

Synergy of Nanotopography and Electrical Conductivity of PEDOT/PSS for Enhanced Neuronal Development

“…Nuclei were counterstained with 10 μg/mL Hoechst 33342 blue dye. Images were acquired with a Nikon A1 confocal laser scanning microscope …”

“…Images were acquired with a Nikon A1 confocal laser scanning microscope. 37 SEM Analysis of Neural Cell Morphology. For SEM evaluations, cells cultured on substrate at different time points were fixed in 4% glutaraldehyde in phosphate buffer (pH 7.4), postfixed in 2% osmium tetroxide, dehydrated in an ascending series of alcohols, and dried with hexamethyldisilazane.…”

Human dental pulp stem cells (hDPSCs) promote the lipofibroblast transition in the early stage of a fibro-inflammatory process

“…Among different MSC types, human dental pulp stem cells (hDPSCs) have been demonstrated to display pericyte-like features, to own a wide differentiation potential in vitro and to contribute to regenerating processes and to the maintenance of tissue homeostasis through direct histointegration, immunomodulatory abilities, and promotion of neoangiogenesis ( Janebodin et al, 2013 ; Nuti et al, 2016 ; Martinez-Sarrà et al, 2017 ; Paganelli et al, 2021 ). Recently, our research group has demonstrated that hDPSCs are able to modulate the immune response by activating different pathways, i.e., PD1/PD-L1 and Fas/FasL, and that the surrounding microenvironment may affect their immunomodulatory/inflammatory phenotype ( Di Tinco et al, 2021 ; Bertani et al, 2023 ). However, little is known about the role of hDPSCs in fibrosis processes.…”