ABSTRACT
Wound healing is a dynamic process that involves a series of molecular and cellular events aimed at replacing devitalized and missing cellular components and/or tissue layers. Recently, extracellular vesicles (EVs), naturally cell-secreted lipid membrane-bound vesicles laden with biological cargos including proteins, lipids, and nucleic acids, have drawn wide attention due to their ability to promote wound healing and tissue regeneration. However, current exploitation of EVs as therapeutic agents is limited by their low isolation yields and tedious isolation processes. To circumvent these challenges, bioinspired cell-derived nanovesicles (CDNs) that mimic EVs were obtained by shearing mesenchymal stem cells (MSCs) through membranes with different pore sizes. Physical characterisations and high-throughput proteomics confirmed that MSC-CDNs mimicked MSC-EVs. Moreover, these MSC-CDNs were efficiently uptaken by human dermal fibroblasts and demonstrated a dose-dependent activation of MAPK signalling pathway, resulting in enhancement of cell proliferation, cell migration, secretion of growth factors and extracellular matrix proteins, which all promoted tissue regeneration. Of note, MSC-CDNs enhanced angiogenesis in human dermal microvascular endothelial cells in a 3D PEG-fibrin scaffold and animal model, accelerating wound healing in vitro and in vivo. These findings suggest that MSC-CDNs could replace both whole cells and EVs in promoting wound healing and tissue regeneration.
ABSTRACT
Mineral trioxide aggregate (MTA) can provide bioactivity to poly-caprolactone (PCL), which is an inert polymer used to print scaffolds. However, testing all combinations of scaffold characteristics (e.g., composition, pore size, and distribution) to optimize properties of scaffolds is time-consuming and costly. The Taguchi's methods can identify characteristics that have major influences on the properties of complex designs, hence decreasing the number of combinations to be tested. The objective was to assess the potential of Taguchi's methods as a predictive tool for the optimization of bioactive scaffold printed using electro-hydro dynamic jetting. A three-level approach assessed the influence of PCL/MTA proportion, pore size, fiber dimension and number of layers in pH, degradation rate, porosity, yield strength, and Young's modulus. Data were analyzed using Tukey's honest significant difference test, analysis of mean and signal-to-noise ratio (S/N) test. Cytocompatibility and differentiation potential were assessed for 5 and 30 days using dental pulp stem cells and analyzed with one-way analysis of variance (proliferation) or Mann-Whitney (qPCR). The S/N ratio and analysis of mean showed that fiber diameter and composition were the most influential characteristics in all properties. The experimental data confirmed that the addition of MTA to PCL increased the pH and scaffold degradation. Only PCL and PCL with 4% MTA allowed cell proliferation. The latter increased the genetic expression of ALP, COL-1, OCN, and MSX-1. The theoretical predictions were confirmed by the experiments. The Taguchi's identified the inputs that can be disregarded to optimize 3D printed meshed bioactive scaffolds.