Inhibition of Rho-Associated Protein Kinase Increases the Angiogenic Potential of Mesenchymal Stem Cell Aggregates via Paracrine Effects

Abstract

The aggregation of multiple cells, such as mesenchymal condensation, is an important biological process in skeletal muscle development, osteogenesis, and adipogenesis. Due to limited in vivo study model systems, a simple and effective in vitro three-dimensional (3D) aggregation system is required to study the mechanisms of multicellular aggregation and its applications. We first generated controlled mesenchymal stem cell (MSC) aggregates using a bioprinting technique to monitor their aggregation and sprouting. We induced the angiogenic potential of the MSCs through chemical inhibition of the Rho/Rho-associated protein kinase (ROCK) pathway, which led to hairy sprouting in the aggregates. The angiogenic potential of this 3D construct was then tested by subcutaneously implanting the Matrigel with 3D MSC aggregates in a rat. Treatment of 3D MSCs with the ROCK inhibitor, Y27632, increased their angiogenic activity in vivo. The gene expressions and histological staining indicated that angiogenesis and neovascularization were mainly regulated by the paracrine factors secreted from human 3D MSC constructs. Our results demonstrate the enhancement of the angiogenic potential of the MSC constructs through the secretion of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) by the inhibition of the Rho/ROCK pathway.

Cell therapy of critical limb ischemia-problems and prospects

“…Platelet lysate was used as a supplement to the medium for cultivating MSCs instead of standard bovine serum [70] as well as a chemical inhibition of Rho/Rho-associated protein kinase [71] in order to increase the SC angiogenic potential. This allowed for demonstrating of an increase in the angiogenic activity of MCS constructs via an increase in VEGF and EGF secretion.…”

The promising rise of bioprinting in revolutionalizing medical science: Advances and possibilities

“…Additionally, hMSCs showed higher expression of angiogenic markers such as CD31 or CD105. Further studies have been conducted by the same research group on ROCK inhibition [ 169 ]. A scaffold-free liver tissue model was constructed using primary hepatocytes, stellate cells, and endothelial cells for experimental assessment of the toxicity of two drugs-levofloxacin and trovafloxacin [ 170 ].…”

The Promising Rise of Bioprinting in Revolutionalizing Medical Science: Advances and Possibilities

“…by the same research group on ROCK inhibition [165]. A scaffold-free liver tissue model was constructed using primary hepatocytes, stellate cells, and endothelial cells for experimental assessment of the toxicity of two drugs-levofloxacin and trovafloxacin [166].…”