Induction of osteogenic differentiation of MSCs by GSK3beta knockdown through GSK3beta siRNAs transfection

Abstract

The development of effective strategies for the treatment of bone defects is based on gene therapy methods aimed at regulating the differentiation of osteoprogenitor cells. One approach is the development of knockdown systems of inhibitory genes of osteogenic cell differentiation using siRNA molecules. In this work, we developed approaches to induce osteogenic differentiation of mesenchymal stem cells (MSCs) by knockdown of GSK3{beta} using siRNAs in cultures of MSCs derived from human adipose tissue (AD-MSCs). For this purpose, we performed a comparative evaluation of the efficacy of lipoplexes and polyplexes formed with one of the 4 siRNA molecules and 5 commercial transfection agents most commonly used in laboratory practice. The most effective transfection agent appeared to be PEI, which demonstrated high cytocompatibility in free form and as part of polyplexes even when maximum concentrations were used. Using the polyplexes formed by siRNA molecule designed for the first time and PEI, we developed a highly efficient GSK3{beta} gene knockdown system, which showed its effectiveness in cultures of AD-MSCs. As a result, we demonstrated the osteoinductive properties of GSK3{beta} siRNA molecules in these cultures. The results obtained can be applied in the development of gene therapy strategies based on siRNA molecules in human bone tissue diseases.