Calcium Response to Fluid Flow - Induced Mechanotransduction in MC3T3-E1 Cells, Mesenchymal Stem Cells and Human Osteoblasts

Abstract

For the utilisation of flow - induced shear stress on MC3T3-E1 cell, human mesenchymal cells and human osteoblasts were all conducted for the study of bone reconstruction. Mechanical stimulus has played an important role in 2D and 3D growth process of cells. In this study, 0.01Pa, 0.02 Pa, and 0.045 Pa of flow-induced shear stress were applied to detect whether there was any changes of intracellular Ca2+ release rate or not. The results showed that even low shear stresses can have a positive stimulation for cell growth on 2D substance. In the role of the three different low shear stresses, the increasing Ca2+ release rates of MC3T3-E1 cell was flow rate dependent. Compared with baseline, the maximum calcium concentration of MC3T3-E1 cells is 5.7-fold (0.01Pa), 10.7-fold (0.02Pa), and 15.4-fold (0.045Pa) in normal medium. Also, the maximum calcium concentration of MC3T3-E1 cells is 5.8-fold (0.01Pa), 5.8-fold (0.02Pa), and 6.3-fold (0.045Pa) in Ca2+-free medium. For MSCs, the calcium concentration peak is 1.8-fold (0.01Pa), 2.4-fold (0.02Pa), and 1.1-fold (0.045Pa) in normal medium; and 2.2-fold (0.01Pa), 1.9-fold (0.02Pa), and 2.1-fold (0.045Pa) in Ca2+-free medium. For HOB cells, the results showed no significant similar pattern with these three different levels of lower flow rate. The intracellular calcium concentration is 2.3-fold (0.01Pa), 1.8-fold (0.02Pa), and 2.3-fold (0.045Pa) in normal medium; and 3.2-fold (0.01Pa), 4.1-fold (0.02Pa), and 2.3-fold (0.045Pa) in Ca2+-free medium. This study suggests that compared to high fluid shear stress and high requirement of experimental equipment, low fluid shear stress for 2D cell mechanotransduction experiment is more simple, convenient and effective.

Date of Award	31 Dec 2014
Original language	English
Awarding Institution	The University of Manchester