Effect of volume flows on the viability of MSCs during injection through a cannula

Abstract

Introduction: Clinical trials currently evaluate the use of mesenchymal stem cells (MSCs) for the treatment of non-union bone fractures. The stem cells are injected directly into the non-union area of a bone via a cannula. During this injection process, pressure and shear forces affect the MSCs which could influence the viability of the cells. One parameter that influences the level of the shear forces is the volume flow. The aim of this study is to show whether the injection process with two different volume flows influences the cell viability. Methods: MSCs were isolated from bone tissue, harvested during arthroplasty. Afterwards, they were diluted to a concentration of 1 million cells/mL and 1 mL of this suspension was injected through a cannula with 200 mm length and 2 mm diameter (14 G) with volume flows of 38 and 100 mL/min. The evaluation was performed by detecting living, apoptotic, and dead cells using flow cytometry. The statistical analysis was performed with a Kruskal-Wallis-test to identify significant differences and with a TOST procedure for significant equivalence. The significance level was set to 5 % and the equivalence margin to 20 %. Results: The cell population of healthy cells was in the control group 85.88±2.98 %. After an injection with 38 mL/min the population of healthy cells was 86.04±2.53 % and with 100 ml/min 85.48±1.64 %. The statistical analysis revealed no significant difference between these groups (p = 0.99), but a significant equivalence between the control group and the two volume flows (38 mL/min: p = 0.002, 100 mL/min: p = 0.001). In addition the results show no increase of apoptotic and dead cells in the population after injection. Conclusion: The results indicate that the injection process through the cannula with these volume flows has no effect on the viability of the MSCs.

Keywords

• mesenchymal stem cells
• cell viability
• nonunion
• volume flows
• shear forces