Int J Clin Exp Pathol. 2015 Nov 1;8(11):14425-32. eCollection 2015.

Effect of ITGA5 down-regulation on the migration capacity of human dental pulp stem cells.

Xu S¹, Cui L², Ma D², Sun W³, Wu B².

Abstract

BACKGROUND:

The purpose of this study was to evaluate the role of integrin-α5 (ITGA5) in regulating the migration capacity of human dental pulpstem cells (hDPSCs), which might provide new evidence for understanding the repair and regeneration mechanisms of dental pulp tissues.

MATERIALS AND METHODS:

The enzyme digestion method was employed to isolate the hDPSCs from dental pulp tissues. The cell surface markers of hDPSCs were detected using flow cytometry analysis. Then the colony forming and multi-differentiation capacity of hDPSCs were evaluated. The lentivirus vector that carried the ITGA5 shRNA was constructed and real-time PCR was used to examine the effectiveness of ITGA5 shRNA lentivirus. Then transwell assay was performed to evaluate the impact of ITGA5 inhibition on the migration capability of hDPSCs.

RESULTS:

Our results showed that the cells we isolated from the dental pulps were positive for mesenchymal stem cells biomarkers. In addition, the cells possessed both colony forming capacity and multi-differentiation potential. ITGA5 shRNA lentivirus could not only infect hDPSCs with high efficiency, but also down-regulate the expression level of ITGA5 mRNA significantly (P<0.01). The transwell assay revealed the number of cells that migrated to the lower chamber was significantly less in the ITGA5 shRNA group compared with that in the scrambled shRNA group (P=0.016).

CONCLUSION:

ITGA5 plays an important role in maintaining and regulating the normal migration capacity of hDPSCs.