Shuaimei Xu1, Li Cui2, Dandan Ma2, Wenjuan Sun3, Buling Wu2. 1. Department of Endodontics, The Stomatological Hospital of Guangdong ProvinceGuangzhou 510280, China; Department of Stomatology, Nanfang HospitalGuangzhou 510515, China. 2. Department of Stomatology, Nanfang Hospital Guangzhou 510515, China. 3. Department of Stomatology, The Third Affiliated Hospital, Sun Yat-sen University Guangzhou, China.
Abstract
BACKGROUND: The purpose of this study was to evaluate the role of integrin-α5 (ITGA5) in regulating the migration capacity of human dental pulp stem 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.
BACKGROUND: The purpose of this study was to evaluate the role of integrin-α5 (ITGA5) in regulating the migration capacity of human dental pulp stem 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.
Authors: Giovanni Marchetti; Sarah Escuin; Arjan van der Flier; Adèle De Arcangelis; Richard O Hynes; Elisabeth Georges-Labouesse Journal: Eur J Neurosci Date: 2010-01-25 Impact factor: 3.386