Gene expression analysis of stem cell migration in local angiogenesis for tissue repair

Nur Syazwani Aziz; Azlina Ahmad; Norhayati Yusop

Nur Syazwani Aziz Basic Sciences and Oral Biology Unit, School of Dental Science, University Sains Malaysia
Azlina Ahmad Basic Sciences and Oral Biology Unit, School of Dental Science, University Sains Malaysia
Norhayati Yusop Basic Sciences and Oral Biology Unit, School of Dental Science, University Sains Malaysia

Abstract

Dental stem cells offer great potential benefit for application in stem cell-based therapy. Stem cells from exfoliated deciduous teeth (SHED) exhibit mesenchymal stem cell characteristics, with the ability to form various cell types of different lineages. Migration of endothelial cells plays a critical role in angiogenesis, to further support blood vessel formation during tissue repair and regeneration. However, the exact cellular characteristics of SHED undergoing angiogenesis, in term of its migratory capacity and gene expression pattern have not been fully understood. Hence, this study aims to assess the differential gene expression of SHED following angiogenesis and migratory induction. SHED migration capacity under different seeding density, and between undifferentiated state and the angiogenically induced group are investigated. Briefly, in-vitro cultured SHED was induced to form endothelial cells by supplementation of angiogenic factors. Scratch test assay was used to estimate the rate of cell migration, whereas transwell migration assay was performed to collect RNA samples on day 1,3,7,10 and 14. Following RNA extraction, the samples were further analysed by RT-PCR for detection of stem cell markers, migration markers and angiogenic markers. The gathered findings indicated that SHED is highly capable of forming endothelial cells. SHED was found to maintain stem cell markers expression (CD73, CD90 and CD 105) throughout the 14 days of angiogenic induction. Meanwhile, the expression of migratory markers; CCR1, CXCR4 and CCL28 were found to be downregulated in comparison to the angiogenic markers; Ang1, IL8 and VE Cadherin. SHED also demonstrated a higher capacity to undergo cell migration under immature state in comparison to the angiogenically induced environment. In conclusion, SHED undergoing angiogenesis is postulated to have much more lower capacity to migrate to the healing site. The knowledge gained from this study can be used to plan a strategic approach for stem cell-based tissue repair.

Keywords: dental; stem cell; repair; angiogenesis; migration; SHED