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Amniotic Membrane Enhance the Effect of Vascular Endothelial Growth Factor on the Angiogenic Marker Expression of Stem Cells from Human Exfoliated Deciduous Teeth.
Applied Biochemistry and Biotechnology ( IF 3 ) Pub Date : 2020-02-25 , DOI: 10.1007/s12010-020-03266-1
Muhammad Fuad Hilmi Yusof 1 , Siti Nurnasihah Md Hashim 1 , Wafa' Zahari 1 , Hamshawagini Chandra 1 , Khairul Bariah Ahmad Amin Noordin 1 , Thirumulu Ponnuraj Kannan 1, 2 , Suzina Sheikh Abdul Hamid 3 , Khairani Idah Mokhtar 4 , Ahmad Azlina 1, 2
Affiliation  

Abstract

Previously, it was reported that human amniotic membrane (AM) induced stem cells from human deciduous exfoliated teeth (SHED) endothelial-like-cell differentiation. This interesting effect of AM matrix on SHED demands further elucidation. Objective of this in vitro work was to study the effect of 24-h VEGF induced on SHED endothelial differentiation when seeded on acellular stromal side (SS) of AM matrix. Stemness of SHED was identified by flow cytometry. Cell attachment and morphological changes towards the matrix was observed by scanning electron microscopy. Protein expression of endothelial marker was examined by Western blot. The expression of stem cells and endothelial-specific gene markers of VEGF-induced SHED cultured on human AM was inspected via reverse transcriptase-polymerase chain reaction. Results showed SHED at both passages retain stemness property. Ang-1 protein was expressed in SHED. Cells treated with VEGF and cultured on AM transformed attached well to AM. VEGF-induced SHED expressed both stem cell and endothelial-specific markers throughout the treatments and timeline. Interestingly, prolonged VEGF treatment increased the expression of Cox-2 and VE-Cadherin genes in all treated groups when compared to SHED. It was concluded that the VEGF-induced SHED showed better expression of endothelial-specific markers when cultured on SS of AM, with prolonged VEGF treatment.



中文翻译:

羊膜增强血管内皮生长因子对人脱落乳牙干细胞血管生成标记表达的影响。

摘要

以前,有报道说人羊膜(AM)可以诱导人乳头脱落牙齿(SHED)内皮样细胞分化产生的干细胞。AM矩阵对SHED的有趣影响需要进一步阐明。这项体外研究的目的是研究接种于AM基质的无细胞基质侧(SS)时诱导的24 h VEGF对SHED内皮细胞分化的影响。通过流式细胞术鉴定SHED的茎。通过扫描电子显微镜观察到细胞向基质的附着和形态变化。通过Western印迹检查内皮标志物的蛋白表达。通过逆转录酶-聚合酶链反应检测VEGF诱导的SHED在人AM上的干细胞表达及内皮特异性基因标记。结果表明,SHED在两个通道均保留茎性。Ang-1蛋白在SHED中表达。用VEGF处理并在AM转化上培养的细胞与AM良好结合。在整个治疗过程和时间表中,VEGF诱导的SHED均表达干细胞和内皮特异性标志物。有趣的是,长时间的VEGF治疗可增加VEGF的表达与SHED相比,所有治疗组的Cox-2VE-钙黏着蛋白基因。结论是,在长时间的VEGF处理下,当在AM的SS上培养时,VEGF诱导的SHED表现出更好的内皮特异性标记物表达。

更新日期:2020-02-25
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