School of Dental Sciences, Universiti Sains Malaysia, Health Campus, 16150, Kubang Kerian, Kelantan, Malaysia.
Human Genome Centre, School of Medical Sciences, Universiti Sains Malaysia, Health Campus, 16150, Kubang Kerian, Kelantan, Malaysia.
Appl Biochem Biotechnol. 2020 May;191(1):177-190. doi: 10.1007/s12010-020-03266-1. Epub 2020 Feb 25.
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.
先前有报道称，人羊膜（AM）诱导人乳牙脱落（SHED）内皮样细胞分化的干细胞。AM 基质对 SHED 具有这种有趣的影响，需要进一步阐明。本体外工作的目的是研究在接种于 AM 基质去细胞基质侧（SS）的情况下，24 小时 VEGF 对 SHED 内皮分化的诱导作用。通过流式细胞术鉴定 SHED 的干性。通过扫描电子显微镜观察细胞在基质上的附着和形态变化。通过 Western blot 检测内皮标志物的蛋白表达。通过逆转录-聚合酶链反应检查 VEGF 诱导的培养在人 AM 上的 SHED 的干细胞和内皮特异性基因标志物的表达。结果表明，第 2 代和第 4 代 SHED 均保持干性。SHED 表达 Ang-1 蛋白。用 VEGF 处理并培养在 AM 上的细胞很好地附着在 AM 上。VEGF 诱导的 SHED 在整个治疗过程和时间线上均表达干细胞和内皮特异性标志物。有趣的是，与 SHED 相比，延长 VEGF 处理可增加所有处理组中 Cox-2 和 VE-Cadherin 基因的表达。结论是，VEGF 诱导的 SHED 在 AM 的 SS 上培养时，内皮特异性标志物的表达更好，而延长 VEGF 处理则可增加 Cox-2 和 VE-Cadherin 基因的表达。
