Lv Yue, Ge Lihong, Zhao Yuming
Pediatric Dentistry Department, Stomatological Hospital of Peking University No. 22 South Street, Zhongguancun, Haidian District, Beijing 100081, China.
Am J Transl Res. 2017 Feb 15;9(2):489-498. eCollection 2017.
To evaluate the effect of stem cells from human exfoliated deciduous teeth (SHED) upon the ulcer wound healing and evaluate the mechanism underlying the role of SHED in Sprague-Dawley rat models with diabetic foot ulcer. The rats with diabetic ulcer were established and treated with SHED, mesenchymal stem cell (MSC) and PBS, respectively. The expression of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), matrix metalloproteinase-2 (MMP-2) and MMP9 at both protein and RNA levels was quantitatively measured. The serum levels of VEGF, IL-1β, TNF-1α and IL-10 were detected by ELISA. The remaining tissues were fixed in 4% chloral hydrate for hematoxylin and eosin (H.E) staining and immunohistochemical staining. MSC and SHED administration could reduce ulceration area and accelerate wound healing at 7 and 14 d after treatment as compared with the control group (all <0.05), which were validated by H.E and immunohistochemical staining. Western blot results revealed that the expression levels of VEGF, eNOS, MMP2 and MMP9 proteins in the MSC and SHED groups were considerably up-regulated compared with those in the control group at different time points (all <0.05). The same trend was also observed in the mRNA expression of these cytokines detected by RT-PCR. At 3-d after treatment, no statistical significance was noted in the IL-10 level among three groups, but the IL-10 concentration in the SHED and MSC groups was significantly down-regulated at 7- and 14-d post-treatment (all <0.05). SHED administration, similar to MSCs, could accelerate wound healing, promote angiogenesis and suppress inflammatory responses in rat models with diabetic ulceration.
评估人乳牙脱落干细胞（SHED）对溃疡伤口愈合的影响，并评估SHED在糖尿病足溃疡的Sprague-Dawley大鼠模型中发挥作用的潜在机制。建立糖尿病溃疡大鼠模型，并分别用SHED、间充质干细胞（MSC）和磷酸盐缓冲液（PBS）进行治疗。定量检测血管内皮生长因子（VEGF）、内皮型一氧化氮合酶（eNOS）、基质金属蛋白酶-2（MMP-2）和MMP9在蛋白质和RNA水平的表达。通过酶联免疫吸附测定（ELISA）检测血清中VEGF、白细胞介素-1β（IL-1β）、肿瘤坏死因子-1α（TNF-1α）和IL-10的水平。其余组织用4%水合氯醛固定，进行苏木精-伊红（H.E）染色和免疫组织化学染色。与对照组相比，在治疗后7天和14天，给予MSC和SHED可减少溃疡面积并加速伤口愈合（均P<0.05），H.E染色和免疫组织化学染色证实了这一点。蛋白质印迹法结果显示，在不同时间点，MSC和SHED组中VEGF、eNOS、MMP2和MMP9蛋白的表达水平与对照组相比显著上调（均P<0.05）。通过逆转录-聚合酶链反应（RT-PCR）检测这些细胞因子的mRNA表达也观察到相同趋势。治疗后3天，三组间IL-10水平无统计学意义，但在治疗后7天和14天，SHED组和MSC组的IL-10浓度显著下调（均P<0.05）。与MSC相似，给予SHED可加速糖尿病溃疡大鼠模型的伤口愈合，促进血管生成并抑制炎症反应。
