气管瘘(TF)可能是危险的,甚至是致命的。目前的治疗方法非常具有挑战性。先前的研究报道,间充质干细胞(MSCs)可用于治疗呼吸道瘘。来自人脱落的乳牙(SHED)的干细胞被认为是MSC样细胞,也可能具有治疗气管瘘的潜力。在这项研究中,我们研究了SHED在大鼠气管瘘模型中的治疗作用。
将80只SD大鼠随机分为5组:假手术组,当地PBS组(L-PBS),静脉注射PBS组(I-PBS),局部SHED治疗组(L-SHED),和静脉SHED治疗组(I-SHED)。L-SHED组和I-SHED组局部应用于瘘管周围或经尾静脉注射1*107SHED,分别,而L-PBS和I-PBS组通过局部或静脉给药给予等量的PBS。2日采用立体显微镜观察瘘管愈合情况,3rd,移植后第5天。第七天,通过免疫荧光观察SHED的存活。通过苏木精和曙红(H&E)和Masson染色观察肺和瘘的病理学。Toll样受体4(TLR4)的表达水平,白细胞介素(IL)-1β,使用免疫组织化学测量IL-33和IL-4。TLR4、高迁移率族蛋白1(HMGB1)的表达水平,使用蛋白质印迹研究了骨髓分化因子88(MYD88)。在第14天,检测并分析大鼠的气道反应性。
移植后,L-SHED和I-SHED组的瘘管愈合快于其各自的PBS组。在第3天,L-SHED和I-SHED组的瘘管直径明显小于L-PBS和I-PBS组。此外,在L-SHED和I-SHED组中,瘘管周围成纤维细胞增殖和新血管生长的现象似乎更为明显.尽管SHED治疗后气道反应性没有明显差异,静脉注射SHED治疗可降低肺部炎症程度.然而,通过局部SHED治疗,肺部炎症没有显著减轻.I-SHED组IL-1β和IL-33的表达水平降低,与I-PBS组相比,IL-4升高。有趣的是,静脉SHED治疗可抑制TF大鼠肺组织中HMGB1/TLR4/MYD88的活化。
SHED移植加速大鼠瘘管愈合速度。静脉SHED治疗可减少肺部炎症。因此,SHED可能在气管瘘的治疗中具有潜力,为TF未来的治疗发展提供了希望。
Tracheal fistulas (TF) can be dangerous and even fatal in patients. The current treatment is really challenging. Previous studies reported that mesenchymal stem cells (MSCs) could be used to treat respiratory tract fistulas. Stem cells from human exfoliated deciduous teeth (SHED) are considered to be MSC-like cells that may also have the potential to treat the tracheal fistulas. In this study, we investigated the therapeutic effects of SHED in rat tracheal fistula models.
A total of 80 SD rats were randomly divided into five groups: a sham-operated group, a local PBS group (L-PBS), an intravenous PBS group (I-PBS), a local SHED treatment group (L-SHED), and an intravenous SHED treatment group (I-SHED). The L-SHED and I-SHED groups were given a topical application around the fistula or an intravenous injection of 1*107 SHED via the tail vein, respectively, while the L-PBS and I-PBS groups were given an equivalent volume of PBS through local or intravenous administration. A stereomicroscope was used to observe fistula healing on the 2nd, 3rd, and 5th days following transplantation. On the 7th day, the survival of SHED was observed by immunofluorescence. The pathology of the lungs and fistulas was observed by hematoxylin and eosin (H&E) and Masson staining. The expression levels of the Toll-like receptor 4 (TLR4), interleukin (IL)-1β, IL-33, and IL-4 were measured using immunohistochemistry. The expression levels of TLR4, high mobility group box 1 (HMGB1), and myeloid differentiation factor 88 (MYD88) were studied using western blotting. On day 14, airway responsiveness of rats was detected and analyzed.
Fistula healing in the L-SHED and I-SHED groups was faster than that in their respective PBS groups after transplantation. The fistula diameters in the L-SHED and I-SHED groups were significantly smaller than those in the L-PBS and I-PBS groups on the 3rd day. Moreover, the phenomenon of fibroblast proliferation and new blood vessel growth around the fistula seemed more pronounced in the L-SHED and I-SHED groups. Although no discernible difference was found in airway responsiveness after SHED treatment, the degree of inflammation in the lungs was reduced by intravenous SHED treatment. However, there was no significant reduction in lung inflammation by local SHED treatment. The expression levels of IL-1β and IL-33 were decreased in the I-SHED group, while IL-4 was elevated compared with the I-PBS group. Interestingly, intravenous SHED treatment inhibited the activation of HMGB1/TLR4/MYD88 in the lung tissues of TF rats.
SHED transplantation accelerated the rate of fistula healing in rats. Intravenous SHED treatment reduced lung inflammation. Thus, SHED may have potential in the treatment of tracheal fistula, providing hope for future therapeutic development for TF.