背景:良性牙源性病变(BOLs)可导致严重的颌骨缺损并损害患者的生活质量。细胞外囊泡(EV)是介导病理生理事件的成熟和多才多艺的参与者。间质空间中的EV(组织衍生的EV或Ti-EV)在疾病相关的生物标志物发现中具有更高的特异性和敏感性。然而,负载Ti-EV的蛋白在介导BOLs发展中的作用仍未被开发。在这里,我们的目的是探讨Ti-EV负载蛋白对BOLs发育的贡献。
方法:从3个牙囊中获得样品,3有牙质囊肿(DC),7与牙源性角化囊肿(OKC),成釉细胞瘤(AM)3例。然后提取组织来源的电动汽车,纯化,并使用超速离心进行了验证,透射电子显微镜,和西方印迹。使用LC-ESI串联质谱法分析来自Ti-EV的蛋白质,并筛选差异表达的蛋白质。然后通过免疫组织化学和免疫荧光分析进行验证。
结果:通过LC-MS分析绘制各组中Ti-EV的蛋白质谱图。BOL衍生的Ti-EV中的前10个丰富蛋白是COL6A3,COL6A1,ALB,HIST1H4A,HBB,ACTB,HIST1H2BD,ANXA2、COL6A2和FBN1。此外,确定了来自各种病变的Ti-EV中的独特蛋白质。此外,粘着斑激酶(FAK)和髓样分化原发反应88(MyD88)在来自OKC和AM的Ti-EV中显示出更高的表达,免疫组织化学和免疫荧光染色证实。
结论:含FAK和MyD88的Ti-EV可能与OKC和AM的发生有关,可能是潜在的治疗靶点。
BACKGROUND: Benign odontogenic lesions (BOLs) can cause severe jaw bone defects and compromise the quality of life of patients. Extracellular vesicles (EVs) are well-established and versatile players in mediating pathophysiological events. EVs in the interstitial space (tissue-derived EVs or Ti-EVs) possess higher specificity and sensitivity in disease-related biomarker discovery. However, the role of Ti-EV-loaded proteins in mediating the development of BOLs has remained untapped. Herein, we aim to explore the contribution of Ti-EV-loaded proteins to the development of BOLs.
METHODS: Samples were obtained from 3 with dental follicle, 3 with dentigerous cyst (DC), 7 with odontogenic keratocyst (OKC), and 3 patients with ameloblastoma (AM). Tissue-derived EVs were then extracted, purified, and validated using ultracentrifugation, transmission electron microscopy, and western blotting. Proteins from Ti-EVs were analyzed using LC-ESI tandem mass spectroscopy and differentially expressed proteins were screened, which was then validated by immunohistochemistry and immunofluorescence assays.
RESULTS: The protein profile of Ti-EVs in each group was mapped by LC-MS analysis. The top 10 abundant proteins in BOL-derived Ti-EVs were COL6A3, COL6A1, ALB, HIST1H4A, HBB, ACTB, HIST1H2BD, ANXA2, COL6A2 and FBN1. Additionally, unique proteins in the Ti-EVs from various lesions were identified. Moreover, focal adhesion kinase (FAK) and myeloid differentiation primary response 88 (MyD88) showed higher expressions in Ti-EVs derived from OKC and AM, which were confirmed by immunohistochemistry and immunofluorescence staining.
CONCLUSIONS: Ti-EVs containing FAK and MyD88 might be related to the development of OKC and AM, which can be potential therapeutic targets.