The State Key Laboratory Breeding Base of Basic Science of Stomatology and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei 430079, P.R. China.
Int J Oncol. 2018 Jun;52(6):1863-1874. doi: 10.3892/ijo.2018.4361. Epub 2018 Apr 5.
The aim of this study was to examine the level and basic characteristics of cell‑derived microparticles (MPs) in the cyst fluids of odontogenic keratocysts (OKCs). For this purpose, MPs from the cyst fluids (CFMPs) of OKCs were purified by a classic differential centrifugation method and characterized by a transmission electron microscope and fluorescence microscope. Flow cytometric analysis was used to determine the size, concentration and cellular origins of the CFMPs. Moreover, the expression level of receptor activator for nuclear factor‑κB ligand in the OKCs was evaluated by immunohistochemical staining and then analyzed for its correlation with the concentration of CFMPs by Spearman's rank correlation test. In addition, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and tartaric‑resistant acid phosphatase (TRAP) staining were performed to examine the osteoclastogenesis of mouse bone marrow‑derived macrophages (BMMs) in response to CFMPs. The results revealed that the levels of total CFMPs were significantly elevated in OKCs compared with dentigerous cysts (DCs) and radicular cysts (RCs). In addition, in vitro experiments further revealed that CFMPs derived from the OKCs of patients could be taken up by BMMs, leading to a significant increase in the mRNA expression levels of nuclear factor of activated T‑cells 1 (NFATc1) and TRAP. Moreover, TRAP‑positive multinucleated osteoclasts were successfully cultured in the presence of macrophage colony‑stimulating factor (M‑CSF) and CFMPs with BMMs. On the whole, our findings indicate that patients with OKCs have higher levels of CFMPs compared with patients with DCs and RCs, which may be associated with the bone resorption of OKCs.
本研究旨在探讨牙源性角化囊肿(OKC)囊液中细胞衍生的微颗粒(MPs)的水平和基本特征。为此,采用经典差速离心法纯化 OKC 囊液中的 MPs(CFMPs),并通过透射电子显微镜和荧光显微镜进行表征。采用流式细胞术分析确定 CFMPs 的大小、浓度和细胞来源。此外,通过免疫组织化学染色评估 OKC 中核因子-κB 配体受体激活剂的表达水平,并通过 Spearman 秩相关检验分析其与 CFMPs 浓度的相关性。此外,还进行了逆转录-定量聚合酶链反应(RT-qPCR)和酒石酸抗性酸性磷酸酶(TRAP)染色,以检测 CFMPs 对小鼠骨髓源性巨噬细胞(BMMs)的破骨细胞生成的影响。结果表明,与含牙囊肿(DCs)和根尖囊肿(RCs)相比,OKC 中的总 CFMPs 水平显著升高。此外,体外实验进一步表明,来自 OKC 患者的 CFMPs 可被 BMMs 摄取,导致 T 细胞活化核因子 1(NFATc1)和 TRAP 的 mRNA 表达水平显著增加。此外,在巨噬细胞集落刺激因子(M-CSF)和 CFMPs 存在的情况下,成功培养了 BMMs 中的 TRAP 阳性多核破骨细胞。总的来说,我们的研究结果表明,与 DCs 和 RCs 患者相比,OKC 患者的 CFMPs 水平更高,这可能与 OKC 的骨质吸收有关。