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1,25-二羟维生素 D3 对人根尖乳头干细胞的影响:黏附、铺展、增殖和成骨分化。

Effect of 1,25-Dihydroxyvitamin D3 on Stem Cells from Human Apical Papilla: Adhesion, Spreading, Proliferation, and Osteogenic Differentiation.

机构信息

Department of Oral and Maxillofacial Surgery, Affiliated Stomatological Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China.

Taizhou Polytechnic College, School of Pharmacy, Bone Tissue Engineering Research Center of Taizhou, Taizhou, Jiangsu 225300, China.

出版信息

Biomed Res Int. 2021 Oct 8;2021:1481215. doi: 10.1155/2021/1481215. eCollection 2021.

Abstract

Currently, it still remains a difficult problem to treat apical insufficiency of young permanent teeth resulted from pulp necrosis or periapical periodontitis. Previous studies have demonstrated that the treatment of revascularization using stem cells from apical papilla (SCAPs) results in increased root length and thickness of traumatized immature teeth and necrotic pulp. In this study, we investigated the role of 1,25-dihydroxyvitamin D3 in regulating the adhesion, spreading, proliferation, and osteogenic differentiation of SCAP, laying the foundation for subsequent clinical drug development. The immature tooth samples were collected in clinical treatment. SCAPs with stable passage ability were isolated and cultured. The multidifferentiation potential was determined by directed induction culture, while the stem cell characteristics were identified by flow cytometry. There were three groups: group A-SCAPs general culture group; group B-SCAPs osteogenesis induction culture group; and group C-SCAPs osteogenesis induction culture+1,25-dihydroxyvitamin D group, and the groups were compared statistically. The proliferation of SCAPs in each groups was detected through CCK-8 assay. RT-qPCR was used to detect the transcription levels of , , , and of SCAPs in each groups. Results exhibited that the isolated SCAPs had multidifferentiation potential and stem cell characteristics. After 24 h culturing, cells in group C spread better than those in groups A and B. The proliferation activity of SCAPs factored by CCK-8 ranked as group C > group B > group A, while the transcription levels of Runx2, ALP, Col I, and OCN leveled as group C > group B > group A. These results suggested that 1,25-dihydroxyvitamin D can significantly promote the adhesion, spreading, and proliferation of SACPs and improve the osteogenic differentiation of SCAPs by means of regulating upward the transcription level of osteogenic differentiation marker.

摘要

目前,治疗因牙髓坏死或根尖周炎导致的年轻恒牙根尖发育不全仍然是一个难题。先前的研究表明,使用根尖乳头干细胞(SCAP)进行血管再生治疗可增加创伤性未成熟牙和坏死牙髓的根长和厚度。在这项研究中,我们研究了 1,25-二羟维生素 D3 在调节 SCAP 黏附、铺展、增殖和成骨分化中的作用,为后续的临床药物开发奠定了基础。从临床治疗中收集未成熟牙样本。分离并培养具有稳定传代能力的 SCAP。通过定向诱导培养确定多向分化潜能,通过流式细胞术鉴定干细胞特征。共分为三组:A 组-SCAP 常规培养组;B 组-SCAP 成骨诱导培养组;C 组-SCAP 成骨诱导培养+1,25-二羟维生素 D 组,对各组进行统计学比较。通过 CCK-8 检测各组 SCAP 的增殖情况。通过 RT-qPCR 检测各组 SCAP 中 、 、 、 的转录水平。结果表明,分离的 SCAP 具有多向分化潜能和干细胞特征。培养 24 小时后,C 组细胞铺展情况优于 A 组和 B 组。CCK-8 检测的 SCAP 增殖活性依次为 C 组>B 组>A 组,而 Runx2、ALP、Col I 和 OCN 的转录水平依次为 C 组>B 组>A 组。这些结果表明,1,25-二羟维生素 D 可通过调节成骨分化标志物的转录水平,显著促进 SACPs 的黏附、铺展和增殖,提高 SCAP 的成骨分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ccfb/8519691/9c1d23d1a304/BMRI2021-1481215.001.jpg

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