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两种不同方法分离的牙髓炎症组织来源的牙髓干细胞的成骨和成牙分化潜能。

Osteogenic and odontogenic differentiation potential of dental pulp stem cells isolated from inflamed dental pulp tissues (I-DPSCs) by two different methods.

机构信息

Department of Preventive and Restorative Dentistry, College of Dental Medicine, University of Sharjah, Sharjah, UAE.

The Sharjah Institute for Medical Research, University of Sharjah, Sharjah, UAE.

出版信息

Acta Odontol Scand. 2020 May;78(4):281-289. doi: 10.1080/00016357.2019.1702716. Epub 2019 Dec 19.

Abstract

The objective of the present study is to isolate stem cells from inflamed dental pulp tissues (I-DPSCs) and study the characteristic such as surface markers, osteo/odontogenic differentiation potential between the outgrowth (OG) and enzymatic digestion (COL) methods. I-DPSCs harvested by both methods were analysed for Mesenchymal Stem Cell marker expression by flow cytometry. The metabolic activity of the isolated cells was assessed by MTT assay. The Alkaline Phosphatase (ALP) and Alizarin red staining was done to analyse the osteogenic potential of isolated cells. The osteo/odontogenic differentiation was done by checking the expression of Dentine Matrix Protein 1 (DMP1), Dentine Sialophosphoprotein (DSPP), ALP and Bone Gamma-Carboxyglutamate Protein (BGLAP) by Real time PCR. The isolated cells were positive for MSC markers such as CD-90, CD-105 and CD-73 and negative for CD-14, CD-45 and STRO-1. MTT assay indicated that the I-DPSCs from OG method showed higher metabolic activity than cells from COL. However, the osteo/odontogenic differentiation was in favour of cells isolated by COL method. Although the cell metabolic rate was more in OG, the osteo/odontogenic differentiation was higher in COL, suggesting that the isolation method and culture conditions do affect the differentiation capacity of isolated cells.

摘要

本研究的目的是从发炎牙髓组织(I-DPSCs)中分离干细胞,并研究两种方法(外生法(OG)和酶消化法(COL))分离的干细胞的表面标志物和骨/牙向分化潜能的特征。采用流式细胞术分析两种方法分离的 I-DPSCs 中间充质干细胞标志物的表达。通过 MTT 法评估分离细胞的代谢活性。通过碱性磷酸酶(ALP)和茜素红染色分析分离细胞的成骨潜能。通过实时 PCR 检查牙本质基质蛋白 1(DMP1)、牙本质涎磷蛋白(DSPP)、ALP 和骨γ-羧基谷氨酸蛋白(BGLAP)的表达来进行骨/牙向分化。分离的细胞呈间充质干细胞标志物阳性,如 CD-90、CD-105 和 CD-73,而 CD-14、CD-45 和 STRO-1 为阴性。MTT 试验表明,OG 法分离的 I-DPSCs 的代谢活性高于 COL 法分离的细胞。然而,COL 法分离的细胞更有利于成骨/牙向分化。尽管 OG 中的细胞代谢率更高,但 COL 中的成骨/牙向分化更高,这表明分离方法和培养条件确实会影响分离细胞的分化能力。

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