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人上颌窦膜中具有成骨潜能的间充质干细胞:一项体外研究。

Mesenchymal stem cells with osteogenic potential in human maxillary sinus membrane: an in vitro study.

作者信息

Berbéri Antoine, Al-Nemer Fatima, Hamade Eva, Noujeim Ziad, Badran Bassam, Zibara Kazem

机构信息

Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Lebanese University, Rafic Hariri Campus, P.O. box 5208-116, Beirut, Lebanon.

ER045, Laboratory of Stem Cells, DSST, PRASE, Lebanese University, Beirut, Lebanon.

出版信息

Clin Oral Investig. 2017 Jun;21(5):1599-1609. doi: 10.1007/s00784-016-1945-6. Epub 2016 Sep 1.

Abstract

OBJECTIVES

The aim of our study is to prove and validate the existence of an osteogenic progenitor cell population within the human maxillary Schneiderian sinus membrane (hMSSM) and to demonstrate their potential for bone formation.

MATERIALS AND METHODS

Ten hMSSM samples of approximately 2 × 2 cm were obtained during a surgical nasal approach for treatment of chronic rhinosinusitis and were retained for this study. The derived cells were isolated, cultured, and assayed at passage 3 for their osteogenic potential using the expression of Alkaline phosphatase, alizarin red and Von Kossa staining, flow cytometry, and quantitative real-time polymerase chain reaction.

RESULTS

hMSSM-derived cells were isolated, showed homogenous spindle-shaped fibroblast-like morphology, characteristic of mesenchymal progenitor cells (MPCs), and demonstrated very high expression of MPC markers such as STRO-1, CD44, CD90, CD105, and CD73 in all tested passages. In addition, von Kossa and Alizarin red staining showed significant mineralization, a typical feature of osteoblasts. Moreover, alkaline phosphatase (ALP) activity was significantly increased at days 7, 14, 21, and 28 of culture in hMSSM-derived cells grown in osteogenic medium, in comparison to controls. Furthermore, osteogenic differentiation significantly upregulated the transcriptional expression of osteogenic markers such as ALP, Runt-related transcription factor 2 (Runx-2), bone morphogenetic protein (BMP)-2, osteocalcin (OCN), osteonectin (ON), and osteopontin (OPN), confirming that hMSSM-derived cells are of osteoprogenitor origin. Finally, hMSSM-derived cells were also capable of producing OPN proteins upon culturing in an osteogenic medium.

CONCLUSION

Our data showed that hMSSM holds mesenchymal osteoprogenitor cells capable of differentiating to the osteogenic lineage.

CLINICAL RELEVANCE

hMSSM contains potentially multipotent postnatal stem cells providing a promising clinical application in preimplant and implant therapy.

摘要

目的

本研究的目的是证明并验证人类上颌窦施耐德氏膜(hMSSM)中存在成骨祖细胞群,并证明其骨形成潜力。

材料与方法

在治疗慢性鼻-鼻窦炎的手术鼻腔入路过程中获取10个约2×2 cm的hMSSM样本,并保留用于本研究。分离、培养所获得的细胞,并在第3代时使用碱性磷酸酶表达、茜素红和冯·科萨染色、流式细胞术以及定量实时聚合酶链反应检测其成骨潜力。

结果

分离出hMSSM来源的细胞,其呈现出均一的纺锤形成纤维细胞样形态,这是间充质祖细胞(MPC)的特征,并且在所有测试传代中均显示出MPC标志物如STRO-1、CD44、CD90、CD105和CD73的高表达。此外,冯·科萨和茜素红染色显示出明显的矿化,这是成骨细胞的典型特征。而且,与对照组相比,在成骨培养基中培养的hMSSM来源细胞在培养第7、14、21和28天时碱性磷酸酶(ALP)活性显著增加。此外,成骨分化显著上调了成骨标志物如ALP、 runt相关转录因子2(Runx-2)、骨形态发生蛋白(BMP)-2、骨钙素(OCN)、骨连接蛋白(ON)和骨桥蛋白(OPN)的转录表达,证实hMSSM来源的细胞起源于成骨祖细胞。最后,hMSSM来源的细胞在成骨培养基中培养时也能够产生OPN蛋白。

结论

我们的数据表明hMSSM含有能够分化为成骨谱系的间充质成骨祖细胞。

临床意义

hMSSM含有潜在的多能产后干细胞,在种植前和种植治疗中具有广阔的临床应用前景。

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