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间充质基质细胞的成骨分化:人皮下脂肪组织与牙髓的比较分析

Osteogenic Differentiation of Mesenchymal Stromal Cells: A Comparative Analysis Between Human Subcutaneous Adipose Tissue and Dental Pulp.

作者信息

D'Alimonte Iolanda, Mastrangelo Filiberto, Giuliani Patricia, Pierdomenico Laura, Marchisio Marco, Zuccarini Mariachiara, Di Iorio Patrizia, Quaresima Raimondo, Caciagli Francesco, Ciccarelli Renata

机构信息

1 Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara , Chieti, Italy .

3 Department of Aging Research Center and Translational Medicine (CeSI-MeT), University of Chieti-Pescara , Chieti, Italy .

出版信息

Stem Cells Dev. 2017 Jun 1;26(11):843-855. doi: 10.1089/scd.2016.0190. Epub 2017 Mar 13.

Abstract

White adipose tissue is a source of mesenchymal stromal/stem cells (MSCs) that are actively studied for their possible therapeutic use in bone tissue repair/remodeling. To better appreciate the osteogenic potential of these cells, we compared some properties of MSCs from human subcutaneous adipose tissue [subcutaneous-adipose stromal cells (S-ASCs)] and dental pulp stem cell (DPSCs) of third-impacted molars, the latter representing a well-established MSC source. Both undifferentiated cell types showed similar fibroblast-like morphology and mesenchymal marker expression. However, undifferentiated S-ASCs displayed a faster doubling time coupled to greater proliferation and colony-forming ability than DPSCs. Also, the osteogenic differentiation of S-ASCs was greater than that of DPSCs, as evaluated by the higher levels of expression of early osteogenic markers Runt-related transcription factor-2 (RUNX2) and alkaline phosphatase at days 3-14 and of extracellular matrix mineralization at days 14-21. Moreover, S-ASCs showed a better colonization of the titanium scaffold. In addition, we investigated whether S-ASC osteogenic commitment was enhanced by adenosine A1 receptor (A1R) stimulation, as previously shown for DPSCs. Although A1R expression was constant during DPSC differentiation, it increased in S-ASC at day 21 from osteogenesis induction. Accordingly, A1R stimulation by the agonist 2-chloro-N-cyclopentyl-adenosine, added to the cultures at each medium change, stimulated proliferation only in differentiating DPSC and enhanced the osteogenic differentiation earlier in DPSCs than in S-ASCs. These effects were counteracted by cell pretreatment with a selective A1R antagonist. Thus, our findings suggest that S-ASCs could be advantageously used in regenerative orthopedics/dentistry, and locally released or exogenously added purines may play a role in bone repair/remodeling, even though this aspect should be more thoroughly evaluated.

摘要

白色脂肪组织是间充质基质/干细胞(MSCs)的一个来源,人们正在积极研究其在骨组织修复/重塑中的潜在治疗用途。为了更好地了解这些细胞的成骨潜力,我们比较了来自人皮下脂肪组织的MSCs[皮下脂肪基质细胞(S-ASCs)]和第三恒磨牙牙髓干细胞(DPSCs)的一些特性,后者是一种成熟的MSCs来源。两种未分化的细胞类型均表现出相似的成纤维细胞样形态和间充质标志物表达。然而,未分化的S-ASCs比DPSCs具有更快的倍增时间,以及更强的增殖和集落形成能力。此外,通过在第3 - 14天早期成骨标志物Runt相关转录因子2(RUNX2)和碱性磷酸酶的更高表达水平,以及在第14 - 21天细胞外基质矿化情况评估,S-ASCs的成骨分化能力大于DPSCs。此外,S-ASCs在钛支架上的定植情况更好。另外,我们研究了腺苷A1受体(A1R)刺激是否会增强S-ASC的成骨定向分化,正如之前在DPSCs中所显示的那样。虽然在DPSC分化过程中A1R表达恒定,但在成骨诱导第21天,S-ASC中的A1R表达增加。相应地,在每次更换培养基时向培养物中添加激动剂2-氯-N-环戊基腺苷对A1R进行刺激,仅在分化的DPSC中刺激增殖,并比在S-ASCs中更早地增强DPSCs的成骨分化。这些作用被用选择性A1R拮抗剂对细胞进行预处理所抵消。因此,我们的研究结果表明,S-ASCs可有利地用于再生骨科/牙科,并且局部释放或外源性添加的嘌呤可能在骨修复/重塑中发挥作用,尽管这方面应进行更全面的评估。

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