Increased extracellular matrix deposition during chondrogenic differentiation of dental pulp stem cells from individuals with neurofibromatosis type 1: an in vitro 2D and 3D study.
Increased extracellular matrix deposition during chondrogenic differentiation of dental pulp stem cells from individuals with neurofibromatosis type 1: an in vitro 2D and 3D study.
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DOI：
10.1186/s13023-018-0843-1
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发表时间：
2018-06-25
影响因子：
3.7
通讯作者：
Cunha KS
中科院分区：
文献类型：
--
作者：
Almeida PN;Barboza DDN;Luna EB;Correia MCM;Dias RB;Siquara de Sousa AC;Duarte MEL;Rossi MID;Cunha KS
神经纤维瘤病1（NF 1）表现出广泛的临床表现，包括骨改变。寻求了解NF 1骨科问题的细胞和分子机制的研究对于更好地了解发病机制和新疗法的发展非常重要。牙髓干细胞（DPSC）被用作几种疾病的体外模型，并作为NF 1的合适模型出现。本研究的目的是评估在体外软骨分化的DPSCs从个人与NF 1使用二维（2D）和三维（3D）文化。为了满足国际细胞治疗学会的标准，DPSC的特征在于表面抗原表达和它们的多潜能性，在2D培养中被诱导向成脂、成骨和成软骨谱系分化。来自具有NF 1的个体的DPSC（NF 1 DPSC）和对照培养物均为CD 90、CD 105、CD 146阳性，而CD 13、CD 14、CD 45和CD 271阴性，并且在方案后成功分化。在2D和3D（颗粒）培养物中评价软骨形成分化，并通过光学显微镜和透射电子显微镜（TEM）进一步评价。在软骨形成分化过程中，与对照相比，2D培养物显示NF 1 DPSC中细胞外基质沉积更多。在半薄切片中，对照细胞团具有大小均匀的细胞内和细胞外基质囊泡，而NF 1培养物具有不同大小的基质囊泡。透射电镜分析显示，与对照培养物相比，NF 1培养物中胶原纤维的量更高。NF 1 DPSCs在软骨形成分化过程中细胞外基质沉积增加，这可能与NF 1个体的骨骼变化有关。本文的在线版本（10.1186/s13023-018-0843-1）包含补充材料，可供授权用户使用。
Neurofibromatosis 1 (NF1) presents a wide range of clinical manifestations, including bone alterations. Studies that seek to understand cellular and molecular mechanisms underlying NF1 orthopedic problems are of great importance to better understand the pathogenesis and the development of new therapies. Dental pulp stem cells (DPSCs) are being used as an in vitro model for several diseases and appear as a suitable model for NF1. The aim of this study was to evaluate in vitro chondrogenic differentiation of DPSCs from individuals with NF1 using two-dimensional (2D) and three-dimensional (3D) cultures. To fulfill the criteria of the International Society for Cellular Therapy, DPSCs were characterized by surface antigen expression and by their multipotentiality, being induced to differentiate towards adipogenic, osteogenic, and chondrogenic lineages in 2D cultures. Both DPSCs from individuals with NF1 (NF1 DPSCs) and control cultures were positive for CD90, CD105, CD146 and negative for CD13, CD14, CD45 and CD271, and successfully differentiated after the protocols. Chondrogenic differentiation was evaluated in 2D and in 3D (pellet) cultures, which were further evaluated by optical microscopy and transmission electron microscopy (TEM). 2D cultures showed greater extracellular matrix deposition in NF1 DPSCs comparing with controls during chondrogenic differentiation. In semithin sections, control pellets hadhomogenous-sized intra and extracelullar matrix vesicles, whereas NF1 cultures had matrix vesicles of different sizes. TEM analysis showed higher amount of collagen fibers in NF1 cultures compared with control cultures. NF1 DPSCs presented increased extracellular matrix deposition during chondrogenic differentiation, which could be related to skeletal changes in individuals with NF1. The online version of this article (10.1186/s13023-018-0843-1) contains supplementary material, which is available to authorized users.
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