Functionalized carbon nanotubes as suitable scaffold materials for proliferation and differentiation of canine mesenchymal stem cells
Functionalized carbon nanotubes as suitable scaffold materials for proliferation and differentiation of canine mesenchymal stem cells
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DOI：
10.2147/ijn.s122945
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发表时间：
2017-01-01
影响因子：
8
通讯作者：
Bag, Sadhan
中科院分区：
文献类型：
--
作者：
Das, Kinsuk;Madhusoodan, A. P.;Bag, Sadhan
在再生医学领域，可以设想间充质干细胞（MSC）的许多潜在应用，因为它们能够根据其生长的基质分化成一系列组织。随着纳米技术的发展，碳纳米管作为细胞培养基质在组织工程中的应用越来越广泛。在这项研究中，犬骨髓来源的MSC被认为是体外研究的细胞模型，以阐明集体的细胞过程，使用三种不同种类的功能化碳纳米管薄膜（COOH-单壁碳纳米管[SWCNTs]，COOH-多壁碳纳米管[MWCNTs]和聚乙二醇[PEG]-SWCNTs），将其喷雾干燥到预热的盖玻片上。细胞在碳纳米管膜上更好地展开，导致更高的细胞表面积和丝状伪足的发生，应力纤维束具有平行取向。与对照相比，犬MSC在所有类型的CNT基底上以较慢的速率增殖，但在研究期间没有注意到细胞数量下降。随着时间的推移，CNT基底上的骨坏死相关基因的表达降低。在膜联蛋白V-异硫氰酸荧光素/碘化丙啶（PI）染色后的流式细胞术上，与PEG官能化的膜相比，COOH官能化的膜中凋亡和坏死细胞的总数保持较低。总的来说，这些结果表明COOH-MWCNT基材提供了低细胞毒性的环境。通过在特定分化条件下培养，进一步诱导犬MSC沿成骨、成软骨和神经元谱系分化沿着。细胞化学和免疫细胞化学染色结果，以及骨标记基因的表达，使我们假设COOH-MWCNT底物作为一个更好的线索，加速成骨分化过程。然而，虽然软骨形成促进COOH-SWCNT，神经元分化促进COOH-SWNCT和COOH-MWCNT。总之，这些发现表明，COOH功能化的碳纳米管代表了一个有前途的支架组件，用于未来利用犬骨髓间充质干细胞在再生医学中的选择性分化。
In the field of regenerative medicine, numerous potential applications of mesenchymal stem cells (MSCs) can be envisaged, due to their ability to differentiate into a range of tissues on the basis of the substrate on which they grow. With the advances in nanotechnology, carbon nanotubes (CNTs) have been widely explored for use as cell culture substrate in tissue engineering applications. In this study, canine bone marrow-derived MSCs were considered as the cellular model for an in vitro study to elucidate the collective cellular processes, using three different varieties of thin films of functionalized carbon nanotubes (COOH-single-walled CNTs [SWCNTs], COOH-multiwalled CNTs [MWCNTs] and polyethylene glycol [PEG]-SWCNTs), which were spray dried onto preheated cover slips. Cells spread out better on the CNT films, resulting in higher cell surface area and occurrence of filopodia, with parallel orientation of stress fiber bundles. Canine MSCs proliferated at a slower rate on all types of CNT substrates compared to the control, but no decline in cell number was noticed during the study period. Expression of apoptosis-associated genes decreased on the CNT substrates as time progressed. On flow cytometry after AnnexinV-fluorescein isothiocyanate/propidium iodide (PI) staining, total number of apoptotic and necrotic cells remained lower in COOH-functionalized films compared to PEG-functionalized ones. Collectively, these results indicate that COOH-MWCNT substrate provided an environment of low cytotoxicity. Canine MSCs were further induced to differentiate along osteogenic, chondrogenic, and neuronal lineages by culturing under specific differentiation conditions. The cytochemical and immunocytochemical staining results, as well as the expression of the bone marker genes, led us to hypothesize that the COOH-MWCNT substrate acted as a better cue, accelerating the osteogenic differentiation process. However, while chondrogenesis was promoted by COOH-SWCNT, neuronal differentiation was promoted by both COOH-SWNCT and COOH-MWCNT. Taken together, these findings suggest that COOH-functionalized CNTs represent a promising scaffold component for future utilization in the selective differentiation of canine MSCs in regenerative medicine.