3D Porous Chitosan Scaffolds Suit Survival and Neural Differentiation of Dental Pulp Stem Cells
3D Porous Chitosan Scaffolds Suit Survival and Neural Differentiation of Dental Pulp Stem Cells
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DOI：
10.1007/s10571-014-0063-8
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发表时间：
2014-08-01
影响因子：
4
通讯作者：
Zhang, Xinhua
中科院分区：
文献类型：
--
作者：
Feng, Xingmei;Lu, Xiaohui;Zhang, Xinhua
细胞替代疗法在脑损伤治疗中的一个关键方面是构建合适的生物材料支架，使其能够有效地携带和运输治疗细胞到靶区。在本研究中，我们通过冷冻干燥法制备了小型三维多孔壳聚糖支架，并证明了这些支架在体外可以支持和促进牙髓干细胞向神经细胞的分化。DPSCs取自成人第三磨牙牙髓。在膨胀率接近4.33+/-A 10.92%时，扫描电子显微镜显示支架具有较高的孔隙率和孔的连通性。细胞计数Kit-8实验证实了壳聚糖支架的生物相容性，支持了DPSCs的生长和存活。RT-PCR、Western blotting和免疫荧光检测DPSCs的神经分化情况。我们发现支架贴壁的DPSCs中巢蛋白的高表达，诱导分化后Nestin的表达急剧下降。神经分子标志物微管相关蛋白2、胶质纤维酸性蛋白和2‘，3’-环核苷酸磷酸二酯酶的表达也增加。本研究表明，颗粒状3D壳聚糖支架无细胞毒性，生物相容性好，为DPSCs的附着、存活和神经分化提供了有利的微环境。这些支架具有巨大的潜力，可以促进未来脑损伤治疗的进展。
A key aspect of cell replacement therapy in brain injury treatment is construction of a suitable biomaterial scaffold that can effectively carry and transport the therapeutic cells to the target area. In the present study, we created small 3D porous chitosan scaffolds through freeze-drying, and showed that these can support and enhance the differentiation of dental pulp stem cells (DPSCs) to nerve cells in vitro. The DPSCs were collected from the dental pulp of adult human third molars. At a swelling rate of similar to 4.33 +/- A 10.92 %, the scaffold displayed high porosity and interconnectivity of pores, as revealed by SEM. Cell counting kit-8 assay established the biocompatibility of the chitosan scaffold, supporting the growth and survival of DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence. We found that the scaffold-attached DPSCs showed high expression of Nestin that decreased sharply following induction of differentiation. Exposure to the differentiation media also increased the expression of neural molecular markers Microtubule-associated protein 2, glial fibrillary acidic protein, and 2',3'-cyclic nucleotide phosphodiesterase. This study demonstrates that the granular 3D chitosan scaffolds are non-cytotoxic, biocompatible, and provide a conducive and favorable micro-environment for attachment, survival, and neural differentiation of DPSCs. These scaffolds have enormous potential to facilitate future advances in treatment of brain injury.