Li Sheng-Tien, Young Tai-Horng, Lin Chih-Feng, Huang Tsung-Wei
Instuitute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan.
Am J Rhinol Allergy. 2017 Sep 1;31(5):289-292. doi: 10.2500/ajra.2017.31.4456.
Olfactory dysfunction significantly influences patients' quality of life. Chitosan has been reported to support neuron and Schwann cell growth and even leads to orient axonal growth. However, researchers have yet to explore whether chitosan solution can promote differentiation of olfactory receptor neurons of the olfactory neuroepithelium and be used for treating olfactory dysfunction.
To evaluate the effect of chitosan solution on the differentiation of olfactory neuroepithelial cells.
Olfactory neuroepithelial cells were isolated from embryonic day 17 of Wistar rats and then cultured with and without soluble chitosan for 9 days. The concentration of chitosan solution was set at 0.1 mg/mL. The effects of treatment were assessed by immunocytochemistry and Western blot after culturing.
The morphologic analysis indicated that olfactory neuroepithelial cells treated with chitosan exhibited bipolar shape with asymmetric processes. In addition, from days 3 to 9, the expression level of βIII tubulin gradually reduced, but the expression level of olfactory marker protein significantly rose at day 9 in the chitosan groups (p < 0.05). Importantly, chitosan-treated olfactory neuroepithelial cells expressed more signal transduction apparatuses, olfactory neuron specific-G protein and adenylate cyclase 3, than those without chitosan treatment at day 9. Western blot analysis also further confirmed the results (p < 0.05).
Experimental results revealed that soluble chitosan promoted differentiation of olfactory neuroepithelial cells based on its role in olfactory receptor neuron differentiation, neurite outgrowth, and signal transduction apparatus expressions.
嗅觉功能障碍显著影响患者的生活质量。据报道，壳聚糖可支持神经元和雪旺细胞生长，甚至可引导轴突生长。然而，研究人员尚未探讨壳聚糖溶液是否能促进嗅神经上皮的嗅觉受体神经元分化并用于治疗嗅觉功能障碍。
评估壳聚糖溶液对嗅神经上皮细胞分化的影响。
从Wistar大鼠胚胎第17天分离嗅神经上皮细胞，然后分别在有无可溶性壳聚糖的条件下培养9天。壳聚糖溶液浓度设定为0.1mg/mL。培养后通过免疫细胞化学和蛋白质免疫印迹法评估处理效果。
形态学分析表明，用壳聚糖处理的嗅神经上皮细胞呈现出具有不对称突起的双极形状。此外，在第3至9天，壳聚糖组中βIII微管蛋白的表达水平逐渐降低，但嗅觉标记蛋白的表达水平在第9天显著升高（p<0.05）。重要的是，在第9天，经壳聚糖处理的嗅神经上皮细胞比未用壳聚糖处理的细胞表达更多的信号转导装置、嗅觉神经元特异性G蛋白和腺苷酸环化酶3。蛋白质免疫印迹分析也进一步证实了该结果（p<0.05）。
实验结果表明，可溶性壳聚糖基于其在嗅觉受体神经元分化、神经突生长和信号转导装置表达方面的作用，促进了嗅神经上皮细胞的分化。
