Electrospun regenerated silk fibroin is a promising biomaterial for the maintenance of inner ear progenitors in vitro
Electrospun regenerated silk fibroin is a promising biomaterial for the maintenance of inner ear progenitors in vitro
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DOI：
10.1177/08853282211051501
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发表时间：
2021-10-28
影响因子：
2.9
通讯作者：
Sun, Shan
中科院分区：
文献类型：
--
作者：
Li, Guangfei;Yin, Yanbo;Sun, Shan
目的探讨电纺再生丝素蛋白(RSF)与内耳祖细胞的生物相容性，特别是其对内耳祖细胞向毛细胞分化的影响。方法收集20只新生小鼠耳蜗组织，连续数天消化后形成球体。然后将从球体中消化的细胞接种到排列的或随机的RSF垫子上，并以涂有层粘连蛋白的盖片作为对照。用原位末端标记法检测内耳祖细胞的死亡率，用扫描电子显微镜检测细胞在RSF垫或盖片上的粘附力。最后，肌球蛋白7a的表达决定了从内耳祖细胞分化的毛细胞的数量。本研究采用非配对学生t检验和单因素方差分析后的Dunnett多重比较检验(p<；0.05)。结果培养5d后，内耳祖细胞在排列的和随机排列的RSF垫上均有良好的粘附性，TUNEL+细胞数与盖片相比差异无统计学意义(p>；0.05)。体外分化培养7天后，对照、排列和随机RSF垫上分化的毛细胞百分率分别为2.5+/-0.5%、2.7+/-0.4%和2.4+/-0.2%，而Myosin7a(+)细胞在任何一种RSF垫上的分化毛细胞与对照组相比均无显著差异(p>；0.05)。结论排列整齐、随机排列的RSF垫片与内耳祖细胞具有良好的生物相容性，有助于维持内耳祖细胞的骨干性。因此，我们的结果表明，RSF垫片为开发内耳组织工程研究的新策略提供了有用的支架。
Objective We sought to determine the biocompatibility of electrospun regenerated silk fibroin (RSF) mats with inner ear progenitors, especially their effect on the differentiation of inner ear progenitors into hair cells. Methods Neonatal mouse cochleae (n = 20) were collected and digested and allowed to form spheres over several days. Cells digested from the spheres were then seeded onto aligned or random RSF mats, with laminin-coated coverslips serving as controls. The inner ear progenitor cell mortality was examined by TUNEL labeling, and the adhesion of cells to the RSF mats or coverslip was determined by scanning electron microscopy. Finally, the number of hair cells that differentiated from inner ear progenitors was determined by Myosin7a expression. Unpaired Student's t-tests and one-way ANOVA followed by a Dunnett's multiple comparisons test were used in this study (p < 0.05). Results After 5 days of culture, the inner ear progenitors had good adhesion to both the aligned and random RSF mats and there was no significant difference in TUNEL+ cells between the mats compared to the coverslip (p > 0.05). After 7 days of in vitro differentiation culture, the percentage of differentiated hair cells on the control, aligned, and random RSF mats was 2.5 +/- 0.5%, 2.7 +/- 0.4%, and 2.4 +/- 0.2%, respectively, and there was no significant difference between Myosin7a(+) cells on either RSF mat compared to controls (p > 0.05). Conclusion The aligned and random RSF mats had excellent biocompatibility with inner ear progenitors and helped the inner ear progenitors maintain their stemness. Our results thus indicate that RSF mats represent a useful scaffold for the development of new strategies for inner ear tissue engineering research.