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牙髓干细胞体外向内耳毛细胞分化能力的研究:治疗感音神经性聋的初步尝试。

Differentiation capacity of dental pulp stem cell into inner ear hair cell using an in vitro assay: a preliminary step toward treating sensorineural hearing loss.

机构信息

Department of Otorhinolaryngology-Head and Neck Surgery, Faculty of Medicine, Universitas Sumatera Utara, Jl. Dr. Mansyur No 5, Medan, Sumatera Utara, 20155, Indonesia.

Division of Oral Biology, Departement of Biochemistry and Molecular Biology, Faculty of Dentistry, Trisakti University, Jakarta, Indonesia.

出版信息

Eur Arch Otorhinolaryngol. 2022 Apr;279(4):1805-1812. doi: 10.1007/s00405-021-06864-9. Epub 2021 May 19.

Abstract

PURPOSE

Sensorineural hearing loss (SNHL) is commonly caused by the death or dysfunction of cochlear cell types as a result of their lack of regenerative capacity. However, regenerative medicine, such as stem cell therapy, has become a promising tool to cure many diseases, including hearing loss. In this study, we determined whether DPSCs could differentiate into cochlear hair cell in vitro.

METHODS

DPSCs derived from human third molar dental pulp were induced into NSCs using a medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) for 7 days, and then into cochlear hair cell using a medium containing EGF and IGF-1 for the next 14 days. We used the neuroepithelial protein marker nestin and cochlear hair cell marker myosin VIIa as the markers for cells differentiation. Cells expressing the positive markers under the microscope were confirmed to have differentiated into cochlear hair cell.

RESULTS

DPSCs were successfully induced to differentiate into NSCs, with mean 24% nestin-positive cells. We found that DPSC-derived NSCs have a great capacity in differentiating into inner ear hair cell-like cells with an average of 81% cells presenting myosin VIIa. Thus, DPSCs have high potential to serve as a good resource for SNHL treatment.

CONCLUSION

We found the high potential of DPSCs to differentiate into NSC. The ability of DPSCs in differentiating into neural lineage cell made them a good candidate for regenerative therapy in neural diseases, such as SNHL.

摘要

目的

感音神经性听力损失(SNHL)通常是由于耳蜗细胞类型因缺乏再生能力而死亡或功能障碍引起的。然而,再生医学,如干细胞疗法,已成为治愈许多疾病的有前途的工具,包括听力损失。在这项研究中,我们确定了 DPSCs 是否可以在体外分化为耳蜗毛细胞。

方法

从人第三磨牙牙髓中分离出 DPSCs,使用含有碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)的培养基将其诱导为 NSCs,持续 7 天,然后使用含有 EGF 和 IGF-1 的培养基将其诱导为耳蜗毛细胞,持续 14 天。我们使用神经上皮蛋白标志物巢蛋白和耳蜗毛细胞标志物肌球蛋白 VIIa 作为细胞分化的标志物。在显微镜下观察到表达阳性标志物的细胞被确认为已分化为耳蜗毛细胞。

结果

DPSCs 成功地被诱导分化为 NSCs,平均有 24%的巢蛋白阳性细胞。我们发现 DPSC 衍生的 NSCs 具有很强的分化为内耳毛细胞样细胞的能力,平均有 81%的细胞表达肌球蛋白 VIIa。因此,DPSCs 具有作为 SNHL 治疗良好资源的巨大潜力。

结论

我们发现 DPSCs 分化为 NSC 的潜力很大。DPSCs 分化为神经谱系细胞的能力使它们成为 SNHL 等神经疾病再生治疗的良好候选者。

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