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人牙髓干细胞移植对患有感音神经性听力损失的斯普拉格-道利大鼠的疗效评估。

Evaluation of the Efficacy of Human Dental Pulp Stem Cell Transplantation in Sprague-Dawley Rats with Sensorial Neural Hearing Loss.

作者信息

Rawiwet Visut, Vijitruth Rattanavijit, Thonabulsombat Chareonsri, Vongsavan Kutkao, Sritanaudomchai Hathaitip

机构信息

Central Animal Facility, Faculty of Science, Mahidol University (MUSC-CAF), Bangkok, Thailand.

Department of Biology, Faculty of Science, Mahidol University, Bangkok, Thailand.

出版信息

Eur J Dent. 2023 Oct;17(4):1207-1214. doi: 10.1055/s-0043-1761190. Epub 2023 Jan 30.

Abstract

OBJECTIVES

The purpose of the present study was to evaluate the efficacy of spiral ganglion neuron (SGN) regeneration after dental pulp stem cell (DPSC) transplantation in a rat sensorineural hearing loss (HL) model.

MATERIALS AND METHODS

Sham or experimental HL was induced in adult Sprague-Dawley rats by cochlear round window surgery. An HL rat model was established with a single 10 mM ouabain intratympanic injection. After 7 days, the rats received DPSCs, stem cells from human exfoliated deciduous teeth (SHED), or culture medium in the sutural area to establish four groups: sham, HL-DPSC, HL-SHED, and HL-medium. Histological analyses were performed at 4, 7, and 10 weeks after transplantation, and the number of SGNs, specific SGN protein expression, and the function of SGNs were evaluated.

STATISTICAL ANALYSIS

Data were statistically by MS Excel and SPSS v.15.0. Intergroup level of significance was determined via a one-way analysis of variance and Duncan's multiple range test with 95% confidence intervals.

RESULTS

New SGN formation was observed in the HL-DPSC and HL-SHED rat groups. The number of SGNs was significantly higher in the HL-DPSC and HL-SHED groups than in the HL-medium group over 4 to 10-week survival period. HL-DPSC rats exhibited higher SGN density compared with that in HL-SHED group, which was statistically significant at week 10. The regenerated SGNs expressed cochlear wiring regulator GATA-binding-protein 3. Moreover, the SGNs from the HL-DPSC group also exhibited a higher expression of synaptic vesicle protein and regulated action potential-dependent neurotransmitter release compared with SGNs from the HL-SHED group.

CONCLUSIONS

Our findings suggest that DPSCs and SHED repair and regenerate SGNs in rat HL model. Dental pulp stem cells represent a promising treatment strategy for restoring damage to the sensory circuits associated with deafness.

摘要

目的

本研究旨在评估在大鼠感音神经性听力损失(HL)模型中,牙髓干细胞(DPSC)移植后螺旋神经节神经元(SGN)的再生效果。

材料与方法

通过耳蜗圆窗手术在成年Sprague-Dawley大鼠中诱导假手术或实验性HL。用单次10 mM哇巴因鼓膜内注射建立HL大鼠模型。7天后,大鼠在缝合区域接受DPSC、人脱落乳牙干细胞(SHED)或培养基,分为四组:假手术组、HL-DPSC组、HL-SHED组和HL-培养基组。在移植后4、7和10周进行组织学分析,评估SGN的数量、特定SGN蛋白表达以及SGN的功能。

统计分析

数据采用MS Excel和SPSS v.15.0进行统计。组间显著性水平通过单因素方差分析和Duncan多重范围检验确定,置信区间为95%。

结果

在HL-DPSC和HL-SHED大鼠组中观察到新的SGN形成。在4至10周的生存期内,HL-DPSC组和HL-SHED组的SGN数量显著高于HL-培养基组。与HL-SHED组相比,HL-DPSC大鼠的SGN密度更高,在第10周具有统计学意义。再生的SGN表达耳蜗布线调节因子GATA结合蛋白3。此外,与HL-SHED组的SGN相比,HL-DPSC组的SGN还表现出更高的突触小泡蛋白表达,并调节动作电位依赖性神经递质释放。

结论

我们的研究结果表明,DPSC和SHED可在大鼠HL模型中修复和再生SGN。牙髓干细胞是恢复与耳聋相关的感觉回路损伤的一种有前景的治疗策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/232b/10756831/8ede7e3a249e/10-1055-s-0043-1761190-i2282340-1.jpg

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