Hearing restoration in a deaf animal model with transplantation of adipose-derived mesenchymal stem cells from guinea pigs

doi:10.3969/j.issn.2095-4344.2014.32.015

Abstract

Abstract:

BACKGROUND: Can adipose-derived mesenchymal stem cells be used to treat sensorineural deafness caused by hair cell degeneration and loss?
OBJECTIVE: To investigate the effect of adipose-derived mesenchymal stem cells from guinea pigs transplanted via the scala tympani on the recovery from sensorineural hearing in an animal model.
METHODS: Intraperitoneal injection of gentamicin was performed to prepare sensorineural deafness models in guinea pigs. Then, adipose-derived mesenchymal stem cells from guinea pig were transplanted via the scala tympani. After 1 and 3 weeks, auditory brainstem responses were detected to observe the hearing variation after adipose-derived mesenchymal stem cell transplantation. Meanwhile, the migration and distribution of EDU-labeled adipose-derived mesenchymal stem cells in the cochlea were traced.
RESULTS AND CONCLUSION: After 1 and 3 weeks of implantation, the results from auditory brainstem response showed that the hearing was gradually improved. At 1 week of implantation, most of the cells were distributed in the perilymph and some cells migrated to the Corti organ and attached to the basement membrane. At 3 weeks of implantation, cells migrated and attached to the basement membrane of Corti organ, furthermore, some cells migrated to the cochlear nerve. The longer the time of implantation, the less cell survived. The results indicate that adipose-derived mesenchymal stem cells from guinea pig that are transplanted via the scala tympani can directly migrate and survive ultimately to improve the hearing.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: adipose tissue, mesenchymal stem cells, hearing loss, sensorineural, cochlea

CLC Number:

R394.2

Wang Xiao-yan, Ji Bin, Li Bing-bing, Bi Xiao-juan, Liu Li-zhong. Hearing restoration in a deaf animal model with transplantation of adipose-derived mesenchymal stem cells from guinea pigs[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(32): 5173-5177.

Figures/Tables 2

2.1 豚鼠感音神经性耳聋模型建立情况豚鼠通过腹腔连续注射庆大霉素进行造模，造模过程豚鼠体质量都有不同程度减轻，近1/5豚鼠出现恶心，食欲减退，毛发脱落，耳郭反射消失等症状。其中感音神经性耳聋组死亡2只，因肠胀气和腹泻死亡。感音神经性耳聋植入脂肪间充质干细胞组无死亡。感音神经性耳聋植入生理盐水组死亡2只，1只因前庭功能障碍反应拒食死亡，另1只因右眼感染引起颅内感染，抗生素治疗无效死亡。 2.2 豚鼠脂肪间充质干细胞的形态原代培养一两天时开始贴壁生长，呈梭形、三角形等，四五天为对数生长期，细胞生长迅速，7-9 d细胞汇合达80%-90%，呈旋涡状生长，传至第3代脂肪间充质干细胞较纯，形态及活力良好，可用于细胞植入，见图1。 2.3 EDU标记的脂肪间充质干细胞第3代脂肪间充质干细胞培养至第4天时加EDU标记细胞，48 h后荧光显色，倒置荧光显微镜下观察EDU标记细胞为红色，核染标记为蓝色，细胞标记率> 95%，可用于标记后细胞植入，见图1。 2.4 脂肪间充质干细胞移植前后听力检测结果为了评估豚鼠脂肪间充质干细胞耳蜗植入后对耳聋动物听力的影响，于细胞植入后1周和3周进行听性脑干诱发反应检测。刺激强度从100 dB到10 dB，记录最大、最稳和最后消失的主波Ⅲ波波形及阈值。听力正常的豚鼠可测到稳定的Ⅰ波至Ⅳ波，重复后波形吻合，不是因干扰所造成，听力在30 dB左右，见图2A。感应神经性耳聋动物听性脑干诱发反应检测时90 dB仍未引出说明耳聋模型造模成功，见图2B。耳聋动物植入豚鼠脂肪间充质干细胞术后1周听性脑干诱发反应检测结果显示听力提高约20 dB，Ⅲ波波形稳定，见图2C。3周再次对植入后的动物进行检测，听性脑干诱发反应阈值显示听力较前明显提高，较治疗前听力提高约40 dB，见图2D。 2.5 EDU追踪移植细胞在耳蜗内的分布对耳聋动物植入生理盐水及植入豚鼠脂肪间充质干细胞1，3周进行耳蜗组织冰冻切片EDU荧光追踪，生理盐水组可见细胞核染蓝色，鼓阶及前庭阶结构完整，从底回到顶回，柯蒂器结构完整，未见细胞及组织破坏现象，说明通过鼓阶微孔技术植入对耳蜗的结构无明显影响，见图3A。植入后1周结果显示EDU标记细胞大部分在外淋巴液中，已有一部分迁移至耳蜗柯蒂器贴附于基底膜上，说明移植的细胞能够迁移到受损毛细胞所在位置，而且能够存活替代受损毛细胞发挥作用，见图3B。植入后3周结果显示，大部分位于外淋巴中未贴附的细胞已死亡，贴服存活的细胞能够定向归巢，且部分细胞出现向蜗神经迁移的趋向，说明植入的细胞不仅对受损毛细胞有一定的替代作用，而且有可能对神经修复也具有一定的作用，见图3C。"

[1]	Dang Yi, Du Chengyan, Yao Honglin, Yuan Nenghua, Cao Jin, Xiong Shan, Zhang Dingmei, Wang Xin. Hormonal osteonecrosis and oxidative stress [J]. Chinese Journal of Tissue Engineering Research, 2023, 27(9): 1469-1476.
[2]	Li Yujiao, Su Kunxia. High-intensity endurance exercise influences browning of white adipose tissue in a mouse model of high-fat diet induced obesity [J]. Chinese Journal of Tissue Engineering Research, 2023, 27(5): 707-713.
[3]	Li Rui, Liu Zhen, Guo Zige, Lu Ruijie, Wang Chen. Aspirin-loaded chitosan nanoparticles and polydopamine modified titanium sheets improve osteogenic differentiation [J]. Chinese Journal of Tissue Engineering Research, 2023, 27(3): 374-379.
[4]	Su Meng, Wang Xin, Zhang Jin, Bei Ying, Huang Yu, Zhu Yanzhao, Li Jiali, Wu Yan. Nanocellular vesicles loaded with curcumin promote wound healing in diabetic mice [J]. Chinese Journal of Tissue Engineering Research, 2023, 27(12): 1877-1883.
[5]	Ma Munan, Xie Jun, Sang Yuchao, Huang Lei, Zhang Guodong, Yang Xiaoli, Fu Songtao. Electroacupuncture combined with bone marrow mesenchymal stem cells in the treatment of chemotherapy-induced premature ovarian insufficiency in rats [J]. Chinese Journal of Tissue Engineering Research, 2023, 27(1): 1-7.
[6]	Liu Wentao, Feng Xingchao, Yang Yi, Bai Shengbin. Effect of M2 macrophage-derived exosomes on osteogenic differentiation of bone marrow mesenchymal stem cells [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(在线): 1-6.
[7]	Wang Jing, Xiong Shan, Cao Jin, Feng Linwei, Wang Xin. Role and mechanism of interleukin-3 in bone metabolism [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(8): 1260-1265.
[8]	Xiao Hao, Liu Jing, Zhou Jun. Research progress of pulsed electromagnetic field in the treatment of postmenopausal osteoporosis [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(8): 1266-1271.
[9]	Tian Chuan, Zhu Xiangqing, Yang Zailing, Yan Donghai, Li Ye, Wang Yanying, Yang Yukun, He Jie, Lü Guanke, Cai Xuemin, Shu Liping, He Zhixu, Pan Xinghua. Bone marrow mesenchymal stem cells regulate ovarian aging in macaques [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 985-991.
[10]	Hou Jingying, Guo Tianzhu, Yu Menglei, Long Huibao, Wu Hao. Hypoxia preconditioning targets and downregulates miR-195 and promotes bone marrow mesenchymal stem cell survival and pro-angiogenic potential by activating MALAT1 [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1005-1011.
[11]	Liang Xuezhen, Yang Xi, Li Jiacheng, Luo Di, Xu Bo, Li Gang. Bushen Huoxue capsule regulates osteogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells via Hedgehog signaling pathway [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1020-1026.
[12]	Wen Dandan, Li Qiang, Shen Caiqi, Ji Zhe, Jin Peisheng. Nocardia rubra cell wall skeleton for extemal use improves the viability of adipogenic mesenchymal stem cells and promotes diabetes wound repair [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1038-1044.
[13]	Zhu Bingbing, Deng Jianghua, Chen Jingjing, Mu Xiaoling. Interleukin-8 receptor enhances the migration and adhesion of umbilical cord mesenchymal stem cells to injured endothelium [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1045-1050.
[14]	Fang Xiaolei, Leng Jun, Zhang Chen, Liu Huimin, Guo Wen. Systematic evaluation of different therapeutic effects of mesenchymal stem cell transplantation in the treatment of ischemic stroke [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1085-1092.
[15]	Guo Jia, Ding Qionghua, Liu Ze, Lü Siyi, Zhou Quancheng, Gao Yuhua, Bai Chunyu. Biological characteristics and immunoregulation of exosomes derived from mesenchymal stem cells [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1093-1101.