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用于声带增强新型填充材料的人鼻中隔软骨脱细胞处理

Decellularization of Human Nasal Septal Cartilage for the Novel Filler Material of Vocal Fold Augmentation.

作者信息

Kang Dae-Woon, Shin Sung-Chan, Jang Jeon-Yeob, Park Hee-Young, Lee Jin-Choon, Wang Soo-Geun, Lee Byung-Joo

机构信息

Department of Otorhinolaryngology-Head and Neck Surgery, Pusan National University School of Medicine and Biomedical Research Institute, Pusan National University, Busan, Republic of Korea.

Department of Otorhinolaryngology-Head and Neck Surgery, Pusan National University School of Medicine and Biomedical Research Institute, Pusan National University Yangsan Hospital, Yangsan, Kyeongnam, Republic of Korea.

出版信息

J Voice. 2017 Jan;31(1):127.e1-127.e6. doi: 10.1016/j.jvoice.2015.12.013. Epub 2016 Feb 5.

Abstract

OBJECTIVES

The clinical application of allogenic and/or xenogenic cartilage for vocal fold augmentation requires to remove the antigenic cellular component. The objective of this study was to assess the effect of cartilage decellularization and determine the change in immunogenicity after detergent treatment in human nasal septal cartilage flakes made by the freezing and grinding method.

METHODS

Human nasal septal cartilages were obtained from surgical cases. The harvested cartilages were treated by the freezing and grinding technique. The obtained cartilage flakes were treated with 1% Triton X-100 or 2% sodium dodecyl sulfate (SDS) for decellularization of the cartilage flakes. Hematoxylin and eosin stain (H&E stain), surface electric microscopy, immunohistochemical stain for major histocompatibility complex I and II, and ELISA for DNA contents were performed to assess the effect of cartilage decellularization after detergent treatment.

RESULTS

A total of 10 nasal septal cartilages were obtained from surgical cases. After detergent treatment, the average size of the cartilage flakes was significantly decreased. With H&E staining, the cell nuclei of decellularized cartilage flakes were not observed. The expression of major histocompatibility complex (MHC)-I and II antigens was not identified in the decellularized cartilage flakes after treatment with detergent. DNA content was removed almost entirely from the decellularized cartilage flakes.

CONCLUSION

Treatment with 2% SDS or 1% Triton X-100 for 1 hour appears to be a promising method for decellularization of human nasal septal cartilage for vocal fold augmentation.

摘要

目的

同种异体和/或异种软骨用于声带增厚的临床应用需要去除抗原性细胞成分。本研究的目的是评估软骨脱细胞的效果,并确定通过冷冻研磨法制备的人鼻中隔软骨薄片经去污剂处理后免疫原性的变化。

方法

从手术病例中获取人鼻中隔软骨。收获的软骨采用冷冻研磨技术处理。将获得的软骨薄片用1% Triton X-100或2%十二烷基硫酸钠(SDS)处理以实现软骨薄片的脱细胞。进行苏木精和伊红染色(H&E染色)、表面电镜检查、主要组织相容性复合体I和II的免疫组化染色以及DNA含量的ELISA检测,以评估去污剂处理后软骨脱细胞的效果。

结果

共从手术病例中获取了10块鼻中隔软骨。去污剂处理后,软骨薄片的平均尺寸显著减小。经H&E染色,未观察到脱细胞软骨薄片的细胞核。用去污剂处理后的脱细胞软骨薄片中未检测到主要组织相容性复合体(MHC)-I和II抗原的表达。DNA含量几乎从脱细胞软骨薄片中完全去除。

结论

用2% SDS或1% Triton X-100处理1小时似乎是一种有前景的用于声带增厚的人鼻中隔软骨脱细胞方法。

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