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应用合成自组装肽纳米纤维支架(PuraMatrix)原位组织工程技术促进大鼠中耳黏膜再生。

In situ tissue engineering with synthetic self-assembling peptide nanofiber scaffolds, PuraMatrix, for mucosal regeneration in the rat middle-ear.

机构信息

Department of Otolaryngology-Head and Neck Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.

出版信息

Int J Nanomedicine. 2013;8:2629-40. doi: 10.2147/IJN.S47279. Epub 2013 Jul 24.

Abstract

Middle-ear mucosa maintains middle-ear pressure. However, the majority of surgical cases exhibit inadequate middle-ear mucosal regeneration, and mucosal transplantation is necessary in such cases. The aim of the present study was to assess the feasibility of transplantation of isolated mucosal cells encapsulated within synthetic self-assembling peptide nanofiber scaffolds using PuraMatrix, which has been successfully used as scaffolding in tissue engineering, for the repair of damaged middle-ear. Middle-ear bullae with mucosa were removed from Sprague Dawley (SD) transgenic rats, transfected with enhanced green fluorescent protein (EGFP) transgene and excised into small pieces, then cultured up to the third passage. After surgical elimination of middle-ear mucosa in SD recipient rats, donor cells were encapsulated within PuraMatrix and transplanted into these immunosuppressed rats. Primary cultured cells were positive for pancytokeratin but not for vimentin, and retained the character of middle-ear epithelial cells. A high proportion of EGFP-expressing cells were found in the recipient middle-ear after transplantation with PuraMatrix, but not without PuraMatrix. These cells retained normal morphology and function, as confirmed by histological examination, immunohistochemistry, and electron microscopy, and multiplied to form new epithelial and subepithelial layers together with basement membrane. The present study demonstrated the feasibility of transplantation of cultured middle-ear mucosal epithelial cells encapsulated within PuraMatrix for regeneration of surgically eliminated mucosa of the middle-ear in SD rats.

摘要

中耳黏膜维持中耳压力。然而,大多数手术病例显示中耳黏膜再生不足,需要进行黏膜移植。本研究旨在评估使用 PuraMatrix 移植包被在合成自组装肽纳米纤维支架内的分离黏膜细胞的可行性,PuraMatrix 已成功用于组织工程中的支架,用于修复受损的中耳。从 Sprague Dawley(SD)转基因大鼠中取出带有黏膜的中耳鼓室,转染增强型绿色荧光蛋白(EGFP)转基因并切成小块,然后培养至第三代。在 SD 受体大鼠的中耳黏膜被手术消除后,将供体细胞包被在 PuraMatrix 内并移植到这些免疫抑制大鼠中。原代培养细胞对泛细胞角蛋白呈阳性,但对波形蛋白呈阴性,并且保留了中耳上皮细胞的特征。在移植 PuraMatrix 后,受体中耳中有很高比例的表达 EGFP 的细胞,但没有 PuraMatrix 则没有。这些细胞保持正常形态和功能,通过组织学检查、免疫组织化学和电子显微镜证实,并增殖形成新的上皮和上皮下层以及基底膜。本研究证明了在 PuraMatrix 内包被培养的中耳黏膜上皮细胞移植用于再生 SD 大鼠中耳手术切除的黏膜的可行性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8e86/3728305/53fc3732dc06/ijn-8-2629Fig1.jpg

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