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首页> 外文期刊>The Journal of heart valve disease >Decellularization of bovine pericardium for tissue-engineering by targeted removal of xenoantigens.
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Decellularization of bovine pericardium for tissue-engineering by targeted removal of xenoantigens.

机译：牛心包的脱细胞用于组织工程化，通过有针对性地清除异种抗原。

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BACKGROUND AND AIM OF STUDY: Assessment of decellularization of xenogeneic biological scaffolds for tissue engineering has relied primarily on histological cellularity, though this may not ensure the removal of known xenogeneic antigens such as galactose-alpha1,3-galactose (alpha-gal) and MHC I. METHODS: Bovine pericardium (BP) underwent standard (Std) decellularization consisting of hypotonic lysis and treatment with DNAse/RNAse. In addition to Std decellularization, tissues were treated for 24 h with either 0.5% Triton X-100, 0.5% sodium deoxycholate (SD), 0.1% sodium dodecyl sulfate (SDS), alpha-galactosidase (5 U/ml) or phospholipase (PL) A2 (150 U/ml). Tissues underwent a 96-h washout under gentle agitation at 27 degrees C, and then evaluated by light microscopy for % cellularity, and by immunohistochemistry and Western blot for alpha-gal, bovine MHC I and smooth muscle alpha-actin. RESULTS: Standard treatment of BP resulted in only partial removal histological cellularity and persistence of alpha-gal, MHC I and alpha-actin. Adding SD treatment resulted in apparent acellularity, but persistence of xenogeneic antigens. Only the addition of SDS resulted in complete histological acellularity and removal of xenogeneic antigens. Treatment with alpha-galactosidase selectively removed alpha-gal from BP. CONCLUSION: Histological cellularity is not an adequate end-point for assuring removal of antigenicity from xenogeneic biological scaffolds. However, known xenogeneic antigens can be targeted for removal by novel decellularization treatments such as alpha-galactosidase.
机译：研究背景和研究目的：用于组织工程的异种生物支架的脱细胞评估主要依赖于组织学细胞学，尽管这可能不能确保去除已知的异种抗原，例如半乳糖-α1,3-半乳糖（α-gal）和MHC I.方法：对牛心包（BP）进行标准（Std）脱细胞，包括低渗裂解和DNAse / RNAse处理。除Std脱细胞外，还用0.5％Triton X-100、0.5％脱氧胆酸钠（SD），0.1％十二烷基硫酸钠（SDS），α-半乳糖苷酶（5 U / ml）或磷脂酶（24％）处理组织24小时。 PL）A2（150 U / ml）。在27摄氏度的轻微搅拌下，对组织进行96小时冲洗，然后通过光学显微镜评估细胞百分率，并通过免疫组织化学和Western blot评估α-gal，牛MHC I和平滑肌α-肌动蛋白。结果：BP的标准治疗仅导致部分去除组织学细胞性和α-gal，MHC I和α-肌动蛋白的持久性。添加SD治疗导致明显的脱细胞，但异种抗原的持久性。仅添加SDS导致完全的组织学脱细胞性和异种抗原的去除。用α-半乳糖苷酶处理选择性地从BP中去除了α-gal。结论：组织细胞学并不是确保从异种生物支架中去除抗原性的适当终点。但是，已知的异种抗原可以通过新型脱细胞处理（例如α-半乳糖苷酶）靶向去除。

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