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去细胞化的心包膜生物支架的成分浸提及其对巨噬细胞反应的影响。

The component leaching from decellularized pericardial bioscaffolds and its implication in the macrophage response.

机构信息

Departamento de Ingenierías Química, Electrónica y Biomédica, DCI Universidad de Guanajuato, León, 37150, GTO, Mexico.

Departamento de Biología, DCNE, Universidad de Guanajuato, Guanajuato, 36050, GTO, Mexico.

出版信息

J Biomed Mater Res A. 2016 Nov;104(11):2810-22. doi: 10.1002/jbm.a.35825. Epub 2016 Jul 18.

Abstract

The extracellular matrix molecules remaining in bioscaffolds derived from decellularized xenogeneic tissues appear to be important for inducing cell functions conducting tissue regeneration. Here, we studied whether decellularization methods, that is, detergent Triton X-100 (TX) alone and TX combined with reversible alkaline swelling (STX), applied to bovine pericardial tissue, could affect the bioscaffold components. The in vitro macrophage response, subdermal biodegradation, and cell infiltration were also studied. The results indicate a lower leaching of fibronectin, but a higher leaching of laminin and sulfated glycosaminoglycans from tissues decellularized with STX and TX, respectively. The in vitro secretion of interleukin-6 and monocyte chemoattractant protein by RAW264.7 macrophages is promoted by decellularized bioscaffold leachates. A lower polymorphonuclear cell density is observed around decellularized bioscaffolds at 1-day implantation; concurrently showing a higher cell infiltration in STX- than in TX-implant. Cells infiltrated into TX-implant show a fibroblastic morphology at 7-day implantation, concurrently the capillary formation is observed at 14-day. Pericardial bioscaffolds suffer biodegradation more pronounced in STX- than in TX-implant. Both TX and STX decellularization methods favor a high leaching of basal lamina components, which presumably promotes a faster macrophage stimulation compared to nondecellularized tissue, and appear to be associated with an increased host cell infiltration in a rat subdermal implantation. Meanwhile, the connective tissue components leaching from TX decellularized bioscaffolds, unlike the STX ones, appear to be associated with an enhanced angiogenesis accompanied by an early-promoted fibroblastic cell transition. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2810-2822, 2016.

摘要

细胞外基质分子在脱细胞异种组织衍生的生物支架中残留,对于诱导细胞功能、促进组织再生似乎很重要。在这里,我们研究了脱细胞方法,即单独使用去污剂 Triton X-100(TX)和 TX 与可逆碱性膨胀(STX)联合应用于牛心包组织,是否会影响生物支架成分。还研究了细胞外巨嗜细胞的反应、皮下生物降解和细胞浸润。结果表明,STX 和 TX 处理的组织脱细胞后,纤维连接蛋白的浸出率较低,但层粘连蛋白和硫酸化糖胺聚糖的浸出率较高。RAW264.7 巨噬细胞的白细胞介素-6 和单核细胞趋化蛋白-1 的体外分泌受到脱细胞生物支架浸出液的促进。在植入 1 天时,脱细胞生物支架周围观察到较少的多形核细胞密度;同时,在 STX 植入物中观察到更高的细胞浸润。在 7 天植入时,细胞浸润到 TX 植入物中呈现出成纤维细胞形态,同时在 14 天观察到毛细血管形成。在 STX 植入物中,心包生物支架的生物降解比 TX 植入物更明显。TX 和 STX 脱细胞方法都有利于基膜成分的高浸出,这可能与非脱细胞组织相比,刺激巨噬细胞的速度更快,并与大鼠皮下植入物中宿主细胞的浸润增加有关。同时,从 TX 脱细胞生物支架中浸出的结缔组织成分,与 STX 脱细胞生物支架不同,似乎与增强的血管生成有关,同时伴随着早期促进的成纤维细胞转化。© 2016 Wiley Periodicals, Inc. J 生物材料 Res 部分 A:104A:2810-2822,2016。

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