Abstract：
BACKGROUND: The necessity to manufacture scaffolds with superior capabilities of biocompatibility and biodegradability has led to the production of extracellular matrix (ECM) scaffolds. Among their advantages, they allow better cell colonization, which enables its successful integration into the hosted tissue, surrounding the area to be repaired and their formulations facilitate placing it into irregular shapes. The ECM from porcine urinary bladder (pUBM) comprises proteins, proteoglycans and glycosaminoglycans which provide support and enable signals to the cells. These properties make it an excellent option to produce hydrogels that can be used in regenerative medicine.
OBJECTIVE: The goal of this study was to assess the biocompatibility of an ECM hydrogel derived from the porcine urinary bladder (pUBMh) in vitro using fibroblasts, macrophages, and adipose-derived mesenchymal stem cells (AD-MCSs), as well as biocompatibility in vivo using Wistar rats.
METHODS: Effects upon cells proliferation/viability was measured using MTT assay, cytotoxic effects were analyzed by quantifying lactate dehydrogenase release and the Live/Dead Cell Imaging assay. Macrophage activation was assessed by quantification of IL-6, IL-10, IL-12p70, MCP-1, and TNF-α using a microsphere-based cytometric bead array. For in vivo analysis, Wistar rats were inoculated into the dorsal sub-dermis with pUBMh. The specimens were sacrificed at 24 h after inoculation for histological study.
RESULTS: The pUBMh obtained showed good consistency and absence of cell debris. The biocompatibility tests in vitro revealed that the pUBMh promoted cell proliferation and it is not cytotoxic on the three tested cell lines and induces the production of pro-inflammatory cytokines on macrophages, mainly TNF-α and MCP-1. In vivo, pUBMh exhibited fibroblast-like cell recruitment, without tissue damage or inflammation.
CONCLUSIONS: The results show that pUBMh allows cell proliferation without cytotoxic effects and can be considered an excellent biomaterial for tissue engineering.
OBJECTIVE: The goal of this study was to assess the biocompatibility of an ECM hydrogel derived from the porcine urinary bladder (pUBMh) in vitro using fibroblasts, macrophages, and adipose-derived mesenchymal stem cells (AD-MCSs), as well as biocompatibility in vivo using Wistar rats.
METHODS: Effects upon cells proliferation/viability was measured using MTT assay, cytotoxic effects were analyzed by quantifying lactate dehydrogenase release and the Live/Dead Cell Imaging assay. Macrophage activation was assessed by quantification of IL-6, IL-10, IL-12p70, MCP-1, and TNF-α using a microsphere-based cytometric bead array. For in vivo analysis, Wistar rats were inoculated into the dorsal sub-dermis with pUBMh. The specimens were sacrificed at 24 h after inoculation for histological study.
RESULTS: The pUBMh obtained showed good consistency and absence of cell debris. The biocompatibility tests in vitro revealed that the pUBMh promoted cell proliferation and it is not cytotoxic on the three tested cell lines and induces the production of pro-inflammatory cytokines on macrophages, mainly TNF-α and MCP-1. In vivo, pUBMh exhibited fibroblast-like cell recruitment, without tissue damage or inflammation.
CONCLUSIONS: The results show that pUBMh allows cell proliferation without cytotoxic effects and can be considered an excellent biomaterial for tissue engineering.
摘要：
背景：制造具有优异的生物相容性和生物降解能力的支架的必要性已导致细胞外基质（ECM）支架的产生。在他们的优势中，它们允许更好的细胞定植，这使得它能够成功地整合到托管组织中，围绕要修复的区域及其配方有助于将其放置成不规则形状。来自猪膀胱的ECM（pUBM）包含蛋白质，蛋白聚糖和糖胺聚糖，为细胞提供支持和信号。这些特性使其成为生产可用于再生医学的水凝胶的绝佳选择。
目的：本研究的目的是使用成纤维细胞在体外评估源自猪膀胱（pUBMh）的ECM水凝胶的生物相容性，巨噬细胞，和脂肪间充质干细胞（AD-MCSs），以及使用Wistar大鼠的体内生物相容性。
方法：使用MTT测定法测量对细胞增殖/活力的影响，通过定量乳酸脱氢酶释放和活/死细胞成像分析来分析细胞毒性作用。巨噬细胞活化通过使用基于微球的细胞计数珠阵列定量IL-6、IL-10、IL-12p70、MCP-1和TNF-α来评估。对于体内分析，用pUBMh将Wistar大鼠接种到背侧真皮下。在接种后24小时处死样本用于组织学研究。
结果：获得的pUBMh显示出良好的一致性和不存在细胞碎片。体外生物相容性测试表明，pUBMh促进细胞增殖，对三种测试细胞系没有细胞毒性，并诱导巨噬细胞产生促炎细胞因子，主要是TNF-α和MCP-1。在体内,pUBMh表现出成纤维细胞样细胞募集，没有组织损伤或炎症。
结论：结果表明，pUBMh允许细胞增殖而没有细胞毒性作用，并且可以被认为是组织工程的出色生物材料。
目的：本研究的目的是使用成纤维细胞在体外评估源自猪膀胱（pUBMh）的ECM水凝胶的生物相容性，巨噬细胞，和脂肪间充质干细胞（AD-MCSs），以及使用Wistar大鼠的体内生物相容性。
方法：使用MTT测定法测量对细胞增殖/活力的影响，通过定量乳酸脱氢酶释放和活/死细胞成像分析来分析细胞毒性作用。巨噬细胞活化通过使用基于微球的细胞计数珠阵列定量IL-6、IL-10、IL-12p70、MCP-1和TNF-α来评估。对于体内分析，用pUBMh将Wistar大鼠接种到背侧真皮下。在接种后24小时处死样本用于组织学研究。
结果：获得的pUBMh显示出良好的一致性和不存在细胞碎片。体外生物相容性测试表明，pUBMh促进细胞增殖，对三种测试细胞系没有细胞毒性，并诱导巨噬细胞产生促炎细胞因子，主要是TNF-α和MCP-1。在体内,pUBMh表现出成纤维细胞样细胞募集，没有组织损伤或炎症。
结论：结果表明，pUBMh允许细胞增殖而没有细胞毒性作用，并且可以被认为是组织工程的出色生物材料。
