过去二十年代,组织工程已迅速发展,以解决器官供体短缺的问题。已经开发了去细胞化技术以减轻移植中的免疫排斥和同种异体反应。然而,有效脱细胞的明确定义仍然难以捉摸。本研究使用人筋膜模型比较了各种去细胞化方案。形态学,结构和细胞毒性/活力分析表明,所有五个测试方案是等效的,符合Crapo成功脱细胞的标准。有趣的是,只有大鼠体内免疫试验显示差异。只有一个方案显示人类白细胞抗原(HLA)含量低于1%残留阈值,在一个月后没有大鼠抗人IgG转换的情况下阻止大鼠免疫的唯一标准(7组中每组N=4个供体,加上阴性和阳性对照,n=28)。通过遵守一套完善的标准,即缺乏可见的核材料,<50ngDNA/mg干重的细胞外基质,和<1%残余HLA含量,潜在的不良宿主反应可以大大减少。总之,这项研究强调了在脱细胞方案中不仅要考虑核成分,还要考虑主要组织相容性复合物的重要性,并提出了新的指南,以促进更安全的临床开发和使用生物工程支架。
Last twenties, tissue engineering has rapidly advanced to address the shortage of organ donors.
Decellularization techniques have been developed to mitigate immune rejection and alloresponse in transplantation. However, a clear definition of effective
decellularization remains elusive. This study compares various
decellularization protocols using the human fascia lata model. Morphological, structural and cytotoxicity/viability analyses indicated that all the five tested protocols were equivalent and met Crapo\'s criteria for successful
decellularization. Interestingly, only the in vivo immunization test on rats revealed differences. Only one protocol exhibited Human Leucocyte Antigen (HLA) content below 1% residual threshold, the only criterion preventing rat immunization with an absence of rat anti-human IgG switch after one month (N=4 donors for each of the 7 groups, added by negative and positive controls, n=28). By respecting a refined set of criteria, i.e. lack of visible nuclear material, <50ng DNA/mg dry weight of extracellular matrix, and <1% residual HLA content, the potential for adverse host reactions can be drastically reduced. In conclusion, this study emphasizes the importance of considering not only nuclear components but also major histocompatibility complex in
decellularization protocols and proposes new guidelines to promote safer clinical development and use of bioengineered scaffolds.