Mu Yufeng, Zhang Yu, Wei Lina, Chen Liang, Hao Feng, Shao Anliang, Qu Shuxin, Xu Liming
Institute for Medical Device Control, National Institutes for Food and Drug Control, Beijing, 102629, China.
School of Materials Science and Engineering, Southwest Jiaotong University, Chengdu, 610031, China.
Mater Today Bio. 2022 Nov 25;18:100505. doi: 10.1016/j.mtbio.2022.100505. eCollection 2023 Feb.
Wild type (WT) animals cannot be used to objectively assess the immunogenicity of animal tissue-derived biomaterials when used as recipients due to difference with human in α-Gal expression. The purpose of this study is to compare the differences of immunological responses between the GGTA1 gene-knockout (GTKO) rabbits and WT rabbits after implantation with animal tissue-derived biomaterials. The porcine-derived decellularized bone matrix (natural bone material, NBM) and fresh porcine cancellous bone (PCB) were implanted in GTKO rabbits and WT rabbits, respectively, and sham operation was used as control (Con). At 2- and 6-week post-implantation, the related immunological items including antibody levels, serum-mediated cell lysis, cytokines, lymphocyte subtypes, and histopathological changes were assessed. GTKO rabbits exhibited more sensitive immune responses than WT rabbits after PCB implantation, resulted from a significant increase of antibodies (except total antibodies) and cytokines levels, cell lysis ratios, CD4/CD8 proportions, and inflammatory cells infiltration. Immunological factors and inflammatory cells infiltrate in GTKO rabbits after NBM implantation were significantly lower than those in the PCB group. Among the three groups, the NBM group showed the highest contents of new bone formation elements. In conclusion, the GTKO rabbit is a more sensitive alternative model than WT rabbit for preclinical study of xenografts via in situ implantation. Studies on multiple gene-edited animals are also necessary for more comprehensively evaluating xenoimmunologen risks of animal tissue-derived biomaterials in the future. Additionally, the immunogenicity of NBM was remarkably decreased compared to PCB.
由于野生型（WT）动物与人类在α - Gal表达上存在差异，当用作受体时，它们不能用于客观评估动物组织来源生物材料的免疫原性。本研究的目的是比较GGTA1基因敲除（GTKO）兔和WT兔在植入动物组织来源生物材料后的免疫反应差异。将猪源脱细胞骨基质（天然骨材料，NBM）和新鲜猪松质骨（PCB）分别植入GTKO兔和WT兔体内，并以假手术作为对照（Con）。在植入后2周和6周，评估包括抗体水平、血清介导的细胞裂解、细胞因子、淋巴细胞亚群和组织病理学变化等相关免疫项目。植入PCB后，GTKO兔比WT兔表现出更敏感的免疫反应，这是由于抗体（除总抗体外）和细胞因子水平、细胞裂解率、CD4/CD8比例以及炎性细胞浸润显著增加所致。NBM植入后GTKO兔的免疫因子和炎性细胞浸润明显低于PCB组。在三组中，NBM组新骨形成元素的含量最高。总之，对于通过原位植入进行异种移植的临床前研究，GTKO兔是比WT兔更敏感的替代模型。未来为了更全面地评估动物组织来源生物材料的异种免疫原风险，对多种基因编辑动物进行研究也是必要的。此外，与PCB相比，NBM的免疫原性显著降低。
