A comparative study of cartilage engineered constructs in immunocompromised, humanized and immunocompetent mice

Emma Cavalli , Philipp Fisch , Florian A. Formica , Ralph Gareus , Thomas Linder , Lee Ann Applegate , Marcy Zenobi-Wong
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引用次数: 10

Abstract

Choosing the best ectopic in vivo model for cartilage engineering studies remains challenging and there is no clear consensus on how different models compare to one another. The use of xenogenic cells can often limit the choice to immunocompromised animals only and thus prevents the understanding of how tissue-engineered grafts perform with potential active inflammatory and immunological responses. The aim of this study was to evaluate the chondrogenic potential of a recently developed hydrogel, hyaluronan transglutaminase (HA-TG), in four mouse strains with varying immune systems: NSG, nude, NSG-SGM3 humanized and C57BL/6. The hyaluronan-based hydrogel was implanted subcutaneously for 4 weeks after an in vitro pre-culture time of 4 weeks. Scaffolds were prepared without cell seeding as the control and in combination with either human auricular chondrocytes (hAUR) or human fetal chondroprogenitor cells (hCC). We have seen that constructs were able to maintain their volumes and resisted vascularization as well as macrophage infiltration in vivo. Both hAUR and hCC maintained and produced ECM in vivo, but hAUR showed higher levels of innate collagen 2 even without mechanical stimulation. Collagen 1 and 2 deposition as well as mechanical properties of the scaffolds were comparable in all mouse strains. The C57BL/6 mouse model consistently displayed higher levels of C-reactive protein (CRP), serum amyloid A (SAA), and serum amyloid protein (SAP) in serum as a reaction to the foreign material and human cells. In addition, the number of CD68 + and CD163+ macrophages as well as CD3+ lymphocytes around the constructs in C57BL/6 mice was significantly higher than in humanized and immunocompromised mouse models. The results show that it is possible to engineer a cartilage-like graft subcutaneously not only in immunocompromised, but also in immunocompetent and humanized mouse models.

Abstract Image

免疫功能低下小鼠、人源化小鼠和免疫功能正常小鼠软骨工程构建体的比较研究
为软骨工程研究选择最佳异位体内模型仍然具有挑战性,并且对于不同模型之间的比较没有明确的共识。异种细胞的使用往往限制了免疫功能低下动物的选择,从而阻碍了对组织工程移植物如何表现潜在的主动炎症和免疫反应的理解。本研究的目的是评估最近开发的水凝胶透明质酸转谷氨酰胺酶(HA-TG)在四种具有不同免疫系统的小鼠品系(NSG,裸鼠,NSG- sgm3人源化和C57BL/6)中的成软骨潜力。体外预培养4周后,将透明质酸基水凝胶皮下植入4周。制备的支架没有细胞播种作为对照,与人耳软骨细胞(hAUR)或人胎儿软骨祖细胞(hCC)结合。我们已经看到,构建物能够保持其体积,抵抗体内血管化和巨噬细胞浸润。hAUR和hCC在体内均维持并产生ECM,但即使没有机械刺激,hAUR也显示出更高的先天胶原2水平。胶原蛋白1和2的沉积以及支架的力学性能在所有小鼠品系中均具有可比性。C57BL/6小鼠模型对外来物质和人类细胞的反应始终显示血清中较高水平的c反应蛋白(CRP)、血清淀粉样蛋白A (SAA)和血清淀粉样蛋白(SAP)。此外,C57BL/6小鼠构建体周围CD68 +和CD163+巨噬细胞以及CD3+淋巴细胞的数量显著高于人源化和免疫功能低下小鼠模型。结果表明,不仅在免疫功能低下的小鼠模型中,而且在免疫功能正常的和人源化的小鼠模型中,都可以进行软骨样移植物的皮下工程。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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