Cell-based resurfacing of large cartilage defects
Cell-based resurfacing of large cartilage defects
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DOI：
10.1002/art.23124
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发表时间：
2008-02-01
影响因子：
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通讯作者：
Schneider, Holm
中科院分区：
文献类型：
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作者：
Gelse, Koja;Muehle, Christiane;Schneider, Holm
Objective.研究转基因活化的骨膜细胞用于永久性修复大部分软骨缺损的潜力。在小型猪，自体骨膜细胞刺激离体骨形态发生蛋白2基因转移，使用脂质体或腺相关病毒（AAV）和腺病毒（Ad）载体的组合，被施加在生物可吸收的支架上的软骨损伤，包括整个内侧一半的髌骨。在移植后6周和26周，通过组织化学和免疫组织化学方法评估所得修复组织。修复组织的生物力学性能，其特征在于纳米压痕测量。植入未经刺激的细胞和未经处理的病变作为对照。所有移植物均表现出令人满意的整合到先前存在的软骨中。移植后6周，AAV/Ad刺激的骨膜细胞在所有层中均具有软骨细胞样表型;新形成的基质富含蛋白多糖和II型胶原，其接触刚度接近健康透明软骨。未受刺激的骨膜细胞和脂质体基因转移激活的细胞仅形成具有轻微接触刚度的纤维软骨修复组织。然而，在移植后6个月内，AAV/Ad刺激的细胞在浅表区倾向于去分化，如由从II型胶原合成转换为I型胶原合成和接触刚度降低所示。在更深的区域，这些细胞保留了它们的软骨细胞表型，与基质中II型胶原的阳性染色相一致。转基因活化骨膜细胞形成透明样软骨可有效修复大面积部分软骨缺损。软骨的长期稳定性似乎取决于仅在软骨组织的较深区域中活跃的物理生物化学因素。
Objective. To investigate the potential of transgene-activated periosteal cells for permanently resurfacing large partial-thickness cartilage defects.Methods. In miniature pigs, autologous periosteal cells stimulated ex vivo by bone morphogenetic protein 2 gene transfer, using liposomes or a combination of adeno-associated virus (AAV) and adenovirus (Ad) vectors, were applied on a bioresorbable scaffold to chondral lesions comprising the entire medial half of the patella. The resulting repair tissue was assessed, 6 and 26 weeks after transplantation, by histochemical and immunohistochemical methods. The biomechanical properties of the repair tissue were characterized by nanoindentation measurements. Implants of unstimulated cells and untreated lesions served as controls.Results. All grafts showed satisfactory integration into the preexisting cartilage. Six weeks after transplantation, AAV/Ad-stimulated periosteal cells had adopted a chondrocyte-like phenotype in all layers; the newly formed matrix was rich in proteoglycans and type II collagen, and its contact stiffness was close to that of healthy hyaline cartilage. Unstimulated periosteal cells and cells activated by liposomal gene transfer formed only fibrocartilaginous repair tissue with minor contact stiffness. However, within 6 months following transplantation, the AAV/Ad-stimulated cells in the superficial zone tended to dedifferentiate, as indicated by a switch from type II to type I collagen synthesis and reduced contact stiffness. In deeper zones, these cells retained their chondrocytic phenotype, coinciding with positive staining for type II collagen in the matrix.Conclusion. Large partial-thickness cartilage defects can be resurfaced efficiently with hyaline-like cartilage formed by transgene-activated periosteal cells. The long-term stability of the cartilage seems to depend on physicobiochemical factors that are active only in deeper zones of the cartilaginous tissue.