Department of Biomedical Engineering, University of Melbourne, Parkville, Victoria, Australia.
Department of Otorhinolaryngology and Department of Orthopaedics, Erasmus MC, University Medical Centre, Rotterdam, the Netherlands.
Cartilage. 2021 Dec;13(2_suppl):476S-485S. doi: 10.1177/19476035211000811. Epub 2021 Mar 22.
To investigate GAG-ECM (glycosaminoglycan-extracellular matrix) interactions in different cartilage types. To achieve this, we first aimed to determine protocols for consistent calculation of GAG content between cartilage types.
Auricular cartilage containing both collagen and elastin was used to determine the effect of lyophilization on GAG depletion activity. Bovine articular, auricular, meniscal, and nasal cartilage plugs were treated using different reagents to selectively remove GAGs. Sulfated glycosaminoglycan (sGAG) remaining in the sample after treatment were measured, and sGAG loss was compared between cartilage types.
The results indicate that dry weight of cartilage should be measured prior to cartilage treatment in order to provide a more accurate reference for normalization. Articular, meniscal, and nasal cartilage lost significant amounts of sGAG for all reagents used. However, only hyaluronidase was able to remove significant amount of sGAG from auricular cartilage. Furthermore, hyaluronidase was able to remove over 99% of sGAG from all cartilage types except auricular cartilage where it only removed around 76% of sGAG. The results indicate GAG-specific ECM binding for different cartilage types and locations.
In conclusion, lyophilization can be performed to determine native dry weight for normalization without affecting the degree of GAG treatment. To our knowledge, this is the first study to compare GAG-ECM interactions of different cartilage types using different GAG extraction methods. Degree of GAG depletion not only varied with cartilage type but also the same type from different anatomic locations. This suggests specific structure-function roles for GAG populations found in the tissues.
研究不同类型软骨中糖胺聚糖-细胞外基质(GAG-ECM)的相互作用。为此,我们首先旨在确定在不同软骨类型之间一致计算 GAG 含量的方案。
使用同时含有胶原和弹性蛋白的耳廓软骨来确定冻干对 GAG 耗竭活性的影响。使用不同的试剂处理牛关节、耳廓、半月板和鼻软骨塞,以选择性去除 GAG。测量处理后样品中残留的硫酸化糖胺聚糖(sGAG),并比较软骨类型之间的 sGAG 损失。
结果表明,在进行软骨处理之前,应该测量软骨的干重,以便为归一化提供更准确的参考。关节、半月板和鼻软骨的所有试剂均显著丢失了 sGAG。然而,只有透明质酸酶能够从耳廓软骨中去除大量的 sGAG。此外,透明质酸酶能够从所有软骨类型中去除超过 99%的 sGAG,除了耳廓软骨,其中仅去除了约 76%的 sGAG。结果表明,不同的软骨类型和位置存在 GAG 特异性细胞外基质结合。
总之,可以进行冻干以确定用于归一化的天然干重,而不会影响 GAG 处理的程度。据我们所知,这是首次使用不同的 GAG 提取方法比较不同类型软骨的 GAG-ECM 相互作用的研究。GAG 耗竭的程度不仅随软骨类型而异,而且同一类型的不同解剖部位也不同。这表明组织中 GAG 群体具有特定的结构-功能作用。