Mechanisms of cartilage growth - Modulation of balance between proteoglycan and collagen in vitro using chondroitinase ABC
Mechanisms of cartilage growth - Modulation of balance between proteoglycan and collagen in vitro using chondroitinase ABC
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DOI：
10.1002/art.22298
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发表时间：
2007-01-01
影响因子：
--
通讯作者：
Sah, Robert L.
中科院分区：
文献类型：
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作者：
Asanbaeva, Anna;Masuda, Koichi;Sah, Robert L.
目标。通过在培养前用软骨素酶处理降低GAG含量来检测糖胺多聚糖(GAG)膨胀压力和胶原网络抑制功能之间平衡的破坏对软骨生长的相关影响。取未成熟的牛关节软骨浅层和中层，在含血清的培养液中孵育13天后立即进行分析。其他外植体用软骨素酶ABC处理以去除组织GAG，并立即或在添加血清的培养液中孵育13天后进行分析。检测处理和孵育过程中组织体积、蛋白多糖和胶原网络成分的含量以及拉伸力学性能的变化。在含血清的培养液中孵育导致膨胀性生长，组织体积显着增加，这与拉伸完整性的降低有关。相反，软骨素酶ABC处理第0天导致GAG含量显著减少，拉伸完整性增强，随后的孵育导致成熟生长，组织体积变化最小，拉伸完整性保持在增强的水平。结果表明，控制关节软骨组织中GAG的含量可以明显改变软骨的生长表型。这可能在组织工程和软骨修复方面具有实用价值。例如，扩张型生长表型可能有助于填充软骨缺损，而成熟型生长表型可能有助于在填充缺损区后诱导基质稳定。
Objective. To examine the cartilage growth-associated effects of a disruption in the balance between the swelling pressure of glycosaminoglycans (GAGs) and the restraining function of the collagen network, by diminishing GAG content prior to culture using enzymatic treatment with chondroitinase ABC.Methods. Immature bovine articular cartilage explants from the superficial and middle layers were analyzed immediately or after incubation in serum-supplemented medium for 13 days. Other explants were treated with chondroitinase ABC to deplete tissue GAG and also either analyzed immediately or after incubation in serum- supplemented medium for 13 days. Treatment- and incubation-associated variations in tissue volume, contents of proteoglycan and collagen network components, and tensile mechanical properties were assessed.Results. Incubation in serum-supplemented medium resulted in expansive growth with a marked increase in tissue volume that was associated with a diminution of tensile integrity. In contrast, chondroitinase ABC treatment on day 0 led to a marked reduction of GAG content and enhancement of tensile integrity, and subsequent incubation led to maturational growth with minimal changes in tissue volume and maintenance of tensile integrity at the enhanced levels.Conclusion. The data demonstrate that a manipulation of GAG content in articular cartilage explants can distinctly alter the growth phenotype of cartilage. This may have practical utility for tissue engineering and cartilage repair. For example, the expansive growth phenotype may be useful to fill cartilage defects, while the maturational growth phenotype may be useful to induce matrix stabilization after filling defect spaces.