A high-throughput model of post-traumatic osteoarthritis using engineered cartilage tissue analogs.
A high-throughput model of post-traumatic osteoarthritis using engineered cartilage tissue analogs.
复制标题
DOI：
10.1016/j.joca.2014.06.032
复制
发表时间：
2014-09
影响因子：
7
通讯作者：
Dodge, G. R.
中科院分区：
文献类型：
--
作者：
Mohanraj, B.;Meloni, G. R.;Mauck, R. L.;Dodge, G. R.
关键词：
已经开发了许多创伤后骨关节炎（PTOA）的体外模型来研究机械过载对调节软骨退化过程的影响。虽然此类框架对于识别治疗靶点至关重要，但现有技术的吞吐能力有限。在这里，我们验证了一个包含工程软骨的高通量机械损伤测试平台。我们利用高通量机械测试平台对工程软骨施加损伤性压缩，并确定其对损伤的应变和应变率依赖性反应。接下来，我们通过对软骨外植体应用相同的损伤条件来验证这种反应。最后，我们对假定的 PTOA 治疗化合物进行了初步筛选。工程软骨对损伤的反应是应变依赖性的，与 50% 应变相比，75% 时 GAG 损失增加了 2 倍。在裂隙附近观察到广泛的细胞死亡，LDH 释放的显着增加证实了膜破裂。对已建立的 PTOA 疗法的测试表明，与单独损伤相比，泛半胱天冬酶抑制剂 (ZVF) 可有效减少细胞死亡，而两亲聚合物 (P188) 和自由基清除剂 (NAC) 可减少 GAG 损失。该工程软骨模型中的损伤反应复制了软骨外植体反应的关键特征，验证了该系统在生理相关损伤性压缩中的应用。这项研究为发现软骨损伤的机制建立了一个新工具，并为鉴定治疗 PTOA 的新分子建立了一个筛选平台。
A number of in vitro models of post-traumatic osteoarthritis (PTOA) have been developed to study the effect of mechanical overload on the processes that regulate cartilage degeneration. While such frameworks are critical for the identification therapeutic targets, existing technologies are limited in their throughput capacity. Here, we validate a test platform for high-throughput mechanical injury incorporating engineered cartilage. We utilized a high throughput mechanical testing platform to apply injurious compression to engineered cartilage and determined their strain and strain rate dependent responses to injury. Next, we validated this response by applying the same injury conditions to cartilage explants. Finally, we conducted a pilot screen of putative PTOA therapeutic compounds. Engineered cartilage response to injury was strain dependent, with a 2-fold increase in GAG loss at 75% compared to 50% strain. Extensive cell death was observed adjacent to fissures, with membrane rupture corroborated by marked increases in LDH release. Testing of established PTOA therapeutics showed that pan-caspase inhibitor (ZVF) was effective at reducing cell death, while the amphiphilic polymer (P188) and the free-radical scavenger (NAC) reduced GAG loss as compared to injury alone. The injury response in this engineered cartilage model replicated key features of the response from cartilage explants, validating this system for application of physiologically relevant injurious compression. This study establishes a novel tool for the discovery of mechanisms governing cartilage injury, as well as a screening platform for the identification of new molecules for the treatment of PTOA.
登录
查看更多内容
影响因子：
7
作者：
Nishimuta, J. F.;Levenston, M. E.
通讯作者：
Levenston, M. E.
影响因子：
--
作者：
DiMicco, MA;Patwari, P;Grodzinsky, AJ
通讯作者：
Grodzinsky, AJ
影响因子：
2.8
作者：
Hurtig, Mark;Chubinskaya, Susan;Rueger, David
通讯作者：
Rueger, David
影响因子：
--
作者：
Green, D. M.;Noble, P. C.;Birdsall, H. H.
通讯作者：
Birdsall, H. H.
影响因子：
3.4
作者：
Moo, Eng Kuan;Amrein, Matthias;Herzog, Walter
通讯作者：
Herzog, Walter