Comparison of mRNA levels for matrix molecules in normal and disrupted human anterior cruciate ligaments using reverse transcription polymerase chain reaction
Comparison of mRNA levels for matrix molecules in normal and disrupted human anterior cruciate ligaments using reverse transcription polymerase chain reaction
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DOI：
10.1002/jor.1100160405
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发表时间：
1998-07-01
影响因子：
2.8
通讯作者：
Frank, CB
中科院分区：
文献类型：
--
作者：
Lo, IKY;Marchuk, LL;Frank, CB
从7例正常人尸体膝关节（16-50岁）中获取前交叉韧带的中间物质样本，并与30例患者（15-40岁）的瘢痕前交叉韧带的中间物质碎片进行比较。从每个韧带中分离RNA，并使用β-肌动蛋白作为管家基因，通过定量逆转录聚合酶链反应评价I型胶原、III型胶原、双糖链蛋白聚糖、核心蛋白聚糖、光蛋白聚糖和金属蛋白酶组织抑制剂-1的表达。损伤韧带的数据作为损伤后时间的函数进一步进行统计学比较，以更好地定义随时间推移的细胞表达模式。我们的假设是，受伤的韧带将显示出最小的细胞活性，并随着时间的推移而降低活性。结果显示，正常和受伤的前交叉韧带都含有表达所有研究分子mRNA的细胞。然而，损伤韧带中的细胞表达的I型胶原和III型胶原的信息量（p < 0.000001）以及双糖链蛋白聚糖（p < 0.02）和金属蛋白酶组织抑制剂-1（p < 0.0003）的量比正常前交叉韧带中的细胞高得多，但仍然成比例。这些水平在受伤后持续升高超过1年。线性回归分析显示双糖蛋白聚糖表达与损伤时间相关（r（2）= -0.69; p = 0.007）。这些结果共同表明，受伤的人前交叉韧带含有表达瘢痕样分子的细胞，并且随着时间的推移，受伤的韧带可能继续重塑基质。此外，他们认为人类前交叉韧带没有愈合失败，由于疤痕形成本身的失败。这个疤痕的质量和数量仍然值得怀疑;然而，它作为人类前交叉韧带愈合策略的可能性不能被排除。
Midsubstance samples of anterior cruciate ligaments from seven normal human cadaver knees (16-50 years old) were harvested and compared with midsubstance pieces of scarred anterior cruciate ligaments from 30 patients (15-40 years old). RNA was isolated from each ligament, and the expression of type-I collagen, type-III collagen, biglycan, decorin, lumican, and tissue inhibitor of metalloproteinase-l was evaluated by quantitative reverse transcription-polymerase chain reaction with use of p-actin as the housekeeping gene. Data for injured ligaments were further compared statistically as a function of time after injury to better define patterns of cellular expression over time. Our hypothesis was that injured ligaments would show minimal cellular activity and decreasing activity over time. The results revealed that both normal and injured anterior cruciate ligaments contain cells that express mRNA for all molecules studied. However, cells in injured ligaments express much higher, but still proportional, quantities of message for type-I collagen and type-III collagen (p < 0.000001) and higher quantities of biglycan (p < 0.02) and tissue inhibitor of metalloproteinase-l (p < 0.0003) than do cells in normal anterior cruciate ligaments. These levels remained elevated for longer than 1 year after injury. Linear regression analysis showed biglycan expression correlated with time from injury (r(2) = -0.69; p = 0.007). These results collectively demonstrate that injured human anterior cruciate ligaments contain cells that express scar-like molecules and that the injured ligaments are likely continuing to remodel matrix over time. Furthermore, they suggest that human anterior cruciate ligaments have not failed to heal due to the failure of scar formation per se. The quality and quantity of this scar remain questionable; however, the possibility of its enhancement as a healing strategy for human anterior cruciate ligaments cannot be dismissed.