Development of fibrocartilage layers in the anterior cruciate ligament insertion in rabbits
Development of fibrocartilage layers in the anterior cruciate ligament insertion in rabbits
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DOI：
10.1186/s12891-019-2614-0
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发表时间：
2019-05-18
影响因子：
2.3
通讯作者：
Sakane, Masataka
中科院分区：
文献类型：
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作者：
Mutsuzaki, Hirotaka;Nakajima, Hiromi;Sakane, Masataka
研究背景对前交叉韧带（ACL）止点的纤维软骨层进行详细的评估，对于考虑止点的再生是必要的。本研究探讨了在ACL胫骨插入兔的纤维软骨层的发展，定量形态计量学评价的基础上，组织学和免疫组化analysis.MethodsMale日本白色兔，因为他们的历史使用的ACL插入的组织形态计量学分析，并消除对ACL的女性激素的影响。在每个年龄（1天和1、2、4、6、8、12和24周）对6只动物实施安乐死;总共使用了48只动物。检测软骨细胞增殖率、凋亡率、Sox 9阳性率和软骨细胞数。还评价了番红O染色的糖胺聚糖（GAG）面积、潮标长度、ACL插入宽度和ACL长度。所有参数进行了比较，与那些在24周龄ages.ResultsHigh水平的软骨细胞增殖和Sox 9的表达持续到4和8周龄，分别，然后逐渐下降。软骨细胞凋亡增加至8周。软骨细胞数量、ACL插入宽度、ACL长度、番红O染色GAG面积和潮标长度在12周龄时逐渐增加。结论软骨细胞增殖和Sox 9表达在12周龄时逐渐增加，与ACL长度和插入宽度的发育一致。GAG产量和潮标长度在12周龄前也呈增加趋势。前交叉韧带附着处的纤维软骨层在12周龄时发育完成。
BackgroundA detailed evaluation focusing on the fibrocartilage layers in the anterior cruciate ligament (ACL) insertion is necessary to consider regeneration of the insertion. This study examined the development of the fibrocartilage layers in the ACL tibial insertion in rabbits by quantitative morphometric evaluations based on histological and immunohistochemical analyses.MethodsMale Japanese white rabbits were used because of their history of use for histomorphometric analyses of the ACL insertion and to eliminate the influence of female hormones on the ACL. Six animals were euthanized at each age (1day and 1, 2, 4, 6, 8, 12, and 24weeks); in total, 48 animals were used. Proliferation rate, apoptosis rate, Sox9-positive rate, and chondrocyte number were evaluated. Safranin O-stained glycosaminoglycan (GAG) areas, tidemark length, ACL insertion width, and ACL length were also evaluated. All parameters were compared with those at age 24weeks of age.ResultsHigh levels of chondrocyte proliferation and Sox9 expression continued until 4 and 8weeks of age, respectively, and then gradually decreased. Chondrocyte apoptosis increased up to 8weeks. The chondrocyte number, ACL insertion width, ACL length, safranin O-stained GAG areas, and tidemark length gradually increased up to 12weeks.ConclusionChondrocytes that displayed chondrocyte proliferation and Sox9 expression increased until 12weeks of age, in accordance with development of the ACL length and its insertion width. The GAG production and tidemark length also increased until 12weeks of age. The development of fibrocartilage layers in the ACL insertion was complete at 12weeks of age.