摘要
背景及目的: Neubert和Barrach在1977年提出一套肢芽软骨评分系统,但该系统仅主要应用于体外培养的肢芽。另外该系统本身有许多不足,并没有考虑到软骨内成骨等因素。我们通过大量的实验,通过连续切取不同时期胚胎后肢芽,充分研究其骨骼、软骨内成骨的形成规律,对Neubert等人的评分系统进行改良完善。本文主要介绍改良后的评分系统和对先天性马蹄内翻足后肢芽发育进行初步的量化评分。
材料和方法:60只SD母鼠,体重220-250g,被随机分成6个实验组和对照组,下午雌雄合笼,第二天早上阴道涂片发现精子为怀孕天数0天。分别在受孕第13天,14天,15天,16天,17天和18天取出胎鼠的后肢芽并经过一系列处理并用改良后的评分系统对制取的后肢芽进行量化评分。
结果:从E13到E18实验组和对照组的软骨量化评分差异有统计学意义(T-检验, P <0.05),发现对照组从第14天起软骨开始发育并在第17天到达顶峰。第13天、16天、17天和18天实验组和对照组后肢芽长度差异有统计学意义(P<0.05)。
结论:改良后的评分系统主要应用于在活体内发育的后肢芽软骨及形态学分化水平的量化评分。
Background and Objective The Limb Morphogenetic Differentiation Scoring system introduced by Neubert and Barrach in 1977 has been used in drug testing as a measure of the degree of cartilage growth inhibition especially for forelimb in vitro. But the in-vitro system is not applicable to in-vivo assessment. Besides, there are some limitations associated with their system. Therefore, in this study, we have investigated the differentiation of cartilage in a series of experiments using the hindlimb buds obtained from different embryonic stages in vivo, and developed a new scoring system for limb morphogenetic differentiation.
Materials and Methods A total of 60 female Sprague-Dawley rats weighing 220–250g were assigned at random to six control groups and six experimental groups on day 0 of embryonic. The experimental groups were treated with all-trans-retinoic acid (ATRA). A new limb morphogenetic differentiation scoring system was developed and used to quantify the degree of development of the hindlimb buds from the fetuses at embryonic days E13 to E18.
Results The differentiation of cartilages assessed by our new scoring system showed a statistically significant difference between the experimental group and the control group from E13 to E18 (T-test, p <0.05). Cartilage growth (the proximodistal length) in the control group increased gradually from E14, reaching its peak at E17, but in the experimental group the growth at E13, E16, E17, and E18, was significantly shorter (p <0.05).
Conclusions Our new limb morphogenetic differentiation scoring system described here in can be used to quantify the degree of inhibition of the hindlimb bud development by teratogenic drugs or materials, and morphogenetic differentiation in vivo.
引文
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