Morphometric characteristics of the structural components of the intervertebral disc in rats at different times of the experimental opioid exposure and after its withdrawal

M. Pankiv, Y. Paltov, Z. Z. Masna, I. Chelpanova
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Abstract

Background. The world and domestic literature from time to time report about the problems of uncontrolled drug abuse. This problem is associated not only with the negative impact on the morphological structure, but also is a serious factor which, if it has been exposing for a long time, leads to disability and death. Despite a significant number of studies in this area, the problem of changes in the morphometric parameters of the intervertebral disc’s structural components under chronic exposure of opioid agents still remains unresolved. That is why the study of morphometric characteristics of the intervertebral disc’s structural components under experimental opioid exposure will be interesting for both - morphologists and practical traumatologists. Objective. To study the morphometric parameters of the structural components of the intervertebral disc in rats at different times of the experimental opioid exposure and after its withdrawal. Methods. The objects of the study were 61 mature outbred male rats, weight - 80-135g, age - 4.5-7.5 months. Animals were injected with nalbuphine intramuscularly once daily (at 10-11 am) for 42 days. Histological specimens were stained with alcyan blue (for the study of the collagen fibers thickness of the anterior and posterior longitudinal ligaments) and PAS-reaction (for the study of the gelatinous nucleus and its capsule thickness). For measurements of collagen fiber thickness, were taken images at x100 magnification, for measurements of the gelatinous nucleus and its capsule thickness - at x40 magnification. Measurements were performed with ImageJver software. 1.51 using the tool "straight" for linear measurements. Results. Thus, changes in the collagen fibers thickness of the anterior and posterior longitudinal ligaments during the experiment were uneven and had different tendencies. The thickness of the posterior longitudinal fibers changed mostly within the central trend of the control group with a significant decrease only at the 2nd, 3rd, and 4th weeks. However, more significant, as for pathogenetic changes, was a sharp decrease in the number of fibers thicker than 25 μm in animals of the experimental group after 2 weeks of the study, and the maximum thickness at 5th and 6th week was only 31.41 μm and 35.24 μm whereas the maximum thickness of the control group was 81.48 μm. In the withdrawal group, only a decrease in sites with nuclear edema can be considered positive, which is displayed by morphometrical decrease in the maximum value, but the central trend remains much lower than in the control group, which also indicates the predominance of decompensatory changes. Conclusion. During the whole experiment, we observed similar changes in the nucleus capsule thickness: non-systemic fluctuations of the central parameters ​​up to the 5th week and a sharp decompensation at the 6th week, which was manifested by a sharp decrease in thickness. Simultaneously, we observed numerous sites of edema and thickening by more than 100 μm on the 2nd week. Despite a general decrease in median and quartile at this time, such sites were also noted at the 6th week. Thickness of the nucleus capsule in the withdrawal group did not demonstrate significant dynamics and remained similar to those at the 6th week. Although, this can be interpreted as the stabilization at a certain level without further negative dynamics.
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实验性阿片类药物暴露不同时间及停药后大鼠椎间盘结构成分的形态特征
背景。国际和国内文献不时报道不受控制的药物滥用问题。这一问题不仅与对形态结构的负面影响有关,而且是一个严重的因素,如果长期暴露,将导致残疾和死亡。尽管在这一领域进行了大量的研究,但慢性阿片类药物暴露下椎间盘结构成分形态计量参数的变化问题仍未得到解决。这就是为什么在实验性阿片类药物暴露下研究椎间盘结构成分的形态特征对形态学家和实际创伤学家来说都是有趣的。目标。研究大鼠在阿片类药物暴露不同时间及停药后椎间盘结构成分的形态学参数。方法。研究对象为61只纯种雄性大鼠,体重- 80-135克,年龄- 4.5-7.5月龄。每天(上午10-11点)肌肉注射纳布啡1次,连续42天。组织学标本进行alcyan blue染色(用于研究前后纵韧带的胶原纤维厚度)和pas反应(用于研究胶质核及其被膜厚度)。为了测量胶原纤维的厚度,在x100的放大倍率下拍摄了图像,为了测量胶质核及其被膜的厚度,在x40的放大倍率下拍摄了图像。使用ImageJver软件进行测量。1.51使用工具“直”进行直线测量。结果。因此,实验过程中前后纵韧带胶原纤维厚度的变化是不均匀的,有不同的趋势。后纵纤维厚度的变化主要在对照组的中心趋势内,仅在第2周、第3周和第4周明显减少。然而,在病理变化方面,更显著的是实验组动物在研究2周后,厚度大于25 μm的纤维数量急剧减少,第5周和第6周的最大厚度仅为31.41 μm和35.24 μm,而对照组的最大厚度为81.48 μm。在戒断组中,只有核性水肿部位减少才可视为阳性,表现为形态学上的最大值减少,但中心趋势仍远低于对照组,这也表明失代偿变化占主导地位。结论。在整个实验过程中,我们观察到类似的核被膜厚度变化:直到第5周中心参数的非全体性波动,在第6周出现急剧失代偿,表现为厚度急剧下降。同时,我们在第2周观察到大量水肿和增厚超过100 μm的部位。尽管此时中位数和四分位数普遍下降,但这些部位在第6周也被注意到。停药组的核囊厚度没有明显的变化,与第6周相似。虽然,这可以解释为稳定在一定水平,没有进一步的负面动态。
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