MORPHOMETRIC ANALYSIS OF NEUROCYTES OF THE CA3 REGION OF THE RAT HIPPOCAMPUS AFTER THERMAL INJURY WITH THE USE OF LYOPHILIZED XENOGRAFT

Zoia Nebesna, Diana Lytvynyuk, Nataliia Ohinska, Svitlana Lytvynyuk
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Abstract

Abstract. The hippocampus has a complex structure, and its work is coordinated with other parts of the brain, helping to perform its main functions: memory, behavior, reaction to the surrounding world. The purpose of the study was to determine the dynamics of morphometric changes in neurocytes of the CA3 region of the rat hippocampus after experimental thermal injury using a cryolyophilized xenodermal substrate. Experimental studies were conducted on 56 sexually mature white male laboratory rats. The thermal injury was infl icted under thiopental- sodium anesthesia with copper plates heated in boiled water.The area of the lesion was 18-20 % of the body surface. Early necrectomy of the damaged skin was performed 1 day after the thermal injury. The resulting wound was closed with a crushed substrate of lyophilized xeno-skin. The animals were decapitated on days 7, 14, and 21. Material for microscopic studies was collected according to the generally accepted method. The hippocampal histological preparations were used to determine the numerical density of CA3 neurons in the hippocampal region, the area of bodies and nuclei of diff erent types of neurons, and their nuclear- cytoplasmic ratio. Statistical processing of digital data was performed using the software «Excel» and «STATISTICA 6.0» using parametric and nonparametric methods of data evaluation. A comparative analysis of the numerical density of neurocytes in the CA3 region of the hippocampus of animals with thermal injury and after correction showed that the number of cells that died was lower on day 7 after the use of lyophilized xenoceramics, but still not signifi cant. On day 14, the number of sharply hypochromic neurocytes with total tigrolysis in its CA3 region was 1.7 times lower and the number of pycnomorphic, shriveled cells, respectively, 3.5 times lower than in the second control group of animals. The number of normochromic neurocytes in the hippocampal fi elds increased by 1.40 times compared to the burned animals. They are characterized by normalization of the basophilic substance content and nuclei condition. Histological studies of hippocampal fi elds on day 21 of the experiment showed that the use of xeno-skin to close burn wounds signifi cantly normalizes their structural components. Microscopically, CA3 region at this time contained many normochromic neurocytes, 12.5 times (p<0.05) more than in animals with burns, respectively, 7.7 times fewer sharply hypochromic cells.Conclusions. Morphometrically, it was found that in the early period after thermal injury under the conditions of lyophilized xenodermal substrate application in the CA3 region of the hippocampus, the population of normochromic neurocytes increased by 1.43 times, the number of sharply hyper- and hypochromic forms decreased by 1.95 and 2.11 times compared to animals of the second group, indicating the onset of restorative, regenerative processes in the neurocyte- gliocyte-hemocapillary complex. Under the conditions of application of the corrective factor in the late period after thermal injury (14, 21 days), neurocyte recovery and normalization of their morphometric parameters were established. The content of normochromic neurocytes increases 1.40 times (14 days) and 2.94 times (21 days) in the CA3 region, compared to the parameters of the group of animals with thermal injury without correction, the content of sharply hypo- and hyperchromic cells signifi cantly decreases, their nuclear- cytoplasmic ratios improve, which indicates the restoration of the morphofunctional state of CA3 neurocytes in the hippocampus.
冻干异种移植物对大鼠海马ca3区神经细胞热损伤的形态学分析
摘要海马体结构复杂,它的工作与大脑的其他部分协调,帮助执行其主要功能:记忆、行为、对周围世界的反应。本研究的目的是利用冻干的异种真皮底物确定实验性热损伤后大鼠海马CA3区神经细胞形态变化的动态。实验研究以56只性成熟雄性白鼠为实验对象。热损伤采用硫喷妥钠麻醉,用沸水加热铜板。病变面积为体表的18- 20%。热伤后1天早期切除损伤皮肤。用冻干的异种皮肤压碎的基质缝合伤口。分别于第7、14、21天斩首。显微研究的材料是按照普遍接受的方法收集的。采用海马组织标本测定海马区CA3神经元的数量密度、不同类型神经元的体、核面积及核质比。使用«Excel»和«STATISTICA 6.0»软件对数字数据进行统计处理,采用参数和非参数方法进行数据评估。对比分析热损伤动物和校正后海马CA3区神经细胞的数值密度显示,使用冻干异种陶瓷后第7天死亡的细胞数量较低,但仍无显著性差异。第14天,与第二对照组相比,CA3区出现完全褪黑的神经细胞数量减少了1.7倍,出现幻形、萎缩的细胞数量减少了3.5倍。海马区正色神经细胞数量较烧伤组增加1.40倍。它们的特征是亲碱性物质含量和细胞核状态的正常化。实验第21天的海马区组织学研究表明,异种皮肤修复烧伤创面后,海马区结构成分明显恢复正常。显微镜下观察,此时CA3区含有较多的正色神经细胞,分别比烧伤大12.5倍(p < 0.05),比烧伤大7.7倍(p < 0.05)。形态学上发现,在热损伤后早期,在海马CA3区应用冻干异种真皮底物的条件下,与第二组动物相比,正色神经细胞的数量增加了1.43倍,急剧高色和低色的数量减少了1.95倍和2.11倍,这表明神经细胞-胶质细胞-血液毛细血管复合体的恢复和再生过程开始了。在热损伤后期(14、21 d)应用校正因子的条件下,观察神经细胞的恢复情况和形态计量参数的归一化。CA3区正色神经细胞含量较未校正热损伤组分别增加1.40倍(14 d)和2.94倍(21 d),急剧低色和高色细胞含量显著减少,核质比提高,表明海马CA3神经细胞形态功能状态恢复。
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