Histochemical assessment of the femora of spontaneously diabetic torii-leprfa (SDT-fa/fa) rats that mimic type II diabetes

IF 2.6 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Biosciences Pub Date : 2024-12-18 DOI:10.1016/j.job.2024.100602

Qi Yao , Kanako Tsuboi , Hiromi Hongo , Mako Sakakibara , Tomomaya Yamamoto , Mai Haraguchi-Kitakamae , Hotaka Ishizu , Xuanyu Liu , Yan Shi , Weisong Li , Jiaxin Cui , Tomohiro Shimizu , Norio Amizuka , Atsuro Yokoyama , Tomoka Hasegawa , Kiwamu Sakaguchi

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Abstract

Objective

To elucidate the mechanisms underlying diabetic osteoporosis, we conducted a comprehensive histological examination of the femora of Spontaneously Diabetic Torii-Lepr^fa (SDT-fa/fa) rats, an established model of obesity-related type 2 diabetes.

Materials and methods

Femora from 12 30-week-old male SDT-fa/fa rats and age-matched Sprague–Dawley (SD) rats (controls) were used for detailed histochemical analyses, including tartrate-resistant acid phosphatase (TRAP), cathepsin K, tissue nonspecific alkaline phosphatase (ALP), phosphoethanolamine/phosphocholine phosphatase 1 (PHOSPHO1), dentin matrix protein 1 (DMP-1), matrix extracellular phosphoglycoprotein (MEPE), sclerostin, osteocalcin staining, silver impregnation, von Kossa staining, and micro-computed tomography (CT).

Results

Micro-CT and hematoxylin-eosin staining demonstrated significantly reduced trabecular bone volume in the femoral metaphyses of SDT-fa/fa rats. Although the number of TRAP-positive osteoclasts per bone surface remained comparable between both groups, SDT-fa/fa rats exhibited reduced areas of ALP-positive and PHOSPHO1-reactive mature osteoblasts/BS. Silver impregnation revealed a well-organized osteocytic lacunar–canalicular system in both groups. However, immunostaining identified aberrant DMP-1 and MEPE expression localized predominantly in the lacunae in SDT-fa/fa rats and in the lacunae and canaliculi of SD rats. Additionally, intense osteocalcin and sclerostin immunoreactivity was detected in osteocytes, along with a higher proportion of osteocalcin-positive osteocytes in SDT-fa/fa rats, distinguishing them from controls.

Conclusion

SDT-fa/fa rats displayed a significant decline in osteoblastic function and distinctive distribution patterns of osteocyte-derived peptides, suggesting that this diabetic model may manifest alterations in osteoblastic activity and the osteocytic lacunar-canalicular network.

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模拟II型糖尿病的自发性糖尿病Torii-Leprfa （SDT-fa/fa）大鼠股骨的组织化学评价

目的：为了阐明糖尿病性骨质疏松的机制，我们对建立肥胖相关2型糖尿病模型的SDT-fa/fa大鼠进行了全面的股骨组织学检查。材料和方法：采用12只30周龄雄性SDT-fa/fa大鼠和年龄匹配的SD大鼠（对照）股骨进行详细的组织化学分析，包括抗酒石酸酸性磷酸酶（TRAP）、组织蛋白酶K、碱性磷酸酶（ALP）、磷酸乙醇胺/磷酸胆碱磷酸酶1 （PHOSPHO1）、牙本质基质蛋白(DMP)-1、基质细胞外磷酸糖蛋白（MEPE）、硬化蛋白、骨钙素染色、银浸渍、von Kossa染色和显微计算机断层扫描（CT）。结果：显微ct和苏木精-伊红染色显示，SDT-fa/fa大鼠股骨干骨小梁体积明显减少。尽管两组之间每个骨表面的trap阳性破骨细胞数量保持相当，但SDT-fa/fa大鼠表现出alp阳性和phospho1反应的成熟成骨细胞/BS面积减少。银浸渍显示两组均有组织良好的骨细胞腔管系统。然而，免疫染色发现DMP-1和MEPE的异常表达主要局限于SDT-fa/fa大鼠的腔隙和SD大鼠的腔隙和小管。此外，在骨细胞中检测到强烈的骨钙素和硬化蛋白免疫反应性，同时在SDT-fa/fa大鼠中检测到更高比例的骨钙素阳性骨细胞，将其与对照组区分开来。结论：SDT-fa/fa大鼠成骨功能明显下降，骨细胞衍生肽分布模式不同，提示该糖尿病模型可能出现成骨细胞活性和骨细胞腔隙网络的改变。

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