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Expression of nerve growth factor in the callus during fracture healing in a fracture model in aged mice
Bio-Medical Materials and Engineering ( IF 1 ) Pub Date : 2021-09-02 , DOI: 10.3233/bme-211284
Hiroyuki Sekiguchi 1, 2 , Gen Inoue 1 , Shintaro Shoji 1 , Ryo Tazawa 1 , Akiyoshi Kuroda 1 , Masayuki Miyagi 1 , Masashi Takaso 1 , Kentaro Uchida 1, 2
Affiliation  

BACKGROUND:Impaired fracture healing results in extensive and prolonged disability and long-term pain. Previous studies reported that nerve growth factor (NGF) was expressed during fracture healing and that anti-NGF antibody improves physical activity associate with facture pain. However, NGF expression levels in delayed or non-union are not fully understood. OBJECTIVE:We compared chronological changes in NGF in the callus of young mice after femur fracture with those in aged mice after femur fracture as a model of bone fracture in the elderly. METHODS:We used young (age 8 weeks) and aged (age 10 months) male C57BL/6J mice. A fracture was generated in the femur. At 5, 7, 10, 14, 17, and 21 days after creation of a fracture, mRNA expression levels of Col2a1, Col10a1, NGF were evaluated using quantitative PCR. We examined NGF protein expression levels and localization in the callus at day 14 using ELISA and immunohistochemistry, respectively. RESULTS:Expression of NGF in the callus after femur fracture in aged mice was significantly greater than that in young mice at days 5, 7, 10, 17, and 21 days. NGF protein levels in the callus of aged mice were also significantly higher than that in young mice. Immunohistochemical staining showed that NGF was heavily expressed in hypertrophic chondrocytes in the callus in aged mice. CONCLUSIONS:It is suggested that delayed Col2a1 and Col10a1 expression reflects delayed chondrocyte formation and delayed chondrocyte maturation in aged mice and that higher NGF expression in aged mice at day 14 may be associated with the presence of remaining hypertrophic chondrocytes in callus with delaying endochondral ossification.

中文翻译:

老年小鼠骨折模型骨折愈合过程中愈伤组织神经生长因子的表达

背景:骨折愈合受损会导致广泛而长期的残疾和长期疼痛。先前的研究报道,神经生长因子 (NGF) 在骨折愈合过程中表达,抗 NGF 抗体可改善与骨折疼痛相关的身体活动。然而,延迟或不愈合的 NGF 表达水平尚不完全清楚。目的:比较年轻小鼠股骨骨折后骨痂与老年小鼠股骨骨折后骨痂中NGF的时间变化,作为老年骨折模型。方法:我们使用年轻(8 周龄)和老年(10 个月龄)雄性 C57BL/6J 小鼠。股骨发生骨折。在骨折产生后 5、7、10、14、17 和 21 天,使用定量 PCR 评估 Col2a1、Col10a1、NGF 的 mRNA 表达水平。我们分别使用 ELISA 和免疫组织化学在第 14 天检查了 NGF 蛋白表达水平和愈伤组织中的定位。结果:在第5、7、10、17、21天,老年小鼠股骨骨折后愈伤组织中NGF的表达明显高于幼鼠。老年小鼠愈伤组织中 NGF 蛋白水平也显着高于年轻小鼠。免疫组织化学染色显示,NGF 在老年小鼠愈伤组织的肥大软骨细胞中大量表达。结论:表明延迟的 Col2a1 和 Col10a1 表达反映了老年小鼠软骨细胞形成延迟和软骨细胞成熟延迟,并且在第 14 天老年小鼠中较高的 NGF 表达可能与愈伤组织中剩余的肥大软骨细胞的存在与延迟软骨内骨化有关。
更新日期:2021-09-03
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