Expression of nerve growth factor in the callus during fracture healing in a fracture model in aged mice.

Hiroyuki Sekiguchi, Gen Inoue, Shintaro Shoji, Ryo Tazawa, Akiyoshi Kuroda, Masayuki Miyagi, Masashi Takaso, Kentaro Uchida

Bio-medical materials and engineering 2022

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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. 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. 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. 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. 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.

Citation

Hiroyuki Sekiguchi, Gen Inoue, Shintaro Shoji, Ryo Tazawa, Akiyoshi Kuroda, Masayuki Miyagi, Masashi Takaso, Kentaro Uchida. Expression of nerve growth factor in the callus during fracture healing in a fracture model in aged mice. Bio-medical materials and engineering. 2022;33(2):131-137