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CaMKK2 Inhibition in Enhancing Bone Fracture Healing

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Technical Report,01 May 2015,30 Apr 2016

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Indiana University Bloomington United States

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Ca2 calmodulin CaM-dependent protein kinase kinase 2 CaMKK2 has roles in the anabolic and catabolic pathways of bone remodeling. We hypothesized that targeting CaMKK2 will result in accelerated fracture healing. We generated unilateral mid-shaft fractures using a three-point bending method first described for use in rats by Bonnarens and Einhorn, 1986 in the right femurs of 10-week old anesthetized male mice after first inserting an intramedullary pin 25 gauge needle, approx. 0.5 mm retrograde through the distal condyle of the femur. Radiographic analyses were performed to confirm the location and quality of the fractures. Since CaMKK2 inhibition is associated with anti-inflammatory phenotype, we wanted to determine the optimal time for STO-609 administration. Thus, fractured animals were divided into three groups a saline only n15, b STO-609 from day 0 n15 and c STO-609 from day 7 n15. Tri-weekly intraperitoneal i.p. injections of saline or STO-609 10 molkg mouse body weight were performed for 6 weeks. Progression of fracture healing was monitored through weekly radiographic examination. Fractured callus and non-fractured contralateral femur diaphysis were analyzed by micro computed tomography, histology, immunohistochemistry and histomorphometry. Preliminary results indicate that treatment with STO-609 results in the formation of a robust callus at the fracture site, indicating accelerated healing of femoral fractures following the pharmacological inhibition of CaMKK2.

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