[99mTc]cFLFLF for Early Diagnosis and Therapeutic Evaluation in a Rat Model of Acute Osteomyelitis.

Purpose: Early diagnosis and therapeutic monitoring of acute osteomyelitis (AO) is challenging. Here, we use a polyethylene glycol (PEG)ylated chemotactic peptide cinnamoyl-F-(D)L-F-(D)L-F (cFLFLF) conjugated with hydrazinonicotinamide (HYNIC) and labeled with Tc-99m ([(99m)Tc]cFLFLF) to image AO in a rat model and to validate its efficacy in early diagnosis and therapeutic evaluation of AO.

Procedures: Forty rats were divided into eight groups of five each. Groups A, B, C, G, and H were AO models, and D, E, and F were sham controls. Groups A and D underwent [(99m)Tc]cFLFLF scintigraphy, groups B and E underwent [(99m)Tc]methylene diphosphonate ([(99m)Tc]MDP) bone scan, and groups C and F underwent 2-deoxy-2-[(18)F]fluoro-D-glucose ([(18)F]FDG) positron emission tomography (PET)/computed tomography (CT) scan. [(99m)Tc]cFLFLF biodistribution was assessed in group G. The response to antibiotic therapy was evaluated using [(99m)Tc]cFLFLF scintigraphy in group H. Conventional radiographs were obtained following scintigraphy. Ratios of infected or sham-operated tibia to the opposite tibia (T/B) were calculated. Immediately after the imaging studies, infected tibias were excised and underwent histopathological analysis and immunohistochemistry staining.

Results: AO was present in all rats of groups A, B, C, G, and H. Total histological scores were not significantly different among groups A, B, and C (F = 0.34, p = 0.71). The biodistribution results revealed significant uptake and excellent retention of [(99m)Tc]cFLFLF in the infected tibia. [(99m)Tc]cFLFLF scintigraphy and [(99m)Tc]MDP bone scan both detected AO. The mean T/B ratio of [(99m)Tc]cFLFLF scintigraphy 1 h postinjection was 2.09-fold higher than that of [(99m)Tc]MDP bone scan (t = 13.81, p <0.001). The mean T/B ratio of [(18)F]FDG PET/CT scan was not significantly different from the control group F (t = 2.17, p = 0.062). [(99m)Tc]cFLFLF scintigraphy revealed a significant attenuation of inflammation in group H following a 3-week antibiotic treatment, which was verified by histopathological analysis and immunohistochemistry staining.

Conclusion: Our results suggest that the specificity and image quality of [(99m)Tc]cFLFLF are superior to those of the [(99m)Tc]MDP and [(18)F]DFG imaging probes currently used for early diagnosis of AO. Furthermore, [(99m)Tc]cFLFLF was able to effectively evaluate the therapeutic response to antibiotic treatment of AO. Our data suggest that [(99m)Tc]cFLFLF is a promising imaging agent for detection of infectious diseases.