Controlled release of Clarithromycin from PLGA microspheres enhances bone regeneration in rabbit calvaria defects

Purpose: To evaluate the controlled release effect of Clarithromycin loaded in PLGA microspheres in a rabbit calvaria defect model. Methods: Clarithromycin-loaded PLGA microspheres (MSPs) were formulated by modified O/W single emulsion/solvent evaporation method. After characterization, in vivo animal experiment was conducted. Four critical size bone defects were created in the calvaria of New Zealand White rabbits (n=21, n=7/time point). The bone defects were randomly designated to 4 groups: Group 1: No augmentation (sham), Group 2: beta-Tricalcium phosphate (β-TCP), Group 3: beta-Tricalcium phosphate (β-TCP) with 0.12 µg clarithromycin, and Group 4: beta-Tricalcium phosphate (β-TCP) with 6.12 µg PLGA microspheres (loaded with 0.12 µg clarithromycin). After 2, 4 and 12 weeks of healing, the levels of bone regeneration were evaluated using micro- computed tomography and histology. Results: The average size of the PLGA microspheres was 26.38 μm that showed 94% encapsulation efficacy with clarithromycin. Clarithromycin release from PLGA microspheres revealed sustained release for around 4 weeks with approximately 50% release of clarithromycin during the first week. In the histological analysis, new bone formation was evident at 2 and 4 weeks of healing in all groups and bone formation increased as a function of healing time in vivo. At 12 weeks, Group 4 showed significantly higher amount of newly formed bone compared to Group 1 (p=0,002). Moreover, during the micro CT exam, Group 4 expressed significantly higher bone formation compared to Group 1 at all time points tested (p=0.00, 0.014, and 0.002 in 2, 4, and 12 weeks, respectively). Conclusions: PLGA microspheres demonstrated initial burst release of clarithromycin followed by a sustained release profile. The in vivo findings showed that β-TCP with clarithromycin-loaded microspheres can enhance bone formation in bone defects