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Bone Defect model

Prior to the surgical intervention, the animals were anaesthetized using sevoflurane inhalation anaesthesia (4 – 5% induction; 2 – 3% maintenance). Then, the skin that covered the skull, around the incision area, was shaved and disinfected with iodopovidone solution.A subcutaneous injection of 0.3– 0.4 mL of 1% lidocaine, used as a local anaesthetic, was administered along the sagittal midline of the skull. A sur- gical blade was then used to perform a midline incision, from the nasofrontal area to an anterior location of the exter- nal occipital protuberance.The skin was then reflected bilaterally and a new midline incision, of around 3 cm, allowed the division of the subcutaneous fascia and the bilateral reflection of the periosteal flaps following blunt dissection, to expose the calvarial bone surface.Careful drilling with an 8 mm diameter tre- phine bur, mounted on a low-speed dental handpiece, was done around the sagittal suture, and a standardized, round, segmental defect was made. During drilling, the area was continuously irrigated with sterile saline solution and extreme care was taken not to damage the underlying blood vessels and sinus.The obtained calvarial disk was carefully removed to avoid tearing of the subjacent cranial structures.