1. INTRODUCTION:
Titanium and its alloys are considered as essential materials for use in biomedical application [1]. Compared with other biometals used as dental implants, titanium and its alloys have noticeable attracted two due to their exceptional biocompatibility, corrosion resistance, and mechanical properties regarding high hardness, low elastic modulus and low density [2]–[9]. The critical factor of an excellent implant device is that its Young’s moduli should be similar to that of the human bone. The reabsorption of adjacent bone tissue occurs provided that there is a great difference in modulus between the implant device and adjacent bone [10]. Heretofore, commercially α+β-type Ti-6Al-4V alloy is the common materials used for dental…show more content… The procedure area was shaved and cleaned using iodine scrubs. Local anesthesia of 1% Lidocaine and 1:100000 epinephrine was added to reduce hemorrhaging under the skin over the calvaria. The operation started with a midsagittal incision through the skin and the periosteum of the calvaria. A flap was reflected, and the incised periosteum was gently lifted and pushed laterally to visualize the calvaria on both sides of the midline. A critical size defect (8mm of diameter) were created by a trephine bur with an inner diameter of 8mm under saline irrigation. Rinse off the defect with saline to remove bone chips. The membrane was placed to enshroud the defect carefully. The raised periosteum was closed over the membrane with bioabsorbable suture. The reflected flap was repositioned over the periosteum, and the skin was sutured with a 4-0 silk suture (4/0 black silk, Ailee). For three days postoperatively, the rats received subcutaneously administered antibiotic (300µL/kg) for infected precaution. The rats were euthanized after five weeks, and biopsies were conducted to take out block specimens that included the membranes, the surrounding soft tissue, and the underlying calvaria…show more content… Histology and Fluorescent Analysis
After completing Micro-CT scanning, two fixed calvaria blocks in each group were immersed in Villanueva solution (Polysciences) for three days and dehydrated in a graded series of ethanols and acetone 100%, and embedded in methyl methacrylate resin. Sagittal sections (approx. 0.5 mm thickness) were cut, ground to 40 µm thickness, and polished to prepare for hard tissue evaluation. The samples were examined under optical microscopy (EZ4D, Leica).
To assess the mineralized bone dynamic, fluorescent labeling was conducted using two fluorescent agents, Alizarin complexion (red) and calcein (green: both from Sigma) with one animal in each group. Alizarin- complexion solution (1.67 mg/kg with Alizarin 15 mg/ml) was administrated subcutaneously at four weeks prior to sacrifice, while calcein solution (1.25 mg/kg with calcein 20 mg/ml) was administrated at two weeks prior to sacrifice. The fluorescent samples were prepared following the preparation of histology specimens, then were examined using a confocal laser scanning