Oral implant osseointegration model in C57Bl/6 mice: microtomographic, histological, histomorphometric and molecular characterization

Abstract Despite the successful clinical application of titanium (Ti) as a biomaterial, the exact cellular and molecular mechanisms responsible for Ti osseointegration remains unclear, especially because of the limited methodological tools available in this field. Objective: In this study, we present a microscopic and molecular characterization of an oral implant osseointegration model using C57Bl/6 mice. Material and Methods: Forty-eight male wild-type mice received a Ti implant on the edentulous alveolar crest and the peri-implant sites were evaluated through microscopic (μCT, histological and birefringence) and molecular (RealTimePCRarray) analysis in different points in time after surgery (3, 7, 14 and 21 days). Results: The early stages of osseointegration were marked by an increased expression of growth factors and MSC markers. Subsequently, a provisional granulation tissue was formed, with high expression of VEGFb and earlier osteogenic markers (BMPs, ALP and Runx2). The immune/inflammatory phase was evidenced by an increased density of inflammatory cells, and high expression of cytokines (TNF, IL6, IL1) chemokines (CXCL3, CCL2, CCL5 and CXC3CL1) and chemokine receptors (CCR2 and CCR5). Also, iNOS expression remained low, while ARG1 was upregulated, indicating predominance of a M2-type response. At later points in time, the bone matrix density and volume were increased, in agreement with a high expression of Col1a1 and Col21a2. The remodelling process was marked by peaks of MMPs, RANKL and OPG expression at 14 days, and an increased density of osteoclasts. At 21 days, intimate Ti/bone contact was observed, with expression of final osteoblast differentiation markers (PHEX, SOST), as well as red spectrum collagen fibers. Conclusions: This study demonstrated a unique molecular view of oral osseointegration kinetics in C57Bl/6 mice, evidencing potential elements responsible for orchestrating cell migration, proliferation, ECM deposition and maturation, angiogenesis, bone formation and remodeling at the bone-implant interface in parallel with a novel microscopic analysis.

Corrosion behavior of zirconia in acidulated phosphate fluoride

ABSTRACT Objective The corrosion behavior of zirconia in acidulated phosphate fluoride (APF) representing acidic environments and fluoride treatments was studied. Material and Methods Zirconia rods were immersed in 1.23% and 0.123% APF solutions and maintained at 37°C for determined periods of time. Surfaces of all specimens were imaged using digital microscopy and scanning electron microscopy (SEM). Sample mass and dimensions were measured for mass loss determination. Samples were characterized by powder X-ray diffraction (XRD) to detect changes in crystallinity. A biosensor based on electrochemical impedance spectroscopy (EIS) was used to detect ion dissolution of material into the immersion media. Results Digital microscopy revealed diminishing luster of the materials and SEM showed increased superficial corrosion of zirconia submerged in 1.23% APF. Although no structural change was found, the absorption of salts (sodium phosphate) onto the surface of the materials bathed in 0.123% APF was significant. EIS indicated a greater change of impedance for the immersion solutions with increasing bathing time. Conclusion Immersion of zirconia in APF solutions showed deterioration limited to the surface, not extending to the bulk of the material. Inferences on zirconia performance in acidic oral environment can be elucidated from the study.