Altered bone development and turnover in transgenic mice over-expressing lipocalin-2 in bone

Lipocalin-2 (LCN2) is a protein largely expressed in many tissues, associated with different biological phenomena such as cellular differentiation, inflammation and cancer acting as a survival/apoptotic signal. We found that LCN2 was expressed during osteoblast differentiation and we generated transgenic (Tg) mice over-expressing LCN2 in bone. Tg mice were smaller and presented bone microarchitectural changes in both endochondral and intramembranous bones. In particular, Tg bones displayed a thinner layer of cortical bone and a decreased trabecular number. Osteoblast bone matrix deposition was reduced and osteoblast differentiation was slowed-down. Differences were also observed in the growth plate of young transgenic mice where chondrocyte displayed a more immature phenotype and a lower proliferation rate. In bone marrow cell cultures from transgenic mice, the number of osteoclast progenitors was increased whereas in vivo it was increased the number of mature osteoclasts expressing tartrate-resistant acid phosphatase (TRAP). Finally, while osteoprotegerin (OPG) levels remained unchanged, the expression of the conventional receptor activator of nuclear factor-κB ligand (RANKL) and of the IL-6 was enhanced in Tg mice. In conclusion, we found that LCN2 plays a role in bone development and turnover having both a negative effect on bone formation, by affecting growth plate development and interfering with osteoblast differentiation, and a positive effect on bone resorption by enhancing osteoclast compartment

In Vivo Regenerative Properties of Coralline-Derived (Biocoral) Scaffold Grafts in Human Maxillary Defects: Demonstrative and Comparative Study with Beta-Tricalcium Phosphate and Biphasic Calcium Phosphate by Synchrotron Radiation X-Ray Microtomography

Background: In recent years, there has been interest on the fabrication of systems using particulates or block-based approach for bone tissue engineering (TE) scaffolds, possessing porous interconnected structures. In fact, these particular morphologies greatly increase the surface area for more chemical and biological reactions to take place. Purpose: This study was designed to demonstrate the unique capability of the synchrotron radiation x-ray microtomography (micro-CT) in offering an advanced characterization of coralline-derived (Biocoral) biomaterials placed in human maxillary defects as it allows, in a nondestructive way, a complete, precise, and high-resolution three-dimensional analysis of their microstructural parameters. Moreover, the comparison between Biocoral and other biomaterials was explored to understand the mechanism of their biological behavior as bone substitute. Materials and Methods: Implant survival, bone regeneration, graft resorption, neovascularization, and morphometric parameters (including anisotropy and connectivity index of the structures) were evaluated by micro-CT in Biocoral and the other biomaterials after 6 to 7 months from implantation in human maxillary bone defects. Results: After the in vivo tests, a huge amount of bone was detected in the retrieved Biocoral-based samples, coupled with a good rate of biomaterial resorption and the formation of a homogeneous and rich net of new vessels. The morphometric parameters were comparable to those obtained in the biphasic calcium phosphate-based control, with the exception of the connectivity index for which this control exhibited the most well-connected structure. This last result, together with those referred to the poor performances of the b-tricalcium phosphate block-based sample, suggests that the particular scaffold morphology may play a role in the hunt the optimal scaffold structure to be implanted. Conclusion: In this limited study, implant success rate seems not strictly dependent on the biomaterial that is used, but on the scaffold morphology. Micro-CT technique was demonstrated to play a fundamental role in advanced characterization of bone TE constructs

Bone Turnover in Wild Type and Pleiotrophin-Transgenic Mice Housed for Three Months in the International Space Station (ISS)

Bone is a complex dynamic tissue undergoing a continuous remodeling process. Gravity is a physical force playing a role in the remodeling and contributing to the maintenance of bone integrity. This article reports an investigation on the alterations of the bone microarchitecture that occurred in wild type (Wt) and pleiotrophin-transgenic (PTN-Tg) mice exposed to a near-zero gravity on the International Space Station (ISS) during the Mice Drawer System (MDS) mission, to date, the longest mice permanence (91 days) in space. The transgenic mouse strain over-expressing pleiotrophin (PTN) in bone was selected because of the PTN positive effects on bone turnover. Wt and PTN-Tg control animals were maintained on Earth either in a MDS payload or in a standard vivarium cage. This study revealed a bone loss during spaceflight in the weight-bearing bones of both strains. For both Tg and Wt a decrease of the trabecular number as well as an increase of the mean trabecular separation was observed after flight, whereas trabecular thickness did not show any significant change. Non weight-bearing bones were not affected. The PTN-Tg mice exposed to normal gravity presented a poorer trabecular organization than Wt mice, but interestingly, the expression of the PTN transgene during the flight resulted in some protection against microgravity’s negative effects. Moreover, osteocytes of the Wt mice, but not of Tg mice, acquired a round shape, thus showing for the first time osteocyte space-related morphological alterations in vivo. The analysis of specific bone formation and resorption marker expression suggested that the microgravity-induced bone loss was due to both an increased bone resorption and a decreased bone deposition. Apparently, the PTN transgene protection was the result of a higher osteoblast activity in the flight mice

Advanced Microstructural Characterizations of Some Biomaterials and Scaffolds for Regenerative Orthopaedics

In the last decades, very significant advances have been made for what concerns bone and joint substitution and in the repair and regeneration of bone defects. Though some strong requirements are still to be met, biomaterials for these purposes have known an impressive evolution, for what concerns their mechanical behaviour, their bioresorbability and finally their capability to generate new bone tissue in a stable way in long periods. The validation of such materials necessarily depends on a suitable characterization of their properties. In this article a brief review of some works in this field, carried out by the authors’ research group, is presented. It was shown in particular how advanced experimental methods, such as synchrotron radiation μCT and synchrotron radiation diffraction can offer very important information, can be not only complementary methods to more standard techniques (electron microscopy, X-ray diffraction), but can also offer the possibility to measure parameters that cannot be obtained otherwise

MicroCT and optical coherence tomography imagistic assessment of the dental roots adhesive

Several obturation methods are available today to study the 3D filling of the root canal. There are also several methods capable to evaluate the ability to seal apically the root canals. However, the common methods of investigation are invasive; they also lead to the destruction of the samples. If the sectioning differs slightly from the desired area, the investigation is non-conclusive regarding the micro-leakages. Also, although the use of Cone-Beam Micro Computer Tomography (CBCT) appears to be most promising for endodontic purposes, its effective radiation doses are higher than with conventional intra-oral and panoramic imaging. In contrast, enface (ef) Optical Coherence Tomography (OCT) proves to be efficient for the investigation of material defects of dental restorations, dental materials, and micro-leakage at the interfaces, where the penetration depth depends on the material. Therefore, ef OCT has been proposed in our studies as a potential tool for in vivo endodontic imaging. Twenty five recently extracted human maxillary molars were selected for the study for caries or periodontal reasons. The pulp chambers were completely opened, the dental pulp was removed, and the root canals were shaped. Silver nanoparticles were used in half of the samples in order to increase the scattering of the adhesive material in comparison with the dental roots walls. The sample teeth were then probed using Time Domain (TD) OCT working at 1300 nm. A synchrotron radiation X-Ray microCT experiment was also performed. The imagistic results pointed out the efficiency of the silver nanoparticle layer used in order to increase the scattering of the root canal adhesive scattering for the OCT non-invasive investigation. MicroCT allowed for obtaining qualitative data related to the depth penetration of the root canal adhesive into the dentin walls. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only