Thermoneutral Housing Did Not Impact the Combined Effects of External Loading and Raloxifene on Bone Morphology and Mechanical Properties in Growing Female Mice

Indiana University-Purdue University Indianapolis (IUPUI)Raloxifene is an FDA-approved selective estrogen receptor modulator (SERM) that improves tissue quality by binding to collagen and increasing the bound water content in the bone matrix in a cell-independent manner. In this thesis, active tissue formation was induced by non-invasive external tibial loading in female mice and combined with raloxifene treatment to assess their combined effect on bone morphology and mechanical properties. Thermoregulation is an important factor that could have physiological consequences on research outcomes, and was introduced as an additional experimental factor in this study. We hypothesized that by removing the mild cold stress under which normal lab animals are housed, a metabolic boost would allow for further architectural and mechanical improvements as a result of the combination of tibial loading and raloxifene treatment. Ten week old female C57BL/6J mice were treated with raloxifene, underwent tibial loading to a strain level of 2050µε and were housed in thermoneutral conditions (32°C) for 6 weeks. We investigated bone morphology through microcomputed tomography (µCT) and mechanical properties via four-point bending and fracture toughness testing. Results indicated a combined improvement by external loading and raloxifene on geometry, particularly in the cancellous region of the bone, and also in bone mechanics leading to greater improvements than either treatment individually. Temperature did not have a robust impact on either bone architecture or mechanical integrity

Morphological and mechanical characterization of bone phenotypes in the Amish G610C murine model of osteogenesis imperfecta

Osteogenesis imperfecta (OI) is a hereditary bone disease where gene mutations affect Type I collagen formation resulting in osteopenia and increased fracture risk. There are several established mouse models of OI, but some are severe and result in spontaneous fractures or early animal death. The Amish Col1a2G610C/+ (G610C) mouse model is a newer, moderate OI model that is currently being used in a variety of intervention studies, with differing background strains, sexes, ages, and bone endpoints. This study is a comprehensive mechanical and architectural characterization of bone in G610C mice bred on a C57BL/6 inbred strain and will provide a baseline for future treatment studies. Male and female wild-type (WT) and G610C mice were euthanized at 10 and 16 weeks (n = 13-16). Harvested tibiae, femora, and L4 vertebrae were scanned via micro-computed tomography and analyzed for cortical and trabecular architectural properties. Femora and tibiae were then mechanically tested to failure. G610C mice had less bone but more highly mineralized cortical and trabecular tissue than their sex- and age-matched WT counterparts, with cortical cross-sectional area, thickness, and mineral density, and trabecular bone volume, mineral density, spacing, and number all differing significantly as a function of genotype (2 Way ANOVA with main effects of sex and genotype at each age). In addition, mechanical yield force, ultimate force, displacement, strain, and toughness were all significantly lower in G610C vs. WT, highlighting a brittle phenotype. This characterization demonstrates that despite being a moderate OI model, the Amish G610C mouse model maintains a distinctly brittle phenotype and is well-suited for use in future intervention studies

The Effect of Single Versus Group μCT on the Detection of Trabecular and Cortical Disease Phenotypes in Mouse Bones

Micro‐computed tomography is a critical assessment tool for bone‐related preclinical research, especially in murine models. To expedite the scanning process, researchers often image multiple bones simultaneously; however, it is unknown if this impacts scan quality and alters the ability to detect differences between experimental groups. The purpose of this study was to assess the effect of multibone scanning on detecting disease‐induced changes in bone microarchitecture and mineral density by group scanning two murine models with known skeletal defects: the Col1a2 G610C/+ model of osteogenesis imperfecta and an adenine‐induced model of chronic kidney disease. Adult male femurs were scanned individually and in random groups of three and eight in a Bruker Skyscan 1172 and 1176, respectively, then assessed for standard trabecular and cortical bone measures. Although scanning methodology altered raw values, with trabecular microarchitecture values more affected than cortical properties, a disease phenotype was still detectable in both group and solo scans. However, tissue mineral density in both trabecular and cortical bone was significantly impacted by group versus solo scanning. Researchers may be able to use small groupings in a single μCT scan to expedite preclinical analyses when the overall bone phenotype is large to decrease costs and increase speed of discoveries; however the details of scanning (single or group) should always be reported