Limited Impacts of Thermoneutral Housing on Bone Morphology and Mechanical Properties in Growing Female Mice Exposed to External Loading and Raloxifene Treatment

Abstract

Thermoregulation is an important factor that could have physiological consequences on pre-clinical research outcomes. Simply housing mice at thermoneutral temperature has been shown to prevent the well-established loss of cancellous bone that is typical in growing mice. In this study, active tissue formation was induced by non-invasive tibial loading in female mice and combined with raloxifene treatment to assess whether temperature could enhance their combined effects on bone morphology and mechanical properties. It was hypothesized that by removing the cold stress under which normal lab mice are housed, a metabolic boost would allow for further architectural and mechanical improvements in mice exposed to a combination of tibial loading and raloxifene. Ten-week old female C57BL/6J mice were treated with raloxifene, underwent tibial loading to a maximum tensile stress of 2050 με, and were housed in thermoneutral conditions (32 °C) for 6 weeks. We investigated bone morphology through microcomputed tomography (μCT), mechanical properties via four-point bending, and fracture toughness testing. Results confirmed previous work showing a combined effect of external loading and raloxifene which led to greater improvements in most properties than either individual treatment. Counter to the hypothesis, temperature had modest effects on body weight, overall bone size, and trabecular architecture, and most effects were detrimental. Thermoneutrality had no impact on mechanical integrity or fracture toughness. In most cases, the magnitude of temperature-based effects were less robust than either RAL treatment or loading.