Predicting altered bone biomechanics in juvenile mice: insights from microgravity simulation, loading interventions, and Raman Spectroscopy.

BACKGROUND: Microgravity, a condition experienced in a spatial environment, poses unique challenges to the skeletal system, particularly in juvenile organisms. This study aimed to investigate alterations in bone biomechanics of juvenile mice due to unloading - that simulates microgravity in the laboratory-and the effects of a bone-loading intervention. We compared bone compositional and mechanical properties between 21-six-week-old C57Bl/6 from a control group (wild type) and a group that underwent a tail-suspension unloading protocol to mimic microgravity (MG). The second group (MG) experienced additional in vivo loading protocol (MG + LDG) on the right hind leg, where dynamic compressive loading was applied to the right knee using a custom-built loading device. RESULTS: Our results show that after two weeks, we successfully induced bone alterations by (i) decreasing the energy dissipated before fracture and (ii) decreasing the yield and maximum stress. In addition, we showed that Mineral to matrix component [ν1PO4/Amide I], Carbonate to Amide [CO3/Amide I], and Crystallinity [1/FWHM(ν1PO4)] are strongly linked in physiological bone but not in microgravity even after loading intervention. While Crystallinity is very sensitive to bone deformation (strain) alterations coming from simulated microgravity, we show that Carbonate to Amide [CO3/Amide I] - a common marker of turnover rate/remodeling activity-is a specific predictor of bone deformation for bone after simulated microgravity. Our results also invalidate the current parameters of the loading intervention to prevent bone alterations entirely in juvenile mice. CONCLUSIONS: Our study successfully induced bone alterations in juvenile mice by using an unloading protocol to simulate microgravity, and we provided a new Raman Spectroscopy (RS) dataset of juvenile mice that contributes to the prediction of cortical bone mechanical properties, where the degree of interrelationship for RS data for physiological bone is improved compared to the most recent evidence.

Subchondral bone alterations in a novel model of intermediate post traumatic osteoarthritis in mice.

In Post Traumatic Osteoarthritis (PTOA), hypomineralization and increased remodeling of the Subchondral bone (SB) are the first stages of tissue alterations. Although these alterations are well depicted and one of the main targets in OA intervention, the link between SB compositional and mechanical properties alterations during OA progression remains scarce in the literature. Here, we hypothesized that SB shows - right after the first sign of gait pattern changes - a decrease in SB tissue formation depicted by (i) a decrease in thickness, (ii) a lower nanoscopic stiffness, and (iii) a decrease in mineral and collagen maturity. To test our hypothesis, we investigated PTOA in female C57Bl6 mice's right knee (n = 13 control group [CL] and n = 27 PTOA group) by using Gait Analysis, Histomorphometry, Nanoindentation, and Raman Spectroscopy (RS). We showed (i) an increased OA histological grade, (ii) a decrease in Cartilage and SB thickness, and (ii) an increase of stance time and stride length on both limbs. The lateral condyle - where the main forces were applied - of mice with PTOA decreased in the degree of mineralization and crystal size and presented a lower Modulus of Elasticity (E). However, while no difference was observed regarding collagen or mineral-related compositional RS properties, we depicted higher crystallinity in the medial condyle than the lateral condyle in the PTOA group, which we did not observe in the control group. Our study depicts an early onset of intermediate PTOA where SB nanoscopic stiffness decreases while the degree of mineralization is not severely altered yet.

Bones of teleost fish demonstrate high fracture strain.

The endoskeleton of teleosts (bony fish) includes a vertebral spine with articulating rib bones (RBs) similar to humans and further encompasses mineralized tissues that are not found in mammals, including intermuscular bones (IBs). RBs form through endochondral ossification and protect the inner organs, and IBs form through intramembranous ossification within the myosepta and play a role in force transmission and propulsion during locomotion. Based on previous findings suggesting that IBs show a much higher ability for fracture strain compared to mammalian bones, this study aims to investigate whether this ability is general to teleost bones or specific to IBs. We analyzed RBs and IBs of 25 North Atlantic Herring fish. RBs were analyzed using micro-mechanical tensile testing and micro-computed tomography, and both RB and IB were additionally analyzed with Raman spectroscopy. Based on our previous results from IB, we found that RBs are more elastically deformable (on average, 50% higher yield strain and 115% higher elastic work) and stronger (55% higher fracture stress) than values reported for IBs. However, these differences were neither associated with a higher Young's modulus nor a higher degree of mineralization in RBs. Astonishingly, RBs and IBs showed similar fracture strains (12-15% on average, reaching up to 20%), reflecting a much higher ability for tensile deformation than reported for mammalian bone, and further highlighting the biomimetic potential of teleost fish bones for inspiring innovative biomaterials.

Microstructure, mineral and mechanical properties of teleost intermuscular bones.

There is an increasing interest in understanding teleost bone biomechanics in several scientific communities, for instance as interesting biomaterials with specific structure-function relationships. Intermuscular bones of teleost fish have previously been described to play a role in the mechanical force transmission between muscle and bone, but their biomechanical properties are not yet fully described. Here, we have investigated intermuscular bones (IBs) of the North Atlantic Herring with regard to their structure and micro-architecture, mineral-related properties, and micro-mechanical tensile properties. A total of 115 IBs from 18 fish were investigated. One cohort of IBs, containing 20 bones from 2 smaller fish and 23 bones of 3 larger fish, was used for mechanical testing, wide-angle X-ray scattering, and scanning electron microscopy. Another cohort, containing 36 bones from 7 smaller fish and 36 bones from 6 larger fish, was used for microCT. Results show some astonishing properties of the IBs: (i) IBs present higher ductility, lower Young's modulus but similar strength and TMD (Tissue Mineral Density) compared to mammalian bone, and (ii) IBs from small fish were 49% higher in Young's modulus than fish bones from larger fish while their TMD was not statistically different and crystal length was 8% higher in large fish bones. Our results revealed that teleost IB presents a hybrid nature of soft and hard tissue that differs from other bone types, which might be associated with their evolution from mineralized tendons. This study provides new data regarding teleost fish bone biomechanical and micro-structural properties.