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Item Assessing the inter- and intra-animal variability of in vivo OsteoProbe skeletal measures in untreated dogs(Elsevier, 2016-12) McNerny, Erin M.B.; Organ, Jason M.; Wallace, Joseph M.; Newman, Christopher L.; Brown, Drew M.; Allen, Matthew R.; Department of Anatomy and Cell Biology, School of MedicineThe OsteoProbe is a second-generation reference point indentation (RPI) device without a reference probe that is designed to simplify RPI testing for clinical use. Successful clinical implementation of the OsteoProbe would benefit from a better understanding of how its output, bone material strength index (BMSi), relates to the material properties of bone and under what conditions it reliably correlates with fracture risk. Large animal models have the potential to help fill this knowledge gap, as cadaveric studies are retrospective and limited by incomplete patient histories (including the potential use of bone matrix altering drugs such as bisphosphonates). The goal of this study was to assess the intra and inter-animal variability of OsteoProbe measures in untreated beagle dogs (n = 12), and to evaluate this variability in comparison to traditional mechanical testing. OsteoProbe measurements were performed in vivo on the left tibia of each dog and repeated 6 months later on the day of sacrifice. Within-animal variation of BMSi (CV of 5–10 indents) averaged 8.9 and 9.0% at the first and second timepoints, respectively. In contrast, inter-animal variation of BMSi increased from 5.3% to 9.1%. The group variation of BMSi was on par with that of traditional 3-point mechanical testing; inter-animal variation was 10% for ultimate force, 13% for stiffness, and 12% for total work as measured on the femur. There was no significant change in mean BMSi after 6 months, but the individual change with time across the 12 dogs was highly variable, ranging from − 12.4% to + 21.7% (mean 1.6%, SD 10.6%). No significant correlations were found between in vivo tibia BMSi and femur mechanical properties measured by ex vivo 3-pt bending, but this may be a limitation of sample size or the tests being performed on different bones. No relationship was found between BMSi and tissue mineral density, but a strong positive correlation was found between BMSi and tibia cortical thickness (ρ = 0.706, p = 0.010). This report shows that while the OsteoProbe device has inter-individual variability quite similar to that of traditional mechanical testing, the longitudinal changes show high levels of heterogeneity across subjects. We further highlight the need for standardization in post-testing data processing and further study of the relationships between OsteoProbe and traditional mechanical testing.Item Geometric Characterization of Local Changes in Tungsten Microneedle Tips after In-Vivo Insertion into Peripheral Nerves(MDPI, 2022) Sergi, Pier Nicola; Jensen, Winnie; Yoshida, Ken; Biomedical Engineering, Purdue School of Engineering and TechnologyPeripheral neural interfaces are used to connect the peripheral nervous system to high-tech robotic devices and computer interfaces. Soft materials are nowadays used to build the main structural part of these interfaces because they are able to mimic the mechanical properties of peripheral nerves. However, if on the one hand soft materials provide effective connections, reducing mechanical mismatch with nervous tissues and creating a close contact between active sites and neural fibers, on the other hand, most of them are not mechanically stable during implantation. As a consequence, tungsten (W) microneedles are used to insert soft neural interfaces, because they are able to pierce the peripheral nervous tissue because of their high stiffness. Nevertheless, this stiffness cannot prevent microneedles from local microscopic structural damage, even after successful insertions. In addition, the nature of this damage is not totally clear. Therefore, this work aimed at quantitatively investigating the phenomenological changes of the microneedles’ tip shape after insertion into the in vivo peripheral nerves. In particular, a quantification of the interactions between peripheral nerves and W microneedles was proposed through the Oliver-Pharr formula, and the interaction force was found to be directly proportional to the power < m > = 2.124 of the normalized indentation depth. Moreover, an experimental correlation between insertion force and the opening tip angle was described together with an assessment of the minimum diameter to effectively puncture the peripheral nervous tissue. Finally, a computational framework was presented to describe the local changes affecting the microneedles’ tip shape. This approach was able to detect a bulging phenomenon along with the microneedle tips with a characteristic amplitude of approximately 100 μm, and a folding phenomenon, with a characteristic mean amplitude of less than 20 μm, affecting the extreme ending sections of the microneedle tips. These geometrical changes were related to the synergistic action of interaction forces likely resulting in compression and elastic instability of the tip.Item Substrate strain mitigates effects of β-aminopropionitrile-induced reduction in enzymatic crosslinking(Springer, 2019-09-03) Canelón, Silvia P.; Wallace, Joseph M.; Biomedical Engineering, School of Engineering and TechnologyEnzymatic crosslinks stabilize type I collagen and are catalyzed by lysyl oxidase (LOX), a step interrupted through β-aminopropionitrile (BAPN) exposure. This study evaluated dose-dependent effects of BAPN on osteoblast gene expression of type I collagen, LOX, and genes associated with crosslink formation. The second objective was to characterize collagen produced in vitro after exposure to BAPN, and to explore changes to collagen properties under continuous cyclical substrate strain. To evaluate dose-dependent effects, osteoblasts were exposed to a range of BAPN dosages (0–10 mM) for gene expression analysis and cell proliferation. Results showed significant upregulation of BMP-1, POST, and COL1A1and change in cell proliferation. Results also showed while the gene encoding LOX was unaffected by BAPN treatment, other genes related to LOX activation and matrix production were upregulated. For the loading study, the combined effects of BAPN and mechanical loading were assessed. Gene expression was quantified, atomic force microscopy was used to extract elastic properties of the collagen matrix, and Fourier Transform infrared spectroscopy was used to assess collagen secondary structure for enzymatic crosslinking analysis. BAPN upregulated BMP-1 in static samples and BAPN combined with mechanical loading downregulated LOX when compared to control-static samples. Results showed a higher indentation modulus in BAPN-loaded samples compared to control-loaded samples. Loading increased the mature to immature crosslink ratios in control samples, and BAPN increased the height ratio in static samples. In summary, effects of BAPN (upregulation of genes involved in crosslinking, mature/immature crosslinking ratios, upward trend in collagen elasticity) were mitigated by mechanical loading.