RESUMEN

Voluntary wheel running (VWR) is widely used to study how exercise impacts a variety of physiologies and pathologies in rodents. The primary activity readout of VWR is aggregated wheel turns over a given time interval (most often, days). Given the typical running frequency of mice (∼4 Hz) and the intermittency of voluntary running, aggregate wheel turn counts, therefore, provide minimal insight into the heterogeneity of voluntary activity. To overcome this limitation, we developed a six-layer convolutional neural network (CNN) to determine the hindlimb foot strike frequency of mice exposed to VWR. Aged female C57BL/6 mice (22 months, n = 6) were first exposed to wireless angled running wheels for 2 h/d, 5 days/wk for 3 weeks with all VWR activities recorded at 30 frames/s. To validate the CNN, we manually classified foot strikes within 4800 1-s videos (800 randomly chosen for each mouse) and converted those values to frequency. Upon iterative optimization of model architecture and training on a subset of classified videos (4400), the CNN model achieved an overall training set accuracy of 94%. Once trained, the CNN was validated on the remaining 400 videos (accuracy: 81%). We then applied transfer learning to the CNN to predict the foot strike frequency of young adult female C57BL6 mice (4 months, n = 6) whose activity and gait differed from old mice during VWR (accuracy: 68%). In summary, we have developed a novel quantitative tool that non-invasively characterizes VWR activity at a much greater resolution than was previously accessible. This enhanced resolution holds potential to overcome a primary barrier to relating intermittent and heterogeneous VWR activity to induced physiological responses.

RESUMEN

The anabolic response of aged bone to skeletal loading is typically poor. Efforts to improve mechanotransduction in aged bone have met with limited success. This study investigated whether the bone response to direct skeletal loading is improved by reducing sympathetic suppression of osteoblastic bone formation via ß2AR. To test this possibility, we treated aged wild-type C57BL/6 mice with a selective ß2AR antagonist, butaxamine (Butax), before each of nine bouts of cantilever bending of the right tibia. Midshaft periosteal bone formation was assessed by dynamic histomorphometry of loaded and contralateral tibias. Butax treatment did not alter osteoblast activity of contralateral tibias. Loading alone induced a modest but significant osteogenic response. However, when loading was combined with Butax pretreatment, the anabolic response was significantly elevated compared with loading preceded by saline injection. Subsequent studies in osteoblastic cultures revealed complex negative interactions between adrenergic and mechanically induced intracellular signaling. Activation of ß2AR by treatment with the ß1, ß2-agonist isoproterenol (ISO) before fluid flow exposure diminished mechanically stimulated ERK1/2 phosphorylation in primary bone cell outgrowth cultures and AKT phosphorylation in MC3T3-E1 pre-osteoblast cultures. Expression of mechanosensitive Fos and Ptgs2 genes was enhanced with ISO treatment and reduced with flow in both MC3T3-E1 and primary cultures. Finally, co-treatment of MC3T3-E1 cells with Butax reversed these ISO effects, confirming a critical role for ß2AR in these responses. In combination, these results demonstrate that selective inhibition of ß2AR is sufficient to enhance the anabolic response of the aged skeleton to loading, potentially via direct effects upon osteoblasts. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

RESUMEN

Nearly all exogenous loading models of bone adaptation apply dynamic loading superimposed upon a time invariant static preload (SPL) in order to ensure stable, reproducible loading of bone. Given that SPL may alter aspects of bone mechanotransduction (eg, interstitial fluid flow), we hypothesized that SPL inhibits bone formation induced by dynamic loading. As a first test of this hypothesis, we utilized a newly developed device that enables stable dynamic loading of the murine tibia with SPLs ≥ -0.01 N. We subjected the right tibias of BALB/c mice (4-month-old females) to dynamic loading (-3.8 N, 1 Hz, 50 cycles/day, 10 s rest) superimposed upon one of three SPLs: -1.5 N, -0.5 N, or -0.03 N. Mice underwent exogenous loading 3 days/week for 3 weeks. Metaphyseal trabecular bone adaptation (µCT) and midshaft cortical bone formation (dynamic histomorphometry) were assessed following euthanasia (day 22). Ipsilateral tibias of mice loaded with a -1.5-N SPL demonstrated significantly less trabecular bone volume/total volume (BV/TV) than contralateral tibias (-12.9%). In contrast, the same dynamic loading superimposed on a -0.03-N SPL significantly elevated BV/TV versus contralateral tibias (12.3%) and versus the ipsilateral tibias of the other SPL groups (-0.5 N: 46.3%, -1.5 N: 37.2%). At the midshaft, the periosteal bone formation rate (p.BFR) induced when dynamic loading was superimposed on -1.5-N and -0.5-N SPLs was significantly amplified in the -0.03-N SPL group (>200%). These data demonstrate that bone anabolism induced by dynamic loading is markedly inhibited by SPL magnitudes commonly implemented in the literature (ie, -0.5 N, -1.5 N). The inhibitory impact of SPL has not been recognized in bone adaptation models and, as such, SPLs have been neither universally reported nor standardized. Our study therefore identifies a previously unrecognized, potent inhibitor of mechanoresponsiveness that has potentially confounded studies of bone adaptation and translation of insights from our field. © 2018 The Authors. JBMR Plus Published by Wiley Periodicals, Inc. on behalf of the American Society for Bone and Mineral Research.

RESUMEN

INTRODUCTION: Locked plating leads to improved fixation in osteoporotic bone. In addition, experimental data suggest that overall construct stiffness is increased. Ideal stiffness may be significantly less than that achieved with these locked constructs, and overly stiff constructs may lead to impaired fracture healing and stress concentration at the ends of the plate. In osteoporotic bone, this stiffness mismatch can be even more pronounced. We hypothesized that substituting slots for holes in the near cortex under a locked plate would lead to predictably lower stiffness without diminishing implant stability. METHODS: Osteoporotic bone substitute segments were used. Locking screws and plates were applied to each specimen using either standard holes or near cortical slots. The slots were designed to allow axial displacement of the screw in the near cortex only, while continuing to provide some torsional stability. Mechanical testing was performed using a progressive dynamic displacement load protocol to determine failure and stiffness. Next, cyclic axial loading was performed with a physiologic load for 10,000 cycles to determine change in stiffness with cycling. Outcomes were compared between groups using Mann-Whitney U tests. RESULTS: In the dynamic displacement tests, the slotted specimens reached both maximum load and failure load at a significantly greater displacement than the non-slot group (p=0.008), indicating later failure. The magnitude of the maximum load achieved was no different between groups. In the cyclic loading tests, the axial stiffness in the slotted group was significantly lower (1199 N/mm) than the non-slotted group (3538 N/mm; p<0.05 at all cycles). Stiffness did not change significantly in either group over the course of cycling. DISCUSSION: The ability to predictably adjust the axial stiffness of locked plating constructs is critical, particularly in osteoporotic bone. The use of near cortical slots decreases axial stiffness of locking plates, while maintaining fixation stability. This may allow the surgeon to more closely tailor the construct stiffness to the clinical situation to minimize stiffness mismatches and complications.

Asunto(s)

RESUMEN

OBJECTIVES: Axial stiffness is a critical mechanical parameter in fracture plating. Standard locked plates allow minimal opportunities for stiffness alteration, and current methods are arbitrary and may lead to stiffness mismatch between the implant and bone. Milling the near cortex into a slot allows for an increase in translation of the screw shaft at the near cortex. The purpose of this proof of concept study was to determine the effects of slots on stiffness and their ability to maintain fixation of locking plates under cyclic loading. METHODS: Using segments of fourth-generation synthetic diaphyseal bone, a simulated fracture with a gap was created and locked plates were applied with 4 bicortical locked screws in each fragment. On one fragment, the 4 near cortex holes were sequentially milled to 5 x 6-mm slots. Axial and torsional stiffnesses were determined for constructs with 0 through 4 slots. Specimens with 4 slots then underwent axial cyclic loading to determine the change in stiffness and loss of fixation. Extraction torque was measured for all screws to assess for screw loosening with cycling. RESULTS: In constructs with 4 slots, axial stiffness decreased by 73% (P < 0.05) relative to the 0-slot constructs. Torsional stiffness of the 3- and 4-slot specimens decreased by 20% (SD, 13%; P < 0.05) and 17% (SD, 13%; P < 0.05), respectively, compared with the 0-slot specimens. With cyclic loading, no failures occurred in any specimen. No change in stiffness had occurred by the end of cycling (106% of initial stiffness; SD, 4%; P = 0.96). No screw loosening occurred during cyclic loading. CONCLUSIONS: Purposeful stiffness modulation in fracture fixation is critical to facilitate uneventful fracture healing. Converting near cortical holes to slots allowed selective axial stiffness adjustment without sacrificing fixation stability under cyclic loading. With further refinement, this simple modification of standard implant application may allow the surgeon to decrease the modulus mismatch between plating constructs and bone to decrease the risk of fixation failure.

Asunto(s)

RESUMEN

OBJECTIVE: The objective of the study was to determine which vehicle factors are significantly related to pelvic injury in side impact collisions. Identification of relevant parameters could aid in the reduction of these injuries. METHOD: Side impact crashes from the CIREN database were separated into those in which the occupant sustained a pelvic fracture and those in which no pelvic fracture occurred, although all occupants had serious injuries. A multibody MADYMO model was created of a USDOT SINCAP (U.S. Department of Transportation Side Impact New Car Assessment Program) test of a vehicle with a large center console. RESULTS: From a study of 113 side impact crashes in the ciren database, nearside occupants with pelvic fractures (n = 78) had (i) more door intrusion (mean, 37 vs. 32 cm, p = 0.02) than those who had serious injuries, but not pelvic fractures (ii) a greater likelihood that the lower border of the door intruded more than the upper part (40% vs. 18%, p < 0.025); and (iii) a greater likelihood that their vehicle had a center console (47 vs. 17%, p < 0.005). Other parameters such as occupant age, weight, gender, vehicle weight, and struck vehicle speed change were not significantly different. MADYMO modeling showed that with a center console, an initial positive pelvic acceleration occurred at about 30 msec, followed at about 45 msec by a second acceleration peak in the opposite direction. Reducing console stiffness reduced the second acceleration but not the initial peak. Allowing the seat to translate laterally when contacted by the door reduced the initial pelvic acceleration by 50% and eliminated the second acceleration peak. CONCLUSIONS: Redesigning the center console using less stiff materials and allowing some lateral translation of the seat could aid in reducing pelvic injuries in side impact collisions.

Asunto(s)

RESUMEN

The material properties of the anterior cruciate ligament (ACL) in female rats with normal estrous cycles were compared to those regulated by oral contraceptive steroids. Forty female Sprague-Dawley rats were divided into two groups: an experimental group received daily ethinylestradiol and levonorgestrel in a dosing model designed to simulate a typical oral contraception regime in humans, while a control group received daily oral placebo. After eight cycles, six rats from each group underwent daily phlebotomy to measure serum estradiol and progesterone levels over the course of a single 5-day estrous cycle. Significant differences between groups were found for the area under the curve of blood progesterone levels versus time over the length of the estrous cycle (P=0.02). After 12 cycles, the rats were euthanized and one femur-ACL-tibia complex from each animal was dynamically loaded to failure. The ACLs from the rats in the experimental group had significantly decreased average and tangent stiffness, (P=0.002 and 0.0001, respectively), and significantly increased elongation (P=0.002) and total energy absorbed (P=0.03), or greater toughness than controls. In rats, it appears that the administration of reproductive hormones designed to simulate typical oral contraception in humans alters the mechanical properties of the rat ACL.

Asunto(s)