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Item Cell Mechanosensitivity to Extremely Low Magnitude Signals is Enabled by a LINCed Nucleus(Wiley, 2015-06) Uzer, Gunes; Thompson, William R.; Sen, Buer; Xie, Zhihui; Yen, Sherwin S.; Miller, Sean; Bas, Guniz; Styner, Maya; Rubin, Clinton T.; Judex, Stefan; Burridge, Keith; Rubin, Janet; Physical Therapy, School of Health and Rehabilitation SciencesA cell's ability to recognize and adapt to the physical environment is central to its survival and function, but how mechanical cues are perceived and transduced into intracellular signals remains unclear. In mesenchymal stem cells (MSCs), high-magnitude substrate strain (HMS, ≥2%) effectively suppresses adipogenesis via induction of focal adhesion (FA) kinase (FAK)/mTORC2/Akt signaling generated at FAs. Physiologic systems also rely on a persistent barrage of low-level signals to regulate behavior. Exposing MSC to extremely low-magnitude mechanical signals (LMS) suppresses adipocyte formation despite the virtual absence of substrate strain (<0.001%), suggesting that LMS-induced dynamic accelerations can generate force within the cell. Here, we show that MSC response to LMS is enabled through mechanical coupling between the cytoskeleton and the nucleus, in turn activating FAK and Akt signaling followed by FAK-dependent induction of RhoA. While LMS and HMS synergistically regulated FAK activity at the FAs, LMS-induced actin remodeling was concentrated at the perinuclear domain. Preventing nuclear-actin cytoskeleton mechanocoupling by disrupting linker of nucleoskeleton and cytoskeleton (LINC) complexes inhibited these LMS-induced signals as well as prevented LMS repression of adipogenic differentiation, highlighting that LINC connections are critical for sensing LMS. In contrast, FAK activation by HMS was unaffected by LINC decoupling, consistent with signal initiation at the FA mechanosome. These results indicate that the MSC responds to its dynamic physical environment not only with "outside-in" signaling initiated by substrate strain, but vibratory signals enacted through the LINC complex enable matrix independent "inside-inside" signaling.Item Characterization and Measurement of Hybrid Gas Journal Bearings(2012-08-28) Lawrence, Tom Marquis; Kemple, Marvin D.; Decca, Ricardo; Joglekar, Yogesh; Petrache, Horia; Akay, Hassan; Krousgrill, Charles MortonThis thesis concentrates on the study of hybrid gas journal bearings (bearings with externally pressurized mass addition). It differs from most work in that it goes back to “basics” to explore the hydrodynamic phenomena in the bearing gap. The thesis compares geometrically identical bearings with 2 configurations of external pressurization, porous liners where mass-addition compensation is varied by varying the liner’s permeability, and bushings with 2 rows of 6 feedholes where the mass-addition compensation is varied by the feedhole diameter. Experimentally, prototype bearings with mass-addition compensation that spans 2 orders of magnitude with differing clearances are built and their aerostatic properties and mass addition characteristics are thoroughly tested. The fundamental equations for compressible, laminar, Poiseuille flow are used to suggest how the mass flow “compensation” should be mathematically modeled. This is back-checked against the experimental mass flow measurements and is used to determine a mass-addition compensation parameter (called Kmeas) for each prototype bushing. In so doing, the methodology of modeling and measuring the mass addition in a hybrid gas bearing is re-examined and an innovative, practical, and simple method is found that makes it possible to make an “apples-to-apples” comparison between different configurations of external pressurization. This mass addition model is used in conjunction with the Reynolds equation to perform theory-based numerical analysis of virtual hybrid gas journal bearings (CFD experiments). The first CFD experiments performed use virtual bearings modeled to be identical to the experimental prototypes and replicate the experimental work. The results are compared and the CFD model is validated. The ontological significance of appropriate dimensionless similitude parameters is re-examined and a, previously lacking, complete set of similitude factors is found for hybrid bearings. A new practical method is developed to study in unprecedented detail the aerostatic component of the hybrid bearings. It is used to definitively compare the feedhole bearings to the porous liner bearings. The hydrostatic bearing efficiency (HBE) is defined and it is determined that the maximum achievable hydrostatic bearing efficiency (MAHBE) is determined solely by the bearing’s mass addition configuration. The MAHBE of the porous liner bearings is determined to be over 5 times that of the feedhole bearings. The method also presents a means to tune the Kmeas to the clearance to achieve the MAHBE as well as giving a complete mapping of the hitherto misunderstood complex shapes of aerostatic load versus radial deflection curves. This method also rediscovers the obscure phenomenon of static instability which is called in this thesis the “near surface effect” and appears to be the first work to present a practical method to predict the range of static instability and quantify its resultant stiffness fall-off. It determines that porous liner type bearings are not subject to the phenomenon which appears for feedhole type bearings when the clearance exceeds a critical value relative to its mass-addition compensation. The standing pressure waves of hydrostatic and hybrid bearings with the 2 configurations of external pressurization as well as a geometrically identical hydrodynamic bearing are studied in detail under the methodology of the “CFD microscope”. This method is used to characterize and identify the development, growth, and movement of the pressure wave extrema with increased hydrodynamic action (either increasing speed or increasing eccentricity). This method is also used to determine the “cause” of the “near surface effect”. A gedanken experiment is performed based on these results which indicates that a bearing with a “stronger aerostatic strength” component should be more stable than one with a low aerostatic strength component. Numerical instability “speed limits” are found that are also related to the hydrostatic strength of the bearing. The local conditions in the standing waves are characterized in terms of their local Mach number, Knudsen number, Reynolds number, and Taylor Number. It is concluded that low eccentricity bearing whirl can be attributed to the off load-line orientation of the bearing load force caused by the overlay of the hydrodynamic bearing standing wave onto the hydrostatic bearing wave of the hybrid bearing, whereas it is hypothesized that aperiodic and random self-excited vibration which occurs at high eccentricity, as reported in the literature, is probably due to shock waves, turbulence, near surface effect, and slip at local areas of the standing wave.Item Effects of vibration forces on maxillary expansion and orthodontic tooth movement(2015) Aldosari, Mohammad Abdullah M.; Liu, Sean Shih-Yao; Chen, Jie; Katona, Thomas R.; Ghoneima, Ahmed A. M.; Chu, Tien-Min GabrielVibration forces (VF) have been shown to alter the formative and resorptive activities of bone. Studies have investigated the use of VF in applications such as the treatment of osteoporosis, bone fracture healing and implant osteointegration with favorable results. In dentistry, orthodontic tooth movement and maxillary suture expansion are common procedures typically requiring prolonged treatment durations with high relapse rates. We hypothesized that local, intermittent VF applications can enhance bone formation during rapid maxillary expansion and accelerate orthodontic tooth movement. Moreover, we also investigated expression of periostin/OSF-2, an adhesion molecule implicated in the formation of bone during maxillary suture expansion. Our results showed that intermittent VF significantly increased bone volume density of the expended palatal bone but limited the amount of palatal expansion and mineral apposition rate at the suture margins. Also, intermittent VF forces did not show statistically significant acceleration of orthodontic tooth movement but significantly enhanced bone volume density of the interradicular bone after tooth movement. Maxillary expansion was also shown to induce the expression of periostin which was proportional to the magnitude of the expansion force with increased bone mineral deposition.Item Enhancing anti-tumor potential: low-intensity vibration suppresses osteosarcoma progression and augments MSCs' tumor-suppressive abilities(Ivyspring, 2024-01-27) Xiong, Xue; Huo, Qingji; Li, Kexin; Cui, Changpeng; Chang, Chunyi; Park, Charles; Ku, BonHeon; Hong, Chin-Suk; Lim, HeeChang; Pandya, Pankita H.; Saadatzadeh, M. Reza; Bijangi-Vishehsaraei, Khadijeh; Lin, Chien-Chi; Kacena, Melissa A.; Pollok, Karen E.; Chen, Andy; Liu, Jing; Thompson, William R.; Li, Xue-Lian; Li, Bai-Yan; Yokota, Hiroki; Biomedical Engineering, School of Engineering and TechnologyRationale: Osteosarcoma (OS), a common malignant bone tumor, calls for the investigation of novel treatment strategies. Low-intensity vibration (LIV) presents itself as a promising option, given its potential to enhance bone health and decrease cancer susceptibility. This research delves into the effects of LIV on OS cells and mesenchymal stem cells (MSCs), with a primary focus on generating induced tumor-suppressing cells (iTSCs) and tumor-suppressive conditioned medium (CM). Methods: To ascertain the influence of vibration frequency, we employed numerical simulations and conducted experiments to determine the most effective LIV conditions. Subsequently, we generated iTSCs and CM through LIV exposure and assessed the impact of CM on OS cells. We also explored the underlying mechanisms of the tumor-suppressive effects of LIV-treated MSC CM, with a specific focus on vinculin (VCL). We employed cytokine array, RNA sequencing, and Western blot techniques to investigate alterations in cytokine profiles, transcriptomes, and tumor suppressor proteins. Results: Numerical simulations validated LIV frequencies within the 10-100 Hz range. LIV induced notable morphological changes in OS cells and MSCs, confirming its dual role in inhibiting OS cell progression and promoting MSC conversion into iTSCs. Upregulated VCL expression enhanced MSC responsiveness to LIV, significantly bolstering CM's efficacy. Notably, we identified tumor suppressor proteins in LIV-treated CM, including procollagen C endopeptidase enhancer (PCOLCE), histone H4 (H4), peptidylprolyl isomerase B (PPIB), and aldolase A (ALDOA). Consistently, cytokine levels decreased significantly in LIV-treated mouse femurs, and oncogenic transcript levels were downregulated in LIV-treated OS cells. Moreover, our study demonstrated that combining LIV-treated MSC CM with chemotherapy drugs yielded additive anti-tumor effects. Conclusions: LIV effectively impeded the progression of OS cells and facilitated the transformation of MSCs into iTSCs. Notably, iTSC-derived CM demonstrated robust anti-tumor properties and the augmentation of MSC responsiveness to LIV via VCL. Furthermore, the enrichment of tumor suppressor proteins within LIV-treated MSC CM and the reduction of cytokines within LIV-treated isolated bone underscore the pivotal tumor-suppressive role of LIV within the bone tumor microenvironment.Item The bone-muscle connection in breast cancer: implications and therapeutic strategies to preserve musculoskeletal health(BMC, 2022-11-23) Ballinger, Tarah J.; Thompson, William R.; Guise, Theresa A.; Medicine, School of MedicineBreast cancer and its therapies frequently result in significant musculoskeletal morbidity. Skeletal complications include bone metastases, pain, bone loss, osteoporosis, and fracture. In addition, muscle loss or weakness occurring in both the metastatic and curative setting is becoming increasingly recognized as systemic complications of disease and treatment, impacting quality of life, responsiveness to therapy, and survival. While the anatomical relationship between bone and muscle is well established, emerging research has led to new insights into the biochemical and molecular crosstalk between the skeletal and muscular systems. Here, we review the importance of both skeletal and muscular health in breast cancer, the significance of crosstalk between bone and muscle, and the influence of mechanical signals on this relationship. Therapeutic exploitation of signaling between bone and muscle has great potential to prevent the full spectrum of musculoskeletal complications across the continuum of breast cancer.Item Vibration therapy as an effective approach to improve bone healing in diabetic rats(Frontiers Media, 2022-08-19) Campos, Maysa S.; Volpon, José B.; Ximenez, João Paulo B.; Franttini, Ana Paula; Dalloul, Christopher E.; Sousa-Neto, Manoel D.; Silva, Raquel A.; Kacena, Melissa A.; Zamarioli, Ariane; Orthopaedic Surgery, School of MedicineObjective: To investigate the effects of vibration therapy on fracture healing in diabetic and non-diabetic rats. Methods: 148 rats underwent fracture surgery and were assigned to four groups: (1) SHAM: weight-matched non-diabetic rats, (2) SHAM+VT: non-diabetic rats treated with vibration therapy (VT), (3) DM: diabetic rats, and (4) DM+VT: diabetic rats treated with VT. Thirty days after diabetes induction with streptozotocin, animals underwent bone fracture, followed by surgical stabilization. Three days after bone fracture, rats began VT. Bone healing was assessed on days 14 and 28 post-fracture by serum bone marker analysis, and femurs collected for dual-energy X-ray absorptiometry, micro-computed tomography, histology, and gene expression. Results: Our results are based on 88 animals. Diabetes led to a dramatic impairment of bone healing as demonstrated by a 17% reduction in bone mineral density and decreases in formation-related microstructural parameters compared to non-diabetic control rats (81% reduction in bone callus volume, 69% reduction in woven bone fraction, 39% reduction in trabecular thickness, and 45% in trabecular number). These changes were accompanied by a significant decrease in the expression of osteoblast-related genes (Runx2, Col1a1, Osx), as well as a 92% reduction in serum insulin-like growth factor I (IGF-1) levels. On the other hand, resorption-related parameters were increased in diabetic rats, including a 20% increase in the callus porosity, a 33% increase in trabecular separation, and a 318% increase in serum C terminal telopeptide of type 1 collagen levels. VT augmented osteogenic and chondrogenic cell proliferation at the fracture callus in diabetic rats; increased circulating IGF-1 by 668%, callus volume by 52%, callus bone mineral content by 90%, and callus area by 72%; and was associated with a 19% reduction in circulating receptor activator of nuclear factor kappa beta ligand (RANK-L). Conclusions: Diabetes had detrimental effects on bone healing. Vibration therapy was effective at counteracting the significant disruption in bone repair induced by diabetes, but did not improve fracture healing in non-diabetic control rats. The mechanical stimulus not only improved bone callus quality and quantity, but also partially restored the serum levels of IGF-1 and RANK-L, inducing bone formation and mineralization, thus creating conditions for adequate fracture repair in diabetic rats.