Micro-Acoustic Holograms for Detachable Microfluidic Devices.

Acoustic microfluidic devices have advantages for diagnostic applications, therapeutic solutions, and fundamental research due to their contactless operation, simple design, and biocompatibility. However, most acoustofluidic approaches are limited to forming simple and fixed acoustic patterns, or have limited resolution. In this study,a detachable microfluidic device is demonstrated employing miniature acoustic holograms to create reconfigurable, flexible, and high-resolution acoustic fields in microfluidic channels, where the introduction of a solid coupling layer makes these holograms easy to fabricate and integrate. The application of this method to generate flexible acoustic fields, including shapes, characters, and arbitrarily rotated patterns, within microfluidic channels, is demonstrated.

Surface area-to-volume ratio, not cellular viscoelasticity, is the major determinant of red blood cell traversal through small channels.

The remarkable deformability of red blood cells (RBCs) depends on the viscoelasticity of the plasma membrane and cell contents and the surface area to volume (SA:V) ratio; however, it remains unclear which of these factors is the key determinant for passage through small capillaries. We used a microfluidic device to examine the traversal of normal, stiffened, swollen, parasitised and immature RBCs. We show that dramatic stiffening of RBCs had no measurable effect on their ability to traverse small channels. By contrast, a moderate decrease in the SA:V ratio had a marked effect on the equivalent cylinder diameter that is traversable by RBCs of similar cellular viscoelasticity. We developed a finite element model that provides a coherent rationale for the experimental observations, based on the nonlinear mechanical behaviour of the RBC membrane skeleton. We conclude that the SA:V ratio should be given more prominence in studies of RBC pathologies.

Bone Measures by Dual-Energy X-Ray Absorptiometry and Peripheral Quantitative Computed Tomography in Young Women With Type 1 Diabetes Mellitus.

Understanding bone fragility in young adult females with type 1 diabetes mellitus (T1DM) is of great clinical importance since the high fracture risk in this population remains unexplained. This study aimed to investigate bone health in young adult T1DM females by comparing relevant variables determined by dual-energy X-ray absorptiometry (DXA), peripheral quantitative computed tomography (pQCT) at the tibia and pQCT-based finite element analysis (pQCT-FEA) between T1DM subjects (n = 21) and age-, height- and weight-matched controls (n = 63). Tibial trabecular density (lower by 7.1%; 228.8 ± 33.6 vs 246.4 ± 31.8 mg/cm3, p = 0.02) and cortical thickness (lower by 7.3%; 3.8 ± 0.5 vs 4.1 ± 0.5 cm, p = 0.03) by pQCT were significantly lower in T1DM subjects than in controls. Tibial shear stiffness by pQCT-FEA was also lower in T1DM subjects than in controls at both the 4% site (by 17.1%; 337.4 ± 75.5 vs 407.1 ± 75.4 kN/mm, p < 0.01) and 66% site (by 7.9%; 1113.0 ± 158.6 vs 1208.8 ± 161.8 kN/mm, p = 0.03). These differences remained statistically significant after adjustment for confounding factors. No difference between groups was observed in DXA-determined variables (all p ≥ 0.08), although there was a trend towards lower aBMD at the lumbar spine in T1DM subjects than in controls after adjustment for confounders (p = 0.053). These novel findings elicited using pQCT and pQCT-FEA suggest a clinically significant impact of T1DM on bone strength in young adult females with T1DM. Peripheral QCT and pQCT-FEA may provide more information than DXA alone on bone fragility in this population. Further longitudinal studies with a larger sample size are warranted to understand the evolution and causes of bone fragility in young T1DM females.

Annexin A2 contributes to lung injury and fibrosis by augmenting factor Xa fibrogenic activity.

In lung injury and disease, including idiopathic pulmonary fibrosis (IPF), extravascular factor X is converted into factor Xa (FXa), a coagulant protease with fibrogenic actions. Extracellular annexin A2 binds to FXa, augmenting activation of the protease-activated receptor-1 (PAR-1). In this study, the contribution of annexin A2 in lung injury and fibrosis was investigated. Annexin A2 immunoreactivity was observed in regions of fibrosis, including those associated with fibroblasts in lung tissue of IPF patients. Furthermore, annexin A2 was detected in the conditioned media and an EGTA membrane wash of human lung fibroblast (LF) cultures. Incubation with human plasma (5% vol/vol) or purified FXa (15-50 nM) evoked fibrogenic responses in LF cultures, with FXa increasing interleukin-6 (IL-6) production and cell number by 270 and 46%, respectively (P < 0.05, n = 5-8). The fibrogenic actions of plasma or FXa were attenuated by the selective FXa inhibitor apixaban (10 µM, or antibodies raised against annexin A2 or PAR-1 (2 µg/ml). FXa-stimulated LFs from IPF patients (n = 6) produced twice as much IL-6 as controls (n = 10) (P < 0.05), corresponding with increased levels of extracellular annexin A2. Annexin A2 gene deletion in mice reduced bleomycin-induced increases in bronchoalveolar lavage fluid (BALF) IL-6 levels and cell number (*P < 0.05; n = 4-12). Lung fibrogenic gene expression and dry weight were reduced by annexin A2 gene deletion, but lung levels of collagen were not. Our data suggest that annexin A2 contributes to lung injury and fibrotic disease by mediating the fibrogenic actions of FXa. Extracellular annexin A2 is a potential target for the treatment of IPF.

A cruciate suture technique for rotator cuff repair.

PURPOSE: Re-establishment of the native footprint during rotator cuff repair is important for maximizing healing potential and fixation strength. The purpose of this study was to evaluate the contact area, contact pressure, stiffness and tensile strength of a new single-row cruciate suture repair and to compare these results to those of the Mason-Allen, double-row and transosseous repairs. METHODS: Infraspinatus tendons from fifty-six lamb shoulders were harvested and randomly assigned to cruciate suture, Mason-Allen, double-row and transosseous repair groups. Repairs were performed over pressure-sensitive film and footprint contact area and pressure measured. Repaired tendon specimens were also loaded in uniaxial tension, and ultimate tensile strength and stiffness measured. RESULTS: The cruciate suture repair established significantly greater footprint contact area compared to the Mason-Allen repair (mean difference = 101 mm(2), p = 0.003). The ultimate tensile strength and stiffness of the double-row repair was significantly higher than that of all other repair groups (p < 0.05). The average footprint contact pressure of the cruciate suture repair (0.78 MPa) was similar to that of the Mason-Allen (0.74 MPa) and double-row repairs (0.79 MPa). The ultimate tensile strength of the cruciate suture repair was significantly greater than that of the transosseous repair (mean difference 62.4 N, p = 0.002). CONCLUSION: The cruciate suture repair may improve strength and healing at the repaired tendon rotator cuff insertion relative to other single-row repair techniques. It may represent a faster, easier and more cost-effective alternative to double-row repairs.

Muscle and joint mechanics during maximum force biting following total temporomandibular joint replacement surgery.

Total temporomandibular joint replacement (TMJR) surgery is the established treatment for severe temporomandibular joint disorders. While TMJR surgery is known to increase mouth-opening capacity, reduce pain and improve quality of life, little is known about post-surgical jaw function during activities of daily living such as biting and chewing. The aim of this study was to use subject-specific 3D bite force measurements to evaluate the magnitude and direction of joint loading in unilateral total TMJR patients and compare these data to those in healthy control subjects. An optoelectronic tracking system was used to measure jaw kinematics while biting a rubber sample for 5 unilateral total TMJR patients and 8 controls. Finite element simulations driven by the measured kinematics were employed to calculate the resultant bite force generated when compressing the rubber between teeth during biting tasks. Subject-specific musculoskeletal models were subsequently used to calculate muscle and TMJ loading. Unilateral total TMJR patients generated a bite force of 249.6 ± 24.4 N and 164.2 ± 62.3 N when biting on the contralateral and ipsilateral molars, respectively. In contrast, controls generated a bite force of 317.1 ± 206.6 N. Unilateral total TMJR patients biting on the contralateral molars had a significantly higher lateral TMJ force direction (median difference: 63.6°, p = 0.028) and a significantly lower ratio of working TMJ force to bite force (median difference: 0.17, p = 0.049) than controls. Results of this study may guide TMJ prosthesis design and evaluation of dental implants.

Computational modeling of revision total hip arthroplasty involving acetabular defects: A systematic review.

Revision total hip arthroplasty (rTHA) involving acetabular defects is a complex procedure associated with lower rates of success than primary THA. Computational modeling has played a key role in surgical planning and prediction of postoperative outcomes following primary THA, but modeling applications in rTHA for acetabular defects remain poorly understood. This study aimed to systematically review the use of computational modeling in acetabular defect classification, implant selection and placement, implant design, and postoperative joint functional performance evaluation following rTHA involving acetabular defects. The databases of Web of Science, Scopus, Medline, Embase, Global Health and Central were searched. Fifty-three relevant articles met the inclusion criteria, and their quality were evaluated using a modified Downs and Black evaluation criteria framework. Manual image segmentation from computed tomography scans, which is time consuming, remains the primary method used to generate 3D models of hip bone; however, statistical shape models, once developed, can be used to estimate pre-defect anatomy rapidly. Finite element modeling, which has been used to estimate bone stresses and strains, and implant micromotion postoperatively, has played a key role in custom and off-the-shelf implant design, mitigation of stress shielding, and prediction of bone remodeling and implant stability. However, model validation is challenging and requires rigorous evaluation and comparison with respect to mid- to long-term clinical outcomes. Development of fast, accurate methods to model acetabular defects, including statistical shape models and artificial neural networks, may ultimately improve uptake of and expand applications in modeling and simulation of rTHA for the research setting and clinic.

Transforming growth factor-ß-induced differentiation of airway smooth muscle cells is inhibited by fibroblast growth factor-2.

In asthma, basic fibroblast growth factor (FGF-2) plays an important (patho)physiological role. This study examines the effects of FGF-2 on the transforming growth factor-ß (TGF-ß)-stimulated differentiation of airway smooth muscle (ASM) cells in vitro. The differentiation of human ASM cells after incubation with TGF-ß (100 pM) and/or FGF-2 (300 pM) for 48 hours was assessed by increases in contractile protein expression, actin-cytoskeleton reorganization, enhancements in cell stiffness, and collagen remodeling. FGF-2 inhibited TGF-ß-stimulated increases in transgelin (SM22) and calponin gene expression (n = 15, P < 0.01) in an extracellular signal-regulated kinase 1/2 (ERK1/2) signal transduction-dependent manner. The abundance of ordered α-smooth muscle actin (α-SMA) filaments formed in the presence of TGF-ß were also reduced by FGF-2, as was the ratio of F-actin to G-actin (n = 8, P < 0.01). Furthermore, FGF-2 attenuated TGF-ß-stimulated increases in ASM cell stiffness and the ASM-mediated contraction of lattices, composed of collagen fibrils (n = 5, P < 0.01). However, the TGF-ß-stimulated production of IL-6 was not influenced by FGF-2 (n = 4, P > 0.05), suggesting that FGF-2 antagonism is selective for the regulation of ASM cell contractile protein expression, organization, and function. Another mitogen, thrombin (0.3 U ml(-1)), exerted no effect on TGF-ß-regulated contractile protein expression (n = 8, P > 0.05), α-SMA organization, or the ratio of F-actin to G-actin (n = 4, P > 0.05), suggesting that the inhibitory effect of FGF-2 is dissociated from its mitogenic actions. The addition of FGF-2, 24 hours after TGF-ß treatment, still reduced contractile protein expression, even when the TGF-ß-receptor kinase inhibitor, SB431542 (10 µM), was added 1 hour before FGF-2. We conclude that the ASM cell differentiation promoted by TGF-ß is antagonized by FGF-2. A better understanding of the mechanism of action for FGF-2 is necessary to develop a strategy for therapeutic exploitation in the treatment of asthma.

The use of finite element models for backface deformation and body armour design: a systematic review.

While injuries sustained from body armour backface deformation (BFD) have not been well-documented in military injury trauma registries, data from US law enforcement officers, animal tests and currently available data pertaining to military combatants has shown that BFD can not only cause minor injuries, but also result in serious trauma. However, the nature and severity of injuries sustained depends on a multitude of factors including the projectile type, the impact location and velocity, and the specific type of body armour worn. The difficulties involved in current measurement techniques for ballistic testing has led researchers to seek alternative techniques to evaluate the level of protection from body armour, such as the finite element (FE) method. In the current study, a systematic review of the open literature was undertaken using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses methodology. The aim was to summarise the literature pertaining to the development and application of FE models to investigate body armour BFD and behind armour blunt trauma (BABT), and included FE models representing the projectile, clay-based mediums, ballistic gelatine and the human torso. Using the keywords 'behind armour*', 'ballistic blunt trauma', 'BABT', 'backface signature', 'backface deformation', 'BFS', 'BFD', 'wound ballistic', 'ballistic impact testing', 'body armour', 'bullet proof vest', 'ballistic vest', 'Finite Element*' and 'FE', an electronic database search of EBSCOhost, Google Scholar, ProQuest, Scopus, Standards, Web of Science and PubMed was conducted, and included peer-reviewed journal articles, review papers, research reports, conference papers, and MSc or PhD theses. While this research demonstrates the potential of FE analysis for recreating realistic blunt impact scenarios and enhancing the current understanding of BABT mechanisms, a common limitation in most studies is the lack of validation. Thus, in order to address this issue, it is proposed that injury predictions from FE models be correlated with trauma data from soldiers who have sustained BABT. Consequently, pressure and energy distributions within the organs can be used to interpret the effects of non-penetrating ballistic impacts on the human torso. Bridging the gap between simulation and real-world data is essential in order to validate FE models and enhance their utility in optimising body armour design and employing injury mitigation strategies.

Kinematics of the jaw following total temporomandibular joint replacement surgery.

Total temporomandibular joint (TMJ) replacement surgery aims to improve mandibular function, reduce pain and enhance quality of life in patients suffering from end-stage TMJ disorders. Traditional post-operative jaw evaluation is carried out using measurement of maximum interincisal opening distance; however, this can correlate poorly to joint function. The present study aimed to evaluate three-dimensional (3D) jaw motion during border movements and chewing in unilateral total TMJ replacement patients and healthy controls. Motion analysis experiments were performed on six unilateral total TMJ replacement patients and ten age-matched healthy controls. Subject-specific motion tracking plates worn by each participant were registered to CT scans of each participant's skull and mandible to enable anatomical mandibular kinematics measurement using an optoelectronic system. Participants performed 15 repetitions of maximal opening, protrusion, lateral excursions, and chewing cycles. Total TMJ replacement patients had significantly smaller incisal displacements at maximum mouth opening relative to the controls (median difference: 7.1 mm, p = 0.002) and decreased anterior translation of the prosthetic condyle (median difference: 10.5 mm, p = 0.002). When TMJ replacement subjects chewed using their contralateral molars, there was a significant increase in inferior condylar translation of the non-working condyle (median difference: 9.7 mm, p = 0.016). This study found that unilateral total TMJ replacement surgery was associated with mouth opening capacity within the range of healthy individuals, but reduced anterior movement of the prosthetic condyle and restricted protrusion and lateral excursions. The results provide future direction for prosthetic TMJ design to enhance postsurgical implant functionality and improve long-term clinical outcomes for prosthesis recipients.

Sound-Speed Modifying Acoustic Metasurfaces for Acoustic Holography.

Acoustic metasurfaces offer unique capabilities to steer and direct acoustic fields, though these are generally composed of complex 3D structures, complicating their fabrication and applicability to higher frequencies. Here, an ultrathin metasurface approach is demonstrated, wherein planarized micropillars in a discretized phase array are utilized. This subwavelength metasurface is easily produced via a single-step etching process and is suitable for megahertz-scale applications. The flexibility of this approach is further demonstrated in the production of complex acoustic patterns via acoustic holography. This metasurface approach, with models used to predict their behavior, has broad potential in applications where robust, high-frequency acoustic manipulation is required, including microfluidics, cell/tissue engineering, and medical ultrasound.

Programmable Acoustic Holography using Medium-Sound-Speed Modulation.

Acoustic holography offers the ability to generate designed acoustic fields to manipulate microscale objects. However, the static nature or large aperture sizes of 3D printed acoustic holographic phase plates limits the ability to rapidly alter generated fields. In this work， a programmable acoustic holography approach is demonstrated by which multiple discrete or continuously variable acoustic targets can be created. Here, the holographic phase plate encodes multiple images, where the desired field is produced by modifying the sound speed of an intervening fluid media. Its flexibility is demonstrated in generating various acoustic patterns, including continuous line segments, discrete letters and numbers, using this method as a sound speed indicator and fluid identification tool. This programmable acoustic holography approach has the advantages of generating reconfigurable and designed acoustic fields, with broad potential in microfluidics, cell/tissue engineering, real-time sensing, and medical ultrasound.

Additively manufactured controlled porous orthopedic joint replacement designs to reduce bone stress shielding: a systematic review.

BACKGROUND: Total joint replacements are an established treatment for patients suffering from reduced mobility and pain due to severe joint damage. Aseptic loosening due to stress shielding is currently one of the main reasons for revision surgery. As this phenomenon is related to a mismatch in mechanical properties between implant and bone, stiffness reduction of implants has been of major interest in new implant designs. Facilitated by modern additive manufacturing technologies, the introduction of porosity into implant materials has been shown to enable significant stiffness reduction; however, whether these devices mitigate stress-shielding associated complications or device failure remains poorly understood. METHODS: In this systematic review, a broad literature search was conducted in six databases (Scopus, Web of Science, Medline, Embase, Compendex, and Inspec) aiming to identify current design approaches to target stress shielding through controlled porous structures. The search keywords included 'lattice,' 'implant,' 'additive manufacturing,' and 'stress shielding.' RESULTS: After the screening of 2530 articles, a total of 46 studies were included in this review. Studies focusing on hip, knee, and shoulder replacements were found. Three porous design strategies were identified, specifically uniform, graded, and optimized designs. The latter included personalized design approaches targeting stress shielding based on patient-specific data. All studies reported a reduction of stress shielding achieved by the presented design. CONCLUSION: Not all studies used quantitative measures to describe the improvements, and the main stress shielding measures chosen varied between studies. However, due to the nature of the optimization approaches, optimized designs were found to be the most promising. Besides the stiffness reduction, other factors such as mechanical strength can be considered in the design on a patient-specific level. While it was found that controlled porous designs are overall promising to reduce stress shielding, further research and clinical evidence are needed to determine the most superior design approach for total joint replacement implants.

High-throughput microfluidic compressibility cytometry using multi-tilted-angle surface acoustic wave.

Cellular mechanical properties (e.g. compressibility) are important biophysical markers in relation to cellular processes and functionality. Among the methods for cell mechanical measurement, acoustofluidic methods appear to be advantageous due to tunability, biocompatibility and acousto-mechanical nature. However, the previous acoustofluidic methods were limited in throughput and number of measurements. In this study, we developed a high-throughput microfluidic compressibility cytometry approach using multi-tilted-angle surface acoustic wave, which can provide thousands of single-cell compressibility measurements within minutes. The compressibility cytometer was constructed to drag microparticles or cells towards the microfluidic channel sidewall at different segments based on their biophysical properties (such as size and compressibility), as a result of the varied balance between acoustics and flow. Mathematical analysis and computational simulation revealed that the compressibility of a cell could be estimated from the position of collision with the sidewall. Microbeads of different materials and sizes were experimentally tested to validate the simulation and to demonstrate the capability to characterise size and compressibility. MDA MB231 cells, of the triple negative breast cancer subtype, were treated with the microtubule disrupting agent colchicine which increased compressibility and treated with the actin disrupting agent cytochalasin B which increased cell size but did not change compressibility. Moreover, the highly metastatic variant MDA MB231 LNm5 cell line showed increased compressibility compared to the parent MDA MB231 cells, indicating the potential utility of high-throughput mechanophenotyping for tumour cell characterisation.

Microfluidic acoustic sawtooth metasurfaces for patterning and separation using traveling surface acoustic waves.

We demonstrate a sawtooth-based metasurface approach for flexibly orienting acoustic fields in a microfluidic device driven by surface acoustic waves (SAW), where sub-wavelength channel features can be used to arbitrarily steer acoustic fringes in a microchannel. Compared to other acoustofluidic methods, only a single travelling wave is used, the fluidic pressure field is decoupled from the fluid domain's shape, and steerable pressure fields are a function of a simply constructed polydimethylsiloxane (PDMS) metasurface shape. Our results are relevant to microfluidic applications including the patterning, concentration, focusing, and separation of microparticles and cells.

Unconventional acoustic approaches for localized and designed micromanipulation.

Acoustic fields are ideal for micromanipulation, being biocompatible and with force gradients approaching the scale of single cells. They have accordingly found use in a variety of microfluidic devices, including for microscale patterning, separation, and mixing. The bulk of work in acoustofluidics has been predicated on the formation of standing waves that form periodic nodal positions along which suspended particles and cells are aligned. An evolving range of applications, however, requires more targeted micromanipulation to create unique patterns and effects. To this end, recent work has made important advances in improving the flexibility with which acoustic fields can be applied, impressively demonstrating generating arbitrary arrangements of pressure fields, spatially localizing acoustic fields and selectively translating individual particles in ways that are not achievable via traditional approaches. In this critical review we categorize and examine these advances, each of which open the door to a wide range of applications in which single-cell fidelity and flexible micromanipulation are advantageous, including for tissue engineering, diagnostic devices, high-throughput sorting and microfabrication.

The relationship between microstructure, stiffness and compressive fatigue life of equine subchondral bone.

Subchondral bone injuries often precede articular cartilage damage in osteoarthritis and are common in thoroughbred racehorses due to the accumulation of fatigue damage from high speed racing and training. Thus, racehorses provide a model to investigate the role of subchondral bone in joint disease. We assessed the association of horse and racing related factors and micro-CT based micromorphology of three separate subchondral bone layers with the initial stiffness and compressive fatigue life of bone plugs. Furthermore, we investigated three different definitions of fatigue failure of subchondral bone during compressive fatigue testing. Initial stiffness was 2,362 ±â€¯443 MPa (mean ±â€¯standard deviation). Median compressive fatigue life during cyclic loading to -78 MPa was 16,879 (range 210 to 57,064). Subchondral bone stiffness increased over a median of 24% (range 3%-42%) of fatigue life to a maximum of 3,614 ±â€¯635 MPa. Compressive fatigue life was positively associated with bone volume fraction in the deeper layers of subchondral bone, maximal stiffness, and the number of cycles to maximal stiffness. Initial stiffness was positively associated with tissue mineral density in the deeper layers and bone volume fraction in the superficial layer. Most specimens with a fatigue life of less than 5,500 cycles fractured grossly before reaching 30% reduction of maximal stiffness. Cycles to 10% reduction of maximal stiffness correlated strongly with cycles to lowest recorded stiffness at gross fracture and thus is a valid alternative failure definition for compressive fatigue testing of subchondral bone. Our results show that subchondral bone sclerosis as a result of high speed exercise and measured as bone volume fraction is positively associated with compressive fatigue life and thus has a protective effect on subchondral bone. Further research is required to reconcile this finding with the common collocation of fatigue damage in sclerotic subchondral bone of racehorses.

Measurement of normal and pathological mandibular and temporomandibular joint kinematics: A systematic review.

Motion of the mandible and temporomandibular joint (TMJ) plays a pivotal role in the function of the dentition and associated hard and soft tissue structures, and facilitates mastication, oral communication and access to respiratory and digestive systems. Quantification of TMJ kinematics is clinically relevant in cases of prosthetic rehabilitations, TMJ disorders, osteoarthritis, trauma, tumour resection and congenital abnormalities, which are known to directly influence mandibular motion and loading. The objective of this systematic review was to critically investigate published literature on historic and contemporary measurement modalities used to quantify in vivo mandibular and TMJ kinematics in six degrees of freedom. The electronic databases of Scopus, Web of Science, Medline, Embase and Central were searched and 109 relevant articles identified. Publication quality was documented using a modified Downs and Black checklist. Axiography and ultrasonic tracking are commonly employed in the clinical setting due to their simplicity and capacity to rapidly acquire low-fidelity mandibular motion data. Magnetic and optoelectronic tracking have been used in combination with dental splints to produce higher accuracy measurements while minimising skin motion artefact, but at the expense of setup time and cost. Four-dimensional computed tomography provides direct 3D measurement of mandibular and TMJ motion while circumventing skin motion artefact entirely, but employs ionising radiation, is restricted to low sampling frequencies, and requires time-consuming image processing. Recent advances in magnetic tracking using miniature sensors adhered to the teeth in combination with intraoral scanning may facilitate rapid and high precision mandibular kinematics measurement in the clinical setting. The findings of this review will guide selection and application of mandibular and TMJ kinematic measurement for both clinical and research applications.

On-chip surface acoustic wave and micropipette aspiration techniques to assess cell elastic properties.

The cytoskeletal mechanics and cell mechanical properties play an important role in cellular behaviors. In this study, in order to provide comprehensive insights into the relationship between different cytoskeletal components and cellular elastic moduli, we built a phase-modulated surface acoustic wave microfluidic device to measure cellular compressibility and a microfluidic micropipette-aspiration device to measure cellular Young's modulus. The microfluidic devices were validated based on experimental data and computational simulations. The contributions of structural cytoskeletal actin filament and microtubule to cellular compressibility and Young's modulus were examined in MCF-7 cells. The compressibility of MCF-7 cells was increased after microtubule disruption, whereas actin disruption had no effect. In contrast, Young's modulus of MCF-7 cells was reduced after actin disruption but unaffected by microtubule disruption. The actin filaments and microtubules were stained to confirm the structural alteration in cytoskeleton. Our findings suggest the dissimilarity in the structural roles of actin filaments and microtubules in terms of cellular compressibility and Young's modulus. Based on the differences in location and structure, actin filaments mainly contribute to tensile Young's modulus and microtubules mainly contribute to compressibility. In addition, different responses to cytoskeletal alterations between acoustophoresis and micropipette aspiration demonstrated that micropipette aspiration was better at detecting the change from actin cortex, while the response to acoustophoresis was governed by microtubule networks.

A novel rat model of heterotopic ossification after polytrauma with traumatic brain injury.

Neurological heterotopic ossification (NHO) is characterized by abnormal bone growth in soft tissue and joints in response to injury to the central nervous system. The ectopic bone frequently causes pain, restricts mobility, and decreases the quality of life for those affected. NHO commonly develops in severe traumatic brain injury (TBI) patients, particularly in the presence of concomitant musculoskeletal injuries (i.e. polytrauma). There are currently no animal models that accurately mimic these combinations of injuries, which has limited our understanding of NHO pathobiology, as well as the development of biomarkers and treatments, in TBI patients. In order to address this shortcoming, here we present a novel rat model that combines TBI, femoral fracture, and muscle crush injury. Young adult male Sprague Dawley rats were randomly assigned into three different injury groups: triple sham-injury, peripheral injury only (i.e., sham-TBI + fracture + muscle injury) or triple injury (i.e., TBI + fracture + muscle injury). Evidence of ectopic bone in the injured hind-limb, as confirmed by micro-computed tomography (µCT), was found at 6-weeks post-injury in 70% of triple injury rats, 20% of peripheral injury rats, and 0% of the sham-injured controls. Furthermore, the triple injury rats had higher ectopic bone severity scores than the sham-injured group. This novel model will provide a platform for future studies to identify underlying mechanisms, biomarkers, and develop evidence based pharmacological treatments to combat this debilitating long-term complication of TBI and polytrauma.