Ventricular anisotropic deformation and contractile function of the developing heart of zebrafish in vivo.

BACKGROUND: The developing zebrafish ventricle generates higher intraventricular pressure (IVP) with increasing stroke volume and cardiac output at different developmental stages to meet the metabolic demands of the rapidly growing embryo (Salehin et al. Ann Biomed Eng, 2021;49(9): 2080-2093). To understand the changing role of the developing embryonic heart, we studied its biomechanical characteristics during in vivo cardiac cycles. By combining changes in wall strains and IVP measurements, we assessed ventricular wall stiffness during diastolic filling and the ensuing systolic IVP-generation capacity during 3-, 4-, and 5-day post fertilization (dpf). We further examined the anisotropy of wall deformation, in different regions within the ventricle, throughout a complete cardiac cycle. RESULTS: We found the ventricular walls grow increasingly stiff during diastolic filling with a corresponding increase in IVP-generation capacity from 3- to 4- and 5-dpf groups. In addition, we found the corresponding increasing level of anisotropic wall deformation through cardiac cycles that favor the latitudinal direction, with the most pronounced found in the equatorial region of the ventricle. CONCLUSIONS: From 3- to 4- and 5-dpf groups, the ventricular wall myocardium undergoes increasing level of anisotropic deformation. This, in combination with the increasing wall stiffness and IVP-generation capacity, allows the developing heart to effectively pump blood to meet the rapidly growing embryo's needs.

Scale space detector for analyzing spatiotemporal ventricular contractility and nuclear morphogenesis in zebrafish.

In vivo quantitative assessment of structural and functional biomarkers is essential for characterizing the pathophysiology of congenital disorders. In this regard, fixed tissue analysis has offered revolutionary insights into the underlying cellular architecture. However, histological analysis faces major drawbacks with respect to lack of spatiotemporal sampling and tissue artifacts during sample preparation. This study demonstrates the potential of light sheet fluorescence microscopy (LSFM) as a non-invasive, 4D (3days + time) optical sectioning tool for revealing cardiac mechano-transduction in zebrafish. Furthermore, we have described the utility of a scale and size-invariant feature detector, for analyzing individual morphology of fused cardiomyocyte nuclei and characterizing zebrafish ventricular contractility.

Assessing Pressure-Volume Relationship in Developing Heart of Zebrafish In-Vivo.

During embryogenesis, the developing heart transforms from a linear peristaltic tube into a multi-chambered pulsatile pump with blood flow-regulating valves. In this work, we report how hemodynamic parameters evolve during the heart's development, leading to its rhythmic pumping and blood flow regulation as a functioning organ. We measured the time course of intra-ventricular pressure from zebrafish embryos at 3, 4, and 5 days post fertilization (dpf) using the servo null method. We also measured the ventricular volume and monitored the opening/closing activity of the AV and VB valves using 4D selective plane illumination microscopy (SPIM). Our results revealed significant increases in peak systolic pressure, stroke volume and work, cardiac output, and power generation, and a total peripheral resistance decrease from zebrafish at 4, 5 dpf versus 3 dpf. These data illustrate that the early-stage zebrafish heart's increasing efficiency is synchronous with the expected changes in valve development, chamber morphology and increasing vascular network complexity. Such physiological measurements in tractable laboratory model organisms are critical for understanding how gene variants may affect phenotype. As the zebrafish emerges as a leading biomedical model organism, the ability to effectively measure its physiology is critical to its translational relevance.

Differences in creep response of GBM cells migrating in confinement.

Using a microfluidic platform to apply negative aspiration pressure (-20, -25, -30, -35 and -40 cm H2O), we compared the differences in creep responses of Glioblastoma Multiforme (GBM) cells while migrating in confinement and at a stationary state on a 2D substrate. Cells were either migrating in a channel of 5 x 5 µm cross-section or stationary at the entrance to the channel. In response to aspiration pressure, we found actively migrating GBM cells exhibited a higher stiffness than stationary cells. Additionally, migrating cells absorbed more energy elastically with a relatively small dissipative energy loss. At elevated negative pressure loads up to - 30 cm H2O, we observed a linear increase in elastic deformation and a higher distribution in elastic storage than energy loss, and the response plateaued at further increasing negative pressure loads. To explore the underlying cause, we carried out immuno-cytochemical studies of these cells and found a polarized actin and myosin distribution at the front and posterior ends of the migrating cells, whereas the distribution of the stationary group demonstrated no specific regional differences. These differences in creep response and cytoskeletal protein distribution demonstrate the importance of a migrating cell's kinematic state to the mechanism of cell migration.

Role of key genetic mutations on increasing migration of brain cancer cells through confinement.

Uncontrolled invasive cancer cell migration is among the major challenges for the treatment and management of brain cancer. Although the genetic profiles of brain cancer cells have been well characterized, the relationship between the genetic mutations and the cells' mobility has not been clearly understood. In this study, using microfluidic devices that provide a wide range of physical confinements from 20 × 5 µm2 to 3 × 5 µm2 in cross sections, we studied the effect of physical confinement on the migratory capacity of cell lines with different types of mutations. Human glioblastoma and genetically modified mouse astrocytes were used. Human glioblastoma cells with EGFRvIII mutation were found to exhibit high degree of migratory capacity in narrow confinement. From mouse astrocytes, cells with triple mutations (p53-/- PTEN-/- BRAF) were found to exhibit the highest level of migratory capacity in narrow confinement compared to both double (p53-/- PTEN-/-) and single (p53-/-) mutant cells. Furthermore, when treating the triple mutant astrocytes with AZD-6244, an inhibitor of the RAF/MEK/ERK pathway, we found significant reduction in migration through the confined channels when compared to that of controls (83% decrease in 5 × 5 µm2 and 86% in 3 × 5 µm2 channels). Our data correlate genetic mutations from different cell lines to their motility in different degrees of confinement. Our results also suggest a potential therapeutic target such as BRAF oncogene for inhibition of brain cancer invasion.

Exploring biomechanical methods to study the human vaginal wall.

AIMS: To critically review studies of the biomechanical properties of connective tissue in the normal and prolapsed human vaginal wall and to identify criteria that are suitable for in vivo measurements which could improve patient management. METHODS: This review covers past and current ex vivo and in vivo instrumentation and analytical methods related to the elastic and viscoelastic properties of vaginal wall connective tissues. RESULTS: Classical methods, including digital evaluation of the vagina, histological and biomechanical studies of fresh and frozen-thawed extracts, and biomechanical cadaveric tissue studies have important limitations and have yielded inconsistent results. Newer biomechanical methods may resolve these inconsistencies. One of the more promising is transient, vacuum-induced tissue expansion and relaxation, via cutometer-like devices. The technique permits noninvasive observation, applicable to longitudinal studies of patients. In vivo and ex vivo biomechanical methods may better match vaginal wall tissue properties to help with the design of surgical mesh materials, thus improving surgical support and healing. CONCLUSION: Methods have been identified to characterize the in vivo biomechanical behavior of the prolapsing vagina which may serve to advance the care of affected women. Neurourol. Urodynam. 36:499-506, 2017. © 2016 Wiley Periodicals, Inc.

Brain Tumor Genetic Modification Yields Increased Resistance to Paclitaxel in Physical Confinement.

Brain tumor cells remain highly resistant to radiation and chemotherapy, particularly malignant and secondary cancers. In this study, we utilized microchannel devices to examine the effect of a confined environment on the viability and drug resistance of the following brain cancer cell lines: primary cancers (glioblastoma multiforme and neuroblastoma), human brain cancer cell lines (D54 and D54-EGFRvIII), and genetically modified mouse astrocytes (wild type, p53-/-, p53-/- PTEN-/-, p53-/- Braf, and p53-/- PTEN-/- Braf). We found that loss of PTEN combined with Braf activation resulted in higher viability in narrow microchannels. In addition, Braf conferred increased resistance to the microtubule-stabilizing drug Taxol in narrow confinement. Similarly, survival of D54-EGFRvIII cells was unaffected following treatment with Taxol, whereas the viability of D54 cells was reduced by 75% under these conditions. Taken together, our data suggests key targets for anticancer drugs based on cellular genotypes and their specific survival phenotypes during confined migration.

Mechanical Interaction of an Expanding Coiled Stent with a Plaque-Containing Arterial Wall: A Finite Element Analysis.

Wall injury is observed during stent expansion within atherosclerotic arteries, related in part to stimulation of the inflammatory process. Wall stress and strain induced by stent expansion can be closely examined by finite element analysis (FEA), thus shedding light on procedure-induced sources of inflammation. The purpose of this work was to use FEA to examine the interaction of a coiled polymer stent with a plaque-containing arterial wall during stent expansion. An asymmetric fibrotic plaque-containing arterial wall model was created from intravascular ultrasound (IVUS) images of a diseased artery. A 3D model for a coil stent at unexpanded state was generated in SolidWorks. They were imported into ANSYS for FEA of combined stent expansion and fibrotic plaque-distortion. We simulated the stent expansion in the plaqued lumen by increasing balloon pressure from 0 to 12 atm in 1 atm step. At increasing pressure, we examined how the expanding stent exerts forces on the fibrotic plaque and vascular wall components, and how the latter collectively resist and balance the expansive forces from the stent. Results show the expanding coiled stent creates high stresses within the plaque and the surrounding fibrotic capsule. Lower stresses were observed in adjacent medial and adventitial layers. High principal strains were observed in plaque and fibrotic capsule. The results suggest fibrotic capsule rupture might occur at localized regions. The FEA/IVUS method can be adapted for routine examination of the effects of the expansion of selected furled stents against IVUS-reconstructed diseased vessels, to improve stent deployment practices.

Coiled polymeric growth factor gradients for multi-luminal neural chemotaxis.

In the injured adult nervous system, re-establishment of growth-promoting molecular gradients is known to entice and guide nerve repair. However, incorporation of three-dimensional chemotactic gradients in nerve repair scaffolds, particularly in those with multi-luminal architectures, remains extremely challenging. We developed a method that establishes highly tunable three-dimensional molecular gradients in multi-luminal nerve guides by anchoring growth-factor releasing coiled polymeric fibers onto the walls of collagen-filled hydrogel microchannels. Differential pitch in the coiling of neurotrophin-eluting fibers generated sustained chemotactic gradients that appropriately induced the differentiation of Pheochromocytoma (PC12) cells into neural-like cells along an increasing concentration of nerve growth factor (NGF). Computer modeling estimated the stability of the molecular gradient within the luminal collagen, which we confirmed by observing the significant effects of neurotrophin gradients on axonal growth from dorsal root ganglia (DRG). Neurons growing in microchannels exposed to a NGF gradient showed a 60% increase in axonal length compared to those treated with a linear growth factor concentration. In addition, a two-fold increment in the linearity of axonal growth within the microchannels was observed and confirmed by a significant reduction in the turning angle ratios of individual axons. These data demonstrate the ability of growth factor-loaded polymeric coiled fibers to establish three-dimensional chemotactic gradients to promote and direct nerve regeneration in the nervous system and provides a unique platform for molecularly guided tissue repair.

Influence of body mass index on the biomechanical properties of the human prolapsed anterior vaginal wall.

INTRODUCTION AND HYPOTHESIS: We report the influence of body mass index (BMI) on the biomechanical properties of human prolapsed anterior vaginal wall (AVW) tissue samples. We hypothesize that women with AVW prolapse would have the same vaginal wall biomechanical properties regardless of their weight. METHODS: Following Institutional Review Board approval, age-comparable postmenopausal women with symptomatic stage II-III AVW prolapse underwent excision of a short vaginal wall sample during transvaginal prolapse repair. Excised samples were subjected to uniaxial tensile testing using an Instron 5655 (Instron, Norwood, MA, USA) within 2 h of harvest to measure intrinsic biomechanical properties. Patients were divided into two groups (A: BMI <25 and B: BMI >25) to compare tissue biomechanical properties after controlling for age and parity. RESULTS: From 2011 to 2013, 28 consecutive women were studied, 13 in group A and 15 in group B. Patients with BMI >25 developed higher tissue stresses, including higher tangent moduli, at selected strain levels than patients with BMI <25. CONCLUSIONS: Contrary to our hypothesis, this study observed a relationship between BMI and human AVW biomechanical properties, with more obese women having stiffer tissue properties.

Viscoelastic properties measurement of the prolapsed anterior vaginal wall: a patient-directed methodology.

OBJECTIVE: In-vivo measurement of the viscoelastic properties of the prolapsed anterior vaginal wall (AVW) in post-menopausal women undergoing cystocele repair. STUDY DESIGN: A BTC-2000 cutometer-like instrument was introduced during vaginal repair of symptomatic stage 2-3 AVW prolapse. Under anesthesia, 10-mm orifice probe was applied to the AVW at the level of the bladder neck. A suction pressure ramp (0 to -147 mmHg in 6s) was delivered causing tissue uplift, followed by immediate release to 0 mmHg, measuring tissue relaxation for 20s. Similar measurements were performed over the suprapubic region (SP) for comparison purpose. The rate of tissue recovery was obtained by fitting a Voigt model to the data and expressing results as the ratio E/η [(spring modulus E)/(dashpot viscosity η)]. The effective strain energy (SE) was calculated from the pressure-uplift data and evaluated from initiation to: (1) maximum storage in tissue at peak vacuum; (2) tissue recovery after vacuum release; (3) net SE loss over the entire loading-unloading cycle. RESULTS: In 22 women, higher AVW peak and residual tissue uplift values, and lower E/η ratios were found compared with SP results. The AVW stored less elastic strain energy at peak vacuum than did the SP, and AVW net energy loss over the uplift-recovery cycle was greater than for SP controls. Not only was the AVW more compliant than the SP, with higher viscous damping, but the tissue was also less able to store recoverable energy upon distension. CONCLUSION: Such in-vivo measurements quantify the biomechanical properties of the prolapsed AVW and may assist in its management.

Biphasic investigation of tissue mechanical response during freezing front propagation.

Cryopreservation of engineered tissue (ET) has achieved limited success due to limited understanding of freezing-induced biophysical phenomena in ETs, especially fluid-matrix interaction within ETs. To further our understanding of the freezing-induced fluid-matrix interaction, we have developed a biphasic model formulation that simulates the transient heat transfer and volumetric expansion during freezing, its resulting fluid movement in the ET, elastic deformation of the solid matrix, and the corresponding pressure redistribution within. Treated as a biphasic material, the ET consists of a porous solid matrix fully saturated with interstitial fluid. Temperature-dependent material properties were employed, and phase change was included by incorporating the latent heat of phase change into an effective specific heat term. Model-predicted temperature distribution, the location of the moving freezing front, and the ET deformation rates through the time course compare reasonably well with experiments reported previously. Results from our theoretical model show that behind the marching freezing front, the ET undergoes expansion due to phase change of its fluid contents. It compresses the region preceding the freezing front leading to its fluid expulsion and reduced regional fluid volume fractions. The expelled fluid is forced forward and upward into the region further ahead of the compression zone causing a secondary expansion zone, which then compresses the region further downstream with much reduced intensity. Overall, it forms an alternating expansion-compression pattern, which moves with the marching freezing front. The present biphasic model helps us to gain insights into some facets of the freezing process and cryopreservation treatment that could not be gleaned experimentally. Its resulting understanding will ultimately be useful to design and improve cryopreservation protocols for ETs.

The influence of thermal treatment on the mechanical characteristics of a PLLA coiled stent.

We studied the effects of thermal treatment on the expansive characteristics of a coil-within-coil Poly(L-lactic acid) (PLLA) fiber stent developed at our institution to improve its mechanical performance and reproducibility. Following fabrication, furled stents were thermally treated at 62 degrees C for 25 min. The mechanical characteristics were measured compared with those of untreated stents when both were expanded via sequential balloon catheter pressure loading up to 12 atm. Treated stents reached full diameter at 3 atm and maintained that diameter despite further pressure increases. Using measurements of pressure, diameter, and axial length, we calculated the sequential mechanical work required to unfurl the stent. The mechanical work for complete unfurling of treated stents was significantly less than that required for untreated controls. Little axial dimensional change was observed for treated stents. Treated stents exhibited higher stiffness than controls at all pressure levels and also demonstrated higher resistance to external pressure-induced collapse, as measured in a special apparatus developed in our laboratory. Differential scanning calorimetry measurements indicated higher crystallinity values for fibers used in treated stents compared with controls. SEM examination of striations revealed that treated stents underwent less twist than controls following balloon-induced unfurling. The results indicate that, thermal treatment improves the reorientation and realignment of fiber crystalline structure, and favorably influences on the fiber stress-strain behavior and the expansive mechanical characteristics of the PLLA fiber stents.

Characterizing the expansive deformation of a bioresorbable polymer fiber stent.

Polymeric vascular stents must employ other strategies than malleable deformation, as generally practiced with metal stents, to expand and withstand compressive stresses in situ. The stent expansion strategy must further consider induced flow perturbations and wall stresses that may injure the vessel wall and promote thrombogenesis. Analyzing the stresses furled stents undergo during balloon-assisted expansion is an important first step in achieving a better understanding of stent-wall mechanical interactions, thereby to improve stent function. To this end, we performed finite element (FE) analysis of the balloon-induced unfurling of an internally coiled, bioresorbable polymeric stent employing a 3D FE solid model of a 120 degrees symmetric stent segment and a large deformation finite strain formulation. Uni-axial tensile testing of stent fiber elastic to plastic yielding provided the mechanical property information, and the von Mises criterion was employed to establish the elastic-plastic transition in the FE model. The model was validated with pressure and deformation measurements obtained during stent expansion tests. The internal coils of this inner coil-outer coil design twisted as the stent expanded, leading to plastic yielding at the point of tangency of the inner and outer coils. The remaining stent fiber portions underwent elastic bending. Cross-sections revealed only the outside surface layer of the coiled fiber underwent plastic yielding. The interior elastic fiber was supported by this plastic shell. The analysis suggests that during balloon-induced expansion, local plastic yielding in torsion "sets" the stent fibers, imparting high radial collapse resistance. The results further suggest that the stent exerts non-uniform mechanical forces on the vessel wall during expansion.

Mechanical characteristics of the mandible after bilateral sagittal split ramus osteotomy: comparing 2 different fixation techniques.

PURPOSE: Using finite element (FE) computer model simulation, we compared the mechanical characteristics of the mandible after bilateral sagittal split ramus osteotomy (BSSRO) through the use of 2 different techniques to stabilize the osteotomy. MATERIALS AND METHODS: Based on the reconstructed geometry from computed tomography scans of dry adult skull with a mandibular deformity requiring surgical correction, we developed 3-dimensional FE models that simulate BSSRO with 2 different techniques to stabilize the osteotomy. Technique 1 uses 3 bicortical titanium screws. Technique 2 uses a curved titanium plate with 4 monocortical screws. Five different load cases were applied to the mandible after the simulated BSSRO with the mandible being constrained at both temporomandibular joints. To evaluate the efficacy of these 2 stabilization techniques, we compared 1) the resulting deflections at the central incisor, 2) the mechanical stresses developed in the bone in the vicinity of the stabilizing implants, and 3) the mechanical stresses developed within the screw/plating system themselves. RESULTS: Technique 1, using 3 bicortical titanium screws, leads to smaller deflections at the central incisor for all 5 load cases, suggesting higher mechanical stability. Technique 1 also leads to lower mechanical stresses in the bone and in the implanted screws, whereas technique 2 is associated with higher values in each of these quantities. CONCLUSIONS: To stabilize osteotomies after a 3-dimensional simulated BSSRO, 3 bicortical screws forming an inverted-L configuration are shown to offer more effective load transmission in the mandibular construct. This technique, when examined in an FE model, leads to higher stability with lower mechanical stresses in the bone near the bicortical screws.