Shiva P. Kotha

Rapid and efficient fabrication of multilevel structured silica micro-/nanofibers by centrifugal jet spinning

A rapid and efficient method consisting of two simple steps, centrifugal jet spinning (CJS) and annealing, is introduced to fabricate multilevel structured silica micro-/nanofibers. Using this technique, which is 500 times faster than electrospinning, silica micro-/nanofibers with a hollow or porous internal structure are formed as a result of non-solvent evaporation induced phase separation in the spinning solution. Silica nanofibers with solid cross sections (364 nm and 781 nm), hollow cross sections (outer and internal diameters of 458 nm and 216 nm respectively), and encapsulated voids (outside diameter of 1.4 μm where bi-continuous nano-pores 118 nm are observed) are fabricated by tuning the amount of non-solvent in the spinning solutions. This technique can be readily extended to large-scale and efficient fabrication of various ceramic materials with multileveled fibrous structures.

Collagen Fibrils in Skin Orient in the Direction of Applied Uniaxial Load in Proportion to Stress while Exhibiting Differential Strains around Hair Follicles

We determined inhomogeneity of strains around discontinuities as well as changes in orientation of collagen fibrils under applied load in skin. Second Harmonic Generation (SHG) images of collagen fibrils were obtained at different strain magnitudes. Changes in collagen orientation were analyzed using Fast Fourier Transforms (FFT) while strain inhomogeneity was determined at different distances from hair follicles using Digital Image Correlation (DIC). A parameter, defined as the Collagen Orientation Index (COI), is introduced that accounts for the increasingly ellipsoidal nature of the FFT amplitude images upon loading. We show that the COI demonstrates two distinct mechanical regimes, one at low strains (0%, 2.5%, 5% strain) in which randomly oriented collagen fibrils align in the direction of applied deformation. In the second regime, beginning at 5% strain, collagen fibrils elongate in response to applied deformation. Furthermore, the COI is also found to be linearly correlated with the applied stress indicating that collagen fibrils orient to take the applied load. DIC results indicated that major principal strains were found to increase with increased load at all locations. In contrast, minimum principal strain was dependent on distance from hair follicles. These findings are significant because global and local changes in collagen deformations are expected to be changed by disease, and could affect stem cell populations surrounding hair follicles, including mesenchymal stem cells within the outer root sheath.

Radiative transfer equation modeling by streamline diffusion modified continuous Galerkin method

Abstract. Optical tomography has a wide range of biomedical applications. Accurate prediction of photon transport in media is critical, as it directly affects the accuracy of the reconstructions. The radiative transfer equation (RTE) is the most accurate deterministic forward model, yet it has not been widely employed in practice due to the challenges in robust and efficient numerical implementations in high dimensions. Herein, we propose a method that combines the discrete ordinate method (DOM) with a streamline diffusion modified continuous Galerkin method to numerically solve RTE. Additionally, a phase function normalization technique was employed to dramatically reduce the instability of the DOM with fewer discrete angular points. To illustrate the accuracy and robustness of our method, the computed solutions to RTE were compared with Monte Carlo (MC) simulations when two types of sources (ideal pencil beam and Gaussian beam) and multiple optical properties were tested. Results show that with standard optical properties of human tissue, photon densities obtained using RTE are, on average, around 5% of those predicted by MC simulations in the entire/deeper region. These results suggest that this implementation of the finite element method-RTE is an accurate forward model for optical tomography in human tissues.

Design and analysis of a novel mechanical loading machine for dynamic in vivo axial loading.

This paper describes the construction of a loading machine for performing in vivo, dynamic mechanical loading of the rodent forearm. The loading machine utilizes a unique type of electromagnetic actuator with no mechanically resistive components (servotube), allowing highly accurate loads to be created. A regression analysis of the force created by the actuator with respect to the input voltage demonstrates high linear correlation (R(2) = 1). When the linear correlation is used to create dynamic loading waveforms in the frequency (0.5-10 Hz) and load (1-50 N) range used for in vivo loading, less than 1% normalized root mean square error (NRMSE) is computed. Larger NRMSE is found at increased frequencies, with 5%-8% occurring at 40 Hz, and reasons are discussed. Amplifiers (strain gauge, linear voltage displacement transducer (LVDT), and load cell) are constructed, calibrated, and integrated, to allow well-resolved dynamic measurements to be recorded at each program cycle. Each of the amplifiers uses an active filter with cutoff frequency at the maximum in vivo loading frequencies (50 Hz) so that electronic noise generated by the servo drive and actuator are reduced. The LVDT and load cell amplifiers allow evaluation of stress-strain relationships to determine if in vivo bone damage is occurring. The strain gauge amplifier allows dynamic force to strain calibrations to occur for animals of different sex, age, and strain. Unique features are integrated into the loading system, including a weightless mode, which allows the limbs of anesthetized animals to be quickly positioned and removed. Although the device is constructed for in vivo axial bone loading, it can be used within constraints, as a general measurement instrument in a laboratory setting.

Combinatorial therapy using negative pressure and varying lithium dosage for accelerated wound healing

In this work, we investigated the effects of negative pressure, applied using a pump designed for Negative Pressure Wound Therapy (NPWT), on the process of wound healing in vitro via initiation of the Wnt signaling pathway. Results indicate that negative pressure enhanced Wnt signaling and migration into a simulated wound in vitro in NIH-3T3 murine fibroblast cells. Increasing doses of lithium (upto 15 mM) increased basal Wnt signaling and enhanced cell migration into the simulated wound site. A combination of negative pressure and increased doses of lithium synergistically increased Wnt signaling and demonstrated further enhanced cell migration into simulated wound sites, with maximal filling of the simulated wound observed at lithium concentrations of at least 10mM.

Measurement of lacunar bone strains and crack formation during tensile loading by digital volume correlation of second harmonic generation images

The maintenance of healthy bone tissue depends upon the ability of osteocytes to respond to mechanical cues on the cellular level. The combination of digital volume correlation and second harmonic generation microscopy offers the opportunity to investigate the mechanical microenvironment of intact bone on the scale of individual osteocytes. Adult human femurs were imaged under tensile loads of 5 and 15MPa and volumes of approximately 492×429×31μm(3) were analyzed, along with an image of a bone microcrack under the same loading conditions. Principal strains were significantly higher in three-dimensional digital volume correlation when compared to two-dimensional digital image correlation. The average maximum principal strain magnitude was 5.06-fold greater than the applied global strain, with peak strains of up to 23.14-fold over global strains measured at the borders of osteocyte lacunae. Finally, a microcrack that initiated at an osteocyte lacunae had its greatest tensile strain magnitudes at the crack expansion front in the direction of a second lacunae, but strain at the crack border was reduced to background strain magnitudes upon breaching the second lacunae. This serveed to demonstrate the role of lacunae in initiating, mediating and terminating microcrack growth.

Dental imaging using laminar optical tomography and micro CT

Dental lesions located in the pulp are quite difficult to identify based on anatomical contrast, and, hence, to diagnose using traditional imaging methods such as dental CT. However, such lesions could lead to functional and/or molecular optical contrast. Herein, we report on the preliminary investigation of using Laminar Optical Tomography (LOT) to image the pulp and root canals in teeth. LOT is a non-contact, high resolution, molecular and functional mesoscopic optical imaging modality. To investigate the potential of LOT for dental imaging, we injected an optical dye into ex vivo teeth samples and imaged them using LOT and micro-CT simultaneously. A rigid image registration between the LOT and micro-CT reconstruction was obtained, validating the potential of LOT to image molecular optical contrast deep in the teeth with accuracy, non-invasively. We demonstrate that LOT can retrieve the 3D bio-distribution of molecular probes at depths up to 2mm with a resolution of several hundred microns in teeth.

Bone strain measurement using 3D digital image correlation of second harmonic generation images

Collagen fibrils contribute to the structural integrity and crack resistance of bone, but their response to stress while in bundles of fibrils is not well known. Digital image correlation (DIC) is a powerful technique for measuring strain by comparing images of deformed samples to non-deformed samples, while second harmonic generation microscopy (SHGM) captures bright, high contrast images of bone with strong signals from collagen fibrils. Combining both techniques allows the 3-dimensional strain environments of bundled collagen fibrils to be characterized. SHGM was performed using confocal microscopy, producing images of human femur bone at 5 MPa and 50 MPa of applied tensional stress with 0.5×0.5×1 µm voxels. A DIC algorithm generated preliminary displacement and strain maps from the images, with the average calculated strain varying from the theoretical strain. The local strain average in the direction of applied stress was 1.09%, compared to a global strain of 0.627%, which was attributed to the small volume of analysis compared to the global image, and the proximity of a lacuna to the analysis region. The strain response was also found to be highly heterogeneous. Future work will analyze larger regions of bone.

Dental Optical Tomography with UCNPs

Upconverting nanoparticles (UCNP) have the unique ability to emit both in the NIR and visible light upon NIR excitation. Herein we investigate experimentally the potential to use UCNPs in dental imaging ex vivo.

Magnitude of loads influences the site of failure of highly curved bones

The structure and material properties of bones along with applied boundary conditions determine the region of peak stresses, where fracture is expected to occur. As the site of peak stresses is not influenced by the magnitude of applied load, the fracture site is not expected to change during fatigue loading of whole bone at different loads. However, in a highly curved bone such as the rat ulna, the magnitude of applied loads was found to influence the fracture site. Fatigue loading was conducted under load control on intact rat forearms and on excised ulnae. The distance to the site of failure from the proximal olecranon process of ulnae was determined. In intact forearms, the site of failure demonstrated a linear progression distally, towards sites with lower moment of inertia (or sites exhibiting lower section modulus). Intact rat forearms and excised ulnae loaded to failure at low loads fractured 2-3mm distal to where they failed when applying high loads. This indicates a shift in the site of failure by approximately 10% of whole bone length just by varying the applied load magnitude. The site of failure in excised ulnae was similar when loading at 2Hz or at 4Hz, suggesting that this was frequency independent in this range and indicating that strain rate was not an important contributing factor. Creep loading of excised ulnae also demonstrated similar changes in the site of failure, indicating that magnitude of loads, and not type of loading were important in determining the site of failure. This has important implications with regards to the volume of bone that undergoes damage under physiological loading, before it fails.