George E. Aninwene

Enhanced osteoblast adhesion to drug-coated anodized nanotubular titanium surfaces.

Current orthopedic implants have functional lifetimes of only 10–15 years due to a variety of reasons including infection, extensive inflammation, and overall poor osseointegration (or a lack of prolonged bonding of the implant to juxtaposed bone). To improve properties of titanium for orthopedic applications, this study anodized and subsequently coated titanium with drugs known to reduce infection (penicillin/streptomycin) and inflammation (dexamethasone) using simple physical adsorption and the deposition of such drugs from simulated body fluid (SBF). Results showed improved drug elution from anodized nanotubular titanium when drugs were coated in the presence of SBF for up to 3 days. For the first time, results also showed that the simple physical adsorption of both penicillin/streptomycin and dexamethasone on anodized nanotubular titanium improved osteoblast numbers after 2 days of culture compared to uncoated unanodized titanium. In addition, results showed that depositing such drugs in SBF on anodized titanium was a more efficient method to promote osteoblast numbers compared to physical adsorption for up to 2 days of culture. In addition, osteoblast numbers increased on anodized titanium coated with drugs in SBF for up to 2 days of culture compared to unanodized titanium. In summary, compared to unanodized titanium, this preliminary study provided unexpected evidence of greater osteoblast numbers on anodized titanium coated with either penicillin/streptomycin or dexamethasone using simple physical adsorption or when coated with SBF; results which suggest the need for further research on anodized titanium orthopedic implants possessing drug-eluting nanotubes.

Lubricin: a novel means to decrease bacterial adhesion and proliferation.

This study investigated the ability of lubricin (LUB) to prevent bacterial attachment and proliferation on model tissue culture polystyrene surfaces. The findings from this study indicated that LUB was able to reduce the attachment and growth of Staphylococcus aureus on tissue culture polystyrene over the course of 24 h by approximately 13.9% compared to a phosphate buffered saline (PBS)-soaked control. LUB also increased S. aureus lag time (the period of time between the introduction of bacteria to a new environment and their exponential growth) by approximately 27% compared to a PBS-soaked control. This study also indicated that vitronectin (VTN), a protein homologous to LUB, reduced bacterial S. aureus adhesion and growth on tissue culture polystyrene by approximately 11% compared to a PBS-soaked control. VTN also increased the lag time of S. aureus by approximately 43%, compared to a PBS-soaked control. Bovine submaxillary mucin was studied because there are similarities between it and the center mucin-like domain of LUB. Results showed that the reduction of S. aureus and Staphylococcus epidermidis proliferation on mucin coated surfaces was not as substantial as that seen with LUB. In summary, this study provided the first evidence that LUB reduced the initial adhesion and growth of both S. aureus and S. epidermidis on a model surface to suppress biofilm formation. These reductions in initial bacteria adhesion and proliferation can be beneficial for medical implants and, although requiring more study, can lead to drastically improved patient outcomes.

Nano-BaSO4: a novel antimicrobial additive to pellethane.

Hospital-acquired infections remain a costly clinical problem. Barium sulfate (BaSO4, in micron particulate form) is a common radiopacifying agent that is added to catheters and endotracheal tubes. Due to the recently observed ability of nanostructured surface features to decrease functions of bacteria without the aid of antibiotics, the objective of this in vitro study was to incorporate nano-barium sulfate into pellethane and determine the antimicrobial properties of the resulting composites. The results demonstrated for the first time that the incorporation of nano-barium sulfate into pellethane enhanced antimicrobial properties (using Staphylococcus aureus and Pseudomonas aeruginosa) compared to currently used pellethane; properties that warrant further investigation for a wide range of clinical applications.

Lubricin as a novel nanostructured protein coating to reduce fibroblast density.

Excessive fibroblast adhesion and proliferation on the surface of medical implants (such as catheters, endotracheal tubes, intraocular lenses, etc) can lead to major postsurgical complications. This study showed that when coated on tissue culture polystyrene, lubricin, a nanostructured mucinous glycoprotein found in the synovial fluid of joints, decreased fibroblast density for up to 2 days of culture compared to controls treated with phosphate buffered saline (PBS). When examining why, similar antifibroblast density results were found when coating tissue culture polystyrene with bovine submaxillary mucin (BSM), an even smaller protein closely related to the central subregion of lubricin. Additionally, results from this study demonstrated that in contrast to BSM or controls (PBS-coated and non-coated samples), lubricin was better at preserving the health of nonadherent or loosely adherent fibroblasts; fibroblasts that did not adhere or loosely adhered on the lubricin-coated tissue culture polystyrene adhered and proliferated well for up to an additional day when they were reseeded on uncoated tissue culture polystyrene. In summary, this study provides evidence for the promise of nanostructured lubricin (and to a lesser extent BSM) to inhibit fibroblast adhesion and growth when coated on medical devices; lubricin should be further explored for numerous medical device applications.

Alterations in Multi‐Scale Cardiac Architecture in Association With Phosphorylation of Myosin Binding Protein‐C

Background The geometric organization of myocytes in the ventricular wall comprises the structural underpinnings of cardiac mechanical function. Cardiac myosin binding protein‐C (MYBPC3) is a sarcomeric protein, for which phosphorylation modulates myofilament binding, sarcomere morphology, and myocyte alignment in the ventricular wall. To elucidate the mechanisms by which MYBPC3 phospho‐regulation affects cardiac tissue organization, we studied ventricular myoarchitecture using generalized Q‐space imaging (GQI). GQI assessed geometric phenotype in excised hearts that had undergone transgenic (TG) modification of phospho‐regulatory serine sites to nonphosphorylatable alanines (MYBPC3AllP−/(t/t)) or phospho‐mimetic aspartic acids (MYBPC3AllP+/(t/t)). Methods and Results Myoarchitecture in the wild‐type (MYBPC3WT) left‐ventricle (LV) varied with transmural position, with helix angles ranging from −90/+90 degrees and contiguous circular orientation from the LV mid‐myocardium to the right ventricle (RV). Whereas MYBPC3AllP+/(t/t) hearts were not architecturally distinct from MYBPC3WT, MYBPC3AllP−/(t/t) hearts demonstrated a significant reduction in LV transmural helicity. Null MYBPC3(t/t) hearts, as constituted by a truncated MYBPC3 protein, demonstrated global architectural disarray and loss in helicity. Electron microscopy was performed to correlate the observed macroscopic architectural changes with sarcomere ultrastructure and demonstrated that impaired phosphorylation of MYBPC3 resulted in modifications of the sarcomere aspect ratio and shear angle. The mechanical effect of helicity loss was assessed through a geometric model relating cardiac work to ejection fraction, confirming the mechanical impairments observed with echocardiography. Conclusions We conclude that phosphorylation of MYBPC3 contributes to the genesis of ventricular wall geometry, linking myofilament biology with multiscale cardiac mechanics and myoarchitecture.

Assessing the multiscale architecture of muscular tissue with Q-space magnetic resonance imaging: Review.

Contraction of muscular tissue requires the synchronized shortening of myofibers arrayed in complex geometrical patterns. Imaging such myofiber patterns with diffusion‐weighted MRI reveals architectural ensembles that underlie force generation at the organ scale. Restricted proton diffusion is a stochastic process resulting from random translational motion that may be used to probe the directionality of myofibers in whole tissue. During diffusion‐weighted MRI, magnetic field gradients are applied to determine the directional dependence of proton diffusion through the analysis of a diffusional probability distribution function (PDF). The directions of principal (maximal) diffusion within the PDF are associated with similarly aligned diffusion maxima in adjacent voxels to derive multivoxel tracts. Diffusion‐weighted MRI with tractography thus constitutes a multiscale method for depicting patterns of cellular organization within biological tissues. We provide in this review, details of the method by which generalized Q‐space imaging is used to interrogate multidimensional diffusion space, and thereby to infer the organization of muscular tissue. Q‐space imaging derives the lowest possible angular separation of diffusion maxima by optimizing the conditions by which magnetic field gradients are applied to a given tissue. To illustrate, we present the methods and applications associated with Q‐space imaging of the multiscale myoarchitecture associated with the human and rodent tongues. These representations emphasize the intricate and continuous nature of muscle fiber organization and suggest a method to depict structural “blueprints” for skeletal and cardiac muscle tissue.

Association between tumor architecture derived from generalized Q-space MRI and survival in glioblastoma

While it is recognized that the overall resistance of glioblastoma to treatment may be related to intra-tumor patterns of structural heterogeneity, imaging methods to assess such patterns remain rudimentary. Methods: We utilized a generalized Q-space imaging (GQI) algorithm to analyze magnetic resonance imaging (MRI) derived from a rodent model of glioblastoma and 2 clinical datasets to correlate GQI, histology, and survival. Results: In a rodent glioblastoma model, GQI demonstrated a poorly coherent core region, consisting of diffusion tracts <5 mm, surrounded by a shell of highly coherent diffusion tracts, 6-25 mm. Histologically, the core region possessed a high degree of necrosis, whereas the shell consisted of organized sheets of anaplastic cells with elevated mitotic index. These attributes define tumor architecture as the macroscopic organization of variably aligned tumor cells. Applied to MRI data from The Cancer Imaging Atlas (TCGA), the core-shell diffusion tract-length ratio (c/s ratio) correlated linearly with necrosis, which, in turn, was inversely associated with survival (p = 0.00002). We confirmed in an independent cohort of patients (n = 62) that the c/s ratio correlated inversely with survival (p = 0.0004). Conclusions: The analysis of MR images by GQI affords insight into tumor architectural patterns in glioblastoma that correlate with biological heterogeneity and clinical outcome.

Generalized Q-space MRI reveals macroscopic patterns of tumor architecture in vivo

Current approaches for studying tumor activity in patients involve molecular characterization in excised tissue or biopsied samples. Recognizing that tumors are composed of heterogeneous arrays of cells and their environment, there is a compelling rationale to explore the macroscopic organization of tumor tissue. We present a novel methodology for probing the micro-structural constituents of tumors in vivo utilizing generalized Q-space MRI. This approach employs varying magnetic field gradients and diffusion sensitivities to yield voxel-scale probability distribution functions of proton diffusivity, and then maps multi-voxel cellular alignment with tractography. Using this methodology, we describe the presence of macroscopic organizational features in patients with head and neck cancers, specifically depicting regional differences between the geometrically coherent periphery and incoherent core region. Such methods may comprise a method for assessing attributes of tumor biology in vivo and for predicting the response of such tumors to various drugs and interventions.

Tissue Engineering In Vivo with Nanotechnology

Nanotechnology has become an important part of the field of tissue engineering. Nanomaterials have several advantages over materials which only have features on the macro scale. Nanomaterials open up the possibility for more surface area for cell-surface interactions, the possibility of making stronger scaffolds for tissue re-growth, and the prospect to self-assemble nanostructures which will resemble cellular structures already found in the body. By leveraging the advantageous properties of nanomaterials, it is possible create more biocompatible implants, accelerate wound healing, facilitate tissue repair, guide nerve re-growth, eradicate infections, and reduce post injury inflammation. The following chapter will discuss several important aspects of the impact of nanotechnology on various tissue engineering applications by focusing on in vivo animal studies, a necessary step forward towards the next generation of nanomedicine.

Nano-BaSO4: A Novel Bacteriostatic Polymer Additive

Hospital acquired infections remain a major costly problem. This study sought to understand how incorporating nano-barium sulfate into pellethane composites affect the physical properties and antimicrobial nature of the resulting polymers. The results of this study showed that the incorporation of nano-barium sulfate into pellethane polymers yielded polymers which had enhanced antimicrobial properties, yet had similar hydrodynamic properties compared to pellethane polymers with standard barium sulfate.