Effects of Alendronate on Particle-induced Osteolysis in a Rat Model

The Journal of Bone and Joint Surgery. American Volume. Feb, 2002  |  Pubmed ID: 11861730

Particle-induced osteolysis is currently a major problem affecting the long-term survivorship of total joint replacements. Alendronate is a third-generation bisphosphonate that blocks osteoclastic bone resorption. The objective of this study was to determine whether alendronate could prevent particle-induced osteolysis or restore (reverse) bone loss in established osteolysis.

Elucidation of a Low Spin Cobalt(II) System in a Distorted Tetrahedral Geometry

Journal of the American Chemical Society. Dec, 2002  |  Pubmed ID: 12487609

We have prepared a series of divalent cobalt(II) complexes supported by the [PhBP(3)] ligand ([PhBP(3)] = [PhB(CH(2)PPh(2))(3)](-)) to probe certain structural and electronic phenomena that arise from this strong field, anionic tris(phosphine) donor ligand. The solid-state structure of the complex [PhBP(3)]CoI (1), accompanied by SQUID, EPR, and optical data, indicates that it is a pseudotetrahedral cobalt(II) species with a doublet ground state-the first of its type. To our knowledge, all previous examples of 4-coordinate cobalt(II) complexes with doublet ground states have adopted square planar structure types. Complex 1 provided a useful precursor to the corresponding bromide and chloride complexes, ([PhBP(3)]Co(mu-Br))(2), (2), and ([PhBP(3)]Co(mu-Cl))(2), (3). These complexes were similarly characterized and shown to be dimeric in the solid-state. In solution, however, the monomeric low spin form of 2 and 3 dominates at 25 degrees C. There is spectroscopic evidence for a temperature-dependent monomer/dimer equilibrium in solution for complex 3. Furthermore, the dimers 2 and 3 did not display appreciable antiferromagnetic coupling that is typical of halide and oxo-bridged copper(II) and cobalt(II) dimers. Rather, the EPR and SQUID data for solid samples of 2 and 3 suggest that they have triplet ground states. Complexes 1, 2, and 3 are extremely oxygen sensitive. Thus, stoichiometric oxidation of 1 by dioxygen produced the 4-coordinate, high spin complex [PhB(CH(2)P(O)Ph(2))(2)(CH(2)PPh(2))]CoI, (4), in which the [PhBP(3)] ligand had undergone a 4-electron oxidation. Reaction of 1 with TlOAr (Ar = 2,6-Me(2)Ph) afforded an example of a 4-coordinate, high spin complex, [PhBP(3)]Co(O-2,6-Me(2)Ph) (5), with an intact [PhBP(3)] ligand. The latter two complexes were spectroscopically and structurally characterized for comparison to complexes 1, 2, and 3. Our data for these complexes collectively suggest that the [PhBP(3)] ligand provides an unusually strong ligand-field to these divalent cobalt complexes that is chemically distinct from typical tris(phosphine) donor ligand sets, and distinct from tridentate borato ligands that have been previously studied. Coupling this strong ligand-field with a pronounced axial distortion away from tetrahedral symmetry, a geometric consequence that is enforced by the [PhBP(3)] ligand, provides access to monomeric [PhBP(3)]CoX complexes with doublet rather than quartet ground states.

Characterization of a Developing Lumbar Arthrodesis in a Sheep Model with Quantitative Instability

The Spine Journal : Official Journal of the North American Spine Society. Jul-Aug, 2002  |  Pubmed ID: 14589474

Mechanical forces have been considered responsible for stress shielding an arthrodesis, but the biology of a developing lumbar fusion has not been well characterized.

Assessment of a Synthetic Anterior Cervical Ligament in a Spinal Fusion Model in Sheep

The Spine Journal : Official Journal of the North American Spine Society. Jul-Aug, 2002  |  Pubmed ID: 14589477

The anterior cervical ligament is routinely excised during cervical interbody fusion. Loss of this ligament may predispose to instability at the fusion site. Anterior plating restores stability but leads to a risk of stress shielding. An alternative approach would be to use a less rigid fixation system that would provide anterior support while allowing micromotion that could enhance bone healing within the fusion site.

Defect Repair in the Rat Mandible with Bone Morphogenic Proteins and Marrow Cells

Archives of Facial Plastic Surgery : Official Publication for the American Academy of Facial Plastic and Reconstructive Surgery, Inc. and the International Federation of Facial Plastic Surgery Societies. Jan-Feb, 2003  |  Pubmed ID: 12533151

To investigate the ability of a bone growth factor mixture and bone marrow cells to repair a critical size defect of the rat mandibular body.

Synthesis and Visualization of a Membrane-permeable MRI Contrast Agent

Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry. Sep, 2003  |  Pubmed ID: 14505078

The study of in vivo developmental events has undergone significant advances with the advent of biological molecular imaging techniques such as computer enhanced light microscopy imaging, positron emission tomography (PET), micro-CT, and magnetic resonance imaging (MRI). MRI has proven to be a particularly powerful tool in clinical and biological settings. Images can be acquired of opaque living animals, with the benefit of tracking events of extended periods of time on the same specimen. Contrast agents are routinely used to enhance regions, tissues, and cells that are magnetically similar but histologically distinct. A principal barrier to the development of MR contrast agents for investigating developmental biological questions is the ability to deliver the agent across cellular membranes. As part of our research, we are investigating a number of small molecules that facilitate transport of charged and uncharged species across cell membranes. Here we describe the synthesis and testing of a Gd(III)-based MR contrast agent conjugated to polyarginine that is able to permeate cell membranes. We confirmed cellular uptake of the agent using two-photon laser microscopy to visualize a Eu(III) derivative of the contrast agent in cell culture, and verified this uptake by T(1) analysis of the Gd(III) agent in cells.

Precision and Accuracy of DXA and PQCT for Densitometry of the Rat Femur

Journal of Clinical Densitometry : the Official Journal of the International Society for Clinical Densitometry. 2003  |  Pubmed ID: 14716052

Measurements of bone mineral density and bone mineral content are key data in the study of osteoporosis and pathologic skeletal disease. Dual-energy X-ray absorptiometry and peripheral quantitative computed tomography are used in human and small animal studies. The purpose of this study was to evaluate the precision, accuracy, and systematic bias of measurement of the rat femur. Comparing machine-measured parameters with standard, nonradiographic measurements, we assessed validation of relative and absolute accuracy. Regression analysis and calculations of percent difference from standard values were used to determine the accuracy of each densitometry technique. Machine-specific and subject-specific precision was evaluated for each densitometer using repeated scans to calculate coefficients of variation. Each of the methods of densitometry examined in this study produced comparable results and was sensitive to small changes following experimental stimuli. Further, our assessment of the precision and accuracy observed between methods of scanning excised rat femurs validates our data acquisition method and serves as a foundation for future densitometry studies.

Technical Feasibility and Precision of Radiostereometric Analysis As an Outcome Measure in Canine Cemented Total Hip Replacement

Journal of Orthopaedic Science : Official Journal of the Japanese Orthopaedic Association. 2004  |  Pubmed ID: 14767707

Radiostereometric analysis (RSA) is an analytical technique in which biplanar radiographs are used to quantify the migration of implants in vivo. RSA is now considered the benchmark for quantifying implant migration in clinical studies of total joint replacement, yet its use in preclinical animal models has not been widely reported. The same attributes that make RSA appealing as an analytical method in humans (i.e., high precision and accuracy, noninvasiveness, objectivity) also make it a promising option for animal studies. The specific aims of this study were to determine the technical feasibility and analytical precision of RSA in a canine model of cemented total hip replacement. The precision of RSA was assessed in (1) a Plexiglas phantom, (2) a canine Sawbone model, and (3) a pilot series of dogs implanted with cemented canine THR implants. In vitro precision values, calculated as the 95% confidence limits for the error between duplicate RSA examinations, ranged from 4.3 to 17.9 microm for translation and from 0.01 degrees to 0.22 degrees for rotation. In vivo precision values ranged from 16.2 to 41.1 microm for translation and 0.17 degrees to 0.44 degrees for rotation. As is the case in humans, RSA appears to provide an order-of-magnitude improvement in technical precision as compared to plain film X-ray. RSA can therefore be considered a practical and potentially valuable noninvasive outcome measure for assessing implant function in canine cemented THR.

Cellular Delivery of MRI Contrast Agents

Chemistry & Biology. Mar, 2004  |  Pubmed ID: 15123259

Magnetic resonance imaging (MRI) is a powerful tool for acquiring images of opaque living animals with the benefit of tracking events over extended periods of time on the same specimen. Contrast agents are used to enhance regions, tissues, and cells that are magnetically similar but histologically distinct. A principal barrier to the development of MRI contrast agents for investigating biological questions is the delivery of agents across cellular membranes. Here, we describe the synthesis and in vitro testing of Gd(III)-based MRI contrast agents containing varying length polyarginine oligomers capable of permeating cell membranes. We examine the effect of the length of oligomer on T(1) enhancement and cellular uptake. Furthermore, the effect of incubation time, concentration, and cell type on uptake is explored. Toxicity and washout studies are performed in addition to MRI phantom studies.

Preclinical Evaluation of a Poly (vinyl Alcohol) Hydrogel Implant As a Replacement for the Nucleus Pulposus

Spine. Mar, 2004  |  Pubmed ID: 15129064

An in vivo investigation into the safety of a novel hydrogel implant designed to replace the diseased nucleus pulposus.

Magnetic Resonance Contrast Agents for Medical and Molecular Imaging

Metal Ions in Biological Systems. 2004  |  Pubmed ID: 15206098

Markers of Bone Metabolism in Dog Breeds of Different Size

Research in Veterinary Science. Feb, 2004  |  Pubmed ID: 14659729

Serum and urinary markers of bone turnover may be of value in animals as noninvasive tools for determining the response of the skeleton to disease and injury. Although normal values for bone markers have been reported for the Beagle, concerns remain that breed to breed differences will complicate the interpretation of bone marker data in dogs. To explore this, we examined serum bone markers in two breeds of vastly different size, Pomeranians and Irish Wolfhounds. Our hypothesis was that serum concentrations of bone markers are similar in toy and giant dog breeds and fall within the same range as those reported for Beagles. Bone alkaline phosphatase (BALP) and carboxy-terminal telopeptide of type I collagen (ICTP), respectively markers of bone formation and bone resorption, were measured in age matched Pomeranians (n=14) and Irish Wolfhounds (n=14). No statistically significant differences between the mean BALP and mean ICTP serum concentrations from Pomeranians and Irish Wolfhounds were found. All BALP and ICTP concentrations were within the reference range reported for Beagles. The results of this study suggest that serum BALP and ICTP concentrations in giant and toy breeds are the same as in Beagles and that these assays may be used for dogs of all sizes.

Bone Turnover Mediates Preferential Localization of Prostate Cancer in the Skeleton

Endocrinology. Apr, 2005  |  Pubmed ID: 15637291

Bone metastasis is a common untreatable complication associated with prostate cancer. Metastatic cells seed in skeletal sites under active turnover containing dense marrow cellularity. We hypothesized that differences in these skeletal-specific processes are among the critical factors that facilitate the preferential localization of metastatic prostate cancer in bone. To test this, athymic mice were administered PTH to induce bone turnover and increase marrow cellularity daily 1 wk before and after intracardiac inoculation of luciferase-tagged PC-3 cells. Tumor localization was monitored by bioluminescence imaging weekly for 5 wk. At the time of tumor inoculation, PTH-treated mice demonstrated significant increases in serum levels of bone turnover markers such as osteocalcin and tartrate-resistant acid phosphatase 5b and in the number of tartrate-resistant acid phosphatase-positive osteoclasts per millimeter of bone when compared with the other groups. Likewise, PTH treatment stimulated a qualitative increase in marrow cellular proliferation as determined by 5-bromo-2'-deoxyuridine immunostaining. Skeletal metastases formed in the hind limb and craniofacial regions of young mice with no difference between groups. In adult mice, however, bioluminescent signals in the hind limb and craniofacial regions were 3-fold higher in PTH-treated mice vs. controls. Fluorochrome labeling revealed increased bone formation activity in trabecular bone adjacent to tumors. When zoledronic acid, a nitrogen-containing bisphosphonate that inhibits osteoclast-mediated bone resorption, was administered concurrently with PTH, a significant reduction in the incidence of bone tumors was observed. Overall, these studies provide new evidence that skeletal sites rich in marrow cellularity under active turnover offer a more congenial microenvironment to facilitate cancer localization in the skeleton.

Color-blind Fluorescence Detection for Four-color DNA Sequencing

Proceedings of the National Academy of Sciences of the United States of America. Apr, 2005  |  Pubmed ID: 15800037

We present an approach called pulsed multiline excitation (PME) for measurements of multicomponent, fluorescence species and demonstrate its application in capillary electrophoresis for DNA sequencing. To fully demonstrate the advantages of PME, a fluorescent dye set has been developed whose absorption maxima span virtually the entire visible spectrum. Unlike emission wavelength-dependent approaches for identifying fluorescent species, the removal of the spectral component in PME confers a number of advantages including higher and normalized signals from all dyes present in the assay, the elimination of spectral cross-talk between dyes, and higher signal collection efficiency. Base-calling is unambiguously determined once dye mobility corrections are made. These advantages translate into significantly enhanced signal quality as illustrated in the primary DNA sequencing data and provide a means for achieving accurate base-calling at lower reagent concentrations.

Solid-phase Synthesis of Polymers Using the Ring-opening Metathesis Polymerization

Journal of the American Chemical Society. Oct, 2005  |  Pubmed ID: 16231882

We report a general method for the solid-phase synthesis of polymers via the ring-opening metathesis polymerization (ROMP). The method involves polymerization in solution to form a block copolymer, immobilization of the polymer via reaction of one block with a resin-bound functional group, modification of the other block, and liberation of the polymer from the resin. We demonstrated the utility of this approach by generating a block copolymer with an N-hydroxysuccinimidyl ester-substituted block (for on-resin functionalization) and a maleimide-substituted block (for conjugation to the resin). We showed that the Diels-Alder reaction can be employed to immobilize the polymers and that amines of diverse structure can be used to modify the resin-bound polymers. The reversibility of the furan-maleimide Diels-Alder adduct was exploited to liberate the polymer from the support. Specifically, treatment of the resin with cyclopentadiene resulted in complete polymer release. The resulting polymers are functional: they were as potent in assays with the lectin concanavalin A as polymers generated by traditional solution routes. We anticipate that this method can be used for the rapid synthesis of diverse polymers via ROMP.

Temporal Changes in Bone Mass and Mechanical Properties in a Murine Model of Tumor Osteolysis

Bone. Mar, 2006  |  Pubmed ID: 16278105

Pathological fracture is a devastating complication of osteolytic bone metastases. The progression of osteolysis and its effect on bone fracture risk are poorly understood. The goal of this study was to determine the temporal changes in bone strength following tumor inoculation in a preclinical model of tumor osteolysis. In addition, a predictive model was developed between non-invasive radiographic measures and bone strength. The right femora of female nude mice were injected with breast cancer cells; the left limb served as a sham-operated control. Radiographs and DEXA scans were obtained at the time of surgery and at 3, 6, and 9 weeks. Groups of mice were euthanized at each time point for mechanical assessment. Micro-CT analysis was performed on a sub-set of mice with advanced state disease to quantify bone loss. Radiographs documented an increase in tumor osteolysis over time, with 58% of the mice showing signs of osteolysis at 3 weeks, 75% at 6 weeks, and 81% at 9 weeks. BMD measurements revealed a 21.6% increase from baseline in the controls whereas tumor-injected femora failed to increase in BMD over the same time course. Tumor-bearing limbs exhibited statistically significant decreases in torque at failure (86%), energy to failure (88%), and initial stiffness (94%) compared to the controls. Both lysis scores and BMD measurements proved to be modest predictors of mechanical strength, accounting for approximately 73% and 41% of variation in torque at failure, respectively. Micro-CT analysis revealed decreases in both total bone volume in the distal femur (31%) and metaphyseal fractional trabecular bone (89%). We have shown that non-invasive radiographic techniques provide a useful tool for monitoring the progression of tumor osteolysis and for predicting the mechanical strength of tumor-bearing bones in this model. By integrating non-invasive measures of tumor osteolysis and fracture risk, we have validated a clinically relevant platform for evaluating new therapeutic approaches for preserving and/or restoring bone affected by metastatic disease.

Tissue Response to in Situ Polymerization of a New Two-solution Bone Cement: Evaluation in a Sheep Model

Journal of Biomedical Materials Research. Part B, Applied Biomaterials. Nov, 2006  |  Pubmed ID: 16615074

A two-solution bone cement (2-SC) was evaluated in a non-load bearing sheep model that simulated insertion of a cemented total joint replacement. A commercial powder-liquid bone cement formulation (Palacos R) was used as the control. The systemic response to the two cements was determined by monitoring changes in arterial blood pressure (ABP) and serum concentrations of methyl methacrylate monomer at intervals after insertion of the cement. The short-term tissue response to the two cements was assessed by quantifying histomorphometric parameters of new bone formation at 2, 4, and 12 weeks postsurgery. Intraoperatively, injection and pressurization of bone cement were well tolerated, with no significant changes in ABP in either group and no detectable circulating monomer in any animal. Several interesting trends were identified in the histomorphometry data. In the trabecular specimens, new bone formation immediately adjacent to the cement mantle was apparently suppressed in the first 2 weeks postsurgery, increased dramatically at 4 weeks, and then returned to baseline values by 12 weeks. This pattern was seen with both Palacos and 2-SC. In the cortical specimens, new bone formation was reduced on the endosteal surface when compared with the periosteal surface, with this effect being more noticeable at 2 and 4 weeks than at 12 weeks. There were no significant histopathological findings in either the bone or the draining lymph nodes. These data indicate that the biological response to 2-SC is substantially equivalent to that of Palacos R. Additional testing in a functional, load-bearing animal model is now recommended to more fully characterize the long-term biological response to 2-SC and to determine the mechanical performance of this new cement in vivo.

Contrast Agents for Magnetic Resonance Imaging Synthesized with Ring-opening Metathesis Polymerization

Journal of the American Chemical Society. May, 2006  |  Pubmed ID: 16704234

A monomer for ring-opening metathesis polymerization (ROMP) has been developed that also functions as a portion of a GdIII chelating moiety for a magnetic resonance imaging contrast agent. An increase in per GdIII relaxivity was shown upon transition from monomer to polymer. Additionally, extremely large molecular relaxivities were achieved through incorporation of multiple GdIII ions per polymer. The nature of ROMP-derived polymers allows for functionalization of the monomer units and termini through orthogonal chemistry. This strategy is the basis for a new class of highly sensitive, targeted imaging agents.

Quantitative Imaging of Cell-permeable Magnetic Resonance Contrast Agents Using X-ray Fluorescence

Molecular Imaging. Oct-Dec, 2006  |  Pubmed ID: 17150161

The inability to transduce cellular membranes is a limitation of current magnetic resonance imaging probes used in biologic and clinical settings. This constraint confines contrast agents to extracellular and vascular regions of the body, drastically reducing their viability for investigating processes and cycles in developmental biology. Conversely, a contrast agent with the ability to permeate cell membranes could be used in visualizing cell patterning, cell fate mapping, gene therapy, and, eventually, noninvasive cancer diagnosis. Therefore, we describe the synthesis and quantitative imaging of four contrast agents with the capability to cross cell membranes in sufficient quantity for detection. Each agent is based on the conjugation of a Gd(III) chelator with a cellular transduction moiety. Specifically, we coupled Gd(III)-diethylenetriaminepentaacetic acid DTPA and Gd(III)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid with an 8-amino acid polyarginine oligomer and an amphipathic stilbene molecule, 4-amino-4'-(N,N-dimethylamino)stilbene. The imaging modality that provided the best sensitivity and spatial resolution for direct detection of the contrast agents is synchrotron radiation x-ray fluorescence (SR-XRF). Unlike optical microscopy, SR-XRF provides two-dimensional images with resolution 10(3) better than (153)Gd gamma counting, without altering the agent by organic fluorophore conjugation. The transduction efficiency of the intracellular agents was evaluated by T(1) analysis and inductively coupled plasma mass spectrometry to determine the efficacy of each chelate-transporter combination.

Concurrent Administration of Zoledronic Acid and Irradiation Leads to Improved Bone Density, Biomechanical Strength, and Microarchitecture in a Mouse Model of Tumor-induced Osteolysis

Journal of Surgical Oncology. Mar, 2008  |  Pubmed ID: 18161868

Bone metastases are typically treated with bisphosphonates as adjuncts to radiation therapy. The goal of this study was to determine whether radiotherapy combined with a bisphosphonate could restore bone density, microarchitecture, and biomechanical strength of a tumor-burdened bone to normal.

Predicting Distal Femur Bone Strength in a Murine Model of Tumor Osteolysis

Clinical Orthopaedics and Related Research. Jun, 2008  |  Pubmed ID: 18404290

Predicting pathologic fractures of long bones caused by metastatic disease continues to be a challenging clinical problem. We assessed the ability of noninvasive imaging and computational techniques to predict the strength of bones with osteolytic lesions. A murine model of induced tumor osteolysis to the distal femur was used as a model system resulting in a wide range of lesion sizes. Microcomputed tomography scans were obtained and specimen-specific, voxel-based, finite element analyses were performed and results were compared with direct measurement of biomechanical strength via axial compressive loading of the distal femur. Additional indirect surrogates of bone strength included dual-energy xray absorptiometry to determine bone mineral density, radiographic scoring, and computed tomography volume/mineral estimates. Predicted bone strength was weakest (r(2) = 0.55) for the dual-energy xray absorptiometry measure and strongest (r(2) = 0.91) for the direct computed tomography voxel-based, finite element analysis. The relative success of the voxel-based, finite element modeling approach to estimate bone strength in a murine osteolytic tumor model indicates this approach, with further development and validation, could serve as a way to nondestructively estimate bone strength in a clinical setting.

The Differential Effects of the Radioprotectant Drugs Amifostine and Sodium Selenite Treatment in Combination with Radiation Therapy on Constituent Bone Cells, Ewing's Sarcoma of Bone Tumor Cells, and Rhabdomyosarcoma Tumor Cells in Vitro

Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society. Nov, 2008  |  Pubmed ID: 18473385

The purpose of this study was to determine the differential effects of therapeutic X-radiation on constituent bone cells relative to the pediatric tumor cells: Ewing's sarcoma of bone and rhabdomyosarcoma. In addition, the radioprotectant drugs amifostine and sodium selenite were administered to constituent bone cells and the two tumor cells to determine if the radioprotectants differentially protect bone cells while not benefiting the tumor cells. These studies are a necessary first step in determining the potential clinical benefit of radioprotective therapy. An established in vitro cell culture model employing both constituent bone cells (osteoblasts, primary bone marrow monocytes, osteoclasts chondrocytes, and endothelial cells) and the tumor cells lines (Ewing's sarcoma of bone and rhabdomyosarcoma) were exposed to irradiation, amifostine, and sodium selenite. Cells were then assayed for changes in cell number, cytotoxicity, mineralization, bone resorption, cell attachment, osteocalcin, caspase-3 expression, clonogenic survival, and alkaline phosphatase expression. Radiation therapy differentially decreased cell number; with osteoblasts being shown to be the least sensitive to irradiation, the tumor cells had an intermediate sensitivity and monocytes were the most sensitive. Both amifostine and sodium selenite protected chondrocytes and osteoblasts from the negative effects of irradiation, while not protecting the tumor cells. The pediatric tumor cell lines were generally more radiosensitive than the bone cells examined. The radioprotectant drugs amifostine and sodium selenite provided significant radioprotection to constituent bone cells while not protecting the tumor cells. Finally, amifostine and sodium selenite therapy provided an additional benefit beyond radioprotection by increasing cytotoxicity in nonirradiated and irradiated tumor cells.

Metastatic Osteosarcoma Gene Expression Differs in Vitro and in Vivo

Clinical Orthopaedics and Related Research. Sep, 2008  |  Pubmed ID: 18516656

An understanding of differential gene expression in highly metastatic osteosarcoma could provide gene targets for treatment of metastases. We compared gene expression profiles of high- (LM7) and low- (LM2) metastatic SaOS2-derived cell lines in an in vitro tissue culture model and examined several differentially regulated genes in vivo in a murine orthotopic xenograft model. We hypothesized an orthotopic inoculation of LM2 and LM7 cells would establish a primary lesion and the gene expression profile of cells grafted in this fashion would resemble the gene expression profile observed in an in vitro model. Thirty-five days after inoculation, animals were euthanized and both tibiae were harvested and rapidly frozen in liquid nitrogen. Human-specific GAPDH mRNA was present in two of four tibias inoculated with LM2 cells and three of four tibias inoculated with LM7 cells. Tibiae displaying the presence of human cells were assayed by semiquantitative reverse transcriptase polymerase chain reaction. We observed poor correspondence of in vitro to in vivo gene expression for either cell line. Accordingly, in vitro osteosarcoma gene expression data must be interpreted with caution until confirmed in vivo. Our orthotopic injection model allowed in vivo study of differential gene expression between these two cell lines but did not show radiographic evidence of an established primary lesion.

High-throughput Solution Processing of Large-scale Graphene

Nature Nanotechnology. Jan, 2009  |  Pubmed ID: 19119278

The electronic properties of graphene, such as high charge carrier concentrations and mobilities, make it a promising candidate for next-generation nanoelectronic devices. In particular, electrons and holes can undergo ballistic transport on the sub-micrometre scale in graphene and do not suffer from the scale limitations of current MOSFET technologies. However, it is still difficult to produce single-layer samples of graphene and bulk processing has not yet been achieved, despite strenuous efforts to develop a scalable production method. Here, we report a versatile solution-based process for the large-scale production of single-layer chemically converted graphene over the entire area of a silicon/SiO(2) wafer. By dispersing graphite oxide paper in pure hydrazine we were able to remove oxygen functionalities and restore the planar geometry of the single sheets. The chemically converted graphene sheets that were produced have the largest area reported to date (up to 20 x 40 microm), making them far easier to process. Field-effect devices have been fabricated by conventional photolithography, displaying currents that are three orders of magnitude higher than previously reported for chemically produced graphene. The size of these sheets enables a wide range of characterization techniques, including optical microscopy, scanning electron microscopy and atomic force microscopy, to be performed on the same specimen.

Practical Chemical Sensors from Chemically Derived Graphene

ACS Nano. Feb, 2009  |  Pubmed ID: 19236064

We report the development of useful chemical sensors from chemically converted graphene dispersions using spin coating to create single-layer films on interdigitated electrode arrays. Dispersions of graphene in anhydrous hydrazine are formed from graphite oxide. Preliminary results are presented on the detection of NO(2), NH(3), and 2,4-dinitrotoluene using this simple and scalable fabrication method for practical devices. Current versus voltage curves are linear and ohmic in all cases, studied independent of metal electrode or presence of analytes. The sensor response is consistent with a charge transfer mechanism between the analyte and graphene with a limited role of the electrical contacts. A micro hot plate sensor substrate is also used to monitor the temperature dependence of the response to nitrogen dioxide. The results are discussed in light of recent literature on carbon nanotube and graphene sensors.

Polylactide-co-glycolide Fiber-reinforced Calcium Phosphate Bone Cement

Archives of Facial Plastic Surgery : Official Publication for the American Academy of Facial Plastic and Reconstructive Surgery, Inc. and the International Federation of Facial Plastic Surgery Societies. Mar-Apr, 2009  |  Pubmed ID: 19289682

To compare the strength of polylactide-co-glycolide fiber-reinforced calcium phosphate bone cement (FRC) with nonreinforced calcium phosphate bone cement (NRC) subjected to simulated dural pulsations in defects larger than 25 cm(2).

Low-temperature Solution Processing of Graphene-carbon Nanotube Hybrid Materials for High-performance Transparent Conductors

Nano Letters. May, 2009  |  Pubmed ID: 19361207

We report the formation of a nanocomposite comprised of chemically converted graphene and carbon nanotubes. Our solution-based method does not require surfactants, thus preserving the intrinsic electronic and mechanical properties of both components, delivering 240 ohms/square at 86% transmittance. This low-temperature process is completely compatible with flexible substrates and does not require a sophisticated transfer process. We believe that this technology is inexpensive, is massively scalable, and does not suffer from several shortcomings of indium tin oxide. A proof-of-concept application in a polymer solar cell with power conversion efficiency of 0.85% is demonstrated. Preliminary experiments in chemical doping are presented and show that optimization of this material is not limited to improvements in layer morphology.

Dynamic Measurements of Aqueous Lanthanide Triflate-catalyzed Reactions Using Luminescence Decay

Journal of the American Chemical Society. May, 2009  |  Pubmed ID: 19385628

There is tremendous interest in water-compatible lanthanide triflate-based catalysts for carbon-carbon bond forming reactions; however, poor understanding of their aqueous mechanism severely limits the ability to increase the utility of these catalysts. Here, we report dynamic measurements of the water-coordination number of lanthanide triflate-based catalysts using luminescence-decay measurements in a range of aqueous systems. This unique characterization method is a reliable, convenient, and fast approach to analyze lanthanide-based catalysts in aqueous systems.

Cemented Total Knee Replacement in 24 Dogs: Surgical Technique, Clinical Results, and Complications

Veterinary Surgery : VS. Jul, 2009  |  Pubmed ID: 19573056

To characterize the performance of cemented total knee replacement (TKR) in dogs.

Chemically Induced Folding of Single and Bilayer Graphene

Chemical Communications (Cambridge, England). Nov, 2009  |  Pubmed ID: 19826696

Here we report chemically induced folding of thin graphene flakes. The folding occurs spontaneously when an intercalating species interrupts the adhesion between graphene and a supporting substrate. The morphology of induced folds suggests that the conjugated pi network is capable of extremely sharp curvature. Adjacent folds are often parallel, suggesting preferential deformation along certain crystallographic planes.

ROMP from ROMP: A New Approach to Graft Copolymer Synthesis

Macromolecules. Jun, 2009  |  Pubmed ID: 20161406

A new strategy is presented for the synthesis of graft copolymers using only the ring-opening metathesis polymerization (ROMP). From a ROMP-derived main chain, pendant maleimide functional groups are converted into norbornene moieties via a Diels-Alder reaction with cyclopentadiene. The norbornene groups serve as sites of initiation, and subsequent ROMP from the main chain yields graft copolymers with both main and side chains derived from ROMP. This strategy offers ready access to defined graft copolymers.

Anabolic and Antiresorptive Drugs Improve Trabecular Microarchitecture and Reduce Fracture Risk Following Radiation Therapy

Calcified Tissue International. Sep, 2010  |  Pubmed ID: 20563797

Many patients with symptomatic bone metastases receive radiation therapy, even though radiation is known to have potential adverse effects on bone. We hypothesized that the concurrent use of a bisphosphonate drug (zoledronic acid, ZA) or a combination of ZA plus an anabolic agent (parathyroid hormone, PTH) would lead to improvements in the microarchitecture and mechanical properties of irradiated bone. Human breast cancer cells were injected into the distal femur of 56 female nude mice, which were then divided into four groups: no treatment (0 Gy), radiation administered 4 weeks postinjection (20 Gy), radiation plus ZA (12.5 microg/kg weekly from weeks 4 to 12) (20 Gy + ZA), and radiation followed by ZA (25 microg/kg weekly from weeks 4 to 8) and PTH(1-34) (100 microg microg/kg daily from weeks 8 to 12) (20 Gy + ZA + PTH). Left limbs served as normal control bones. Bone loss over the 12-week study was tracked with serial radiography and bone densitometry. At the end of the study, micro-computed tomography and mechanical testing were used to quantify bone microarchitecture and bone strength. Radiation alone failed to prevent tumor-induced decreases in bone mineral density (BMD), trabecular bone volume, and bone strength. Treatment with 20 Gy + ZA or 20 Gy + ZA + PTH as adjuncts to radiation was effective at preserving trabecular bone architecture and bone strength at normal levels. ZA reduced the risk of mechanical fragility following irradiation of a lytic bone lesion. Supplemental use of PTH did not result in further increases in bone strength but was associated with significant increases in BMD and bone mass, suggesting that it may be beneficial in enhancing bone architecture following radiation therapy.

Local Irradiation Alters Bone Morphology and Increases Bone Fragility in a Mouse Model

Journal of Biomechanics. Oct, 2010  |  Pubmed ID: 20655052

Insufficiency fracture following radiation therapy (RTx) is a challenging clinical problem and typical bone mass measures fail to predict these fractures. The goals of this research were to develop a mouse model that results in reduced bone strength following focal irradiation, quantify morphological and strength changes occurring over time, and determine if a positive correlation between bone morphology and strength is retained after irradiation. Right hind limbs of 13 week-old female Balb/c mice were irradiated (5 or 20 Gy) using a therapeutic X-ray unit. Left limbs served as control. Animals were euthanized at 2, 6, 12, or 26 weeks. Axial compression tests of the distal femur were used to quantify whole bone strength. Specimen-specific, non-linear finite element (FE) analyses of the mechanical tests were performed using voxel-based meshes with two different material failure models: a linear bone density-strength relationship and a non-linear 'embrittled' relationship. Radiation resulted in a dose dependent increase in cortical bone density and marked loss of trabecular bone, measured using micro-CT. An early (2 week) increase in bone volume was associated with an increase in bone strength following irradiation; at 12 weeks there was a loss of bone strength despite higher bone volume for irradiated limbs. There was a positive correlation between bone volume bone and strength in control (r²=0.63) but not irradiated femora (r²=0.08). FE analysis with a constant strain failure model resulted in improved prediction of bone strength for irradiated limbs (r²=0.34) and this was improved further with the embrittled material model (r²=0.46). In summary, focal irradiation leads to substantial changes in bone morphology and strength with time, where there is a decreased bone strength following irradiation in the face of increasing bone mass; FE models with a non-linear embrittled material model were most successful in simulating these experimental findings.

A New Class of Ligands for Aqueous, Lanthanide-catalyzed, Enantioselective Mukaiyama Aldol Reactions

Journal of the American Chemical Society. Sep, 2010  |  Pubmed ID: 20806902

The development of aqueous methods for generating enantiopure β-hydroxy carbonyl compounds is an important goal because these subunits compose many bioactive compounds and the ability to synthesize these groups in water has environmental and cost benefits. In this communication, we report a new class of ligands for aqueous, lanthanide-catalyzed, asymmetric Mukaiyama aldol reactions for the synthesis of chiral β-hydroxy ketones. Furthermore, we have used luminescence-decay measurements to unveil mechanistic information regarding the catalytic reaction via changes in water-coordination number. The precatalysts presented here yielded β-hydroxy carbonyls from aliphatic and aryl substrates with outstanding syn:anti ratios and enantiometric excesses of up to 49:1 and 97%, respectively.

Oxidatively Stable, Aqueous Europium(II) Complexes Through Steric and Electronic Manipulation of Cryptand Coordination Chemistry

Angewandte Chemie (International Ed. in English). Nov, 2010  |  Pubmed ID: 20927788

Honeycomb Carbon: a Review of Graphene

Chemical Reviews. Jan, 2010  |  Pubmed ID: 19610631

Continuity of Graphene on Polycrystalline Copper

Nano Letters. Jan, 2011  |  Pubmed ID: 21117698

The atomic structure of graphene on polycrystalline copper substrates has been studied using scanning tunneling microscopy. The graphene overlayer maintains a continuous pristine atomic structure over atomically flat planes, monatomic steps, edges, and vertices of the copper surface. We find that facets of different identities are overgrown with graphene's perfect carbon honeycomb lattice. Our observations suggest that growth models including a stagnant catalytic surface do not apply to graphene growth on copper. Contrary to current expectations, these results reveal that the growth of macroscopic pristine graphene is not limited by the underlying copper structure.

Physeal Bystander Effects in Rhabdomyosarcoma Radiotherapy: Experiments in a New Xenograft Model

Sarcoma. 2011  |  Pubmed ID: 21559211

Radiotherapy used in the treatment of pediatric musculoskeletal sarcomas may result in crippling defects of skeletal growth. Several radioprotective strategies have shown potential for preserving function of the irradiated epiphysis but have not been evaluated in a tumor-bearing animal model. We developed two bioluminescent human rhabdomyosarcoma cell lines that were used to establish xenograft tumors in skeletally immature mice. Bioluminescence imaging and radiography allowed serial evaluation of tumor growth and tibial elongation following localized radiotherapy. High-dose (10 Gy) radiotherapy significantly reduced tumor growth velocity and prolonged the median survival of tumor-bearing mice but also resulted in a significant 3.3% shortening of the irradiated limb. Exposure to a lower, 2 Gy dose resulted in 4.1% decrease in limb length but did not extend survival. This new model provides a clinically relevant means to test the efficacy and safety of novel radioprotectant and radiorecovery strategies for use in this context.

Influence of Locking Bolt Location on the Mechanical Properties of an Interlocking Nail in the Canine Femur

Veterinary Surgery : VS. Jul, 2011  |  Pubmed ID: 21615431

To determine whether the fatigue properties of an interlocking nail (ILN) construct are influenced by metaphyseal or diaphyseal location of the locking bolt.

Kinematic Motion Patterns of the Cranial and Caudal Canine Cervical Spine

Veterinary Surgery : VS. Aug, 2011  |  Pubmed ID: 21770978

To define the kinematic motion patterns of the canine cervical spine, with a particular emphasis on identifying differences between the cranial (C(2)-C(4)) and caudal (C(5)-C(7)) segments, and to determine the significance of coupled motions (CM) in the canine cervical spine.

Graphene Flash Memory

ACS Nano. Oct, 2011  |  Pubmed ID: 21854056

Graphene's single atomic layer of sp(2) carbon has recently garnered much attention for its potential use in electronic applications. Here, we report a memory application for graphene, which we call graphene flash memory (GFM). GFM has the potential to exceed the performance of current flash memory technology by utilizing the intrinsic properties of graphene, such as high density of states, high work function, and low dimensionality. To this end, we have grown large-area graphene sheets by chemical vapor deposition and integrated them into a floating gate structure. GFM displays a wide memory window of ∼6 V at significantly low program/erase voltages of ±7 V. GFM also shows a long retention time of more than 10 years at room temperature. Additionally, simulations suggest that GFM suffers very little from cell-to-cell interference, potentially enabling scaling down far beyond current state-of-the-art flash memory devices.

A Novel Hydrogel-coated Polyester Mesh Prevents Postsurgical Adhesions in a Rat Model

The Journal of Surgical Research. May, 2011  |  Pubmed ID: 20031154

The specific aim of this study was to determine the whether a novel, hydrogel-coated polyester mesh (Scout) can be used to reduce the incidence and severity of adhesion formation in vivo.

Eu(II)-containing Cryptates As Contrast Agents for Ultra-high Field Strength Magnetic Resonance Imaging

Chemical Communications (Cambridge, England). Dec, 2011  |  Pubmed ID: 22046588

The relaxivity (contrast-enhancing ability) of Eu(II)-containing cryptates was found to be better than a clinically approved Gd(III)-based agent at 7 T. These cryptates are among a few examples of paramagnetic substances that show an increase in longitudinal relaxivity, r(1), at ultra-high field strength relative to lower field strengths.

Evaluation of the Safety of a Novel Knee Load-bypassing Device in a Sheep Model

The Journal of Bone and Joint Surgery. American Volume. Jan, 2012  |  Pubmed ID: 22218385

Unloading treatments for osteoarthritis of the knee, such as valgus braces, have been shown to reduce medial joint-loading and pain. The purpose of this preclinical study was to investigate the tissue response to an extracapsular implantable device (KineSpring System; Moximed, Hayward, California) that is designed to reduce loading across the medial compartment of the knee.

Effects of Extensive Circumferential Periosteal Stripping on the Microstructure and Mechanical Properties of the Murine Femoral Cortex

Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society. Apr, 2012  |  Pubmed ID: 21928431

Extensive periosteal stripping (PS) is a risk factor for post-radiation pathologic fracture following surgery for extremity soft tissue tumors. The purpose of this study was to determine the effects of PS on bone structure and mechanical properties. Thirty-one skeletally mature mice underwent PS, with circumferential removal of periosteum from an 8-mm segment of the mid-diaphysis of the left femur. Thirty-one control mice underwent sham surgery in which the femur was isolated without manipulation of the periosteum. At 2, 6, 12, or 26 weeks following surgery, the left femora were examined by micro-CT to quantify cortical thickness (CtTh), cross-sectional area (CSA), bone volume (BV), and polar moment of inertia (PMI). Three-point mechanical bend testing was performed and peak load, stiffness, and energy to failure were determined. PS resulted in significantly decreased CtTh, CSA, BV, and PMI at all time points. Peak load, stiffness, and energy to failure were significantly reduced at 2, 6, and 12 weeks. There were no significant differences in mechanical properties at 26 weeks. In this mouse model, extensive circumferential PS resulted in sustained changes in bone structure that were still evident after 6 months, accompanied by reductions in bone strength that persisted for at least 3 months. © 2011 Orthopaedic Research Society. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 30:561-568, 2012.