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PURPOSE: The purpose of the present study was to apply noninvasive methods for monitoring regeneration and mechanical properties of tissue-engineered cartilage in vivo at different growth stages using MR elastography (MRE). METHODS: Three types of scaffolds, including silk, collagen, and gelatin seeded by human mesenchymal stem cells, were implanted subcutaneously in mice and imaged at 9.4T where the shear stiffness and transverse MR relaxation time (T2 ) were measured for the regenerating constructs for 8 wk. An MRE phase contrast spin echo-based sequence was used for collecting MRE images. At the conclusion of the in vivo study, constructs were excised and transcript levels of cartilage-specific genes were quantitated using reverse-transcription polymerase chain reaction. RESULTS: Tissue-engineered constructs showed a cartilage-like construct with progressive tissue formation characterized by increase in shear stiffness and decrease in T2 that can be correlated with increased cartilage transcript levels including aggrecan, type II collagen, and cartilage oligomeric matrix protein after 8 wk of in vivo culture. CONCLUSION: Altogether, the outcome of this research demonstrates the feasibility of MRE and MRI for noninvasive monitoring of engineered cartilage construct's growth after implantation and provides noninvasive biomarkers for regeneration, which may be translated into treatment of tissue defects.
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Materiales Biocompatibles , Cartílago Articular/fisiología , Diagnóstico por Imagen de Elasticidad/métodos , Imagen por Resonancia Magnética/métodos , Ingeniería de Tejidos/métodos , Animales , Células Cultivadas , Humanos , Células Madre Mesenquimatosas/citología , Ratones , Andamios del TejidoRESUMEN
Scattering from a cavity in a soft elastic medium, such as silicone rubber, resembles scattering from an underwater bubble in that low-frequency monopole resonance is obtainable in both cases. Arrays of cavities can therefore be used to reduce underwater sound transmission using thin layers and low void fractions. This article examines the role of cavity shape by microfabricating arrays of disk-shaped air cavities into single and multiple layers of polydimethylsiloxane. Comparison is made with the case of equivalent volume cylinders which approximate spheres. Measurements of ultrasonic underwater sound transmission are compared with finite element modeling predictions. The disks provide a deeper transmission minimum at a lower frequency owing to the drum-type breathing resonance. The resonance of a single disk cavity in an unbounded medium is also calculated and compared with a derived estimate of the natural frequency of the drum mode. Variation of transmission is determined as a function of disk tilt angle, lattice constant, and layer thickness. A modeled transmission loss of 18 dB can be obtained at a wavelength about 20 times the three-layer thickness, and thinner results (wavelength/thickness â¼ 240) are possible for the same loss with a single layer depending on allowable hydrostatic pressure.
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Primary brain tumors present significant challenges for surgical resection because of their location and the frequent occurrence of malignant projections extending beyond the primary tumor. Visualization of the tumor margins during surgery is critical for a favorable outcome. We report the use of IRDye 800CW chlorotoxin (CLTX) as a targeted imaging agent for brain tumors in a spontaneous mouse model of medulloblastoma, ND2:SmoA1. Specificity and functionality of the targeted agent were confirmed in cell-based assays. Tumors were detected by magnetic resonance imaging and IRDye 800CW CLTX administered to individual animals for optical imaging at 1-month increments. The integrity of the blood-brain barrier (BBB) was measured by Evan's Blue perfusion prior to sacrifice. Results show that IRDye 800CW CLTX specifically targeted tumor tissue. The extravasation of Evan's Blue was observed in all tumors, suggesting that the presence of the tumors can introduce alterations in the permeability of the BBB. Because increased vascular permeability was observed early in the disease model, larger dye-labeled imaging agents that exceed current BBB size restrictions may warrant renewed consideration as candidates for tumor detection and surgical resection. Our study provides data characterizing in vitro and in vivo use of IRDye 800CW CLTX as a broadly applicable tumor imaging agent.
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Bencenosulfonatos/química , Neoplasias Cerebelosas/diagnóstico , Indoles/química , Meduloblastoma/diagnóstico , Venenos de Escorpión/química , Animales , Barrera Hematoencefálica/metabolismo , Línea Celular Tumoral , Neoplasias Cerebelosas/metabolismo , Neoplasias Cerebelosas/patología , Humanos , Metaloproteinasa 2 de la Matriz/metabolismo , Meduloblastoma/metabolismo , Meduloblastoma/patología , Ratones , Venenos de Escorpión/metabolismoRESUMEN
The objective of tissue engineering (TE) is to create functional replacements for various tissues; the mechanical properties of these engineered constructs are critical to their function. Several techniques have been developed for the measurement of the mechanical properties of tissues and organs; however, current methods are destructive. The field of TE will benefit immensely if biomechanical models developed by these techniques could be combined with existing imaging modalities to enable noninvasive, dynamic assessment of mechanical properties during tissue growth. Specifically, MR elastography (MRE), which is based on the synchronization of a mechanical actuator with a phase contrast imaging pulse sequence, has the capacity to measure tissue strain generated by sonic cyclic displacement. The captured displacement is presented in shear wave images from which the complex shear moduli can be extracted or simplified by a direct measure, termed the shear stiffness. MRE has been extended to the microscopic scale, combining clinical MRE with high-field magnets, stronger magnetic field gradients and smaller, more sensitive, radiofrequency coils, enabling the interrogation of smaller samples, such as tissue-engineered constructs. The following topics are presented in this article: (i) current mechanical measurement techniques and their limitations in TE; (ii) a description of the MRE system, MRE theory and how it can be applied for the measurement of mechanical properties of tissue-engineered constructs; (iii) a summary of in vitro MRE work for the monitoring of osteogenic and adipogenic tissues originating from human adult mesenchymal stem cells (MSCs); (iv) preliminary in vivo studies of MRE of tissues originating from mouse MSCs implanted subcutaneously in immunodeficient mice with an emphasis on in vivo MRE challenges; (v) future directions to resolve current issues with in vivo MRE in the context of how to improve the future role of MRE in TE.
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Diagnóstico por Imagen de Elasticidad/métodos , Imagen por Resonancia Magnética/métodos , Modelos Teóricos , Ingeniería de Tejidos/métodos , Animales , Diagnóstico por Imagen de Elasticidad/instrumentación , Humanos , Implantes Experimentales , Imagen por Resonancia Magnética/instrumentación , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Trasplante de Células Madre , Estrés MecánicoRESUMEN
Microscopic magnetic resonance elastography is a high-resolution method for visualizing shear waves and assessing the biomechanical viscoelastic properties of small biological samples. In this work, we used error propagation to develop a simple analytical model that relates the signal-to-noise ratio of MR magnitude images to the variance in shear-wave maps collected using gradient-echo and spin-echo phase-contrast pulse sequences. Our model predicts results for shear-wave images in phantoms, which match the experimentally observed phase variance within 8%. This model can be used to optimize MR pulse sequences for elastography studies, as well as other phase-difference techniques in MRI.
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Imagen por Resonancia Magnética/métodos , Fenómenos Biomecánicos , Elasticidad , Procesamiento de Imagen Asistido por Computador , Imagen por Resonancia Magnética/estadística & datos numéricos , Modelos Estadísticos , Fantasmas de Imagen , Procesamiento de Señales Asistido por Computador , ViscosidadRESUMEN
Magnetic resonance microscopy (MRM) is used to monitor osteogenesis in tissue-engineered constructs. Measurements of the developing tissue's MR relaxation times (T(1) and T(2)), apparent diffusion coefficient (ADC) and elastic shear modulus were conducted over a 4-week growth period using an 11.74 T Bruker spectrometer with an imaging probe adapted for MR elastography (MRE). Both the relaxation times and the ADC show a statistically significant decrease after only one week of tissue development while the tissue stiffness increases progressively during the first two weeks of in vitro growth. The measured MR parameters are correlated with histologically monitored osteogenic tissue development. This study shows that MRM can provide quantitative data with which to characterize the growth and development of tissue-engineered bone.
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Microscopía/métodos , Espectrofotometría/métodos , Ingeniería de Tejidos/métodos , Fosfatasa Alcalina/metabolismo , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Huesos/patología , Proliferación Celular , Humanos , Imagen por Resonancia Magnética , Células Madre Mesenquimatosas/metabolismo , Modelos Estadísticos , Osteogénesis , Factores de TiempoRESUMEN
Docetaxel (Dtxl) is currently the most common therapeutic option for prostate cancer (PC). However, adverse side effects and problems associated with chemo-resistance limit its therapeutic outcome in clinical settings. A targeted nanoparticle system to improve its delivery to and activity at the tumor site could be an attractive strategy for PC therapy. Therefore, the objective of this study was to develop and determine the anti-cancer efficacy of a novel docetaxel loaded, prostate specific membrane antigen (PSMA) targeted superparamagnetic iron oxide nanoparticle (SPION) (J591-SPION-Dtxl) formulation for PC therapy. Our results showed the SPION-Dtxl formulation exhibits an optimal particle size and zeta potential, which can efficiently be internalized in PC cells. SPION-Dtxl exhibited potent anti-cancer efficacy via induction of the expression of apoptosis associated proteins, downregulation of anti-apoptotic proteins, and inhibition of chemo-resistance associated protein in PC cell lines. J591-SPION-Dtxl exhibited a profound uptake in C4-2 (PSMA(+)) cells compared to PC-3 (PSMA(-)) cells. A similar targeting potential was observed in ex-vivo studies in C4-2 tumors but not in PC-3 tumors, suggesting its tumor specific targeting. Overall, this study suggests that a PSMA antibody functionalized SPION-Dtxl formulation can be highly useful for targeted PC therapy.
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Dextranos/química , Nanopartículas de Magnetita/química , Terapia Molecular Dirigida , Nanopartículas/química , Antígeno Prostático Específico/metabolismo , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/metabolismo , Taxoides/uso terapéutico , Animales , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citosol/efectos de los fármacos , Citosol/metabolismo , Docetaxel , Endocitosis/efectos de los fármacos , Humanos , Imagen por Resonancia Magnética , Masculino , Ratones , Taxoides/farmacología , Tubulina (Proteína)/metabolismoRESUMEN
Tissue engineering (TE) has been introduced for more than 25 years without a boom in clinical trials. More than 70 TE-related start-up companies spent more than $600 million/year, with only two FDA-approved tissue-engineered products. Given the modest performance in clinically approved organs, TE is a tenaciously promising field. The TE community is advocating the application of clinically driven methodologies in large animal models enabling clinical translation. This challenge is hindered by the scarcity of tissue biopsies and the absence of standardized evaluation tools, but can be negated through non-invasive assessment of growth and integration, with reduced sample size and low cost. Solving this issue will speed the transition to cost-efficient clinical studies. In this paper we: (a) introduce magnetic resonance elastography to the tissue-engineering and regenerative medicine (TERM) community; (b) review recent MRE applications in TERM; and (c) discuss future directions of MRE in TERM. We have used MRE to study engineered tissues both in vitro and in vivo, where the mechanical properties of mesenchymally derived constructs were progressively monitored before and after tissues were implanted in mouse models. This study represents a stepping stone toward the applications of MRE in directing clinical trials with low cost and likely expediting the translation to more relevantly large animal models and clinical trials.
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Diagnóstico por Imagen de Elasticidad/métodos , Imagen por Resonancia Magnética/métodos , Medicina Regenerativa/métodos , Ingeniería de Tejidos/métodos , Adipocitos/citología , Animales , Biopsia , Calcio/química , Gelatina/química , Glicerol/química , Humanos , Células Madre Mesenquimatosas/citología , Mesodermo/patología , Ratones , Modelos Animales , Osteogénesis , Resistencia al Corte , Estrés MecánicoRESUMEN
The tissue engineering community has been vocal regarding the need for noninvasive instruments to assess the development of tissue-engineered constructs. Medical imaging has helped fulfill this role. However, specimens allocated to a test tube for imaging cannot be tested for a prolonged period or returned to the incubator. Therefore, samples are essentially wasted due to potential contamination and transfer in a less than optimal growth environment. In turn, we present a standalone, miniature, magnetic resonance imaging-compatible incubator, termed the e-incubator. This incubator uses a microcontroller unit to automatically sense and regulate physiological conditions for tissue culture, thus allowing for concurrent tissue culture and evaluation. The e-incubator also offers an innovative scheme to study underlying mechanisms related to the structural and functional evolution of tissues. Importantly, it offers a key step toward enabling real-time testing of engineered tissues before human transplantation. For validation purposes, we cultured tissue-engineered bone constructs for 4 weeks to test the e-incubator. Importantly, this technology allows for visualizing the evolution of temporal and spatial morphogenesis. In turn, the e-incubator can filter deficient constructs, thereby increasing the success rate of implantation of tissue-engineered constructs, especially as construct design grows in levels of complexity to match the geometry and function of patients' unique needs.
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Reactores Biológicos , Técnicas de Cultivo de Célula , Imagen por Resonancia Magnética , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Técnicas de Cultivo de Célula/instrumentación , Técnicas de Cultivo de Célula/métodos , Células Cultivadas , Humanos , Imagen por Resonancia Magnética/instrumentación , Imagen por Resonancia Magnética/métodosRESUMEN
Interaction of serum proteins and nanoparticles leads to a nanoparticle-protein complex formation that defines the rational strategy for a clinically relevant formulation for drug delivery, hyperthermia, and magnetic resonance imaging (MRI) applications in cancer nanomedicine. Given this perspective, we have examined the pattern of human serum protein corona formation with our recently engineered magnetic nanoparticles (MNPs). The alteration in particle size, zeta potential, hemotoxicity, cellular uptake/cancer cells targeting potential, and MRI properties of the MNPs after formation of human serum (HS) protein corona were studied. Our results indicated no significant change in particle size of our MNPs upon incubation with 0.5-50 wt/v% human serum, while zeta potential of MNPs turned negative due to human serum adsorption. When incubated with an increased serum and particle concentration, apolipoprotein E was adsorbed on the surface of MNPs apart from serum albumin and transferrin. However, there was no significant primary or secondary structural alterations observed in serum proteins through Fourier transform infrared spectroscopy, X-ray diffraction, and circular dichroism. Hemolysis assay suggests almost no hemolysis at the tested concentrations (up to 1 mg/mL) for MNPs compared to the sodium dodecyl sulfate (positive control). Additionally, improved internalization and uptake of MNPs by C4-2B and Panc-1 cancer cells were observed upon incubation with human serum (HS). After serum protein adsorption to the surface of MNPs, the close vicinity within T1 (â¼1.33-1.73 s) and T2 (â¼12.35-13.43 ms) relaxation times suggest our MNPs retained inherent MRI potential even after biomolecular protein adsorption. All these superior clinical parameters potentially enable clinical translation and use of this formulation for next generation nanomedicine for drug delivery, cancer-targeting, imaging and theranostic applications.
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Proteínas Sanguíneas/química , Fenómenos Químicos , Nanopartículas de Magnetita/química , Adsorción , Animales , Materiales Biocompatibles/farmacología , Proteínas Sanguíneas/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Endocitosis/efectos de los fármacos , Humanos , Imagen por Resonancia Magnética , Nanopartículas de Magnetita/toxicidad , Ratones Desnudos , Tamaño de la Partícula , Unión Proteica/efectos de los fármacos , Electricidad EstáticaRESUMEN
Myocarditis is an inflammation of the myocardium, but only -10% of those affected show clinical manifestations of the disease. To study the immune events of myocardial injuries, various mouse models of myocarditis have been widely used. This study involved experimental autoimmune myocarditis (EAM) induced with cardiac myosin heavy chain (Myhc)-α 334-352 in A/J mice; the affected animals develop lymphocytic myocarditis but with no apparent clinical signs. In this model, the utility of magnetic resonance microscopy (MRM) as a non-invasive modality to determine the cardiac structural and functional changes in animals immunized with Myhc-α 334-352 is shown. EAM and healthy mice were imaged using a 9.4 T (400 MHz) 89 mm vertical core bore scanner equipped with a 4 cm millipede radio-frequency imaging probe and 100 G/cm triple axis gradients. Cardiac images were acquired from anesthetized animals using a gradient-echo-based cine pulse sequence, and the animals were monitored by respiration and pulse oximetry. The analysis revealed an increase in the thickness of the ventricular wall in EAM mice, with a corresponding decrease in the interior diameter of ventricles, when compared with healthy mice. The data suggest that morphological and functional changes in the inflamed hearts can be non-invasively monitored by MRM in live animals. In conclusion, MRM offers an advantage of assessing the progression and regression of myocardial injuries in diseases caused by infectious agents, as well as response to therapies.
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Enfermedades Autoinmunes/patología , Imagen por Resonancia Magnética/métodos , Miocarditis/patología , Miocardio/patología , Animales , Enfermedades Autoinmunes/fisiopatología , Femenino , Corazón/fisiopatología , Ratones , Ratones Endogámicos A , Miocarditis/fisiopatologíaRESUMEN
Traumatic brain injury (TBI) presents a variety of causes and symptoms, thus making the development of reliable diagnostic methods and therapeutic treatments challenging. Magnetic resonance elastography (MRE) is a technique that allows for a noninvasive assessment of the mechanical properties of soft biological tissue, such as tissue stiffness, storage modulus, and loss modulus. Importantly, by quantifying the changes in the stiffness of tissue that is often associated with disease, MRE is able to detect tissue pathologies at early stages. Recent improvements in instrumentation have allowed for the investigation of small samples with microscopic resolution (µMRE). We hypothesize that µMRE can sensitively detect variations in micromechanical properties in the brain caused by the compressive and shearing forces sustained during TBI. To test this hypothesis, we randomized 13 C57BL mice to receive a controlled cortical impact at a 0.5 mm or 0.75 mm depth, with both sham and naïve mice as controls. Our objective was to propagate mechanical shear waves throughout the brain for in vivo TBI µMRE imaging. The mechanical properties of the injured brain tissue were determined at days 0, 1, 7, and 28 post-injury. For both groups, we observed a significant drop in the stiffness of the impacted region immediately following the injury; the 0.75 mm animals experienced increased tissue softness that lasted longer than that for the 0.5 mm group. Although the shear stiffness, storage modulus, and loss modulus parameters all followed the same trend, the tissue stiffness yielded the most statistically significant results. Overall, this article introduces a transformative technique for mechanically mapping the brain and detecting brain diseases and injury.
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Lesiones Encefálicas/diagnóstico por imagen , Corteza Cerebral/diagnóstico por imagen , Corteza Cerebral/lesiones , Diagnóstico por Imagen de Elasticidad/tendencias , Elasticidad , Animales , Lesiones Encefálicas/fisiopatología , Corteza Cerebral/fisiología , Elasticidad/fisiología , Diagnóstico por Imagen de Elasticidad/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Distribución Aleatoria , Factores de Tiempo , ViscosidadRESUMEN
Millions of cases of bone injury or loss due to trauma, osteoporosis, and cancer occur in the United States each year. Because bone is limited in its ability to regenerate, alternative therapy approaches are needed. Bone tissue engineering has the potential to correct musculoskeletal disorders through the development of cell-based substitutes for osteogenic tissue replacement. Multiple medical imaging techniques such as magnetic resonance microscopy (MRM) were investigated recently; these techniques are able to provide useful information on the anatomical and structural changes of developing bone. However, there is a need for noninvasive approaches to evaluate biochemical constituents and consequent compositional development associated with growing osteogenic constructs. In this study, near-infrared (NIR) optical imaging with a bone-specific NIR-targeted probe, IRDye(®) 800CW BoneTag™ (800CW BT), was applied in this study to longitudinally visualize regions of mineralization of tissue-engineered bone constructs in vivo. A fluorescent cell-based assay was performed to confirm the preferential binding of 800CW BT to the mineralized matrix of differentiated osteogenically driven human mesenchymal stem cells (hMSCs) in vitro. The hMSCs were seeded onto a biocompatible gelatin scaffold, allowed to develop, and implanted into a mouse model. Engineered constructs were examined in vivo using NIR imaging for bone mineralization, paired with MRM for verification of developing tissue. Results showed that NIR imaging with 800CW BT labeling can effectively assess the calcification of the developing osteogenic constructs, which is consistent with the analysis of excised tissue using NIR microscopy and histology. In conclusion, this study evaluated bone-like function of regenerating bone through tracking calcium deposition via NIR optical imaging with a fluorophore-labeled probe in a noninvasive manner.
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Traditional mechanical testing often results in the destruction of the sample, and in the case of long term tissue engineered construct studies, the use of destructive assessment is not acceptable. A proposed alternative is the use of an imaging process called magnetic resonance elastography. Elastography is a nondestructive method for determining the engineered outcome by measuring local mechanical property values (i.e., complex shear modulus), which are essential markers for identifying the structure and functionality of a tissue. As a noninvasive means for evaluation, the monitoring of engineered constructs with imaging modalities such as magnetic resonance imaging (MRI) has seen increasing interest in the past decade. For example, the magnetic resonance (MR) techniques of diffusion and relaxometry have been able to characterize the changes in chemical and physical properties during engineered tissue development. The method proposed in the following protocol uses microscopic magnetic resonance elastography (µMRE) as a noninvasive MR based technique for measuring the mechanical properties of small soft tissues. MRE is achieved by coupling a sonic mechanical actuator with the tissue of interest and recording the shear wave propagation with an MR scanner. Recently, µMRE has been applied in tissue engineering to acquire essential growth information that is traditionally measured using destructive mechanical macroscopic techniques. In the following procedure, elastography is achieved through the imaging of engineered constructs with a modified Hahn spin-echo sequence coupled with a mechanical actuator. As shown in Figure 1, the modified sequence synchronizes image acquisition with the transmission of external shear waves; subsequently, the motion is sensitized through the use of oscillating bipolar pairs. Following collection of images with positive and negative motion sensitization, complex division of the data produce a shear wave image. Then, the image is assessed using an inversion algorithm to generate a shear stiffness map. The resulting measurements at each voxel have been shown to strongly correlate (R(2)>0.9914) with data collected using dynamic mechanical analysis. In this study, elastography is integrated into the tissue development process for monitoring human mesenchymal stem cell (hMSC) differentiation into adipogenic and osteogenic constructs as shown in Figure 2.
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Diagnóstico por Imagen de Elasticidad/métodos , Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodos , Adipocitos/citología , Diferenciación Celular/fisiología , Diagnóstico por Imagen de Elasticidad/instrumentación , Humanos , Osteocitos/citologíaRESUMEN
BACKGROUND: The next generation magnetic nanoparticles (MNPs) with theranostic applications have attracted significant attention and will greatly improve nanomedicine in cancer therapeutics. Such novel MNP formulations must have ultra-low particle size, high inherent magnetic properties, effective imaging, drug targeting, and drug delivery properties. To achieve these characteristic properties, a curcumin-loaded MNP (MNP-CUR) formulation was developed. METHODS: MNPs were prepared by chemical precipitation method and loaded with curcumin (CUR) using diffusion method. The physicochemical properties of MNP-CUR were characterized using dynamic light scattering, transmission electron microscopy, and spectroscopy. The internalization of MNP-CUR was achieved after 6 hours incubation with MDA-MB-231 breast cancer cells. The anticancer potential was evaluated by a tetrazolium-based dye and colony formation assays. Further, to prove MNP-CUR results in superior therapeutic effects over CUR, the mitochondrial membrane potential integrity and reactive oxygen species generation were determined. Magnetic resonance imaging capability and magnetic targeting property were also evaluated. RESULTS: MNP-CUR exhibited individual particle grain size of ~9 nm and hydrodynamic average aggregative particle size of ~123 nm. Internalized MNP-CUR showed a preferential uptake in MDA-MB-231 cells in a concentration-dependent manner and demonstrated accumulation throughout the cell, which indicates that particles are not attached on the cell surface but internalized through endocytosis. MNP-CUR displayed strong anticancer properties compared to free CUR. MNP-CUR also amplified loss of potential integrity and generation of reactive oxygen species upon treatment compared to free CUR. Furthermore, MNP-CUR exhibited superior magnetic resonance imaging characteristics and significantly increased the targeting capability of CUR. CONCLUSION: MNP-CUR exhibits potent anticancer activity along with imaging and magnetic targeting capabilities. This approach can be extended to preclinical and clinical use and may have importance in cancer treatment and cancer imaging in the future. Further, if these nanoparticles can functionalize with antibody/ligands, they will serve as novel platforms for multiple biomedical applications.
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Neoplasias de la Mama/tratamiento farmacológico , Curcumina/administración & dosificación , Nanopartículas de Magnetita/administración & dosificación , Antineoplásicos/administración & dosificación , Antineoplásicos/farmacocinética , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/metabolismo , Línea Celular Tumoral , Curcumina/farmacocinética , Sistemas de Liberación de Medicamentos , Femenino , Humanos , Imagen por Resonancia Magnética , Nanopartículas de Magnetita/ultraestructura , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Microscopía Electrónica de Transmisión , Nanomedicina , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Traumatic brain injury (TBI) is a major cause of death and disability for which there is no cure. One of the issues inhibiting clinical trial success is the lack of targeting specific patient populations due to inconsistencies between clinical diagnostic tools and underlying pathophysiology. The development of reliable, noninvasive markers of TBI severity and injury mechanisms may better identify these populations, thereby improving clinical trial design. Magnetic resonance elastography (MRE), by assessing tissue mechanical properties, can potentially provide such marker. MRE synchronizes mechanical excitations with a phase contrast imaging pulse sequence to noninvasively register shear wave propagation, from which local values of tissue viscoelastic properties can be deduced. The working hypothesis of this study is that TBI involves a compression of brain tissue large enough to bring the material out of its elastic range, sufficiently altering mechanical properties to generate contrast on MRE measurements. To test this hypothesis, we combined microscopic MRE with brain tissue collected from adult male rats subjected to a controlled cortical impact injury. Measurements were made in different regions of interest (somatosensory cortex, hippocampus, and thalamus), and at different time points following the injury (immediate, 24 h, 7 days, 28 days). Values of stiffness in the somatosensory cortex were found to be 23-32% lower in the injured hemisphere than in the healthy one, when no significant difference was observed in the case of sham brains. A preliminary in vivo experiment is also presented, as well as alternatives to improve the faithfulness of stiffness recovery.
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Lesiones Encefálicas/fisiopatología , Diagnóstico por Imagen de Elasticidad/métodos , Miniaturización/métodos , Animales , Lesiones Encefálicas/patología , Modelos Animales de Enfermedad , Diagnóstico por Imagen de Elasticidad/instrumentación , Hipocampo/lesiones , Hipocampo/patología , Hipocampo/fisiopatología , Masculino , Ratones , Ratas , Ratas Sprague-Dawley , Corteza Somatosensorial/lesiones , Corteza Somatosensorial/patología , Corteza Somatosensorial/fisiopatología , Tálamo/lesiones , Tálamo/patología , Tálamo/fisiopatologíaRESUMEN
We have developed a multi-layer approach for the synthesis of water-dispersible superparamagnetic iron oxide nanoparticles for hyperthermia, magnetic resonance imaging (MRI) and drug delivery applications. In this approach, iron oxide core nanoparticles were obtained by precipitation of iron salts in the presence of ammonia and provided ß-cyclodextrin and pluronic polymer (F127) coatings. This formulation (F127250) was highly water dispersible which allowed encapsulation of the anti-cancer drug(s) in ß-cyclodextrin and pluronic polymer for sustained drug release. The F127250 formulation has exhibited superior hyperthermia effects over time under alternating magnetic field compared to pure magnetic nanoparticles (MNP) and ß-cyclodextrin coated nanoparticles (CD200). Additionally, the improved MRI characteristics were also observed for the F127250 formulation in agar gel and in cisplatin resistant ovarian cancer cells (A12780CP) compared to MNP and CD200 formulations. Furthermore, the drug-loaded formulation of F127250 exhibited many folds of imaging contrast properties. Due to the internalization capacity of the F127250 formulation, its curcumin-loaded formulation (F127250-CUR) exhibited almost equivalent inhibition effects on A2780CP (ovarian), MDA-MB-231 (breast), and PC-3 (prostate) cancer cells even though curcumin release was only 40%. The improved therapeutic effects were verified by examining molecular effects using Western blotting and transmission electron microscopic (TEM) studies. F127250-CUR also exhibited haemocompatibility, suggesting a nanochemo-therapeutic agent for cancer therapy.
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Magnetismo , Nanopartículas/uso terapéutico , Neoplasias/tratamiento farmacológico , Western Blotting , Línea Celular Tumoral , Humanos , Hipertermia Inducida , Imagen por Resonancia Magnética , Nanopartículas/química , beta-Ciclodextrinas/químicaRESUMEN
Assessment of tissue regeneration is essential to optimize the stages of tissue engineering (cell proliferation, tissue development and implantation). Optical and X-ray imaging have been used in tissue engineering to provide useful information, but each has limitations: for example, poor depth penetration and radiation damage. Magnetic resonance imaging (MRI) largely overcomes these restrictions, exhibits high resolution (approximately 100 microm) and can be applied both in vitro and in vivo. Recently, MRI has been used in tissue engineering to generate spatial maps of tissue relaxation times (T(1), T(2)), water diffusion coefficients, and the stiffness (shear moduli) of developing engineered tissues. In addition, through the use of paramagnetic and superparamagnetic contrast agents, MRI can quantify cell death, assess inflammation, and visualize cell trafficking and gene expression. After tissue implantation MRI can be used to observe the integration of a tissue implant with the surrounding tissues, and to check for early signs of immune rejection. In this review, we describe and evaluate the growing role of MRI in the assessment of tissue engineered constructs. First, we briefly describe the underlying principles of MRI and the expected changes in relaxation times (T(1), T(2)) and the water diffusion coefficient that are the basis for MR contrast in developing tissues. Next, we describe how MRI can be applied to evaluate the tissue engineering of mesenchymal tissues (bone, cartilage, and fat). Finally, we outline how MRI can be used to monitor tissue structure, composition, and function to improve the entire tissue engineering process.
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Imagen por Resonancia Magnética/métodos , Ingeniería de TejidosRESUMEN
BACKGROUND: Magnetic resonance microscopy (MRM) is an important experimental tool in the identification of early cartilage lesions. METHODS: Normal and degenerated cartilage samples were imaged at 11.74 T using a standard spin echo sequence. Quantitative MR measurements for T1, T2, and ADC were obtained and mapping for T2 and ADC was performed. The bi-exponential model for T2 relaxation was also explored. Histology was carried out for comparison with MR images. RESULTS: MR images of cartilage samples displaying early stages of degeneration were positively correlated to their histological appearance in 23-microm high-resolution images and also with much shorter imaging times at 47-microm resolution. T2 maps enable delineation of the actual cartilage zones, distinguishing the superficial zone in particular. The bi-exponential model can reflect cartilage components at different stages of degeneration. INTERPRETATION: At 11.74 T, with 23-microm resolution or with 47-microm resolution and shorter imaging times, MRM provides images that allow visualization of early stages of cartilage degeneration, including superficial fibrillation. This has not been shown previously. The images also allow quantitative measurements (T1, T2, and ADC) in each cartilage region, which can be indicative of different stages of cartilage degeneration.
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Cartílago Articular/patología , Imagen por Resonancia Magnética/métodos , Cadáver , Cartílago Articular/lesiones , HumanosRESUMEN
Tissue engineering has the potential to treat bone loss, but current bone restoration methods, including osteogenesis from mesenchymal stem cells (MSCs), require three to four weeks for bone formation to occur. In this study, we stimulated the formation of engineered bone tissue with low-intensity ultrasound, which has been proven to accelerate bone healing in vivo. One group of engineered bone constructs received ultrasound stimulation 20 minutes per day over a 3-week growth period. We monitored the growth of all the engineered constructs quantitatively and noninvasively using magnetic resonance microscopy (MRM), where the T2 relaxation times of all the constructs were measured, on a weekly basis, using an 11.74 T Bruker spectrometer. Histological and immunocytochemical sections were obtained for all constructs and correlated with the MR results. This study shows that ultrasound can accelerate osteogenesis in vitro for tissue engineered bone, the growth and development of which can be monitored using MRM.