Biomedical subjects

Boris Rubinsky

Publications and source records attributed to Boris Rubinsky.

At least 19 recordsLinked to original sources

PubMed2006

In vivo electrical impedance measurements during and after electroporation of rat liver.

Electroporation is used for in vivo gene therapy, drug therapy and minimally invasive tissue ablation. Applying electrical pulses across cells can have a variety of outcomes; from no effect to reversible electroporation to irreversible electroporation. Recently, it has been proposed that measuring the passive electrical properties of electroporated tissues could provide real time feedback on the outcome of the treatment. Here we describe the results from the impedance characterization (single dispersion Cole model) for up to 30 min of the electroporation process in in vivo rat livers (n=8). The electroporation sequence consisted of 8 pulses of 100 micros with a period of 100 ms. Half of the animals were subjected to field magnitudes considered to have reversible effects (R group, E=450 V/cm) whereas for the other half irreversible field amplitudes were applied (I group, E=1500 V/cm). As expected, there was an immediate increase of conductivity (R group Deltasigma/sigma(t=0)=9+/-3%; I group Deltasigma/sigma(t=0)=43+/-1%). However, the overall long term pattern of change in conductivity after electroporation is complex and different between reversible and irreversible groups. This suggests the superposition of different phenomena which together affect the electrical properties.

Animals↗

PubMed2006

A theoretical study on magnetic induction frequency dependence of phase shift in oedema and haematoma.

The spectroscopic distribution of induction phase shift as a function of the relative volume of oedema and haematoma in the brain, lung and muscle in the bulk of tissue was produced from the available tissue data and a simple mathematical model of electromagnetic induction in tissue. The results show that the phase shift is sensitive to the relative volume of oedema/haematoma at frequencies higher than approximately 10 MHz. The effect of oedema/haematoma on brain, lung and muscle tissues is tissue-type specific. Increasing the volume of tissue has the effect of lowering the frequency at which the phase shift becomes sensitive to the volume of oedema/haematoma. The results indicate that bulk induction measurement of the phase shift has the potential for becoming a simple means for non-contact detection of formation of oedema and haematoma in brain, lung and muscle tissues.

Computer Simulation↗

PubMed2006

Analysis of isochoric subcooling.

Because ice-I is less dense than water, the formation of an ice nucleus in an isochoric (constant volume) chamber will cause an increase in pressure. This analysis shows that the energy required to overcome such a pressure increase makes homogeneous ice nucleation thermodynamically improbable in an isochoric system at temperatures above -109 degrees C. By suppressing ice nucleation, isochoric cooling is expected to significantly promote vitrification. Because water has a higher freezing temperature and a lower glass-transition temperature than physiological solutions, this analysis represents a scenario for avoiding ice crystallization during the preservation of biological substances. While isochoric cryopreservation has not yet been put into practice, this theoretical, first-order analysis suggests that if attainable it could make organ preservation significantly more effective and practical.

Cold Temperature↗

PubMed2006

Detecting cryoablation with EIT and the benefit of including ice front imaging data.

Imaging has made cryosurgery, the destruction of unwanted tissue through freezing, valuable. Electrical impedance tomography (EIT) has been explored as a method to determine the volume of tissue that is frozen during the procedure. However, studies have shown that tissue near the edge of the frozen zone often survives since in this region it may only be the extra-cellular space that is frozen. This threatens the usefulness of cryosurgery for cancer therapy since inaccurate ablation either allows the cancer to survive or increases the chances of complications. Since low-frequency conductivity of tissue increases due to cell membrane impairment, and ablated tissue implies impaired membranes, EIT has the capability to recover images of tissue viability. Cryosurgery is a technique that can benefit from this: EIT scans before freezing and after thawing can show changes in conductivity and hence viability due to treatment. Assuming unfrozen tissue will survive treatment, we explore the use of differential EIT in combination with intra-operative ice front imaging modes that are currently in clinical practice to recover enhanced-resolution images of cryosurgical treatment efficacy in a set of simulated experiments. We also investigate the sensitivity to violation of this assumption and predict tolerable levels of measurement noise.

Algorithms↗

PubMed2006

The detection of brain oedema with frequency-dependent phase shift electromagnetic induction.

The spectroscopic distribution of inductive phase shift in the brain as a function of the relative volume of oedema was evaluated with theoretical and experimental methods in the frequency range 1 to 8 MHz. The theoretical study employed a simple mathematical model of electromagnetic induction in tissue and brain tissue data available from the literature to calculate the phase shift as a function of oedema in the bulk of the brain. Experimental data were generated from bulk measurements of ex vivo homogenized pig brain tissue mixed with various volumes of physiological saline in a volume sample typical of the human brain. There is good agreement between the analytical and the experimental results. Detectable changes in phase shift begin from a frequency of about 3 MHz to 4 MHz in the tested compositions and volume. The phase shift increases with frequency and fluid content. The results suggest that measuring phase shift in the bulk of the brain has the potential for becoming a robust means for non-contact detection of oedema in the brain.

Animals↗

PubMed2006

Electrical and thermal characterization of nanochannels between a cell and a silicon based micro-pore.

Micro and nano fabrication techniques have facilitated the production of new devices for manipulation of single cells on a chip, such as the planar micro-pore electroporation technology. To characterize this technology we have studied the seal that forms at the interface between an individual cell and the micro-pore, in which the cell normally resides, as a function of an electrical field applied across the cell and temperature. Mathematical analysis of non-electroporative electrical fields in experiments with Madin-Darby canine kidney (MDCK) cells suggests that nanoscale channels form between the exterior of the cell and the pore wall. The results indicate that the electrical currents through these channels need to be considered when using planar micro-pores in general and performing micro-pore electroporation in particular. Our results show that the size of these channels is strongly temperature dependent and the cell to pore wall distance can increase by as much as 60% when the temperature of the system is lowered from 35 to 0( composite function)C. Temperature appears to be an important factor in the use of devices for cells on a chip and our results suggest that physiological temperatures should yield better seal formation, thus improved feedback sensitivity, than the traditional use of room temperature in planar micro-pore electroporation devices.

Animals↗

PubMed2006

In vivo results of a new focal tissue ablation technique: irreversible electroporation.

This paper reports results of in vivo experiments that confirm the feasibility of a new minimally invasive method for tissue ablation, irreversible electroporation (IRE). Electroporation is the generation of a destabilizing electric potential across biological membranes that causes the formation of nanoscale defects in the lipid bilayer. In IRE, these defects are permanent and lead to cell death. This paper builds on our earlier theoretical work and demonstrates that IRE can become an effective method for nonthermal tissue ablation requiring no drugs. To test the capability of IRE pulses to ablate tissue in a controlled fashion, we subjected the livers of male Sprague-Dawley rats to a single 20-ms-long square pulse of 1000 V/cm, which calculations had predicted would cause nonthermal IRE. Three hours after the pulse, treated areas in perfusion-fixed livers exhibited microvascular occlusion, endothelial cell necrosis, and diapedeses, resulting in ischemic damage to parenchyma and massive pooling of erythrocytes in sinusoids. However, large blood vessel architecture was preserved. Hepatocytes displayed blurred cell borders, pale eosinophilic cytoplasm, variable pyknosis and vacuolar degeneration. Mathematical analysis indicates that this damage was primarily nonthermal in nature and that sharp borders between affected and unaffected regions corresponded to electric fields of 300-500 V/cm.

Animals↗

PubMed2005

Improved viability and reduced apoptosis in sub-zero 21-hour preservation of transplanted rat hearts using anti-freeze proteins.

BACKGROUND: Freeze-tolerant fish survive sub-zero temperatures by non-colligatively lowering the freezing temperature of their body fluids using anti-freeze proteins (AFPs). We sought to evaluate and compare the effects of prolonged sub-zero cryopreservation of transplanted rat hearts using AFP I or AFP III. METHODS: Two heterotopic rat heart transplantation protocols were used. In Protocol 1 (n = 104), hearts (n = 8/group) were preserved for 12, 18 and 24 hours in University of Wisconsin solution (UW) at 4 degrees C, UW at -1.3 degrees C, UW/AFP I at -1.3 degrees C and UW/AFP III at -1.3 degrees C, with and without nucleation. Post-operative evaluation consisted of visual viability scoring of the hearts after 60 minutes. Protocol 2 (n = 58) involved evaluation of 24-hour post-transplant viability, echocardiography (fractional shortening [FS], left ventricular end-systolic and -diastolic diameter [ESD, EDD] and anterior and posterior wall systolic and diastolic thickness [AWT-S, AWT-D, PWT-S, PWT-D]), TUNEL staining and electron microscopy (EM) findings for hearts preserved for 18, 21 and 24 hours in UW at 4 degrees C or UW/AFP III at -1.3 degrees C. RESULTS: Hearts preserved in UW at -1.3 degrees C with nucleation froze and died. Three of 8 hearts preserved in UW at 4 degrees C for 24 hours died, whereas all hearts preserved at -1.3 degrees C survived. Hearts preserved in UW/AFP for 18 and 24 hours at -1.3 degrees C had superior viability scores compared with those in UW at 4 degrees C. Hearts in AFP III at -1.3 degrees C displayed greater AWT-S and AWT-D (3.5 +/- 0.2 vs 2.4 +/- 0.2, p < 0.05, and 3.5 +/- 0.2 vs 2.2 +/- 0.2, p < 0.05, respectively) after 18-hour preservation. In the 21-hour preservation group, AFP-treated hearts displayed improved echocardiographic systolic contraction indices, including: improved FS (27 +/- 3.7 vs 15 +/- 4, p = 0.04); diminished ESD (0.28 +/- 0.57 vs 0.47 +/- 0.6, p < 0.05); greater AWT-S (3.4 +/- 0.18 vs 2.8 +/- 0.2, p < 0.05); and fewer positively TUNEL-stained nuclei per specimen (35 +/- 14 vs 5.3 +/- 2.7, p = 0.04). Also, improved EM scores were noted compared with UW at 4 degrees C. CONCLUSIONS: In prolonged sub-zero cryopreservation, AFPs protect the heart from freezing, improve survival and hemodynamics, and reduce apoptotic cell death.

Adenosine↗

PubMed2005

Front-tracking image reconstruction algorithm for EIT-monitored cryosurgery using the boundary element method.

The effectiveness of cryosurgery, treatment of tumors by freezing, is highly dependent on knowledge of transient freezing extent, and therefore relies heavily on real-time imaging techniques for monitoring. Electrical impedance tomography (EIT) holds much promise for this application. In cryosurgery there is a three order of magnitude change in impedance across the freezing boundary and there is a priori knowledge of the freezing origin. Furthermore, an EIT image of the tissue can be done prior to the cryosurgery. In this study, we have developed an EIT front tracking reconstruction algorithm which takes advantage of these particular attributes of cryosurgery. The method tracks the freezing interface rather than the impedance distribution in the freezing tissue. In addition to drastically reducing the number of parameters needed to define the image, the computational complexity is further reduced by using the more appropriate boundary element method (BEM) for solution to the forward problem. The front-tracking method was found to converge rapidly and accurately to a variety of simulated phantom images.

Algorithms↗

PubMed2005

The thermodynamic principles of isochoric cryopreservation.

The goal of this study is to introduce the fundamental thermodynamic principles of isochoric (constant volume) cryopreservation for low temperature preservation of biological materials. Traditionally, cryopreservation is performed in an isobaric process (constant pressure) at 1 atm, because this is our natural environment and it is most convenient experimentally. More than half a century of studies on cryopreservation shows that the major mechanism of damage during isobaric cryopreservation is the increase in intracellular ionic concentration during freezing, which presumably causes chemical damage to the components of cells. Cryoprotectants as well as hyperbaric pressures have been developed as methods to reduce the extent of chemical damage during freezing. The theoretical studies in this paper show that in isochoric cryopreservation, the increase in solution concentration during freezing is lower at each temperature by almost an order of magnitude from that in isobaric cryopreservation. This suggests that isochoric cryopreservation could be a preferential alternative to isobaric cryopreservation. The technology for isochoric cryopreservation is very simple; freezing in a constant volume chamber. Using a simple isochoric cryopreservation device, we confirm the theoretical thermodynamic predictions.

Cryopreservation↗

PubMed2005

Temperature dependence of tissue impedivity in electrical impedance tomography of cryosurgery.

The temperature-dependent impedivity of rat liver, transverse abdominal muscle and full skin was determined in vitro as a function of frequency across the temperature range 5 degrees C to 37 degrees C and from 100 Hz to 10 kHz. This study was motivated by an increasing interest in using electrical impedance tomography (EIT) for imaging of cryosurgery and a lack of applicable data in the hypothermic range. Using a controlled-temperature impedance analyzer, it was found that as the temperature is reduced the resulting increase in tissue impedivity is more pronounced at low frequencies and that the beta dispersion, resulting from cell membrane polarization, shifts to lower frequencies. With these new data a simple case study of EIT of liver cryosurgery was examined, using a finite-element model incorporating the Pennes bio-heat equation, to determine the impact of this behavior on imaging accuracy. Overestimation of the ice-front position was found to occur if the EIT system ignored the effects of the low-temperature zone surrounding the frozen tissue. This error decreases with increasing blood perfusion and with higher measurement frequencies.

Abdominal Muscles↗

PubMed2005

Cancer cells ablation with irreversible electroporation.

In this study we perform in vitro irreversible electroporation (IRE) experiments with human hepatocarcinoma cells (HepG2) to investigate IRE as a new technique for undesirable tissue ablation. Irreversible electroporation (IRE) is the irreversible permeabilization of the cell membrane through the application of microsecond through millisecond electrical pulses. Until now IRE was studied only as an undesirable condition during the use of reversible electroporation in gene therapy and electrochemotherapy. There was a possibility that the IRE ablation domain is mostly superimposed on the electrical pulses induced Joule heating thermal ablation domain. This study demonstrates that there is a real and substantial domain of electrical parameters for IRE ablation of cancer that is distinct from the thermal domain and which results in complete cancer cell ablation. Experiments show that the application of 1500 V/cm in three sets of ten pulses of 300 microseconds each can produce complete cancer cell ablation. We also find that the use of multiple pulses appears to be more effective for cancer cell ablation than the application of the same energy in one single pulse.

Carcinoma, Hepatocellular↗

PubMed2004

Prolonged 24-hour subzero preservation of heterotopically transplanted rat hearts using antifreeze proteins derived from arctic fish.

BACKGROUND: Arctic fish survive subzero temperatures by producing a family of antifreeze proteins (AFPs) that noncolligatively lower the freezing temperature of their body fluids. We report 24-hour storage of mammalian hearts for transplantation at subzero temperatures using AFPs derived from arctic fish. METHODS: Forty-two heterotopic transplantations were performed in isoimmune Sprague-Dawley rats. Harvested hearts were retrogradely infused with cold 4 degrees C University of Wisconsin (UW) solution and were preserved in a specialized cooling bath at two target temperatures, 4 degrees C and -1.3 degrees C for 12,18, and 24 hours (6 experiments/group). Preservation solutions were UW alone for the 4 degrees C group, and UW with 15 mg/mL AFP III for the -1.3 degrees C group. After hypothermic storage the hearts were heterotopically transplanted into isoimmune rats. Viability was assessed and graded on a scale of 0 to 6 (0 = no contractions to 6 = excellent contractions). Transplanted hearts were then fixed in vivo and were subject to electron microscopy and histopathologic examination. RESULTS: None of the hearts preserved at -1.3 degrees C in UW/AFP III solution froze. All control hearts preserved at -1.3 degrees C without AFP protection froze and died at reperfusion. Viability of hearts preserved at -1.3 degrees C in UW/AFP III solution was significantly better after 18 hours of preservation, 30 and 60 minutes after reperfusion (median, 5 versus 3 and 6 versus 3, respectively; p < 0.05) and after 24 hours of preservation 30 and 60 minutes after reperfusion (median, 4.5 versus 1.5 and 5 versus 2, respectively; p < 0.05). Histologic and electron microscopy studies demonstrated better myocyte structure and mitochondrial integrity preservation with UW/AFP III solution. CONCLUSIONS: Antifreeze proteins prevent freezing in subzero cryopreservation of mammalian hearts for transplantation. Subzero preservation prolongs ischemic times and improves posttransplant viability.

Adenosine↗

PubMed2004

Subzero nonfreezing cryopresevation of rat hearts using antifreeze protein I and antifreeze protein III.

The purpose of the present study was to evaluate whether AFPs protect the heart from freezing and improve survival and viability in subzero cryopreservation. Hearts were subject to 5 preservation protocols; University of Wisconsin solution (UW) at 4 degrees C, UW at -1.3 degrees C without nucleation, UW at -1.3 degrees C with nucleation, UW AFP I (15 mg/cm(3)) at -1.3 degrees C with nucleation, and in UW AFP III (15 mg/cm(3)) at -1.3 degrees C with nucleation. Hearts were preserved for 24, 28, and 32 h, rewarmed and connected to the working isolated perfusion system. Data [heart rate (HR), coronary flow (CF), and developed pressure (dP)] was collected 30 and 60 min after reperfusion. Hearts preserved at -1.3 degrees C without AFPs froze, while hearts preserved with AFP did not freeze when nucleation was initiated and survived. Survival and dP of hearts preserved for 24h at -1.3 degrees C using AFP III was better than those preserved at 4 degrees C, (dP; 1.4 vs. 0.8, p<0.05). Four of six hearts and six of six hearts died when preserved at 4 degrees C for 28 and 32 h, respectively, all of the hearts that were preserved at -1.3 degrees C with or without AFPs survived after 28 h (n=18) and 32 h (n=18). CF was higher in UW -1.3 degrees C group without attempted nucleation than in AFP I and AFP III groups after 28 and 32 h (3.4 vs. 1.7, p<0.05, and 3.4 vs. 1.7, p<0.05, respectively). In conclusion, AFPs were found to protect the heart from freezing and improve survival and dP (AFP III) in prolonged subzero preservation.

Adenosine↗

PubMed2004

Electrical impedance tomography for imaging tissue electroporation.

Electroporation is a method to introduce molecules, such as gene constructs or small drugs, into cells by temporarily permeating the cell membrane with electric pulses. In molecular medicine and biotechnology, tissue electroporation is performed with electrodes placed in the target area of the body. Currently, tissue electroporation, as with all other methods of molecular medicine, is performed without real-time control or near-term information regarding the extent and degree of electroporation. This paper expands the work from our previous study by implementing new ex vivo experimental data with "front-tracking" analysis for the image reconstruction algorithm. The experimental data is incorporated into numerical simulations of electroporation procedures and images are generated using the new reconstruction algorithm to demonstrate that electrical impedance tomography (EIT) can produce an image of the electroporated area. Combining EIT with electroporation could become an important biotechnological and medical technique to introduce therapeutic molecules into cells in tissue at predetermined areas of the body.

Algorithms↗

PubMed2004

Electrical impedance tomography of cell viability in tissue with application to cryosurgery.

Tissue damage that is associated with the loss of cell membrane integrity should alter the bulk electrical properties of the tissue. This study shows that electrical impedance tomography (EIT) should be able to detect and image necrotic tissue inside the body due to the permeabilization of the membrane to ions. Cryosurgery, a minimally invasive surgical procedure that uses freezing to destroy undesirable tissue, was used to investigate the hypothesis. Experimental results with liver tissue demonstrate that cell damage during freezing results in substantial changes in tissue electrical properties. Two-dimensional EIT simulations of liver cryosurgery, which employ the experimental data, demonstrate the feasibility of this application.

Animals↗

PubMed2004

Distributed network imaging and electrical impedance tomography of minimally invasive surgery.

Minimally invasive surgery has become highly dependent on imaging. For instance, the effectiveness of cryosurgery in treating cancer is dependent on knowledge of freezing extent, and relies on real-time imaging techniques for monitoring. However, medical imaging is often very expensive and therefore not available to most of the world population. Here we propose the concept of distributed network imaging (DNI) which could make medical imaging and minimally invasive surgery available to all who need these advanced medical modalities. We demonstrate the concept through electrical impedance tomography (EIT) of cryosurgery. The central idea is to develop an inexpensive measurend (data collection hardware) at a remote site and then to connect the measurend apparatus to an advanced image reconstruction server, which can serve a large number of distributed measurends at remote sites, using existing communication conduits (Ethernet, telephone, satellite, etc.). These conduits transfer the raw data from the measurend to the server and the reconstructed image from the server to the measurend. Electrical impedance tomography (EIT) is an imaging modality which utilizes tissue impedance variation to construct an image. The EIT measurend which consists of electrodes, a power supply, and means to measure voltage is inexpensive, and therefore suitable for DNI. EIT is also very well-suited to imaging cryosurgery since frozen tissue impedance is much higher than that of unfrozen tissue. In this study, we first develop numerical models to illustrate the theoretical ability of EIT to image cryosurgery. We begin with a simplified two dimensional model, and then extend the study to the more appropriate three dimensional model. Our simulated finite element phantoms and pixel-based Newton-Raphson reconstruction algorithms were able to produce easily identifiable images of frozen regions within tissue. Then, we demonstrate the feasibility of the DNI concept though a case study using EIT to image an in vitro liver cryosurgery procedure through a modem. We find that the acquired raw data packets are less than 5KB per image and the images, using compression, do not exceed 50KB per image.

Algorithms↗

PubMed2003

Theoretical analysis of the thermal effects during in vivo tissue electroporation.

Tissue electroporation is a technique that facilitates the introduction of molecules into cells by applying a series of short electric pulses to specific areas of the body. These pulses temporarily increase the permeability of the cell membrane to small drugs and macromolecules. The goal of this paper is to provide information on the thermal effects of these electric pulses for consideration when designing electroporation protocols. The parameters investigated include electrode geometry, blood flow, metabolic heat generation, pulse frequency, and heat dissipation through the electrodes. Basic finite-element models were created in order to gain insight and weigh the importance of each parameter. The results suggest that for plate electrodes, the energy from the pulse may be used to adequately estimate the heating in the tissue. However, for needle electrodes, the geometry, i.e. spacing and diameter, and pulse frequency are critical when determining the thermal distribution in the tissue.

Animals↗