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Ensemble representations have been considered as one of the strategies that the visual system adopts to cope with its limited capacity. Thus, they include various statistical summaries such as mean, variance, and distributional properties and are formed over many stages of visual processing. The present study proposes a population-coding model of ensemble perception to provide a theoretical and computational framework for these various facets of ensemble perception. The proposed model consists of a simple feature layer and a pooling layer. We assumed ensemble representations as population responses in the pooling layer and decoded various statistical properties from population responses. Our model successfully predicted averaging performance in orientation, size, color, and motion direction across different tasks. Furthermore, it predicted variance discrimination performance and the priming effects of feature distributions. Finally, it explained the well-known variance and set-size effects and has a potential for explaining the adaptation and clustering effects. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
Assuntos
Percepção Visual , Humanos , Percepção Visual/fisiologia , Modelos TeóricosRESUMO
Contextual cueing refers to finding a target more efficiently in repeated displays than in novel displays. However, conflicting results have been reported regarding whether target absent judgments can also be efficient in repeated displays. To resolve this controversy, we first tested 3 factors that might influence the strength of distractor-distractor associations and then investigated how such associations produced faster responses on repeated target absent trials by measuring the patterns of eye movements. The factors were the number of distractors, number of repeated configurations, and diversity of the distractors' properties. In Experiments 1 and 2, we found that the diversity of the distractors was the only factor producing contextual cueing without a target. In Experiment 3, we recorded eye movements during a search task and found that the contextual cueing effect in the target absent condition was due to the lower number of fixations and larger mean saccadic amplitudes. Overall, these results suggest that the distractor-distractor associations, strengthened by the diversity of the distractors' properties, widened the attentional window. This enlarged window in turn helps people to reject more distractors at once and enables them to terminate a visual search faster in repeated target absent trials. (PsycInfo Database Record (c) 2020 APA, all rights reserved).
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Cryopreservation represents one if not the only long-term option for tissue and perhaps future organ banking. In one particular approach, cryopreservation is achieved by completely avoiding ice formation (or crystallization) through a process called vitrification. This "ice-free" approach to tissue banking requires a combination of high-concentration cryoprotective additives such as M22 (9.4 M), VS55 (8.4 M), or DP6 (6 M) and sufficiently fast rates of cooling and warming to avoid crystallization. In this article, we report the temperature-dependent specific heat capacity of the above-mentioned cryoprotective additives in small volumes (10 mg sample pans) at rates of 5°C/min and 10°C/min using a commercially available differential scanning calorimetry (TA Instruments Q1000), in the temperature range of -150°C to 30°C. This data can be utilized in heat-transfer models to predict thermal histories in a cryopreservation protocol. More specifically, the effects of temperature dependence of specific heat due to the presence of three different phases (liquid, ice, and vitreous phase) can dramatically impact the thermal history and therefore the outcome of the cryopreservation procedure. The crystallization potential of these cryoprotectants was also investigated by studying cases of maximal and minimal crystallization in VS55 and DP6, where M22 did not crystallize under any rates tested. To further reduce crystallization in VS55 and DP6, a stabilizing sugar (sucrose) was added in varying concentrations (0.15 M and 0.6 M) and was shown to further reduce crystallization, particularly in VS55, at modest rates of cooling (1°C/min, 5°C/min, and 10°C/min).
Assuntos
Criopreservação/métodos , Crioprotetores/química , Cristalização/métodos , Dimetil Sulfóxido/química , Formamidas/química , HEPES/química , Temperatura Alta , Propilenoglicóis/química , Sacarose/químicaRESUMO
Atrial fibrillation (AF) currently affects millions of people in the U.S. alone. Focal therapy is an increasingly attractive treatment for AF that avoids the debilitating effects of drugs for disease control. Perhaps the most widely used focal therapy for AF is heat-based radiofrequency (heating), although cryotherapy (cryo) is rapidly replacing it due to a reduction in side effects and positive clinical outcomes. A third focal therapy, irreversible electroporation (IRE), is also being considered in some settings. This study was designed to help guide treatment thresholds and compare mechanism of action across heating, cryo, and IRE. Testing was undertaken on HL-1 cells, a well-established cardiomyocyte cell line, to assess injury thresholds for each treatment method. Cell viability, as assessed by Hoechst and propidium iodide (PI) staining, was found to be minimal after exposure to temperatures ≤-40 °C (cryo), ≥60 °C (heating), and when field strengths ≥1500 V/cm (IRE) were used. Viability was then correlated to protein denaturation fraction (PDF) as assessed by Fourier transform infrared (FTIR) spectroscopy, and protein loss fraction (PLF) as assessed by bicinchoninic acid (BCA) assay after the three treatments. These protein changes were assessed both in the supernatant and the pellet of cell suspensions post-treatment. We found that dramatic viability loss (≥50%) correlated strongly with ≥12% protein change (PLF, PDF or a combination of the two) in every focal treatment. These studies help in defining both cellular thresholds and protein-based mechanisms of action that can be used to improve focal therapy application for AF.
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The use of thermal based therapies for treatment of atrial fibrillation is increasing. While numerous reports are available in the literature regarding the efficacy of cryotherapy on pulmonary vein survival, there are no reports specifically at the cellular level that establish thermal thresholds and mechanisms of cellular destruction. The current article reports on the response of HL-1 cardiomyocytes to cooling rates and end temperatures during cryothermal exposure. The focus is on establishment of in vitro thresholds while also establishing mechanisms of action due to biophysical events (i.e. intracellular ice formation and water transport).
Assuntos
Fibrilação Atrial/terapia , Temperatura Baixa/efeitos adversos , Criocirurgia/métodos , Miócitos Cardíacos/fisiologia , Linhagem Celular , HumanosRESUMO
There is an urgent need for sensors deployed during focal therapies to inform treatment planning and in vivo monitoring in thin tissues. Specifically, the measurement of thermal properties, cooling surface contact, tissue thickness, blood flow and phase change with mm to sub mm accuracy are needed. As a proof of principle, we demonstrate that a micro-thermal sensor based on the supported "3ω" technique can achieve this in vitro under idealized conditions in 0.5 to 2 mm thick tissues relevant to cryoablation of the pulmonary vein (PV). To begin with "3ω" sensors were microfabricated onto flat glass as an idealization of a focal probe surface. The sensor was then used to make new measurements of 'k' (W/m.K) of porcine PV, esophagus, and phrenic nerve, all needed for PV cryoabalation treatment planning. Further, by modifying the sensor use from traditional to dynamic mode new measurements related to tissue vs. fluid (i.e. water) contact, fluid flow conditions, tissue thickness, and phase change were made. In summary, the in vitro idealized system data presented is promising and warrants future work to integrate and test supported "3ω" sensors on in vivo deployed focal therapy probe surfaces (i.e. balloons or catheters).
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Criocirurgia/instrumentação , Desenho de Equipamento , Monitorização Fisiológica/instrumentação , Condutividade Térmica , Animais , Camundongos , Microtecnologia , SuínosRESUMO
Accurate knowledge of the thermal conductivity (k) of biological tissues is important for cryopreservation, thermal ablation, and cryosurgery. Here, we adapt the 3ω method-widely used for rigid, inorganic solids-as a reusable sensor to measure k of soft biological samples two orders of magnitude thinner than conventional tissue characterization methods. Analytical and numerical studies quantify the error of the commonly used "boundary mismatch approximation" of the bi-directional 3ω geometry, confirm that the generalized slope method is exact in the low-frequency limit, and bound its error for finite frequencies. The bi-directional 3ω measurement device is validated using control experiments to within ±2% (liquid water, standard deviation) and ±5% (ice). Measurements of mouse liver cover a temperature ranging from -69 °C to +33 °C. The liver results are independent of sample thicknesses from 3 mm down to 100 µm and agree with available literature for non-mouse liver to within the measurement scatter.
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Equipamentos e Provisões Elétricas , Condutividade Térmica , Animais , Simulação por Computador , Gelo , Fígado/química , Camundongos , Modelos Biológicos , Temperatura , Água/químicaRESUMO
The outcome of systemic and local therapies (e.g. chemotherapy, radiotherapy, surgery, focal ablation) for prostate cancer can be significantly improved by using tumor-specific adjuvants prior to treatment ("preconditioning"). We propose to use dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) to monitor the in vivo response of a mouse model of prostate cancer treated with a vascular disruptive agent, TNF-α, delivered on a gold nanoparticle (NP-TNF). Six male nude mice bearing 4-5 week old LNCaP tumors were scanned at 9.4 T. DCE-MRI was performed two days before and 4-5 h after treatment with NP-TNF. An intraperitoneal (i.p.) bolus of gadolinium-DTPA (Gd) was administered and contrast enhancement was measured for 90 min. Concentration-time curves of Gd were calculated and the area under the Gd curve (AUGC) was determined pre- and post-treatment. NP-TNF treatment caused an increase in contrast uptake in tumors. Interestingly, the early concentration (10 min post Gd bolus i.p.) was similar in both untreated and treated conditions; however, 90 min after injection, [Gd] was 3.4 times higher than before treatment. AUGC doubled from (11 ± 6) [Gd] × min before treatment to (22 ± 9) [Gd] × min after treatment. An increase in signal enhancement was also observed in the muscle but to a lesser degree. We also evaluated the kinetics of intravenous Gd administration in mice bearing a jugular vein catheter to mimic the delivery method used in clinical trials. The overall treatment effects were independent of the delivery pathway of the contrast agent. In conclusion, we show that DCE-MRI is suitable to detect changes associated with a vascular disruptive agent in a mouse model of prostate cancer. The ability to characterize the effects of nanoparticle therapy in vivo with non-destructive methods is important, as such compounds, in combination with treatment strategies, are progressing towards clinical trials.
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Imageamento por Ressonância Magnética/métodos , Nanopartículas/administração & dosagem , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/patologia , Fator de Necrose Tumoral alfa/administração & dosagem , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/uso terapêutico , Quimioterapia Adjuvante/métodos , Meios de Contraste/farmacocinética , Monitoramento de Medicamentos/métodos , Gadolínio DTPA/farmacocinética , Masculino , Camundongos , Camundongos Nus , Nanopartículas/química , Prognóstico , Resultado do TratamentoRESUMO
Tumor hypoxia is an important factor in assessment of both cancer progression and cancer treatment efficacy. This has driven a substantial effort toward development of imaging modalities that can directly measure oxygen distribution and therefore hypoxia in tissue. Although several approaches to measure hypoxia exist, direct measurement of tissue oxygen through an imaging approach is still an unmet need. To address this, we present a new approach based on in vivo application of photoacoustic lifetime imaging (PALI) to map the distribution of oxygen partial pressure (pO2) in tissue. This method utilizes methylene blue, a dye widely used in clinical applications, as an oxygen-sensitive imaging agent. PALI measurement of oxygen relies upon pO2-dependent excitation lifetime of the dye. A multimodal imaging system was designed and built to achieve ultrasound (US), photoacoustic, and PALI imaging within the same system. Nude mice bearing LNCaP xenograft hindlimb tumors were used as the target tissue. Hypoxic regions were identified within the tumor in a combined US/PALI image. Finally, the statistical distributions of pO2 in tumor, normal, and control tissues were compared with measurements by a needle-mounted oxygen probe. A statistically significant drop in mean pO2 was consistently detected by both methods in tumors.
Assuntos
Hipóxia Celular/fisiologia , Neoplasias/metabolismo , Neoplasias/patologia , Imagem Óptica/métodos , Oxigênio/metabolismo , Técnicas Fotoacústicas/métodos , Animais , Linhagem Celular Tumoral , Humanos , Camundongos , Camundongos Nus , Imagem Óptica/instrumentação , Imagens de Fantasmas , Técnicas Fotoacústicas/instrumentaçãoRESUMO
Surgery, radiation and chemotherapy remain the mainstay of current cancer therapy. However, treatment failure persists due to the inability to achieve complete local control of the tumor and curtail metastatic spread. Vascular disrupting agents (VDAs) are a class of promising systemic agents that are known to synergistically enhance radiation, chemotherapy or thermal treatments of solid tumors. Unfortunately, there is still an unmet need for VDAs with more favorable safety profiles and fewer side effects. Recent work has demonstrated that conjugating VDAs to other molecules (polyethylene glycol, CNGRCG peptide) or nanoparticles (liposomes, gold) can reduce toxicity of one prominent VDA (tumor necrosis factor alpha, TNF-α). In this report, we show the potential of a gold conjugated TNF-α nanoparticle (NP-TNF) to improve multimodal cancer therapies with VDAs. In a dorsal skin fold and hindlimb murine xenograft model of prostate cancer, we found that NP-TNF disrupts endothelial barrier function and induces a significant increase in vascular permeability within the first 1-2 h followed by a dramatic 80% drop in perfusion 2-6 h after systemic administration. We also demonstrate that the tumor response to the nanoparticle can be verified using dynamic contrast-enhanced magnetic resonance imaging (MRI), a technique in clinical use. Additionally, multimodal treatment with thermal therapies at the perfusion nadir in the sub- and supraphysiological temperature regimes increases tumor volumetric destruction by over 60% and leads to significant tumor growth delays compared to thermal therapy alone. Lastly, NP-TNF was found to enhance thermal therapy in the absence of neutrophil recruitment, suggesting that immune/inflammatory regulation is not central to its power as part of a multimodal approach. Our data demonstrate the potential of nanoparticle-conjugated VDAs to significantly improve cancer therapy by preconditioning tumor vasculature to a secondary insult in a targeted manner. We anticipate our work to direct investigations into more potent tumor vasculature specific combinations of VDAs and nanoparticles with the goal of transitioning optimal regimens into clinical trials.
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Antineoplásicos/administração & dosagem , Nanoconjugados/administração & dosagem , Neoplasias da Próstata/tratamento farmacológico , Fator de Necrose Tumoral alfa/administração & dosagem , Animais , Linhagem Celular Tumoral , Terapia Combinada , Ouro , Humanos , Hipertermia Induzida , Masculino , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/química , Camundongos , Camundongos Nus , Nanoconjugados/química , Neoplasias da Próstata/irrigação sanguínea , Neoplasias da Próstata/terapia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
While cryosurgery has proven capable in treating of a variety of conditions, it has met with some resistance among physicians, in part due to shortcomings in the ability to predict treatment outcomes. Here we attempt to address several key issues related to predictive modeling by demonstrating methods for accurately characterizing heat transfer from cryoprobes, report temperature dependent thermal properties for ultrasound gel (a convenient tissue phantom) down to cryogenic temperatures, and demonstrate the ability of convective exchange heat transfer boundary conditions to accurately describe freezing in the case of single and multiple interacting cryoprobe(s). Temperature dependent changes in the specific heat and thermal conductivity for ultrasound gel are reported down to -150 °C for the first time here and these data were used to accurately describe freezing in ultrasound gel in subsequent modeling. Freezing around a single and two interacting cryoprobe(s) was characterized in the ultrasound gel phantom by mapping the temperature in and around the "iceball" with carefully placed thermocouple arrays. These experimental data were fit with finite-element modeling in COMSOL Multiphysics, which was used to investigate the sensitivity and effectiveness of convective boundary conditions in describing heat transfer from the cryoprobes. Heat transfer at the probe tip was described in terms of a convective coefficient and the cryogen temperature. While model accuracy depended strongly on spatial (i.e., along the exchange surface) variation in the convective coefficient, it was much less sensitive to spatial and transient variations in the cryogen temperature parameter. The optimized fit, convective exchange conditions for the single-probe case also provided close agreement with the experimental data for the case of two interacting cryoprobes, suggesting that this basic characterization and modeling approach can be extended to accurately describe more complicated, multiprobe freezing geometries. Accurately characterizing cryoprobe behavior in phantoms requires detailed knowledge of the freezing medium's properties throughout the range of expected temperatures and an appropriate description of the heat transfer across the probe's exchange surfaces. Here we demonstrate that convective exchange boundary conditions provide an accurate and versatile description of heat transfer from cryoprobes, offering potential advantages over the traditional constant surface heat flux and constant surface temperature descriptions. In addition, although this study was conducted on Joule-Thomson type cryoprobes, the general methodologies should extend to any probe that is based on convective exchange with a cryogenic fluid.
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Convecção , Criocirurgia/instrumentação , Temperatura Alta , Ultrassom/instrumentação , Congelamento , Géis , Modelos Teóricos , Imagens de FantasmasRESUMO
The current study presents a new and novel analysis of heat release signatures measured by a differential scanning calorimeter (DSC) associated with water transport (WT), intracellular ice formation (IIF) and extracellular ice formation (EIF). Correlative cryomicroscopy experiments were also performed to validate the DSC data. The DSC and cryomicroscopy experiments were performed on human dermal fibroblast cells (HDFs) at various cytocrit values (0-0.8) at various cooling rates (0.5-250 °C/min). A comparison of the cryomicroscopy experiments with the DSC analysis show reasonable agreement in the water transport (cellular dehydration) and IIF characteristics between both the techniques with the caveat that IIF measured by DSC lagged that measured by cryomicroscopy. This was ascribed to differences in the techniques (i.e. cell vs. bulk measurement) and the possibility that not all IIF is associated with visual darkening. High and low rates of 0.5 °C/min and 250 °C/min were chosen as HDFs did not exhibit significant IIF or WT at each of these extremes respectively. Analysis of post-thaw viability data suggested that 10 °C/min was the presumptive optimal cooling rate for HDFs and was independent of the cytocrit value. The ratio of measured heat values associated with IIF (q(IIF)) to the total heat released from both IIF and water transport or from the total cell water content in the sample (q(CW)) was also found to increase as the cooling rate was increased from 10 to 250 °C/min and was independent of the sample cytocrit value. Taken together, these observations suggest that the proposed analysis is capable of deconvolving water transport and IIF data from the measured DSC latent heat thermograms in cell suspensions during freezing.
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Varredura Diferencial de Calorimetria , Derme/citologia , Fibroblastos/citologia , Gelo/análise , Água/metabolismo , Transporte Biológico , Linhagem Celular , Sobrevivência Celular , Criopreservação/métodos , Fibroblastos/química , Fibroblastos/metabolismo , Congelamento , Humanos , Espaço Intracelular/química , Espaço Intracelular/metabolismo , Microscopia , Água/análiseRESUMO
Solid tumors such as hepatocellular carcinoma are very often not amenable to chemotherapy and radiotherapy. Local ablation methods, including chemical ablation with absolute ethanol, are therefore an option for treatment but lack of information about the mechanism of devitalization leading to cell death is a hindrance to further adoption. Systemic toxicity also has limited the amount of ethanol that can be used in a single treatment session. Therefore we evaluated the mechanism of urea, a denaturant with little or no systemic toxicity, for potential use in chemical ablation. In this study we report on the use of three methods to analyze the effects in cell culture with a view towards eventual clinical application. Human hepatoma HuH-7 cells were analyzed at several time points after treatment using FTIR, DSC, and Raman microspectroscopy based on MTT and PI-exclusion viability assays. Time course fractional denaturation data plotted against viability show that a 50% viability drop occurs after only a 10-20% drop in overall protein denaturation. Other methods of cell death such as apoptosis may also be operative, but this result implies that protein denaturation is one of the major mechanisms of cell death. This is in line with what has been previously suggested for purely thermal methods, and opens the way to mechanism-based improvements in chemical ablation of solid tumors.
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Técnicas de Ablação , Sobrevivência Celular/efeitos dos fármacos , Desnaturação Proteica , Ureia/farmacologia , Varredura Diferencial de Calorimetria , Linhagem Celular Tumoral , Humanos , Neoplasias/terapia , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral Raman , TemperaturaRESUMO
The laser-induced breakdown spectroscopy (LIBS) technique was applied to quantify nutrients (Mg, Ca, Na, and K) in spinach and rice and to discriminate pesticide-contaminated products in a rapid manner. Standard reference materials (spinach leaves and unpolished rice flour) were used to establish a relationship between LIBS intensity and the concentration of each element (Mg, Ca, Na, and K) (i.e., calibration line). The limits of detection (LODs) for Mg, Ca, Na, and K were found to be 29.63, 102.65, 36.36, and 44.46 mg/kg in spinach and 7.54, 1.76, 4.19, and 6.70 mg/kg in unpolished rice, respectively. Concentrations of those nutrient elements present in spinach and unpolished rice from a local market were determined by using the calibration lines and compared with those measured with ICP-OES, showing good agreement. The data also suggested that the LIBS technique with the chemometric method (PLS-DA) could be a great tool to distinguish pesticide-contaminated samples from pesticide-free samples in a rapid manner even though they have similar elemental compositions. Misclassification rates were found to be 0 and 2% for clean spinach and pesticide-contaminated spinach, respectively, by applying the PLS-DA model established from the training set of data to predict the classes of test samples.
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Contaminação de Alimentos/análise , Oryza/química , Praguicidas/análise , Análise Espectral/métodos , Spinacia oleracea/química , Oligoelementos/análise , Calibragem , Alimentos , Limite de Detecção , Análise Espectral/normasRESUMO
While studies on the freezing of cells in suspension have been carried out extensively, corresponding studies with cells in the attached state and in tissue or tissue-equivalents are less developed. As attachment is a hallmark of the tissue state it is important to understand its impact on biophysics and viability to better apply freezing towards tissue preservation. The current study reports on observed biophysical response changes observed during freezing human dermal fibroblasts in suspension, attached cell, and fibrin tissue-equivalent models. Specifically, intracellular ice formation is shown to increase and dehydration is inferred to increase from suspension to attached systems. Biophysical model parameters fit to these experimental observations reflect the higher kinetics in the attached state. Post-thaw viability values from fast cooling rates were higher for suspension systems, and correlated well with the amount of IIF observed. On the other hand, viability values from slow cooling rates were higher for attached systems, although the degree of dehydration was predicted to be comparable to suspension cells. This disconnect between biophysics and viability predictions at slow rates clearly requires further investigation as it runs counter to our current understanding of dehydration injury in cells. This may suggest a possible protective effect of the attachment state on cell systems.
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Fibroblastos/fisiologia , Congelamento , Preservação de Tecido/métodos , Animais , Biofísica , Bovinos , Adesão Celular , Sobrevivência Celular , Células Cultivadas , Temperatura Baixa , Dessecação , Fibrinogênio/metabolismo , Fibroblastos/citologia , Humanos , Gelo , Microscopia , Modelos Teóricos , Trombina/metabolismoRESUMO
Cryopreservation is a convenient method for long-term preservation of natural and engineered tissues in regenerative medicine. Homogeneous loading of tissues with CPAs, however, forms one of the major hurdles in tissue cryopreservation. In this study, computer tomography (CT) as a non-invasive imaging method was used to determine the effective diffusion of Me2SO in tissue-engineered collagen scaffolds. The dimensions of the scaffolds were 30 x 30 x 10 mm3 with a homogeneous pore size of 100 microm and a porosity of 98%. CT images were acquired after equilibrating the scaffolds in phosphate buffered saline (PBS) and transferring them directly in 10% (v/v)Me2SO. The Me2SO loading process of the scaffold could thus be measured and visualized in real time. The experimental data were fitted using a diffusion equation. The calculated effective diffusion constant for Me2SO in the PBS loaded scaffold was determined from experimental diffusion studies to be 2.4 x 10(-6) cm2/s at 20 degrees C.
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Colágeno , Dimetil Sulfóxido , Engenharia Tecidual , Alicerces Teciduais , Tomografia Computadorizada por Raios X , Materiais Biocompatíveis , Criopreservação , Crioprotetores/química , Difusão , Dimetil Sulfóxido/química , Processamento de Imagem Assistida por Computador , Modelos Biológicos , Porosidade , Engenharia Tecidual/métodosRESUMO
It is well accepted in cryobiology that the temperature history and cooling rates experienced in biomaterials during freezing procedures correlate strongly with biological outcome. Therefore, heat transfer measurement and prediction in the cryogenic regime is central to the field. Although direct measurement of temperature history (i.e. heat transfer) can be performed, accuracy is usually achieved only for local measurements within a given system and cannot be readily generalized to another system without the aid of predictive models. The accuracy of these models rely upon thermal properties which are known to be highly dependent on temperature, and in the case of significant cryoprotectant loading, also on crystallized fraction. In this work, we review the available thermal properties of biomaterials in the cryogenic regime. The review shows a lack of properties for many biomaterials in the subzero temperature domain, and especially for systems with cryoprotective agents. Unfortunately, use of values from the limited data available (usually only down to -40 degrees C) lead to an underestimation of thermal property change (i.e. conductivity rise and specific heat drop due to ice crystallization) with lower temperatures. Conversely, use of surrogate values based solely on ice thermal properties lead to an overestimation of thermal property change for most biomaterials. Additionally, recent work extending the range of available thermal properties to -150 degrees C has shown that the thermal conductivity will drop in both PBS and tissue (liver) due to amorphous/glassy phases (versus crystalline) of biomaterials with the addition of cryoprotective additives such as glycerol. Thus, we investigated the implications of using approximated or constant property values versus measured temperature-dependent values for predicting temperature history during freezing in PBS (phosphate-buffered saline) and porcine liver with and without cryoprotectants (glycerol). Using measured property values (thermal conductivity, specific heat, and latent heat of phase change) of porcine liver, a standard was created which showed that values based on surrogate ice properties under-predicted cooling times, while constant properties (i.e. based on limited data reported near the freezing point) over-predicted cooling times. Additionally, a new iterative numerical method that accommodates non-equilibrium cooling effects as a function of time and position (i.e. crystallization versus amorphous phase) was used to predict temperature history during freezing in glycerol loaded systems. Results indicate that in addition to the increase in cooling times due to the lowering of thermal diffusivity with more glycerol, non-equilibrium effects such as the prevention of maximal crystallization (i.e. amorphous phases) will further increase required cooling times. It was also found that the amplified effect of non-equilibrium cooling and crystallization with system size prevents the thermal history to be described with non-dimensional lengths, such as was possible under equilibrium cooling. These results affirm the need to use accurate thermal properties that incorporate temperature dependence and crystallized fraction. Further studies are needed to extract thermal properties of other important biomaterials in the subzero temperature domain and to develop accurate numerical methods which take into account non-equilibrium cooling events encountered in cryobiology when partial or total vitrification occurs.