Objective To evaluate which is better method zymogen or low temperature frozen in removing vascular endothelial cell so as to lay a foundation for creating a kind of brace which is not to be rejected and the same as own blood vessel. Methods Fresh and not damaged umbilical blood vessel was collected from natural labour women, human umbilical blood vessel was remove carefully from normal foetus, then was put into disinfectant at 37℃ for 24 hours. They were divided into 3 groups:normal group(NG),zymogen group(ZG) and low temperature frozen group(LG). ZG: 0.1% collagenⅡ enzyme was addedin umbilical blood vessel and closed the both sides and the vascular endothelialcell was removed in 37℃ water. LG:Umbilical blood vessel was put into liquidnitrogen for 24 hours after frozened step by step, and then it was put into 37℃ water for 30-60 s and the vascular endothelial cells were washed away by normal saline. NG:Umbilical blood vessel was kept into 4℃ Kerb’s liquid. The bacteria were culturedin each group. The samples were stained by HE,elastic fiber and collagen fiberwere observed by light and scanning electron microscope. The difference of compliance was compared. Human leukocyte antigen ABC(HLA-ABC) and HLA-DR were observed by immunohistochemical method and the expression of antigen of umbilical blood vessel was analysed. Results In LG, umbilical vascular endothelial cells were removed completely; artery showed vertical smooth muscle and vein showed elastic membrane. InZG, umbilical vascular endothelial cells were removed completely after 20 minutes;artery showed vertical smooth muscle cells and vein showed lower endothelial layer. The vascular compliance in LG was higher than that in NG, and the latter was also higher than that in ZG,but showing no significant differences (Pgt;0.05). The compliance of umbilical vein was 2-3 times as much asthat of umbilical artery.The expression of HLA-ABC and HLA-DR in LG andZG were lower than that in NG, showing significant differences (Plt;0.01). Conclusion Low temperature frozen methodand zymogen method(0.1% collagen Ⅱ enzyme for 20 min) can remove vascular endothelial cells of human umbilical blood vessel completely.Low temperature frozenmethod was better than zymogen method.
The freeze-drying is a technology that preserves biological samples in a dry state, which is beneficial for storage, transportation, and cost saving. In this study, the bovine pericardium was treated with a freeze-drying protectant composed of polyethylene glycol (PEG) and trehalose (Tre), and then freeze-dried. The results demonstrated that the mechanical properties of the pericardium treated with PEG + 10% w/v Tre were superior to those of the pericardium fixed with glutaraldehyde (GA). The wet state water content of the rehydrated pericardium, determined using the Karl Fischer method, was (74.81 ± 1.44)%, which was comparable to that of the GA-fixed pericardium. The dry state water content was significantly reduced to (8.64 ± 1.52)%, indicating effective dehydration during the freeze-drying process. Differential scanning calorimetry (DSC) testing revealed that the thermal shrinkage temperature of the pericardium was (84.96 ± 0.49) ℃, higher than that of the GA-fixed pericardium (83.14 ± 0.11) ℃, indicating greater thermal stability. Fourier transform infrared spectroscopy (FTIR) results showed no damage to the protein structure during freeze-drying. Hematoxylin and eosin (HE) staining demonstrated that the freeze-drying process reduced pore formation, prevented ice crystal growth, and resulted in a tighter arrangement of tissue fibers. The frozen-dried bovine pericardium was subjected to tests for cell viability and hemolysis rate. The results revealed a cell proliferation rate of (77.87 ± 0.49)%, corresponding to a toxicity grade of 1. Additionally, the hemolysis rate was (0.17 ± 0.02)%, which is below the standard of 5%. These findings indicated that the frozen-dried bovine pericardium exhibited satisfactory performance in terms of cytotoxicity and hemolysis, thus meeting the relevant standards. In summary, the performance of the bovine pericardium treated with PEG + 10% w/v Tre and subjected to freeze-drying could meet the required standards.
The temperature during the brain tumor therapy using high-intensity focused ultrasound (HIFU) should be controlled strictly. This research aimed at realizing uniform temperature distribution in the focal region by adjusting driving signals of phased array transducer. The three-dimensional simulation model imitating craniotomy HIFU brain tumor treatment was established based on an 82-element transducer and the computed tomography (CT) data of a volunteer's head was used to calculate and modulate the temperature distributions using the finite difference in time domain (FDTD) method. Two signals which focus at two preset targets with a certain distance were superimposed to emit each transducer element. Then the temperature distribution was modulated by changing the triggering time delay and amplitudes of the two signals. The results showed that when the distance between the two targets was within a certain range, a focal region with uniform temperature distribution could be created. And also the volume of focal region formed by one irradiation could be adjusted. The simulation results would provide theoretical method and reference for HIFU applying in clinical brain tumor treatment safely and effectively.
The nondestructive reconstruction of three-dimensional (3D) temperature field in biological tissue is always an important problem to be resolved in biomedical engineering field. This paper presents a novel method of nondestructive reconstruction of 3D temperature field in biological tissue based on multi-island genetic algorithm (MIGA). By this method, the resolving of inverse problem of bio-heat transfer is transformed to be a solving process of direct problem. An experiment and its corresponding simulation were carried out to verify the feasibility and reliability. In the experiment a high purity polypropylene material, whose thermophysical parameters were similar to the fat tissue being tested, were adopted so that it could avoid the negative results created by the other factors. We set the position P(x, y, z) as the point heat source in the biological tissue and its temperature t as optimization variable, got the experimental temperature values of the points in a module surface, subtracted them from the corresponding simulating temperature values in the same module surface, and then took the sum of absolute value. We took it as the objective function of successive iteration. It was found that the less the target value was, the more optimal the current variables, i.e. the heat source position and the temperature values, were. To improve the optimization efficiency, a novel establishment method of objective function was also provided. The simulating position and experimental position of heat source were very approximate to each other. When the optimum values are determined, the corresponding 3D temperature field is also confirmed, and the temperature distribution of arbitrary section can be acquired. The MIGA can be well applied in the reconstruction of 3D temperature field in biological tissue. Because of the differences between the MIGA and the traditional numerical methods, we do not have to acquire all the data of surface. It is convenient and fast, and shows a prosperous application future.
ObjectiveTo study the method of rapid and accurate measurement of body temperature in dense population during the coronavirus disease 2019 pandemic.MethodsFrom January 27th to February 8th, 2020, subjects were respectively measured with two kinds of non-contact infrared thermometers (blue thermometer and red one) to measure the temperature of forehead, neck, and inner side of forearm under the conditions of 4–6℃ (n=152), 7–10℃ (n=103), and 11–25℃ (n=209), while the temperature of axillary was measured with mercury thermometer under the same conditions. Taking the mercury thermometer temperature as the gold standard, the measurement results with non-contact infrared thermometers were compared.ResultsAt 7–10℃, there was no statistical difference among the forehead temperatures measured by the two non-contact infrared thermometers and the axillary temperature (P>0.05); there was no difference among the temperature measured by blue thermometer on forehead, neck, and inner side of forearm (P>0.05); no difference was found between the temperature measured by the red thermometer on forehead and inner side of forearm (P>0.05), while there was statistical difference between the temperatures measured by the red thermometer on forehead and neck (P<0.05). Under the environment of 11−25℃, there was no statistical difference among the forehead temperatures measured by the two infrared thermometers and the axillary temperature (P>0.05); the difference between the temperatures of forehead and inner side of forearm measured by the blue thermometer was statistically significant (P<0.05), while no difference appeared between the forehead and neck temperatures measured by the blue thermometer (P>0.05); there was no statistical difference among the temperatures of three body regions mentioned above measured by the red thermometer (P>0.05). According to the manual, the allowable fluctuation range of the blue thermometer was 0.3℃, and that of the red one was 0.2℃. The mean differences in measured values between different measured sites of the two products were within the allowable fluctuation range. Therefore, the differences had no clinical significance in the environment of 7–25℃. Under the environment of 4–6℃, the detection rate of blue thermometer was 2.2% and that of the red one was 19.1%.ConclusionsThere is no clinical difference between the temperature measured by mercury thermometer and the temperature measured by temperature guns at 7–10 or 11–25℃, so temperature guns can be widely used. In order to maintain the maximum distance between the measuring and the measured persons and reduce the infection risk, it is recommended to choose the inner forearm for temperature measurement. Under the environment of ambient temperature 4–6℃, the detection rate of non-contact electronic temperature gun is low, requiring taking thermal measures for the instrument.
In recent years, target temperature management (TTM) has been increasingly applied to cardiac arrest patients, and programs and strategies for TTM are in a constant state of update and refinement. This paper analyzes and proposes relevant strategies from the concept of TTM, its clinical application status for cardiac arrest patients in domestic and international medical institutions, its deficiencies in the clinical practice, and factors affecting the development of TTM, with a view to providing a realistic basis for the development of high-quality TTM in medical institutions.
A multiple-stimuli-responsive drug-conjugated cross-linked micelles was prepared by radical copolymerization. The chemical structure, morphology, and size of the cross-linked micelles were characterized, and the drug loading of the micelle was calculated. The experimental results indicated that the hydrodynamic size of the drug-loaded micelles were about 100 nm, and the as prepared micelles could be degraded and swelled in presence of reducing glutathione (GSH). The low critical solution temperature (LCST) of the micelle was around 39.4℃. According to the experimental results, the micelles will shrink at temperature above the LCST. Subsequently, the accumulative drug release rate was up to 91.78% under acidic (pH 5.0), reductive (GSH 10 mmol/L) and high temperature (42.0℃) conditions mimicking the tumor microenvironment, while a relatively low release rate of 1.12% was observed without stimulation. The drug-conjugated cross-linked micelles showed a strong cell uptake behavior. In the cytotoxicity assay, the micelles exhibited effective anti-cancer activity and excellent biocompatibility. In brief, the experimental results show that the as-prepared drug-conjugated cross-linked micelle exhibits multiple stimuli-responsiveness, which holds great promise for anti-cancer drug delivery.
ObjectiveTo manufacture a polycaprolactone (PCL)/type Ⅰ collagen (COL Ⅰ) tissue engineered meniscus scaffold (hereinafter referred to as PCL/COL Ⅰ meniscus scaffold) by three-dimensional (3D) printing with low temperature deposition technique and to study its physicochemical properties.MethodsFirst, the 15% PCL/4% COLⅠ composite solution and 15% PCL simple solution were prepared. Then, 15% PCL/4% COL Ⅰmeniscus scaffold and 15% PCL meniscal scaffold were prepared by using 3D printing with low temperature deposition techniques. The morphology and microstructure of the scaffolds were observed by gross observation and scanning electron microscope. The compression modulus and tensile modulus of the scaffolds were measured by biomechanical test. The components of the scaffolds were analyzed by Fourier transform infrared spectroscopy (FTIR). The contact angle of the scaffold surface was measured. The meniscus cells of rabbits were cultured with the two scaffold extracts and scaffolds, respectively. After cultured, the cell proliferations were detected by cell counting kit 8 (CCK-8), and the normal cultured cells were used as controls. Cell adhesion and growth of scaffold-cell complex were observed by scanning electron microscope.ResultsAccording to the gross and scanning electron microscope observations, two scaffolds had orientated 3D microstructures and pores, but the surface of the PCL/COLⅠ meniscus scaffold was rougher than the PCL meniscus scaffold. Biomechanical analysis showed that the tensile modulus and compression modulus of the PCL/COL Ⅰ meniscus scaffold were not significantly different from those of the PCL meniscus scaffold (P>0.05). FTIR analysis results showed that COL Ⅰ and PCL were successful mixed in PCL/ COL Ⅰ meniscus scaffolds. The contact angle of PCL/COLⅠ meniscus scaffold [(83.19±7.49)°] was significantly lower than that of PCL meniscus scaffold [(111.13±5.70)°] (t=6.638, P=0.000). The results of the CCK-8 assay indicated that with time, the number of cells cultured in two scaffold extracts showed an increasing trend, and there was no significant difference when compared with the control group (P>0.05). Scanning electron microscope observation showed that the cells attached on the PCL/ COL Ⅰ meniscus scaffold more than that on the PCL scaffold.ConclusionPCL/COLⅠmeniscus scaffolds are prepared by 3D printing with low temperature deposition technique, which has excellent physicochemical properties without cytotoxicity. PCL/COLⅠmeniscus scaffold is expected to be used as the material for meniscus tissue engineering.
By studying the relationship between fingertip temperature changes and arterial function during vascular reactivity test, we established a new non-invasive method for detecting vascular function, in order to provide an assistance for early diagnosis and prevention of cardiovascular diseases. We customized three modules respectively for blood occlusion, measurement of finger temperature and blood oxygen acquisition, and then we established the hardware of data acquisition system. And the software was programmed with Labview. Healthy subjects [group A, n=24, (44.6±9.0) years] and subjects with cardiovascular diseases [group B, n=33, (57.2±9.9) years)] were chosen for the study. Subject's finger temperature, blood oxygen and occlusion pressure of block side during and after unilateral arm brachial artery occlusion were recorded, as well as some other regular physiological indexes. By time-domain analysis, we extracted 12 parameters from fingertip temperature signal, including the initial temperature (Ti), temperature rebound (TR), the time of the temperature recovering to initial status (RIt) and other parameters from the finger temperature signal. We in the experiment also measured other regular physiological body mass index (BMI), systolic blood pressure (SBP), diastiolic blood pressure (DBP) and so on. Results showed that 8 parameters difference between the two group of data were significant. based on the statistical results. A discriminant function of vascular function status was established afterwards. We found in the study that the changes of finger temperature during unilateral arms brachial artery occlusion and open were closely related to vascular function. We hope that the method presented in this article could lay a foundation of early detection of vascular function.
ObjectiveTo study the preparation and cytocompatibility of bone tissue engineering scaffolds by combining low temperature three dimensional (3D) printing and vacuum freeze-drying techniques. MethodsCollagen (COL)and silk fibroin (SF) were manufactured from fresh bovine tendon and silkworm silk. SolidWorks2014 was adopted to design bone tissue engineering scaffold models with the size of 9 mm×9 mm×3 mm and pore diameter of 500μm. According to the behavior of composite materials that low temperature 3D printing equipment required, COL, SF, and nano-hydroxyapatite (nHA)at a ratio of 9:3:2 and low temperature 3D printing in combination with vacuum freeze-drying techniques were accepted to build COL/SF/nHA composite scaffolds. Gross observation and scanning electron microscope (SEM) were applied to observe the morphology and surface structures of composite scaffolds. Meanwhile, compression displacement, compression stress, and elasticity modulus were measured by mechanics machine to analyze mechanical properties of composite scaffolds. The growth and proliferation of MC3T3-E1 cells were evaluated using SEM, inverted microscope, and MTT assay after cultured for 1, 7, 14, and 21 days on the composite scaffolds. The RT-PCR and Western blot techniques were adopted to detect the gene and protein expressions of COL I, alkaline phosphatase (ALP), and osteocalcin (OCN) in MC3T3-E1 cells after 21 days. ResultsCOL/SF/nHA composite scaffolds were successfully prepared by low temperature 3D printing technology and vacuum freeze-drying techniques; the SEM results showed that the bionic bone scaffolds were 3D polyporous structures with macropores and micropores. The mechanical performance showed that the elasticity modulus was (344.783 07±40.728 55) kPa; compression displacement was (0.958 41±0.000 84) mm; and compression stress was (0.062 15±0.007 15) MPa. The results of inverted microscope, SEM, and MTT method showed that a large number of cells adhered to the surface with full extension and good cells growth inside the macropores, which demonstrated a satisfactory proliferation rate of the MC3T3-E1 cells on the composite scaffolds. The RT-PCR and Western blot electrophoresis revealed gene expressions and protein synthesis of COL I, ALP, and OCN in MC3T3-E1 cells. ConclusionLow temperature 3D printing in combination with vacuum freeze-drying techniques could realize multi-aperture coexisted bionic bone tissue engineering scaffolds and control the microstructures of composite scaffolds precisely that possess good cytocompatibility. It was expected to be a bone defect repair material, which lays a foundation for further research of bone defect.