| Organism, microorganism, cell or biological substance is performed as the sensitive elements in biosensor to sense physical or chemical stimulation outside the world. And the response of various organism parameters or physical and chemical parameters were caused by the stimulation. At the same time, the biosensor receives and senses these responses and makes evaluation. Biosensor and its measurement technique can be not only applied to detection the microorganism, metabolism as well as biology study, but also provide scientific and effective method for pharmacology and toxicologic study. Therefore, biosensor has been widely used in clinical diagnosis, function and toxicity analysis of drug, environmental toxicant monitoring, food safety analysis and biologics quality control and other relevant fields.On the series piezoelectric biosensor research, we have spent many years on it and have achieved some progress as well as some accumulation. Specific elements of SPQC include exploring electrodes, quartz crystal, oscillation circuit, and other related ancillary components of the sensor. The core of response signal generation is quartz crystal and function implementation element is detection probe, namely series electrodes. SPQC retains advantages of piezoelectric quartz crystal sensor such as low cost, simple operation, and makes crystal oscillator get rid of the influence of liquid phase loss and significantly improves the stability. It has been well applied in environmental microbial monitoring, rapid detection of clinical bacteria, susceptibility testing and other fields. To satisfy simultaneous on-line NDT for multiple cell samples, provide new scientific and effective method for cellular pharmacology and toxicology, and develop living cells in vitro dynamic monitoring platform for PQC sensor, the paper conducted the following researches:(1) Combined in vitro cell culture technology and PQC sensor and established a real-time online detection platform for ITO-PQC living cell taking ITO electrode as basal cell sensor. The frequency response curves can reflect the process of cell growth sufficiently with anti-background electrolyte disturbance ability thus to maintain good frequency stability. Compared to traditional biological detection means, ITO-PQC sensor is a non-intrusive, cell sensing approach not requiring marking. It can substitute traditional tissue culture plate experiment to online record adherency, growth, proliferation and other growth status automatically and to provide real time dynamic analysis data of cells.(2) Utilized the ITO-PQC real time sensing monitoring platform on monitoring and quantitative analysis of adherency, spreading and proliferation for different inoculation concentration of MG63 cell. Studied impact of low oxygen simulatio n chemical reagent Co Cl2 on biological characteristics of MG63 cell. It is detected that accompanying with complicated influence of cell proliferation and apoptosis, the concentration and processing time is closely related to Co Cl2 chemical simulation low oxygen process. Cells show different biological characteristics under hypoxia state. ITO-MSPQC piezoelectric cell-based sensor provides a simple and cheap method for in vitro evaluation of chemicals cytotoxicity, and can be used on analysis of action mode of drugs on cells.(3) To improve the detection signal strength, perform on line detection of sensitiveness on cell growth. We modified electrode of sensor and introduced coplanar microelectrode to piezoelectric cell sensing detection system, and established new type of Micro Au-PQC sensor suitable for on line monitoring of cell. Spiral rectangular micro gold electrode will consider electrode size according to cell shape and size. Cell adhesion and growth can perform bridge connection for two adjacent electrode bands. The steady state current density between microelectrodes is high. Mass transfer velocity between electrodes is rapid. The respond time is short. More signals can be collected on electrode surface which can reflect minor response of cell growth on electrode. Consequently, sensitivity has obvious improvement. Micro Au-PQC sensor is a new type of non-intrusion on-line monitoring technique of living cell requiring no marking.(4) Performed real time on-line monitoring of human HUVEC cellular damage induced by LPS and pharmacological action of VC/VE antioxidant vitamins on such damage with the newly built Micro Au-PQC series piezoelectric cell-based sensors. The frequency response curve of sensor indicates that low-concentration of LPS can promote proliferation of HUVEC cells of low-concentration in short time and will show proliferation inhibition and apoptosis inducing after a certain time. In addition, it has been detected that drug combination of VC and VE can play an effect on reducing LPS damage excellently at low concentration. The results of sensor monitoring are in consistency with the results of MTT test and microscope observation.(5) In order to better simulate physiological conditions in the body, and study the effects of stimulating factors on endothelial barrier function, we adopted ITO working electrode with biological compatibility conductive polymer film modified sensor, and established BIO-PQC sensor with biocompatibility conductive film modified electrodes. Electrochemical impedance spectroscopy(EIS) have been characterized by different steps of polymer membrane and HUVEC preparation of cell monolayer. Detect the impact of LPS, histamine and other exogenous or endogenous inflammatory substances on endothelial barrier function by adopting sensor. The experimental results indicate that HUVEC cell layer piezoelectric frequency shift response shows dependency to stimulus dose and verifies that BOI-PQC sensor is a valuable method to evaluate damage of endothelial barrier function on inflammatory mediator. The research can further promote the application of piezoelectricity biosensor on cytotoxic drugs test.
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