ABSTRACT
Objective:Mineral trioxide aggregate (MTA),a wildly used pulp capping material,could affect the proliferation and differentiation of dental pulp cells.The aim of this study is to study the roles of Notch signalling and autophagy in MTA induced human dental pulp cells (hDPCs) proliferation promotions.Methods:Healthy human third molars were collected and hDPCs were isolated by a combined digestion of collagenase Ⅰ and dispase Ⅱ.MTA extracts of different concentrations (0.5,1.0,2.0,5.0,10.0 mg/mL) were used to test the cytotoxicity by cells counting kit (CCK-8) assays and to select the optimum concentration for hDPCs survival..Expressions of Notch1,Hes1,LC3Ⅱ / Ⅰ and p62 in wild type and MTA treated hDPCs were detected by western blotting.Results:MTA extracts in a concentration of 1.0 mg/mL exerted most profoundly promotion effects on the proliferation of hDPCs among all concentrations tested.MTA of high concentration (10 mg/mL) was toxically to cells.Compared with that of wild type hDPCs,the expressions of Notch1 and Hes1(P＜0.05),or p62 and LC3Ⅱ/Ⅰ (P＜0.01) in MTA treated hDPCs were significantly increased.Much lower expression of Notch1 was detected in hDPCs when autophagy was induced by Earle's balanced salt solution (EBSS) starvation for 24 h.Conclusions:MTA could up-regulated hDPCs proliferation with highly relevant in stimulating Notch1-Hes1 signalling and inhibition of autophagy.The study is supposed to provide new insight in unrevealing the mechanisms of MTA mediated dental pulp cells proliferation.
ABSTRACT
The detection method of gene chip based on SPR principle is a potential high-throughput microanalysis method without labelling. With the use of Self-assembled monolayer (SAM) technology, the gene chip of Neisseria gonorrhoeae probe lattice has been prepared, detected and analyzed using the Surface plasmon resonance (SPR) and SPR imaging (SPRI) gene chip detection system here-in provided for research in the hybridizatin reaction on the probe lattice of gene chip. The result indicates that there is an obvious resonance assimilate peak on the SPR resonance curve. And after hybridization, the refractive index and resonance as well as molecular weight of the probe have increased. So whether a hybridization takes place or whether the wanted ingredient is in the sample under examination can be determined by using SPR to watch the detecting interface or the resonance curve. The SPRI detection system is available for observing the happening of a hybridization on the probe of gene chip in real-time and straighforwardly. The SPR and SPRI system can do analysis qualitatively and quantitatively.
Subject(s)
DNA Probes , Genetics , Neisseria gonorrhoeae , Genetics , Oligonucleotide Array Sequence Analysis , Methods , Surface Plasmon Resonance , Methods , Surface PropertiesABSTRACT
Objective To study application of surface plasmon resonance(SIR)system in detection of clinical pathogen with a gene chip.Methods 27 clinical samples were detected by SPR-based gene chip system.These samples were composed by 8 positive blood samples,3 positive pyoid samples,9 positive leucorrhea samples and positive reproductive tract pyoid samples,1 positive biopsy sample and 6 negative biopsy samples.Specific primers and probes for target pathogens were designed by bioinformatics methods and validated by PCR and enzyme-labelled chemiluminescence,respectively.SPR-based gene chip was prepared and utilized to detect clinical samples by SPR system.Results The primers and probes showed good specificity and accuracy,which can be applied to perform PCR and application of the gene chip.Compared with the clinical analysis,gene chip analysis of 26 clinical samples showed the consistent results.Conclusions SPR detection system proved to be accurate and reliable.The chip will have a promising prospect in application.