The factors that influence magnesium(Mg)corrosion in vitro are systematically evaluated from a review of the relevant literature. We analysed the influence of the following factors on Mg biocorrosion in vitro:(i)...The factors that influence magnesium(Mg)corrosion in vitro are systematically evaluated from a review of the relevant literature. We analysed the influence of the following factors on Mg biocorrosion in vitro:(i) inorganic ions,including both anions and cations,(ii) organic components such as proteins, amino acids and vitamins, and(iii) experimental parameters such as temperature, p H, buffer system and flow rate. Considerations and recommendations towards a standardised approach to in vitro biocorrosion testing are given. Several potential simulated body fluids are recommended. Implementing a standardised approach to experimental parameters has the potential to significantly reduce variability between in vitro biocorrosion tests, and to help build towards a methodology that accurately and consistently mimics in vivo corrosion. However, there are also knowledge gaps with regard to how best to characterise the in vivo environment and corrosion mechanism. The assumption that blood plasma is the correct bodily fluid upon which to base in vitro methodologies is examined, and factors that influence the corrosion mechanism in vivo, such as specimen encapsulation, bear consideration for further studies.展开更多
Objective: To construct the multi probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD 2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells. Methods: The...Objective: To construct the multi probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD 2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells. Methods: The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method. Results: The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD 2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1 3 hours after 100 ng/ml LPS stimulation. Conclusions: These new RPA multi probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD 2 mRNAs in both constitutive and inducible types.展开更多
基金supported by the Australian Federal Government through an Australian Government Research Training Program Scholarshipsupport of the Australian Research Council (ARC) (DP170102557 "Biodegradable magnesium alloy scaffolds for bone tissue engineering")support of the ARC Research Hub for Advanced Manufacturing of Medical Devices
文摘The factors that influence magnesium(Mg)corrosion in vitro are systematically evaluated from a review of the relevant literature. We analysed the influence of the following factors on Mg biocorrosion in vitro:(i) inorganic ions,including both anions and cations,(ii) organic components such as proteins, amino acids and vitamins, and(iii) experimental parameters such as temperature, p H, buffer system and flow rate. Considerations and recommendations towards a standardised approach to in vitro biocorrosion testing are given. Several potential simulated body fluids are recommended. Implementing a standardised approach to experimental parameters has the potential to significantly reduce variability between in vitro biocorrosion tests, and to help build towards a methodology that accurately and consistently mimics in vivo corrosion. However, there are also knowledge gaps with regard to how best to characterise the in vivo environment and corrosion mechanism. The assumption that blood plasma is the correct bodily fluid upon which to base in vitro methodologies is examined, and factors that influence the corrosion mechanism in vivo, such as specimen encapsulation, bear consideration for further studies.
文摘Objective: To construct the multi probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD 2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells. Methods: The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method. Results: The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD 2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1 3 hours after 100 ng/ml LPS stimulation. Conclusions: These new RPA multi probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD 2 mRNAs in both constitutive and inducible types.