Impact of Implant Surface Micropatterns on Epithelial Cell Behavior.

PURPOSE: This study investigated the effect of topography on cell behavior by screening polydimethylsiloxane (PDMS) molds with different nanoscale micropatterns to determine the ideal surface characteristics for attachment of human epithelial cells. MATERIALS AND METHODS: A soft PDMS mold with regular dot arrays was fabricated based on an aluminum oxide template with ordered nanotube arrays and used as a substrate for cell culture. Cell proliferation, spread, and morphology, as well as features of the extracellular matrix and the actin cytoskeleton were assessed. DISCUSSION: Cells grown on 100-nm regular dot arrays had the highest proliferation rate and spread, with the longest pseudopodia; they showed robust actin distribution relative to the control group. CONCLUSION: Three-dimensional PDMS microstructures with 100 nm regular dot arrays were the most effective surface for epithelial cell attachment. These findings can aid in the manufacture of superior materials for use in implants to better integrate into recipient tissue.

Synthetic symbiotic bacteria reduces the toxicity of mercury ingested via contaminated food.

Mercury contamination in food poses a significant threat to human health. In this article, we propose a novel approach to solve this problem by enhancing the function of gut microbiota against mercury using a synthetically engineered bacterial strain. An engineered Escherichia coli biosensor MerR with mercury binding function was introduced into the intestines of mice for colonization, whereafter the mice were challenged with oral mercury. Compared with the control mice and mice colonized with unengineered Escherichia coli, the mice with biosensor MerR cells in their gut showed significantly stronger mercury resistance. Furthermore, mercury distribution analysis revealed that biosensor MerR cells promoted the excretion of oral mercury with feces, thereby blocking the entry of mercury into the mice, decreasing the concentration of mercury in the circulatory system and organs, and, thus, attenuating the toxicity of mercury to the liver, kidneys and intestines. Colonization with the biosensor MerR did not result in significant health problems in the mice, nor were genetic circuit mutations or lateral transfers identified during the experiments, thus demonstrating the safety of this approach. This study elucidates the remarkable promise of synthetic biology for modulating gut microbiota function.

A gas reporting whole-cell microbial biosensor system for rapid on-site detection of mercury contamination in soils.

As optical reporting elements, fluorescent proteins are extensively used in whole-cell microbial biosensors. However, the use of these optical reporters is limited in opaque media such as soil. This study described a method utilizing gas as a reporting signal that could be used for the rapid on-site detection of mercury in soil. In this biosensor, the MerR protein could capture mercury ions and then bind the promoter of the efe gene to initiate the synthesis of the ethylene (C2H4)-forming enzyme that produced the gas. The research showed that the mercury ion concentrations could be converted into C2H4 gas signals, which were quantified using a handheld C2H4 sensor. By optimizing the biosensor to improve its anti-interference ability in the system, it could detect mercury ion concentrations in the soil ranging from 0.2 to 20 mg/kg within 45 min, effectively reflecting whether the mercury pollution in the soil exceeded the limit standard. This study provides a simple, inexpensive, and portable method for the on-site detection of soil pollutants.

A test strip platform based on a whole-cell microbial biosensor for simultaneous on-site detection of total inorganic mercury pollutants in cosmetics without the need for predigestion.

Mercury pollutants such as mercuric chloride (HgCl2), mercurous chloride (Hg2Cl2) and mercuric ammonium chloride (Hg(NH2)Cl) are often found in cosmetics. Previous attempts at the on-site detection of mercury were hindered by the complicated and dangerous pretreatment procedure of converting various forms of mercury to Hg (II) ions. In this study, a test strip platform was developed based on a whole-cell microbial biosensor for the simultaneous detection of soluble and insoluble inorganic mercury pollutants in cosmetics without the need for predigestion. The genetic circuits with constitutively expressed MerR as sensor proteins and inducible red fluorescent protein (RFP) as the reporter were introduced into Escherichia coli to construct the mercury detection biosensor. The RFP fluorescence intensity of this biosensor showed a excellent linear relationship (R2 = 0.9848) with the Hg (II) concentration ranging from 50 nM to 10 µM in Luria-Bertani (LB) broth. Further research indicated that this biosensor could respond not only to Hg (II) ions but also to insoluble Hg2Cl2 and Hg2Cl2. The transcriptomic results confirmed the mercury conversion ability of the whole-cell biosensor from a gene expression perspective. This biosensor was embedded on filter paper to form a test strip, which could be used to determine whether the total inorganic mercury pollutants in cosmetics exceeded 1 mg/kg. Therefore, this strip provided a low cost, easy-to-use, and instrument-independent method for the detection of mercury pollution in cosmetics, while this study revealed the unique advantages of microbial biosensors in the automatic bioconversion of targets.

Using the promoters of MerR family proteins as "rheostats" to engineer whole-cell heavy metal biosensors with adjustable sensitivity.

BACKGROUND: Whole cell biosensors provide a simple method for the detection of heavy metals. However, previous designs of them rely primarily on simulation of heavy metal resistance systems of bacteria. RESULTS: This study proposes a strategy for the rational design of metal detection circuits based on sensor proteins of the MerR family. Our results indicate the expression level of sensor protein can be used as a "rheostat" for tuning detection sensitivity with parabola curves to represent the relationships between the detection slopes and the sensor protein levels. This circuits design strategy (named as "Parabola Principle"), is used as a guide for the discovery of optimum metal detection circuits, and the design of biosensors with specific metal detection characteristics. For example, visible qualitative Hg (II) biosensors with a threshold of 0.05 mg/L are successfully constructed. CONCLUSIONS: These results indicate the feasibility of developing a sensor that is much more tunable than what is presented.

Feedback regulation mode of gene circuits directly affects the detection range and sensitivity of lead and mercury microbial biosensors.

Whole cell biosensors offer high potential for the detection of heavy metals in a manner that is simple, rapid and low-cost. However, previous researchers have paid little attention to the impacts of construction models on the performance of these biosensors, thereby limiting the achievement of rational design and the optimization of detection characteristics. Herein, for the first time, three basic models of lead and mercury detection circuits, namely feedback coupled, uncoupled and semi-coupled models, have been constructed and compared to explore the effects of uncoupling the topology of sensing circuits on the reporter signals. The results demonstrated that the uncoupled model had better sensitivity for both lead (50 nM) and mercury (1 nM), while the feedback coupled circuits had a wider detection range for mercury (10 nM - 7.5 µM). Introducing the semi-coupled model into the comparison revealed that both the type and location of promoters for regulatory protein genes were key factors for sensitivity. Moreover, the detection characteristics of the uncoupled biosensors were robust, as conditions such as induction time, the concentration of microbial cells, and the concentration of antibiotics had little interference on the performance of the microbial biosensors. This study also established a novel and simple pre-treatment method for sample detection by biosensors. When the uncoupled microbial biosensor was put into practice, the concentration levels of mercury in milk and lead in sewage were determined quickly and accurately. Our study, therefore, provides a strategy for the rational design of whole cell heavy metal biosensors and has developed the potential of their application.

[Influence of MgO and TiO2 on mechanical properties of zirconia toughened alumina ceramics formed by gel-casting technique].

OBJECTIVE: The objective of this study is to investigate the influence of mechanical properties and sintering performance by adding 5% weight percentage aids to nano-compound zirconia toughened alumina (ZTA) ceramics. METHODS: Micrometer Al2O3 and nanometer ZrO2 (quality ratio 4:1) were used to get 55% volume percentage slurry. Magnesium oxide and titanium oxide were taken as aids which were 5% weight percentage of the Al2O3 and ZrO2 powder. Five groups (number 0, 1, 2, 3, 4 group) were divided according to different proportion of aids. After gel-casting, the porcelain pieces were sintered at 1150, 1200, 1300, 1400, 1450, 1500, 1600 degrees C for 2 hours. Static three-point flexure strength, line shrinkage, relative density were measured and scanning electron microscopy (SEM) was used to observe section. RESULTS: Number 1 (MgO 1%, TiO2 4%) group had the highest bending strength. It was (401.78+/-19.50) MPa after sintering at 1600 degrees C for 2 hours and was higher than 0 group (380.64+/-44.50) MPa. Bending strength became lower than 0 group when MgO was more than 2% or more than that weight percentage of ZTA powder. When MgO content was higher than 2% or more than that weight percentage, there was no difference in relative density raising rate between each sintering assistants groups. When the sintering temperature was higher than 1200 degrees C, all groups showed obvious line-shrinkage and the groups which contained sintering assistants were all was higher than 0 group. CONCLUSION: Adding MgO and TiO2 aids from 1% to 4% weight percentage of ZTA will promote fritting and increase ZTA nano-compound ceramics mechanical properties. Adding 2% MgO aids or more than that weight percent will has no obvious help to increase the relative density raising rate of ZTA nano-compound ceramics and will degrade the mechanical properties of ZTA nano-compound ceramics.