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Rapid preparation of fresh frozen tissue-engineered bone sections for histological, histomorphological and histochemical analyses.

Fresh frozen sections are the best materials to assess tissue-engineered bone using cells/ceramic complexes. However, there are a lot of technical difficulties in obtaining serial sections suitable for microscopic examinations. Kawamoto et al. developed a method for the production of fresh frozen sections using new adhesive tape, and showed that sections were very useful for histological and histochemical studies. However, no study reported that the method was useful for tissue-engineered bone from histochemical and histomorphological points of view. This study aimed to determine the efficacy of fresh frozen sectioning for evaluating tissue-engineered bone. We revealed that fresh frozen sections retained the original morphology of tissue-engineered bone, and their biochemical characteristics. Therefore, rapid preparation of fresh frozen sections using adhesive tape is extremely useful for research of tissue-engineered bone, and serial sections can be assessed from both histomorphological and biochemical point of views. It is expected that this method will become a powerful tool in tissue-engineering of hard tissues.

Animals↗

Teaching medical informatics to biomedical engineering students: experiences over 15 years.

The Departments of Biomedical Engineering and Medical Informatics at Linköping University in Sweden were established in 1972-1973. The main purpose was to develop and offer courses in medicine, biomedical engineering and medical informatics to students in electrical engineering and computer science, for a specialization in biomedical engineering and medical informatics. The courses total about 400 hours of scheduled study in the subjects of basic cell biology, basic medicine (terminology, anatomy, physiology), biomedical engineering and medical informatics. Laboratory applications of medical computing are mainly taught in biomedical engineering courses, whereas clinical information systems, knowledge based decision support and computer science aspects are included within the medical informatics courses.

Biomedical Engineering↗

Teaching introductory engineering: a problem based learning experience.

A freshman introductory engineering course is being taught on an experimental basis to approximately 40 cadets per semester at the United States Air Force Academy. The purpose of the course is to better address seven educational outcomes desired in Academy graduates. Problem Based Learning is used to present engineering as a problem solving process that involves a variety of interdisciplinary issues. The setting is an Air Force System Program Office responsible for planning a manned mission to Mars. Students are guided by a mentor to identify the relevant engineering requirements. Traditional instruction is used only to present skills and tools that enhance students' learning. An integrated assessment program is being used to evaluate this instructional approach and how well the educational outcomes are being met. The quantitative assessment findings are inconclusive and indicate further work is needed to develop reliable assessment instruments. Qualitative findings, however, show that the students learned important engineering fundamentals, liked the course, and enjoyed the Mars scenario. They also developed an appreciation for the interdisciplinary nature of engineering, developed confidence in their ability to make decisions for problem-solving, and they acquired self esteem not usually obtained from traditional engineering courses. This paper presents highlights from the three years this course has been offered.

Engineering↗

Oxidative DNA damage and cytogenetic effects in flight engineers exposed to cosmic radiation.

This study set out to analyze biomarkers for genotoxic events, e.g., oxidative DNA damage, chromosomal damage and hprt mutations, among flight personnel, who are known to be occupationally exposed to ionizing radiation of cosmic origin. Twenty-three flight engineers were recruited while ground personnel served as a matched control group. Cumulative radiation doses during flight were calculated on the basis of subjects' flight records assuming an exposure rate of 6 microSv per hour of flight. Oxidative DNA damage in peripheral lymphocytes from flight engineers appeared significantly increased in comparison with controls and was associated with cumulative exposure to cosmic radiation. Frequencies of peripheral lymphocyte chromosome aberrations, micronuclei and hprt mutations appeared also to be increased in flight engineers, but not significantly. It was also observed that DNA damage was higher in flight engineers with a relatively shorter flight history in comparison with flight engineers with higher cumulative exposures to radiation, suggesting adaptation to DNA damage caused by ionizing radiation. DNA repair activities measured as unscheduled DNA synthesis were clearly increased in the higher-exposed subgroup of flight engineers, and appeared significantly correlated with cumulative radiation dose, as well as inversely with oxidative DNA damage. The implications for cancer risk assessment in relation to exposure to cosmic radiation are discussed.

Aerospace Medicine↗

Synthetic biology--putting engineering into biology.

Synthetic biology is interpreted as the engineering-driven building of increasingly complex biological entities for novel applications. Encouraged by progress in the design of artificial gene networks, de novo DNA synthesis and protein engineering, we review the case for this emerging discipline. Key aspects of an engineering approach are purpose-orientation, deep insight into the underlying scientific principles, a hierarchy of abstraction including suitable interfaces between and within the levels of the hierarchy, standardization and the separation of design and fabrication. Synthetic biology investigates possibilities to implement these requirements into the process of engineering biological systems. This is illustrated on the DNA level by the implementation of engineering-inspired artificial operations such as toggle switching, oscillating or production of spatial patterns. On the protein level, the functionally self-contained domain structure of a number of proteins suggests possibilities for essentially Lego-like recombination which can be exploited for reprogramming DNA binding domain specificities or signaling pathways. Alternatively, computational design emerges to rationally reprogram enzyme function. Finally, the increasing facility of de novo DNA synthesis-synthetic biology's system fabrication process-supplies the possibility to implement novel designs for ever more complex systems. Some of these elements have merged to realize the first tangible synthetic biology applications in the area of manufacturing of pharmaceutical compounds.

Animals↗

The use of tissue-engineered skin (Apligraf) to treat a newborn with epidermolysis bullosa.

BACKGROUND: Inherited epidermolysis bullosa (EB) is a mechanobullous disorder. The Dowling-Meara variant, a subtype of EB, is characterized by widespread blister formation that may include the oral cavity and nails. Many patients with the Dowling-Meara phenotype are at increased risk of sepsis and death during infancy. The treatment of EB is generally supportive. The tissue-engineered skin used (Apligraf) is a bilayered human skin equivalent developed from foreskin. It is the only Food and Drug Administration-approved skin equivalent of its kind. It is approved for the treatment of venous ulcers of the lower extremities. It has also been used to treat acute wounds, such as graft donor sites and cancer excision sites. OBSERVATION: To our knowledge, we describe the first case in which a newborn with EB, Dowling-Meara variant, was treated with bilayered tissue-engineered skin. The areas treated with the tissue-engineered skin healed faster than the areas treated with conventional therapy. Most of the areas treated with tissue-engineered skin have remained healed, without developing new blisters. These areas appear to be more resistant to trauma. CONCLUSIONS: Our early success with tissue-engineered skin in this patient may have a significant impact on the future treatment of neonates with EB simplex. Future studies are needed to determine if the beneficial effects of tissue-engineered skin are reproducible in other neonates with EB simplex and in patients of all ages with different subtypes of EB.

Collagen↗

Autogenous tissue-engineered cartilage: evaluation as an implant material.

OBJECTIVES: To determine whether autogenous tissue-engineered cartilage grafts can be synthesized in predetermined shapes, to compare tissue-engineered cartilage with native cartilage with respect to histological characteristics and biomechanical properties, and to demonstrate how multiple transplantations affect tissue-engineered cartilage. DESIGN: Controlled, prospective animal study. SUBJECTS: Twenty New Zealand white rabbits, 3 weeks old. INTERVENTIONS: Autogenous chondrocytes were seeded onto biodegradable polyglycolic acid-poly-L-lactic acid copolymer templates in 1 of 3 shapes (cross, nasal tip graft, or auricle). Grafts and controls of sculpted cartilage were divided among 3 groups: short-term implantation (4 or 8 weeks), long-term implantation (6 or 12 months), and a reimplantation group. The gross morphological features, histological findings, and tensile strength of grafts were assessed. RESULTS: Production of tissue-engineered cartilage was confirmed in 30 of 31 implants. Histological evaluation demonstrated characteristic cartilaginous matrix, but with prominent vascular and fibrous tissue ingrowth. In long-term implantation grafts (n=4), foci of osteoid were evident by 6 months. In the subset of transplanted grafts (n=7), 5 of 7 demonstrated significant loss of cartilage viability. Tensile strength measurements demonstrated values 24% and 41% of those of controls at 4 and 8 weeks, respectively. CONCLUSIONS: Tissue-engineered autogenous cartilage can be reliably produced, and predetermination of graft shape is possible. Histologically, grafts represent composites of mature cartilage infiltrated by vasculature and fibrous tissue, with delayed osteoid formation. Graft viability is compromised by early transplantation, and tensile strength is less than that of native cartilage. These results demonstrate the feasibility of tissue-engineered cartilage as a future graft material.

Animals↗

Tumours of the respiratory tract in rats and hamsters following chronic inhalation of engine exhaust emissions.

The potential carcinogenic effect of inhaled automobile exhaust emissions was examined in rodents. Both rats and hamsters were exposed to the emissions from (1) a gasoline engine, (2) a gasoline engine fitted with a three-way catalytic converter, (3) a diesel engine and (4) a diesel engine with particle filtration. Exposures were for 16 hours per day, 5 days per week, for 2 years. All hamsters were sacrificed at the end of the 2-year exposure period, whereas the rats surviving after 2 years of exposure were maintained for a further 6-month observation period without additional exposure to emissions. Some of the hamsters in each treatment group were pretreated with diethylnitrosamine to induce respiratory tract tumours. No statistically significant changes were seen in the incidence of respiratory tract tumours in emission-exposed hamsters compared to controls. This lack of a treatment-related effect was seen in both the nitrosamine pretreated and the non-pretreated hamsters. There was no increase in the incidence of lung tumours in rats exposed to filtered diesel exhaust or to the exhaust from the gasoline or gasoline-catalyst engines. There was a statistically significant increase in the incidence of lung tumours in rats exposed to diesel engine emissions compared to controls. A clear dose response was evident in both males and females, although the incidence of lung tumours was markedly higher in females (96% in rats surviving beyond 2 years) than in males (44% in rats surviving beyond 2 years). An increased incidence of lung tumours was observed only in rats exposed to mean concentrations of diesel soot particles of either 2200 or 6600 micrograms/m3.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Engineering cells for cell culture bioprocessing--physiological fundamentals.

In the past decade, we have witnessed a tremendous increase in the number of mammalian cell-derived therapeutic proteins with clinical applications. The success of making these life-saving biologics available to the public is partly due to engineering efforts to enhance process efficiency. To further improve productivity, much effort has been devoted to developing metabolically engineered producing cells, which possess characteristics favorable for large-scale bioprocessing. In this article we discuss the fundamental physiological basis for cell engineering. Different facets of cellular mechanisms, including metabolism, protein processing, and the balancing pathways of cell growth and apoptosis, contribute to the complex traits of favorable growth and production characteristics. We present our assessment of the current state of the art by surveying efforts that have already been undertaken in engineering cells for a more robust process. The concept of physiological homeostasis as a key determinant and its implications on cell engineering is emphasized. Integrating the physiological perspective with cell culture engineering will facilitate attainment of dream cells with superlative characteristics.

Animals↗

[Influence of the running conditions of a diesel engine on the mutagenic effects of its emissions].

A direct exposure method to detect mutagenicity of car exhausts was applied to study the influence of the running conditions of a diesel engine functioning on an assay platform. Biological and physico-chemical approaches were carried out together to appreciate the quality of diesel emissions. Exhausts, emitted by a diesel engine (Renault, 2068 cm3) were sampled from a dilution tube connected to a constant volume sampler. Regulated pollutants (CO, NOx, HC and particles) and some non-regulated pollutants (monoaromatic hydrocarbons (MAH) and polyaromatic hydrocarbons (PAH), aldehydes) were analysed when the engine ran at different operating conditions of load (full, 3/4, 2/4 and 1/4) and of engine speed (from 1000 to 4000 rev./min). The mutagenicity (Ames test) of diesel exhaust was assessed by a direct exposure method which does not need extraction of the particles. Bacteria are pre-seeded onto the agar plate and exposed directly to the diesel exhausts parallelly to the measurements of pollutants. The engine speed and the load influence the level, the composition and the induced mutagenicity of the diesel emissions and the results show that: when engine speed increases all the gaz phase indicators increase independently of the load, those of the particulate phase and mutagenicity increase at low load but decrease at full load; when load increase, all the indicators of both phases decrease except NOx which increases.

Carbon Monoxide↗

Evaluation of emission toxicity of urban bus engines: compressed natural gas and comparison with liquid fuels.

Emissions from a spark-ignition (SI) heavy-duty (HD) urban bus engine with a three-way catalyst (TWC), fuelled with compressed natural gas (CNG), were chemically analyzed and tested for genotoxicity. The results were compared with those obtained in a previous study on an equivalent diesel engine, fuelled with diesel oil (D) and a blend of the same with 20% vegetable oil (B20). Experimental procedures were identical, so that emission levels of the CNG engine were exactly comparable to the ones of the diesel engine. The experimental design was focused on carcinogenic compounds and genotoxic activity of exhausts. The results obtained show that the SI CNG engine emissions, with respect to the diesel engine fuelled with D, were nearly 50 times lower for carcinogenic polycyclic aromatic hydrocarbons (PAHs), 20 times lower for formaldehyde, and more than 30 times lower for particulate matter (PM). A 20-30 fold reduction of genotoxic activity was estimated from tests performed. A very high reduction of nitrogen oxides (NO(X)) was also measured. The impact of diesel powered transport on urban air quality, and the potential benefits deriving from the use of CNG for public transport, are discussed.

Air Pollutants↗

Monitoring of arsenic exposure with speciated urinary inorganic arsenic metabolites for ion implanter maintenance engineers.

For wafer fabrication in the semiconductor industry, maintenance engineers are potentially exposed to hazards during their work of disassembling machine components for cleanup. One special concern is the presence of arsenic or arsenic compounds in the working environment. This study analyzed speciated urinary inorganic arsenic metabolites of the maintenance engineers using high-performance liquid chromatography-hydride generation atomic absorption spectrometry to study the potential arsenic exposure during their maintenance work. In total, from six wafer fabrication facilities, 30 maintenance engineers were recruited as the exposed group and another 12 office-based engineers served as the control group. First morning-voided urine samples of each study subject were collected for 7 consecutive days. The levels of total urinary inorganic arsenic metabolites for the exposed group were 1.7+/-1.4, 1.4+/-1.1, 6.2+/-6.7, 20.2+/-14.1, and 29.5+/-17.2 micro g/L for As3+, As5+, monomethylarsonic acid, dimethylarsinic acid, and total inorganic arsenic, respectively. Both the concentration of monomethylarsonic acid and its percentage in total urinary inorganic arsenic metabolites showed significantly ascending trends for the control group, for the engineers without preventative maintenance work prior to their urine sampling, and for the engineers with such work prior to their urine sampling (P<0.05 and P<0.0005, respectively). The data also suggested that, at low-level occupational arsenic exposure, the concentration of total urinary inorganic arsenic metabolites might be misleading due to the confounding effect resulting from intake of seafood, such as arsenosuger. Nevertheless, monitoring of urinary arsenic species by using the percentage change of monomethylarsonic acid in total urinary inorganic arsenic metabolites as an indicator for the verification of arsenic exposure is helpful and appropriate in such cases.

Adult↗

Future perspectives in reconstructive surgery using tissue engineering.

Whenever there is a lack of native urologic tissue, reconstruction usually is performed with native nonurologic tissues, such as gastrointestinal segments, skin, or mucosa from multiple body sites. The use of native nonurologic tissues in the genitourinary tract is associated with adverse effects. Tissue engineering efforts currently are underway for almost every type of tissue and organ within the urinary system including bladder, ureter, urethra, and genitalia. Most of the efforts expended to engineer genitourinary tissues have occurred within the last decade. Tissue engineering techniques require a cell culture facility designed for human application. Personnel who have mastered the techniques of cell harvest, culture and expansion, and polymer design are essential for the successful application of this technology. The first human application of cell-based tissue engineering technology for urologic applications recently occurred with the injection of autologous cells for the correction of vesicoureteral reflux in children and urinary incontinence in adults. Trials involving bladder replacement using tissue engineering techniques currently are being arranged. Recent progress suggests that engineered urologic tissues may have clinical applicability.

Adult↗

Biomaterial developments for bone tissue engineering.

The development of bone tissue engineering is directly related to changes in materials technology. While the inclusion of materials requirements is standard in the design process of engineered bone substitutes, it is also critical to incorporate clinical requirements in order to engineer a clinically relevant device. This review presents the clinical need for bone tissue-engineered alternatives to the present materials used in bone grafting techniques, a status report on clinically available bone tissue-engineering devices, and recent advances in biomaterials research. The discussion of ongoing research includes the current state of osseoactive factors and the delivery of these factors using bioceramics and absorbable biopolymers. Suggestions are also presented as to the desirable design features that would make an engineered device clinically effective.

Biocompatible Materials↗

Exhaust emissions from engines of the Detroit Diesel Corporation in transit buses: a decade of trends.

In the U.S.A., exhaust emissions from city buses fueled by diesel are not characterized well because current emission standards require engine tests rather than tests of whole vehicles. Two transportable chassis dynamometer laboratories developed and operated by West Virginia University (WVU) have been used extensively to gather realistic emission data from heavy-duty vehicles, including buses, tested in simulated driving conditions. A subset of these data has been utilized for a comprehensive introspection into the trends of regulated emissions from transit buses over the last 7 years, which has been prompted by continuously tightening restrictions on one hand, along with remarkable technological progress, on the other hand. Two widely used models of diesel engines manufactured by the Detroit Diesel Corporation (DDC) have been selected as a case-study for such an overview, based on full-scale, on-site testing of actual city buses, driven in accordance with the SAE J1376 standard of a Commercial Business District (CBD) cycle. The results provide solid, quantitative evidence that most regulated emissions from engines produced by DDC have declined over the years, especially with the transition from the 6V-92TA to the Series 50 models. This improvement is remarkable mainly for the emissions of particulate matter (PM), that are lower by over 70%, on average, for the Series 50 engines, though the emissions of nitrogen oxides (NOx) exhibit a reversed trend, showing a degradation of about 6%, on average, with the transition from 6V-92TA to the Series 50 engines. The expected trend of decreasing emission levels with the model year of the engine is clear and consistent for particulate matter (PM), hydrocarbons (HC), carbon monoxide (CO), and nitrogen oxides (NOx), starting with the 1990 models, although it is not conclusive for carbon dioxide (CO2) emissions.

Air Pollution↗

Brassica carinata as an alternative oil crop for the production of biodiesel in Italy: engine performance and regulated and unregulated exhaust emissions.

A comparison of the performance of Brassica carinata oil-derived biodiesel with a commercial rapeseed oil-derived biodiesel and petroleum diesel fuel is discussed as regards engine performance and regulated and unregulated exhaust emissions. B. carinata is an oil crop that can be cultivated in coastal areas of central-southern Italy, where it is more difficult to achieve the productivity potentials of Brassica napus (by far the most common rapeseed cultivated in continental Europe). Experimental tests were carried out on a turbocharged direct injection passenger car diesel engine fueled with 100% biodiesel. The unregulated exhaust emissions were characterized by determining the SOOT and soluble organic fraction content in the particulate matter, together with analysis of the content and speciation of polycyclic aromatic hydrocarbons, some of which are potentially carcinogenic, and of carbonyl compounds (aldehydes, ketones) that act as ozone precursors. B. carinata and commercial biodiesel behaved similarly as far as engine performance and regulated and unregulated emissions were concerned. When compared with petroleum diesel fuel, the engine test bench analysis did not show any appreciable variation of output engine torque values, while there was a significant difference in specific fuel consumption data at the lowest loads for the biofuels and petroleum diesel fuel. The biofuels were observed to produce higher levels of NOx concentrations and lower levels of PM with respect to the diesel fuel. The engine heat release analysis conducted shows that there is a potential for increased thermal NOx generation when firing biodiesel with no prior modification to the injection timing. It seems that, for both the biofuels, this behavior is caused by an advanced combustion evolution, which is particularly apparent at the higher loads. When compared with petroleum diesel fuel, biodiesel emissions contain less SOOT, and a greater fraction of the particulate was soluble. The analysis and speciation of the soluble organic fraction of biodiesel particulate suggest that the carcinogenic potential of the biodiesel emissions is probably lower than that of petroleum diesel. Its better adaptivity and productivity in clay and sandy-type soils and in semiarid temperate climate and the fact that the performance of its derived biodiesel is quite similar to commercial biodiesel make B. carinata a promising oil crop that could offer the possibility of exploiting the Mediterranean marginal areas for energetic purposes.

Air Pollution↗

Diesel engine exhaust emission: oxidative behavior and microstructure of black smoke soot particulate.

Soot particulate collected from a Euro III heavy duty diesel engine run under black smoke conditions was investigated using thermogravimetry, transmission electron microscopy, electron energy loss spectroscopy, and X-ray photoelectron spectroscopy. The characterization results are compared with those of commercial carbon black. The onset temperature toward oxidation of the diesel engine soot in 5% O2 is 150 degrees C lower than that for carbon black. The burn out temperature for the diesel engine soot is 60 degrees C lower than that of the carbon black. The soot primary particles exhibit a core-shell structure. The shell of the soot particles consists of homogeneously stacked basic structure units. The commercial carbon lamp black is more graphitized than the diesel engine soot, whereas the diesel engine soot contains more carbon in aromatic nature than the carbon black and is highly surface-functionalized. Our findings reveal that technical carbon black is not a suitable model for the chemistry of the diesel engine soot.

Air Pollutants↗

[Tissue engineering: artificial tissue replacement containing vital components].

Tissue engineering as a new field of research has gained increasing importance in recent years. The interdisciplinary field combines, biomaterials cell biology, and cell culture bio-engineering technology. The main focus of tissue engineering is the synthesis of artificial constructs or tissues based on vital cells or cell matrix. Biomaterials provide a three-dimensional structure to shape or guide tissue development. Isolated cartilage cells from a patient can form new tissues when suspended in non-woven resorbable polymers for reconstructive surgery. To achieve sufficient amounts of autologous cells for transplant formation, cells from biopsies have to be multiplied in monolayer culture. Dedifferentiated and undifferentiated mesenchymal cells may be used for bone and cartilage engineering. High cell densities in three-dimensional cultures require perfusion techniques to stabilize culture conditions. Morphogenetic factors such as BMP (bone morphogenetic protein) are thought to play a key role in inducing and controlling phenotypic tissue formation. In conclusion, modern in vitro approaches open new avenues for the development of vital tissue replacements for the clinic. Tissues can be repaired with the patient's own cells eventually leaving no residual artificial materials. Tissue engineering further provides new approaches for in vitro models of the extracellular matrix or diseases which mainly affect this matrix such as rheumatoid arthritis or osteoarthritis. This article describes recent developments in connective tissue engineering and discusses the potential for human tissue repair and reconstructive surgery.

Animals↗