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Fluid mechanics of the stagnation point flow chamber and its platelet deposition.

The interaction of flow and thrombus generation often is a crucial question for the engineer working in the field of artificial organs. However, this interaction is only incompletely known, and quantitative data under well-defined experimental conditions are especially rare. These can be attained with the stagnation point flow chamber. This flow model applies platelet-rich plasma (PRP) as fluid. Its flow conditions are assessed with the help of computational fluid mechanics. In addition, the concept of the boundary layer is introduced, which permits assessment of the platelet flow along the wall. The results of the experiment indicate that platelets are deposited at a defined shear rate.

Animals↗

Characterization of an artificial valve flow using the numerical dye washout visualization technique: application to the monoleaflet valve with purged flow.

Until today, no ideal heart valve prosthesis for the replacement of a diseased natural valve or for use in ventricular assist devices exists. Valves still cause thromboembolic complications originating from thrombus formations in the valve's stagnant zones. Optimization of valve design involves avoiding stagnation zones and zones of high shear stresses. This requires detailed flow field investigations. Usually, the regions which are more prone to thrombus formation can be estimated using a dye washout experiment. The method allows an assessment of regions with a high or low residence time that may in turn predict regions with a corresponding thrombus risk. This successful experimental method was simulated using numerical methods with a combination of the computational fluid dynamics program FLUENT (Fluent Inc., Lebanon, NH, USA) and of the visualization tool AMIRA (TGS Inc., San Diego, CA, USA). The numerical dye washout visualization was applied to four monoleaflet valves with varying valve housing geometries. The results show a significant difference in the washout processes of the examined valves. The dye washout was characterized by a time course of the gray value averaged over a defined region of interest. Finally, these curves were quantified by a half dye time. The half dye time in the best optimized valve was only 0.2753 s. The same time in the original valve was 0.6834 s. This study shows that the proposed numerical method of dye washout visualization can be used as an additional tool of the flow characterization in artificial organs.

Dye Dilution Technique↗

Historical perspectives of hybrid hepatic assist devices: tissue sourcing, immunoisolation, and clinical trial.

A hybrid hepatic assist device using canine liver tissues was developed and clinically applied 38 years ago. However, for many years practical hybrid hepatic assist devices were not clinically introduced owing to the many difficulties encountered in employing cultured hepatocytes. These problems include: (1) maintenance of viable cultured cells, (2) maintenance of normal hepatocyte function with these cells, (3) elimination of toxic substances generated by non-viable cultured and/or stored cells, (4) elimination of immunological factors generated by cultured cells and by the patient, and (5) difficulties of the biocompatible immunological barrier for cells against the patient. Fortunately, recent progress in apheresis and biomaterial technologies enable us to isolate cultured cells immunologically and yet maintain effective metabolic functions for the patient. These technologies generate an immunological barrier of a hybrid hepatic assist device for the patients. Proper adsorption columns developed for apheresis procedures enable us to remove the toxic substances released by non-viable cells. Recent development of oxygen-carrying macromolecules enable us to provide sufficient oxygen supply to the cultured cells and to maintain their normal cellular function, not only during the cultured period of time, but also during their actual clinical application. Together with the advancement of cell culture technologies, including the proper cultured environments and cellular seeding environments, these technologies, primarily developed for therapeutic apheresis, should be able to provide more effective and safe hybrid artificial organs.

Animals↗

Standards and guidelines for biopolymers in tissue-engineered medical products: ASTM alginate and chitosan standard guides. American Society for Testing and Materials.

The American Society for Testing and Materials (ASTM) is making a concerted effort to establish standards and guidelines for the entire field of tissue-engineered medical products (TEMPS). Safety, consistency, and functionality of biomaterials used as matrices, scaffolds, and immobilizing agents in TEMPS are a concern. Therefore, the ASTM has established a number of task groups to produce standards and guidelines for such biomaterials. Alginate is a naturally occurring biomaterial used for immobilizing living cells to form an artificial organ, such as encapsulated pancreatic islets. In order to aid in successful clinical applications and to help expedite regulatory approval, the alginate used must be fully documented. The ASTM alginate guide gives information on selection of testing methodologies and safety criteria. Critical parameters such as monomer content, molecular weight, and viscosity, in addition to more general parameters, such as dry matter content, heavy metal content, bioburden, and endotoxin content are described in the ASTM document. In a like manner, the characterization parameters for chitosan, a bioadhesive polycationic polysaccharide, are described in a separate guide. For chitosan, the degree of deacetylation is of critical importance. Control of protein content and, hence, potential for hypersensitivity, endotoxin content, and total bioburden are important in chitosan preparations for TEMPS. Together these two guides represent part of the effort on behalf of the ASTM and other interested parties to ensure quality and standardization in TEMPS.

Alginates↗

Biomaterials and scaffolds in reparative medicine.

Most approaches currently pursued or contemplated within the framework of reparative medicine, including cell-based therapies, artificial organs, and engineered living tissues, are dependent on our ability to synthesize or otherwise generate novel materials, fabricate or assemble materials into appropriate 2-D and 3-D forms, and precisely tailor material-related physical and biological properties so as to achieve a desired clinical response. This paper summarizes the scientific and technological opportunities within the fields of biomaterials science and molecular engineering that will likely establish new enabling technologies for cellular and molecular therapies directed at the repair, replacement, or reconstruction of diseased or damaged organs and tissues.

Animals↗

Unique ability of the Proteus mirabilis capsule to enhance mineral growth in infectious urinary calculi.

Struvite (MgNH4PO4.6H2O) calculi are a common complication of Proteus mirabilis urinary tract infections. Although urease is a major virulence factor in calculus formation, the polysaccharide capsule (CPS) of this organism also enhances struvite crystallization and growth in vitro (L. Clapham, R. J. C. McLean, J. C. Nickel, J. Downey, and J. W. Costerton, J. Crystal Growth 104:475-484, 1990). We obtained purified CPS, of known structure and varying anionic character, from P. mirabilis ATCC 49565 and several other organisms. Artificial urine was added to CPS, and the pH was elevated from 5.8 to 8.5 by the addition of urease or titration with 0.25 M NH4OH to induce struvite crystallization. Crystallization was measured by particle counting (Coulter counter), and the morphology (crystal habit) was examined by phase-contrast microscopy. In the presence of partially anionic P. mirabilis CPS, struvite formation occurred at a lower pH than in the absence of CPS or in the presence of other neutral, partially anionic, or anionic CPS. At pH 7.5 to 8.0, significantly more struvite crystals formed in the presence of P. mirabilis CPS than under other experimental conditions. With the exception of one polymer (curdlan) which did not bind Mg2+, enhancement of struvite formation by CPS polymers was inversely proportional to their Mg2+ binding ability. We speculate that the structure and partial anionic nature of P. mirabilis CPS enable it to enhance struvite formation by weakly concentrating Mg2+ ions during struvite crystal formation. This illustrates a new virulence aspect of bacterial CPS during infection.

Bacterial Capsules↗

Pierre M. Galletti: a personal reflection.

Pierre Galletti, my friend and colleague, passed away on March 8, 1997, having left his mark on the emerging field of biomedical engineering. He was a pioneering researcher, making his impact in such fields as heart-lung bypass, artificial organs, and tissue engineering. He was a dedicated teacher and a mentor to many. He not only provided leadership in the establishment of the medical school at Brown University, but also helped start Morehouse School of Medicine in Atlanta. He was an entrepreneur and an individual who realized that ultimately basic science only impacts patient care when new technology is made available to the public. He served the bioengineering community in many ways, later in life becoming active in public policy, and as the second president of the American Institute for Medical and Biological Engineering, more than anyone focused this organization on its public policy role. He was the consummate biomedical engineer, a person of great vision, a man for all seasons.

Animals↗

Cell mechanics and mechanotransduction: pathways, probes, and physiology.

Cells face not only a complex biochemical environment but also a diverse biomechanical environment. How cells respond to variations in mechanical forces is critical in homeostasis and many diseases. The mechanisms by which mechanical forces lead to eventual biochemical and molecular responses remain undefined, and unraveling this mystery will undoubtedly provide new insight into strengthening bone, growing cartilage, improving cardiac contractility, and constructing tissues for artificial organs. In this article we review the physical bases underlying the mechanotransduction process, techniques used to apply controlled mechanical stresses on living cells and tissues to probe mechanotransduction, and some of the important lessons that we are learning from mechanical stimulation of cells with precisely controlled forces.

Cell Physiological Phenomena↗

Microangiopathic changes of subepidermal capillaries in end-stage renal failure.

Skin biopsies were taken from 63 patients with end-stage renal failure (ESRF) and 5 normal volunteers, 53 of whom were on maintenance hemodialysis (HD) and the other 10 without HD. They were examined with light and electron microscopy and subepidermal capillaries were subjected to image analysis. Thickening of the basement membrane (BM) with multilamination of the basal lamina, endothelial activation and chronic inflammatory cell infiltration with participation of mast cells were evident. The extent of these BM changes was correlated with the duration of HD, but not significantly correlated with the clinical findings indicative of arteriosclerosis. There was significant correlation between the duration of HD and mast cell proliferation or eosinophilia. We suspect that the subepidermal capillary changes represent an inflammatory reaction provoked by uremic toxins or by the repeated use of an artificial organ.

Adolescent↗

'Niche Selection' and the evolution of a complex behavior in a changing environment--a simulation.

One of the key problems in theoretical biology is the identification of the mechanisms underlying the evolution of complexity. This paper suggests that some difficulties in current models could be avoided by taking account of"niche selection"as proposed by Waddington [21] and subsequent authors [2]. Computer simulations, in which an evolving population of artificial organisms"selects"the niche(s) that maximize their fitness, are compared with a Control Model in which"Niche Selection"is absent. In the simulations the Niche Selection Model consistently produced a greater number of"fit"organisms than the Control Model; although the Niche Selection Model tended, in general, to produce organisms occupying simple niches, it was nonetheless more effective than the Control Model in producing well-adapted organisms inhabiting complex niches. It is shown that the production of these organisms is critically dependent on the rate of environmental change: Slow change leads to fit but undifferentiated populations, dominated by organisms occupying simple niches; differentiated populations, including well-adapted organisms living in complex niches, require rates of environmental change lying just beyond a mathematically well-defined critical value. In simulation"Niche Selection,"unlike conventional"Natural Selection,"provides a permanent selective bias in favor of simplicity. This tendency is counterbalanced by statistical forces favoring shifts from rare"simple niches"to commoner niches of greater complexity. Fit organisms inhabiting complex niches only emerge in conditions where the rate of environmental change is high enough to avoid the concentration of the population in very simple niches, but slow enough to permit step-by-step adaptation to niches of gradually increasing complexity. This result appears to be robust to changes in simulation parameters and assumptions, and leads to interesting conjectures about the real world behavior of biological organisms (and other complex adaptive systems). It is suggested that some of these conjectures might be relatively easy to test.

Animals↗

A simple common contexts explanation for the development of abstract letter identities.

Abstract letter identities (ALIs) are an early representation in visual word recognition that are specific to written language. They do not reflect visual or phonological features, but rather encode the identities of letters independent of case, font, sound, and so forth. How could the visual system come to develop such a representation? We propose that because many letters look similar regardless of case, font, and other characteristics, these provide common contexts for visually dissimilar uppercase and lowercase forms of other letters (e.g., e between k and y in key and E in the visually similar context K-Y). Assuming that the distribution of words' relative frequencies is comparable in upper- and lowercase (that just as key is more frequent than pew, KEY is more frequent than PEW), these common contexts will also be similarly distributed in the two cases. We show how this statistical regularity could lead Hebbian learning to produce ALIs in a competitive architecture. We present a self-organizing artificial neural network that illustrates this idea and produces ALIs when presented with the most frequent words from a beginning reading corpus, as well as with artificial input.

Form Perception↗

Unconstrained physiological monotoring in daily living for health care.

This paper deals with the recent development of two types of non-invasive physiological monitoring systems for possible application in an unconstrained manner to normal subjects for health care as well as to patients and/or outpatients with disorders or with life support systems (artificial organs and organ transplantations). One is an ambulatory monitoring system which allows automatic acquisition of blood pressure, cardiac output and other cardiovascular hemodynamic parameters on a beat-by-beat basis using the volume-compensation and transthoracic electrical admittance method. The other is a home monitoring system installed in a lavatory which can measure body and excreta weight together with the ballistocardiogram as an index of cardiac ejecting function in an unaware fashion without attachment of any sensors to the subject's body and without special operations for measurement during toilet use. Outlines of these two systems and monitoring results of laboratory and field testings are presented, and these suggest that the ambulatory and non-conscious physiological monitoring techniques described herein appear promising as a valuable and helpful means for use in research as well as in the practical field of health monitoring at home during daily living.

Activities of Daily Living↗

Is the Vroman effect of importance in the interaction of blood with artificial materials?

The successive displacement of plasma proteins adsorbed to artificial surfaces (biomaterials) is well documented, mostly by specially designed experiments that stretch out the effect in time and space. Analysis of displacement has been focused principally on molecular events on the adsorbing surface. In this paper attention is directed rather to the antecedent transport phenomena necessary to deliver successive proteins to a surface. The different limitations on protein arrival fixed by the total quantity present and by the rates of transport of super-sufficient quantities are distinguished. The transport perspective is then used to ask, and partly answer, the question: Can protein displacement be responsible for patterns of thrombus formation and cellular adhesion that are seen on the blood-wetted surfaces of devices found in medical practice: artificial organs and vascular prostheses? Calculations and a small amount of preliminary data suggest that such patterns may form when blood is introduced into these devices, particularly in the neighborhood of boundary shapes that cause separated flows.

Adsorption↗

Sulfonated poly(ethylene oxide)-grafted polyurethane copolymer for biomedical applications.

Sulfonated poly(ethylene oxide) (PEO-SO3)-grafted polyurethane (PU) copolymer (PU-PEO-SO3) was developed for utilization as a material for biomedical applications. Its bulk properties and biocompatibility were investigated and the results were compared to those of PU and PU-PEO (not sulfonated). PU-PEO-SO3 copolymer showed higher water uptake than PU and PU-PEO, indicative of increased hydrophilicity due to the grafting of PEO-SO3. Compared with PU, PU-PEO-SO3 displayed lower glass transition temperature (Tg) and melting endotherm from DSC analysis, suggesting the increase of microphase separation and the suppression of hard-segment packing. PU-PEO-SO3 had practically similar mechanical properties to PU, indicating that the chemical modification of PU was accomplished without reducing its bulk properties. From the results of biocompatibility tests, PU-PEO-SO3 copolymer showed significantly lower adhesion of platelets and S. epidermidis than PU control and PU-PEO. In addition, the platelet factor 4 (PF4) released from platelets was the lowest in PU-PEO-SO3 among the test materials. PU-PEO-SO3 copolymer, which has excellent mechanical properties and biocompatibility, is expected to be useful as coating, molding, and blending materials for artificial organs and medical devices.

Alkanesulfonates↗

Characterization of platelet adhesion under flow using microscopic image sequence analysis.

A method for quantitative analysis of platelet deposition under flow is discussed here. The model system is based upon perfusion of blood platelets over an adhesive substrate immobilized on a glass coverslip acting as the lower surface of a rectangular flow chamber. The perfusion apparatus is mounted onto an inverted microscope equipped with epifluorescent illumination and intensified CCD video camera. Characterization is based on information obtained from a specific image analysis method applied to continuous sequences of microscopical images. Platelet recognition across the sequence of images is based on a time-dependent, bidimensional, gaussian-like pdf. Once a platelet is located,the variation of its position and shape as a function of time (i.e., the platelet history) can be determined. Analyzing the history we can establish if the platelet is moving on the surface, the frequency of this movement and the distance traveled before its resumes the velocity of a non-interacting cell. Therefore, we can determine how long the adhesion would last which is correlated to the resistance of the platelet-substrate bond. This algorithm enables the dynamic quantification of trajectories, as well as residence times, arrest and release frequencies for a high numbers of platelets at the same time. Statistically significant conclusions on platelet-surface interactions can then be obtained. An image analysis tool of this kind can dramatically help the investigation and characterization of the thrombogenic properties of artificial surfaces such as those used in artificial organs and biomedical devices.

Algorithms↗

Hematological aspects of biocompatibility--review article.

The development and improvement of medical devices and artificial organs is critically dependent on the realisation of the importance of the interactions between materials and body tissues. In this regard, the evaluation of biocompatibility assumes paramount importance. This paper reviews the hematological aspects of biocompatibility--the responses evoked by the material as well as the methods for their detection. The paper also mentions a few methods of improvement of material compatibility.

Animals↗

Estimation of shunting, systemic and pulmonary output of the heart, and regional blood flow distribution in unanaesthetized lizards (Varanus exanthematicus) by injection of radioactively labelled microspheres.

Circulatory parameters in a lizard (Varanus exanthematicus) were determined using the microsphere method. Microspheres (MS) (slightly larger than the erythrocytes and labelled with different gamma-emitting isotopes) were injected into a pulmonary vein or the left atrium for determination of the left-to-right (L-R) shunt and the regional distribution of the ventricular systemic output. Injections were also made into the sinus venosus for determination of the right-to-left (R-L) shunt. The relative blood flow was obtained as the ratio of the MS activity found in the various tissues over the total activity injected. Absolute calibration of the method was performed by introduction of an 'artificial organ' into the circulatory system (Hales, 1973). Ventricular systemic output (VSO), in five animals, averaged 121 ml/ (min.kg) and ventricular pulmonary output 119 ml/(min. kg). The value of VSO was significantly higher than those observed in other lizard species. In all experimental animals both R-L as well as L-R shunting of various extent occurred. The reliability of the microsphere method as applied in lizards is discussed and is considered to be relatively accurate even under conditions of incomplete mixing of shunted and unshunted blood in systemic heart output.

Animals↗