Biomedical Engineering and Bioengineering Commons^™

Cell Adhesion To Crystal Surfaces: A Model For Initial Stages In The Attachment Of Cells To Solid Substrates, D. Hanein, B. Geiger, L. Addadi

Cells and Materials

This study addresses the mechanism of the chirally-restricted, ROD-independent adhesion of A6 epithelial cells to the {011} faces of calcium {R,R)-tartrate tetrahydrate crystals. The extensive and rapid adhesion of the cells to these surfaces, in the presence or absence of serum proteins, is distinctly different from the extracellular matrix-mediated adhesion to conventional tissue culture surfaces or to the {101} faces of the same crystals. The differences are manifested by insensitivity to ATP depletion, to disruption of microfilaments and microtubules and even to formaldehyde fixation of the cells. Furthermore, trypsin pretreatment does not affect cell attachment to the {011} faces, nor …

Room Temperature Aging Of Pd-Cu-Ga Dental Alloys, William A. Brantley, Zhuo Cai, Qiang Wu, Alan B. Carr, John C. Mitchell

Cells and Materials

Specimens of three Pd-Cu-Ga dental alloys cast five years ago and subsequently stored at room temperature were reexamined and observed to have lower amounts of the eutectic constituents in the near-surface region than originally present, along with other microstructural changes. This previously unreported room temperature aging behavior of these alloys is attributed to the presence of high-diffusivity paths in the non-equilibrium ascast eutectic structures and to the essential role of the surface for the vacancy diffusion mechanism. These results may have important clinical significance for the ill vivo corrosion resistance and long-term biocompatibility of the Pd-Cu-Ga alloys.

Biomaterials Associated Infections: The Scale Of The Problem, David J. Stickler, Robert J. C. Mclean

Cells and Materials

The biomaterials used in the manufacture of implanted prosthetic devices profoundly impair the host's ability to opsonise and phagocytose invading microbes. As a result, while these devices generally provide effective relief from painful, crippling and life-threatening disorders, they can also induce vulnerability to infection in the recipients. The surfaces of the implants are susceptible to colonisation by microbial biofilms. The cells in the biofilm.s are further protected against opsonophagocytosis and are also resistant to antibacterials. Device associated infections thus tend to be refractile to antibiotic therapy and in many cases the device has to be removed before the infection will …

Synthetic Hydrogel As An Artificial Vitreous Body. A One-Year Animal Study Of Its Effects On The Retina, T. V. Chirila, I. J. Constable, Y. Hong, S. Vijayasekaran, M. F. Humphrey, P. D. Dalton, S. G. Tahija, M. A. L. Maley, M. J. H. Cuypers, C. Sharp, S. R. Moore, M. J. Davies

Cells and Materials

A hydrogel with a high water content was assessed in vitro and in vivo as a possible vitreous substitute. From a large series of polymers produced by the aqueous polymerization of methyl acrylamidoglycolate methyl ether (MAGME), a gel synthesized in 80% water was selected for an animal study. The gel was injected intravitreally into rabbit eyes and followed clinically by ophthalmoscopy, tonometry, and fundus photography. The gel was clinically well tolerated, but after 6 months ophthalmoscopy revealed progressive pallor of the optic nerve head. The eyes were enucleated one year after injection of polymer. Histopathological examination by light microscopy of …

Three-Dimensional Morphology And Platelet Adhesion On Pyrolytic Carbon Heart Valve Materials, S. L. Goodman, K. S. Tweden, R. M. Albrecht

Cells and Materials

Low-temperature isotropic pyrolytic carbon (LTIC) is the preferred material for mechanical heart valve prosthetics due to its durability and good thromboresistance, although thromboembolic complications remain a significant clinical problem. LTlC morphology has been previously studied using scanning (SEM) and transmission electron microscopy (TEM), and scanning tunneling microscopy (STM). However, these microscopies have limitations with imaging rough surfaces. In this study, LTIC valve leaflets from CarboMedics, Inc. and St. Jude Medical, Inc. were prepared and polished exactly as used in clinical prosthetics, and examined at magnifications up to macromolecular resolution using stereo-pair low-voltage SEM (LV -SEM). LV -SEM reveals that LTIC …

A Morphological Assessment Of Bovine Chondrocytes Cultured On Poly(Ethyl Methacrylate)/Tetrahydrofurfuryl Methacrylate, R. M. Sawtell, M. V. Kayser, S. Downes

Cells and Materials

A heterocyclic methacrylate polymer system, PEMA/THFMA, has shown potential as a biomaterial for cartilage repair in a rabbit model and has properties making it suitable for use clinically. The ability of the polymer system, PEMA/THFMA, and a related system, PEMA/HEMA, to support chondrocytes in vitro was assessed by scanning electron microscopy. Chondrocytes adhered to the surface of the PEMA/THFMA by day one, having a rounded morphology and clustered appearance opposed to the Thermanox® control, where the cells had spread out and become fibroblastic in appearance. The chondrocytes divided rapidly on the PEMA/ THFMA system by day three and had completely …

Adsorption Of Bovine Serum Albumin Onto Octacalcium Phosphate And Its Hydrolyzates, O. Suzuki, H. Yagishita, M. Yamazaki, T. Aoba

Cells and Materials

Octacalcium phosphate (OCP) has been advocated to be a precursor of biomineral s. Tn this report , we aimed at investigating OCP hydrolysis and the interaction of OCP and its hydrolyzates (apatitic products) with bovine serum albumin (BSA). A batch of synthetic OCP was allowed to hydrolyze either (a) in deionized water at 70°C or (b) in Tris buffer (pH 7.4) containing 2 ppm fluoride at 37°C. OCP hydrolysis was completed within reasonable experimental periods: after 48 hours at 70°C and after 10 days at 37°C in the presence of fluoride. The adsorption isotherms of BSA onto the original OCP …

Adsorption Of Proteins At Solid-Liquid Interfaces, Willem Norde

Cells and Materials

This paper concisely reviews the general principles underlying protein adsorption from aqueous solution onto a solid surface. The discussion includes the various stages of the adsorption process, i. e., transport of the protein molecules towards the surface, the absorbed amount under equilibrium conditions, desorption and re-adsorption. Among the interactions that determine the overall protein adsorption process (1 ) redistribution of charged groups in the interfacial layer, (2) changes in the hydration of the sorbent and the protein surface, and (3) structural rearrangements in the protein molecule play major roles. Special attention is given to the relation between the structural stability …

Reliability Of Human Fresh And Frozen Gingiva Explant Culture In Assessing Dental Materials Cytocompatibility, R. Warocquier-Clerout, G. C. Hachom-Nitcheu, M. F. Sigot-Luizard

Cells and Materials

An explant culture of human gingival epithelium has been set up in order to provide a valuable test for evaluating the cytocompatibility of dental material s. In an attempt to supply a bank of gingiva ex plants, frozen and freshly excised specimens were cultured in parallel. Optical and scanning electron microscopy showed an early release of cuboidal cells forming a dense layer around the explants. Afterwards, cultures evolved differently . Spread cells grew and migrated more rap idly in fresh than in frozen explant cultures but their adhesion to substratum increased earlier in frozen ones. Epithelial phenotype of cells had …

Biphasic Sodium Fluoride Effects On Bone And Bone Mineral: A Review, P. -T. Cheng, S. M. Bader, M. D. Grynpas

Cells and Materials

This paper reviews the clinical and experimental findings on the effects of sodium fluoride (NaF) on human and animal bone. NaF has been shown to cause a significant increase in axial skeletal bone mass. However, there is concern that the new bone may not provide the desired increase in bone strength. Yet, NaF remains the most commonly used agent capable of stimulating bone formation in most patients (30% non-responders). But whether NaF reduces vertebral fracture rate (VFR) remains controversial. For a given treatment duration, the effect of F on bone quality appears to depend on dose: there is a marked …

Natural History Of Bone Response To Hydroxyapatite-Coated Hip Prostheses Implanted In Humans, P. Frayssinet, D. Hardy, J. S. Hanker, B. L. Giammara

Cells and Materials

A series of 15 autopsied femurs containing hydroxyapatite- coated (HA-coated) prostheses was analysed histologically. Their implantation time ranged from 5 days up to 3 years. The coating thickness of some prostheses and the percentage of the coating in contact with bone at different levels were evaluated using an image analysis device. After the newly formed bone tissue had became mature, several bone morphotypes were identified at the coating contact. From the proximal to the distal part of the prosthesis, bone morphotype was denser and the percentage of the coating surface in contact with bone increased. Several stages in the prosthesis …

Optical Bone Chambers As Tools For Studying The Bone-Implant Interface: A Review, Howard Winet

Cells and Materials

Bone chamber intravital microscopy combines the control volume of in vitro models and the chemical complexity of in vivo models to optimize the study of bone physiology in situ. As an optical tool it provides a window to dynamic events at the tissue level of magnification. In particular, it allows measures of microvascular events: (1) in space by magnifying local perfused vasculature and microcirculation at any instant, and (2) in time by providing the same volume of tissue for weekly viewing of an evolving process such as bone healing. This quartz-bearing titanium screw has revealed a consistent order for …

Co-Culture Of Endothelial Cells And Smooth Muscle Cells In A Flow Environment: An Improved Culture Model Of The Vascular Wall?, Thierry Ziegler, Keith A. Robinson, R. Wayne Alexander, Robert M. Nerem

Cells and Materials

Numerous studies have demonstrated that the neighboring smooth muscle cells (SMC) influence the morphology, cytoskeleton and growth of co-cultured endothelial cells (EC). Also, flow-induced laminar shear stress has been shown to induce cell elongation, F-actin reorganization and growth inhibition in cultured EC. We investigated the effect of neighboring SMC and collagen matrix on the response of EC to shear stress. The co-culture system was made by growing porcine aortic SMC in a gel of collagen type I and then seeding porcine aortic EC (P AEC) on the top surface. Then the co-culture was exposed to steady, laminar shear stress of …

Bone Absorptiometry In Metabolic Bone Disease: Baseline Values And Long-Term Treatment With Calcitriol (Post-Menopausal Osteoporosis Versus Osteomalacia), A. Caniggia, B. Frediani

Cells and Materials

Total body bone absorptiometry reveals low mineral density in both postmenopausal osteoporosis and osteomalacia patients. The method was used to investigate the effect of calcitriol administration on patients suffering from one of these conditions. In osteomalacia, the administration of calcitriol resulted in a dramatic improvement in bone mineral density (sometimes up to 50% in 12 months), indicating the rapid mineralization of previously uncalcified bone tissue as a result of the normalization of the Ca x P product. In osteoporosis a similar treatment was seen to halt the progressive decrease in bone mineral levels and sometimes resulted in minor increases in …

Scanning Electron Microscopy Studies Of Ceramic Layers And Interfacial Regions For Calcium Phosphate-Coated Titanium Dental Implants, W. A. Brantley, E. Tufecki, J. C. Mitchell, D. W. Foreman, E. A. Mcglumphy

Cells and Materials

The surface and cross-section characteristics of the plasma-sprayed calcium phosphate coatings, along with the microstructures and elemental compositions near the titanium alloy interface, were investigated by scanning electron microscopy for two different dental implants of proprietary compositions: Integra!® (Calcitek) and BioVent ® (Dentsply). Elemental concentrations (Ca, P, Ti, AI, and V) near the interfaces were obtained by X-ray energy-dispersive spectroscopy. Coating surfaces exhibited a splat deposition topography of greatly differing microstructural scale for the two implants, along with other features characteristic of the plasma-spraying process; cross-sections of the coatings revealed minimal porosity. Some interdiffusion of principally titanium and calcium was …

Some Biological And Physical Properties Of Laser Deposited Hydroxyapatite Based Films, Tatjana Dostalova, Lucie Himmlova, Miroslav Jelinek, Jirina Bartova

Cells and Materials

The preliminary results of biological and physical tests of hydroxyapatite thin films deposited on dental implants by a new technology with the KrF excimer laser ablation method were evaluated. Biological and physical properties were studied and analyzed by the lymphocyte proliferation test and scanning electron microscopy (SEM), X-ray analysis, Rutherford backscattering analysis (RBS) and particle induced X-ray emission (PIXE) methods. Ten bioceramic films from 45 samples had very good physical and biological properties. Creation of hydroxyapatite thin films with laser ablation can have a positive effect on adhesion of the film and protection for corrosion.

Some Observations On The Structure Of Encrusting Biofilms Of Proteus Mirabilis On Urethral Catheters, C. Winters, D. J. Stickler, N. S. Howe, T. J. Williams, N. Wilkinson, C. J. Buckley

Cells and Materials

A physical model of the bladder has been used to study the process of urinary catheter encrustation. Techniques have been devised for the preparation of sections through these encrustations in situ on the luminal surfaces of catheters and for mapping the distribution of calcium and magnesium in the biofilms. Transmission electron microscopy on these sections showed struvite-like crystals lying in the matrix and in direct contact with the catheter surface. Calcium phosphate ("bioapatite") was distributed throughout the film and many of these amorphous particles appear to have cells at their cores. Freeze-substituted sections of biofilms also showed electron dense materials …

Heating Of Calcium Phosphate Crystals: Morphological Consequences And Biological Implications, W. Bohne, J. A. Pouezat, L. Peru, G. Daculsi

Cells and Materials

Sintering hydroxyapatite (HA) and ß-tricalcium phosphate (ß-TCP) affects the chemical composition, the crystallinity, and the morphological features as demonstrated by means of X-ray diffraction (XRD) , infrared spectroscopy (IR), and scanning electron microscopy (SEM). When heated to 1230°C, 16.7% of HA had decomposed to ß-TCP. SEM investigations showed homogeneous, sharp angular polyhedric blocks of 30 to 50 µm with rare surface pores. On heating at 1230°C, ß-TCP had entirely transformed to a-TCP. During sintering, the size of the powder grains increased and progressive bridging between the grains was observed. At 1230°C, a network within round-shaped polyhedric blocks of 50 to …

Bone Growth On Sol-Gel Calcium Phosphate Thin Films In Vitro, Q. Qiu, P. Vincent, B. Lowenberg, M. Sayer, J. E. Davies

Cells and Materials

Thin, sub-micron, films of calcium phosphate were fabricated on either glass or quartz supports by a colloidal suspension sol-gel method. These films, which varied in both surface chemistry and topography were then employed as culture substrata for osteogenic rat bone marrow cells. During an 18 day culture period, the cells elaborated a morphologically distinguishable bone matrix on all substrata which was similar to that reported earlier on tissue culture polystyrene. Selected samples of the culture substrata were fractured, critical point dried, and observed by scanning electron microscopy. Particular attention was paid to the morphologies of the interface between the so1-gellayer …

Mineralized Matrix Synthesis By Isolated Mouse Odontoblast-Like Cells In Vitro, P. B. Andrews, A. R. Ten Cate, J. E. Davies

Cells and Materials

First mandibular molar tooth germs were dissected from 17 day mouse embryos. The dental papilla was isolated using both mechanical separation and enzymatic digestion. The cells of the papilla were then enzymatically disaggregated and cultured in 35 mm polystyrene dishes containing alpha minimum essential medium supplemented with 15 % fetal calf serum, 50 μglml ascorbic acid, 1 o-8 M dexamethasone and 10 mM Na-13-glycerophosphate. The cultures were maintained for 23 days. The cultured cells initially appeared as large flat cells having numerous cell processes. Multilayered cell nodules, distributed randomly in the cultures, were apparent after 5 days. Matrix was visible …

Ultrastructural, Cytochemical, And Immunocytochemical Studies On Bone And Its Interfaces, M. D. Mckee, A. Nanci

Cells and Materials

Bone cells possess the ability to synthesize, secrete and direct the assembly and maintenance of extracellular matrix to form a functionally rigid and/or weight-bearing mineralized tissue complex, the skeleton. The skeleton not only supports and protects the organs and tissues of the body, but also acts as a mineral ion reservoir for maintaining systemic calcium balance (calcium homeostasis). The remarkable biological precision necessary for the formation, turnover and constant adaptation of bone to external mechanical forces is a dynamic process requiring a coordinated cellular effort relying on a variety of cell-cell and cell-matrix/mineral interactions. Indeed, cell-matrix interfaces found in a …

Multinucleated Giant Cells-Hydroxyapatite Interactions: A Time-Related Quantitative Study In A Rat Skull Defect, J. M. Dersot, A. Llorens, J. L. Saffar

Cells and Materials

The implantation of calcium phosphate ceramics in the bone environment elicits the differentiation of multinucleated giant cells (MNGCs). MNGC interactions with both crushed and standard hydroxyapatite (HA, Bioapatite ®) particles (cHA and sHA) were studied in rat skull defects. Light microscopy (LM) of undemineralized material after 21 and 42 days of implantation showed that the number of particles in the defect significantly decreased (about SO%) with the two HA forms. Concomitantly, average cHA particle size increased significantly due to the elimination of the smallest particles by the MNGCs. The number of MNGCs per mm2 of defect decreased significantly between 21 …

Evaluation Of Drug Delivery Systems By Electron Microscopy Techniques, Y. V. Pathak, V. D. Labhashetwar

Cells and Materials

This review covers the applications of electron microscopic techniques to the characterization and evaluation of drug delivery systems. Various electron microscopic (EM) related techniques such as transmission electron microscopy, scanning electron microscopy (SEM), SEM with energy dispersive X-ray microanalysis, and freeze-fracture electron microscopy are extensively used for this purpose. Microcapsules, microspheres, nanocapsules, liposomes , polymeric carriers and other drug delivery systems are characterized using EM related techniques for their surface topography, size and shape analysis, biodegradation, in vitro - in vivo evaluation and drug excipient interactions and characterization. Electron microscopy methods are very useful in understanding the mechanism of drug …

A Comparative Study Of The Interactions Of Two Calcium Phosphates, Peo/Pbt Copolymer (Polyactive) And A Silicone Rubber With Bone And Fibrous Tissue, C. A. Van Blitterswijk, J. R. De Wijn, H. Leenders, J. V.D. Brink, S. C. Hesseling, D. Bakker

Cells and Materials

In this study, hydroxyapatite, tetracalcium phosphate, HPEO/PBT 55145 copolymer, PEO/PBT 55!45 copolymer (Polyactive) and silicone rubber were implanted as dense blocks, subcutaneously and. into the tibia of rats. Biocompatibility and degradation were investigated but most attention was directed to .the bone/biomaterial interactions. None of the materials showed any significant adverse tissue reactions. With exception of the silicone rubber, all materials sho~ed bone bonding phenomena based on both morphological and mechanical evaluations. (H)PEO/PBT 55145 copolymer is the first polymer reported to be bonded by bone and thus widens the spectrum of bone bonding materials with a low modulus, degradable, elastomer in …

Immunohistochemical Investigation Of Tracks Left By The Migration Of Fibroblasts On Titanium Surfaces, Y. Abiko, D. M. Brunette

Cells and Materials

Fibronectin, a major attachment protein, has been thought to be involved in pathway guidance, a process in which cells migrate along specific pathways within a tissue during development. Fibroblasts exhibit the phenomenon of contact guidance, the tendency of cells to be guided in their direction of migration by the shape of the substratum. The purpose of our study was to test the hypothesis that fibronectin tracks are deposited by fibroblasts moving on smooth and grooved titanium surfaces.

The study was carried out on human gingival fibroblasts which were plated onto both smooth and grooved titanium substrata using medium containing either …

Experimental Induction Of Odontoblast Differentiation And Stimulation During Preparative Processes, H. Lesot, C. Begue-Kirn, M. D. Kubler, J. M. Meyer, A. J. Smith, N. Cassidy, J. V. Ruch

Cells and Materials

In vivo implantation experiments have shown that ethylenediaminetetraaceticacid(EDTA)-soluble frac tions of dentin stimulate reparative dentinogenesis . When isolated embryonic dental papillae were cultured in the presence of these dentin constituents, odontoblast cytological and functional differentiation could be initiated and maintained in the absence of an enamel organ. These effects were attributed to the presence of TGF-/1- related molecules [TGF-/11 or bone morphogenetic protein -2a (BMP-2a)] which had to be used in combination with an EDT A-soluble fraction of dentin in order to specifically affect competent preodontoblasts . These EDT A-soluble constituents present in dentin could be replaced by heparin or …

Calcification Of Liposomes And Red Cell Ghosts In Vitro And In Vivo, Kookmin M. Kim

Cells and Materials

To study the role of membranous phospholipids in calcification , liposomes made of phosphatidylserine and red blood cell ghosts (RBCG) prepared from rat and canine blood were incubated in minimal essential medium (MEM) with 2 .5 mM (mmol/l) calcium and 1.3 mM phosphate, pH 7.4 (MEM-2.5), and homologous (rat) or autologous (canine) plasma filtrate (serum) at 37°C for up to 1 week. Calcification was determined by electron probe microanalysis , electron diffraction, and depletions of calcium and phosphate from MEM-2.5 and sera. Liposomes and rat-RBCG incubated in MEM-2.5 and serum calcified in a week, whereas isolated collagen and elastin did …

Lysine Effect On Ruthenium Red And Alcian Blue Preservation And Staining Of The Staphylococcal Glycocalyx, Theresa A. Fassel, James R. Sanger, Charles E. Edmiston

Cells and Materials

The effect of lysine on ruthenium red (RR) and alcian blue (AB) preservation arid/or staining for transmission electron microscopy of glycocalyces was evaluated for three species of the gram-positive, coagulase-negative staphylococci. A RR and an AB procedure were compared with and without lysine included in the glutaraldehyde prefixation. Minimal or no glycocalyx was preserved or stained by glutaraldehyde fixation only. For all species, the inclusion of lysine increased visualization of fibrous material. RR treatment without lysine, results in sparse or limited fibrous material or artifactual condensations which contrasts with enhanced fibrous material seen when lysine is included with RR. The …

Influence Of Crystal Structure On The Establishment Of The Bone-Calcium Phosphate Interface In Vitro, J. D. De Bruijn, C. P. A. T. Klein, K. De Groot, C. A. Van Blitterswijk

Cells and Materials

An in vitro rat bone marrow cell system was used to examine the interfacial ultrastructure established between various calcium phosphates and mineralized tissue. The investigated calcium phosphates comprised hydroxyapatite (HA), fluorapatite (FA), tricalcium phosphate (TCP), tetracalcium phosphate (TECP) and magnesium whitlockite (MWL). Both scanning and transmission electron microscopy were used to examine the elaborated interface. The time in which a mineralized extracellular matrix was formed on the various materials differed from 2 weeks on HA, TCP and TECP, to 8 weeks on FA. It was only occasionally observed in some areas on MWL, which might have been due to aluminum …

Osteoclastic Resorption Of Calcium Phosphate Ceramic Thin Films, J. E. Davies, G. Shapiro, B. F. Lowenberg

Cells and Materials

Sub-micron calcium phosphate ceramic thin films were formed by vertically dipping transparent quartz plates in a particulate sol-gel suspension. Primary adult rat bone marrow cell populations were cultured on the ceramic thin films in conditions known to allow the differentiation of cells of the osteoclast lineage. Monitoring the cultures for periods of 11 to 28 days revealed the creation of resorption lacunae in the thin films by multinucleate cells. Some cultures were heated at 42 °C overnight to remove adherent cells; using bright field light microscopy (LM), after staining with silver nitrate, the degree of resorption could be easily assessed. …