Biomedical Engineering and Bioengineering Commons^™

Interaction Of Liposomal Drug Delivery Systems With Cells And Tissues: Microscopic Studies, M. Foldvari, G. T. Faulkner, C. Mezei, M. Mezei

Cells and Materials

Liposomes , as drug carriers, can be administered into the body by several routes e.g. intravenously, intraperitoneally, intramuscularly, intratracheally and topically among others. Radiolabelled markers are suitable to monitor the distribution and elimination of liposomes, but the tissue deposition of intact liposomes, the mode and sites of drug release from the liposomes and liposome-cell interactions cannot be investigated morphologically. Microscopic techniques could provide information regarding the intact state of liposomes and possibly the dynamics of liposomes in tissues provided that they can be identified with certainty in vivo. This is a formidable problem and in spite of several attempts, there …

Microstructure Of Mica Glass-Ceramics And Interface Reactions Between Mica Glass-Ceramics And Bone, W. Holand, W. Gotz, G. Carl, W. Vogel

Cells and Materials

This review paper characterizes glass-ceramics containing mica as main crystal phase. The phase formation reactions in dependence of the chemical composition and the microstructure are shown. Microstructure of mica glass-ceramics has been studied by electron replica and scanning electron microscopic (SEM) techniques.

Mica glass-ceramics have previously been developed in Si02-B20rA120rMgO-F--base glasses. The material is machinable because of the precipitation of micas of fluorophlogopite-type. Also, a machinable glass-ceramic for dental applications was developed based on KMg2_5(Si40 10)F2-micas. We developed mica glass-ceramics in the Si02-Al20rMgO-NaiO-K20-F glass system. Phase formation within these glasses was observed by SEM. A double controlled nucleation and crystallization …

The Effect Of Polymethylmethacrylate Bone Cement Vibration On The Bone-Cement Interface, A. M. C. Thomas, D. J. W. Mcminn, M. Haddaway, I. W. Mccall

Cells and Materials

Low frequency vibration of polymethylmethacrylate (PMMA) bone cement reduces the viscosity of the cement by shear thinning. The effect of this low frequency vibration on the bone-cement interface was studied using microfocal radiography (MFR) and scanning electron microscopy (SEM). Effects were studied in-vitro and in-vivo. In-vitro, samples of Palacos low viscosity PMMA were placed on blocks of Kiel bone and vibrated. MFR and SEM demonstrated an improvement in the appearance of the bone-cement interface. In-vivo, PMMA was injected into the upper tibia of the dog. An assessment of the effect of high and low pressure injection, and the effect of …

Ultrastructural Assessment Of Lesion Development In The Collared Rabbit Carotid Artery Model, J. E. Beesley, A. C. Honey, J. F. Martin

Cells and Materials

Cellular reactions associated with the formation of lesions generated in the carotid artery of rabbits fed either normal or high cholesterol diets by the placement of a flexible, silastic collar around the artery, were studied by electron microscopy.

Endothelial cells remained as a monolayer throughout lesion development. The endothelial cell surface in both experimental and sham operated carotids, 4 hours and 8 hours after the initiation of the experiments, were covered with platelets and leukocytes. Neutrophils were present until 7 days in the arteries from within the collar of animals maintained on a normal diet, but only to 1 day …

Applications Of Fourier Transform Infrared (Ft-Ir) Microscopy To The Study Of Mineralization In Bone And Cartilage, Adele L. Boskey, Nancy Pleshko, Stephen B. Doty, Richard Mendelsohn

Cells and Materials

Knowledge of the phase, composition, and crystallite size and perfection of the mineral in normal and abnormally calcified tissues provides insight into the mechanism by which this mineral was deposited. These data also can be used to develop rational therapies for pathological conditions characterized by abnormal mineral deposition. As illustrated in this review, coupling of an optical microscope with a Fourier transform infrared (FT-IR) spectrophotometer permits the mapping at 20 μm spatial resolution of changes in mineral characteristics (content, particle size, composition) in the growth plate, in bone biopsies, in mineralizing cell culture systems, and in soft tissue calcifications. Based …

Clinical Applications Of Recombinant Gene Technology: Bone And Cartilage Repair, Thomas A. Einhorn

Cells and Materials

Over the past decade, the field of molecular biology has given rise to the development of the applied discipline of molecular medicine. Based on recent developments using recombinant gene technology, genetic mapping analysis and other investigational tools, the physician and surgeon is now ready to begin using those instruments in the diagnosis and treatment of musculoskeletal disease. This article reviews the history of scientific exploration in recombinant gene technology as it applies to bone and cartilage repair. Clinical cases are presented to show how the fruits of scientific knowledge may be brought to bear on some of the more challenging …

Scanning Electron Microscope Observation Of Lubricious Polymer Surface For Medical Use, Hiroyuki Inoue, Yoshikimi Uyama, Emiko Uchida, Yoshito Ikada

Cells and Materials

A peculiar textured structure was found by scanning electron microscopic (SEM) observation on polymer films surface-grafted with water-soluble polyacrylamide and poly(N,N-dimethylacrylamide) after exposure to ozone. The grafted surface became very hydrophilic and highly lubricated when brought into contact with water, due to hydration of the grafted surface layer. Unless water was present, the surface did not become slippery at all. SEM of the grafted surface revealed different profiles, depending on the graft density as well as the process of drying prior to the SEM observation. There was no significant difference between the virgin and the surface-grafted film, when air-dried under …

Evaluation Of Bacterial Glycocalyx Preservation And Staining By Ruthenium Red, Ruthenium Red-Lysine And Alcian Blue For Several Methanotroph And Staphylococcal Species, Theresa A. Fassel, James E. Van Over, Christopher C. Hauser, Lorie E. Buchholz, Charles E. Edmiston, James R. Sanger, Charles C. Remsen

Cells and Materials

The cationic reagents, ruthenium red (RR), alcian blue (AB) and ruthenium red -lysine were compared for ultrastructural preservation and staining of the bacterial glycocalyx. En bloc RR, AB and RR-lysine procedures, and a glutaraldehyde/OsO4 fixation withou t cationic reagent, were evaluated by transmission electron microscopy (TEM) for several methanotroph and staphylococcal species. The glutaraldehyde/OsO4 fixation was totally ineffective in preserving or staining the glycocalyx material in all species studied. The RR procedure was more effective than the glutaraldehyde/OsO4 fixation but was generally less so than the RR-lysine procedure. By the RR-lysine procedure, extensive layers that were often filamentous or fibrous …

A Model For A Model: Pressure Jackets On Nude Mice With Implants Of Hypertrophic Scars, C. Ward Kischer, Jana Pindur

Cells and Materials

The hypertrophic scar, a consequence of deep surface injury, is peculiar to humankind. Subcutaneous implants of these tissues in nude mice provide a quasi - animal model for experimental use. An efficacious treatment to promote acceleration of maturation of the hypertrophic scar is controlled topical pressure produced by flexible dacron fabric wraps. Implants in the nude mouse provide a model for study of the effects of pressure. Preliminary results indicate that those implants in nude mice, responding favorably to pressure jackets, show changes consistent with those previously obtained in human studies. The collagen matrix shows reduced density and parallel layering …

Quantitative Scanning Electron Microscopy (Sem) To Study The Adhesion And Spreading Of Human Endothelial Cells To Surface-Modified Poly(Carbonate Urethane)S, C. James Kirkpatrick, Tillmann Otterbach, Dirk Anderheiden, Johannes Schiefer, Horst Richter, Hartwig Hartwig Hocker, Christian Mittermayer, Albert Dekker

Cells and Materials

The polymers currently in use as vascular prostheses are, in the native state, poor substrates for endothelial cell (EC) adhesion and growth. This has a negative effect on the success of pre-seeding regimes. One attempt to improve the polymer substrate is to covalently couple reactive molecules to the surface, which can be used as anchorage points for EC or serve as spacer molecule to couple biological signal molecules such as oligopeptides. We have used a digitized image analysis system coupled to a scanning electron microscope to study the adhesion and spreading of human venous EC to unmodified poly(carbonate urethane) (uPCU), …

Comparative Assessment Of The Cytotoxicity Of Various Substrates In Organ Culture And Cell Culture: A Scanning Electron Microscopy Study, J. L. Duval, R. Warocquier-Clerout, M. F. Sigot-Luizard

Cells and Materials

A comparative study of the behavior of chick embryo endothelial cells grown on various substrates was performed in order to establish the reliability and the limitation of both cell and organ culture methods. Following substrates were analyzed to compare these two different culture techniques: bovine serum albumin, pigskin gelatin and albumin + gelatin cross-linked by glutaraldehyde or carbodiimide, fibrin glue and negative control (Thermanox*). Parameters of cell growth and adhesion were calculated and compared with electron microscopic observations of cell morphology and of the extracellular matrix. Both culture methods provided complementary results and led to a similar classification of the …

Scanning Electron Microscopy Of The Bone Interface With Titanium, Titanium Alloy And Hydroxyapatite, R. D. Orr, J. D. De Bruijn, J. E. Davies

Cells and Materials

Screw implants of commercially pure titanium and titanium alloy together with rods of slip-cast dense hydroxyapatite were implanted transfemorally in young adult rats. The femora were harvested after fixation and freeze-fractured in liquid nitrogen to create two tissue fragments of each specimen. The first contained the implant while the second enabled examination of the tissue immediately adjacent to the implant. In all cases, the tissue abutting the implant was a cement line-like layer which separated the implant from the surrounding bone proper. Greater adherence of this interfacial tissue was demonstrated to hydroxyapatite samples compared with the titanium screws, while no …

The Tissue Responses To Tricalcium Phosphate Ceramics In Bone And Marrow, R. S. Archer, S. Downes, M. V. Kayser, S. Y. Ali

Cells and Materials

Calcium phosphate ceramics are described as "bioactive". Evidence of biological activity associated with calcium phosphate implanted in bone and marrow can be seen as changes in the morphology of the interfaces. In this in vivo model the sequence of progressive bone growth and bone merger with a dense calcium phosphate ceramics were examined by light and transmission electron microscopy. A spectrum of tissue responses to dense tricalcium phosphate ceramics was observed at intervals over a six week period. It is now well established that calcium phosphate ceramics permit bone ingrowth with little or no immunological response; however, a layer of …

Fine Structure Of Remodeling Sites On Iliac Cancellous Bone In Senile Osteoporosis: A Study By Scanning Electron Microscopy Using An Improved Organic Specimen Preparation Method, Toshihiko Takita, Tomonori Naguro, Kichizo Yamamoto

Cells and Materials

We improved on the organic specimen preparation method in order to investigate the details of the functional state of bone in osteoporosis at high magnification level. The usefulness and possibilities of this method are discussed showing fine structural images of remodeling sites. Iliac bones of senile osteoporotic patients obtained by transilial biopsies were studied by scanning electron microscopy (SEM) using the improved preparation method. Excepting the region near the corticoendosteal surface, most of the iliac cancellous bone was composed of thin rod-shaped trabeculae. Remodeling sites on trabeculae appeared as concavities of various sizes and were easily distinguished from inert surfaces. …

Microstructural Changes In Retrieved Amalgam Restorations, Grayson W. Marshall Jr., D. D. Mccoy, John H. Kinney, Hugo Letzel, Sally J. Marshall

Cells and Materials

Many blended or dispersed phase high Cu amalgams show excellent clinical performance in long term clinical trials. However, little is known concerning the microstructural changes that occur during clinical use of these systems. This investigation sought to characterize some of the changes that occur in the Ag-Cu particles and their surrounding reaction zones based on a study by backscattered scanning electron microscopy (BSEM), energy dispersive spectroscopy and a new rnicroprobe technique called Colorad which assigns various colors to elements and element combinations and indicates changes in elemental concentration by changes in color value (brightness). Restorations were retrieved from clinical trials …

Investigation On The Collagen Fiber Network In Human Dental Tissues - Transmission And Scanning Electron Microscopy, C. Marchetti, P. Menghini, C. Piacentini

Cells and Materials

The collagen fiber component of pulp, dentine and cementum, has been studied by isolating it from the inorganic component and the organic non-collagen matrix. This study was carried out on healthy molar and premolar adult teeth. Following demineralization all of the non-collagenous components of the tooth were removed by sequential treatment with H202, with different concentrations of trypsin and with EDTA. The transmission (TEM) and scanning (SEM) electron microscopy investigation confirmed that such a technique is an accurate method for isolating the collagen fibers while preserving their position and their ultrastructure in the dental tissue. The use of TEM and …

The Role Of Implant Surface Geometry On Mineralization In Vivo And In Vitro; A Transmission And Scanning Electron Microscopic Study, B. Chehroudi, J. Ratkay, D. M. Brunette

Cells and Materials

The purpose of th.is study was to examine the effect of substratum surface topography on bone formation in vivo and in vitro. Precise control over substratum topography was achieved using micromachining, a technique developed from the fabrication of microelectronic components. In the in vivo studies, titanium-coated epoxy replicas of micromachined surfaces were implanted subcutaneously in the parietal area of rats. After 6 weeks, bone-like tissue was found adjacent to some micromachined surfaces. Detailed observation of this tissue with the transmission electron microscope revealed osteoblast/osteocyte-like cells and a fully or partially mineralized collagenous matrix. Mineralized matrix and collagen bundles were …

Reactogenicity Of Biomaterials As Studied By Biochemical, Morphological And Ultrastructural Techniques, Leonid I. Slutskii, Natalya A. Sevastjanova, Ivetta L. Ozolanta, Irina V. Kuzmina, Laimdota E. Dombrovska

Cells and Materials

Reactogenicity is a characteristic of biocompatible materials that provokes the reparative and proliferative reaction of connective tissues, a compulsory stage of which is inflammation. Thus, reactogenicity studies should include experiments in vivo. A quantitative assessment of reactogenicity can be obtained by subcutaneous implantation of standard olive-shaped specimens covered by the biomaterial under study followed by biochemical, histological and scanning electron microscopical studies of the capsule developing around the implant. Reactogenicity of surgical threads is evaluated by semiquantitative histological analysis of the wound healing process after suture application. Biomaterial reactogenicity can be modified by changing the structure of the surface and …

Bone Regeneration Materials For The Mandibular And Craniofacial Complex, Jeffrey Hollinger, Anshumali Chaudhari

Cells and Materials

Due to different functional demands and vascularity, the mandibular and craniofacial complex require bone regenerating materials with physical and chemical properties that are different from long bone. However, certain common requirements to both locales must be addressed. Furthermore, reasons why the autograft and allogeneic bank bones are successful in regenerating bone need to be understood if the researcher is to develop satisfactory alternatives. The purpose of this paper is to review some of the requirements needed for bone regenerating materials for mandibular and craniofacial bone repair. In that effort, we have briefly described the autograft and allogeneic bank bones, animal …

Patency Of Small Caliber, Superhydrophobic E-Ptfe Vascular Grafts; A Pilot Study In The Rabbit Carotid Artery, J. M. Schakenraad, I. Stokroos, H. Bartels, H. J. Busscher

Cells and Materials

e-PTFE vascular grafts (Gore, internal diameter 1.5 mm, pore size 30 μm) were luminally treated by ionbeam etching followed by oxygen glow-discharge, to obtain a superhydrophobic luminal surface. Water contact angles of thus modified sheet material were 140-150°, which is significantly higher than on non-treated sheet PTFE (109°). These modified vascular grafts were implanted in the rabbit carotid artery, using an end to end anastomotic technique. After two hours, the grafts were still patent and the wounds were closed. One week after the implantation, the prostheses were still patent. Subsequently, the grafts were harvested and prepared for light microscopy, transmission …

Effect Of Surface Treatments On Dentin Bonding Shear Strength, James L. Drummond, D. Racean, J. Van Scoyoc, A. Steinberg

Cells and Materials

The purpose of this study was to determine whether four different treatments on wet or dry teeth, on the buccal and occlusal surfaces, affected the shear bond strength. The surface treatments consisted of a dentin conditioner (pH = 4.0), 10% phosphoric acid (pH = 1.6), 2.5 % nitric acid (pH = 1.0), and 2.5 % citric acid (pH = 1.0). One dentin bonding system and one dental composite was used for all of the specimens. The teeth were mounted in acrylic, ground and etched with the appropriate agent for 15 seconds. The dentin adhesive and composite were applied and the …

Scanning Electron Microscopy Study Of Biofilms On Silicone Voice Prosthesis, T. R. Neu, F. Dijk, G. J. Verkerke, H. C. Van Der Mei, H. J. Busscher

Cells and Materials

Patients after laryngectomy often receive silicone made voice prostheses fot speech rehabilitation. The prosthesis is inserted in a shunt between the trachea and the digestive tract. As the prosthesis is placed in a nonsterile environment it becomes rapidly colonized by microorganisms eventually leading to failure and frequent exchange of the implant. In this study, explanted "Groningen Button" silicone voice prostheses were used to investigate by scanning electron microscopy the biofilm developing on the implant. Two main types of microbial colonization forms could be distinguished. Firstly, macroscopically visible, single colonies dominating on the esophagus side of the prosthesis were found, which …

Preservation Of The Cell-Biomaterial Interface At The Ultrastructural Level, J. M. Schakenraad, J. A. Oosterbaan, E. H. Blaauw

Cells and Materials

Studying the tissue-biomaterial interface at the ultrastructural level is not without problems. Dissolution of the biomaterial in one of the dehydration or embedding media causes holes and shatter during sectioning or dislodgement of the biomaterial. The fine tuning of the hardness of both biomaterial and embedding medium, as well as the introduction of butyl-2,3- epoxypropylether as an intermediate between the dehydration series and the Epon resin , improving the impregnation , will solve many of the problems mentioned. With this improved technique good results were obtained with materials ranging from teflon, poly(Lactic acid) and polyurethanes to tissue culture polystyrene. No …

Deposition And Resorption Of Calcified Matrix In Vitro By Rat Marrow Cells, J. E. Davies, R. Chernecky, B. Lowenberg, A. Shiga

Cells and Materials

Rat bone marrow derived cells were cultured using a-Minimal Essential Medium supplemented with antibiotics, ascorbic acid and !3-glycerphosphate in the presence of 10-8M dexamethasone, on polystyrene and hydrophilic fluorocarbon substrata for periods of 2 - 4 weeks. During this time, a large yield of bone nodules was achieved and the elaborated tissue was examined by both scanning and transmission electron microscopy. The matrix produced by the cells contacting the underlying substratum was an afibrillar, globular, calcified material which formed a layer approximately 0.5μm thick. The calcium and phosphorus content of this material was confirmed by energy dispersive X-ray dot mapping …

Analysis Of Protein Adsorption On Retrieved Human Vascular Grafts Using Immunogold Labelling With Silver Enhancement, N. P. Ziats, N. S. Topham, D. A. Pankowsky, J. M. Anderson

Cells and Materials

The adsorption of proteins on biomedical materials such as vascular grafts is important in modulating thrombosis, one eventual cause of vascular graft failure. We have identified proteins expressed on the surfaces of retrieved vascular grafts to determine if certain proteins were present at the end stage of vascular failure. Scanning electron microscopic analysis of protein adsorption on the surfaces of retrieved vascular prostheses was determined using antibodies to human blood proteins fibrinogen, fibronectin, Hageman factor (factor XII) and factor VIII/von Willebrand Factor. The detection of these proteins on the blood contacting surface was evaluated by immunogold labelling with protein A-gold …

Peripheral Nerve Bridges For Central Nervous System Regeneration, Silvio Varon, Theo Hagg, H. Lee Vahlsing, Marston Manthorpe

Cells and Materials

A neuronotrophic hypothesis for the adult central nervous system (CNS) has been increasingly supported by in vivo studies of Nerve Growth Factor (NGF) impacts on adult rat CNS cholinergic neurons. We have extended these investigations to NGF effects on cholinergic axonal regeneration in an adult rat septo-hippocampal model. A fimbriafornix lesion deprives the dorsal hippocampal formation (HF) of its cholinergic afferents and provides a cavity into which sciatic nerve segments can be implanted to serve as regeneration bridges from septum to HF. Septa! cholinergic fibers enter and course through fresh cell-containing nerve bridges to attain a maximal number by the …

Evaluation Of The Images Of Specular Microscopy Of The Cells On Implanted Intraocular Lenses In Vivo, Kiyoshi Okada, Keiko Takahashi, Hiroaki Sagawa, Kuniomi Abe

Cells and Materials

Specular microscopy , an optical method to study the anterior segment of the eye, was applied to observe the cells on implanted intraocular lenses in vivo. The images from specular microscopy were evaluated by comparing the morphology of the same cells on intraocular lenses implanted in rabbit eyes by specular microscopy , light microscopy and scanning electron microscopy (SEM). The time lapse study in living human eyes and the optical evaluation of jts interference images were performed to elucidate its image formation.

The same giant cells, as observed in vivo by specular microscopy, proved to be multinucleated cells. It …

Interactions Of Bacteria And Amoebae With Ocular Biomaterials, Thomas John

Cells and Materials

The use of biomaterials in periocular and intraocular sites has resulted in some ocular inflammations and infections which can result in vision-threatening ocular disease. This review addresses bacterial interactions with, and adherence to ocular biomaterials such as soft contact lenses, surgical suture materials, and intraocular lenses. In addition, adherence of Acanthamoeba to soft contact lenses is described, and the role of these lenses in the development of Acanthamoeba keratitis is discussed.

Dusts Produced During Cutting Of Modern Splinting Bandages, R. Wytch, D. W. Gregory, R. Clayton, D. Wardlaw

Cells and Materials

Previous work carried out by the authors (Wytch et al ., 1988a) has shown that airborne dust generated during removal of orthopaedic casts with a power saw could represent a respiratory hazard. Further work has been carried out on all types of splinting bandage currently available in the United Kingdom to assess the potential dust hazard when sawing these materials. It has been shown that although plaster of Paris bandage produces significant concentrations of airborne dust, most of the polyurethane-impregnated fabric bandages generate extremely low levels of dust and in the case of the non-glass fabrics, the level is insignificant. …

Scanning Electron Microscopic Evaluation Of The Fractured Surfaces Of Canine Calculi From Substrata With Different Surface Free Energy, H. M. W. Uyen, W. L. Jongebloed, H. J. Busscher

Cells and Materials

The strength of adhesion between dental calculus and enamel or dentin surfaces determines the ease with which the calculus can be removed by brushing or professional dental treatment. In this study, we examined the adhesion of canine calculi formed on substrata with different surface free energies (sfe) and roughness by means of scanning electron microscopy. In 4 beagle dogs fenestrated crowns were made on the upper fourth premolars. Subsequently, facings of glass (sfe = 120 mJ. m-2), bovine enamel (sfe = 85 mJ.m-2), bovine dentin (sfe = 92 mJ.m-2), polymethyl-methacrylate (PMMA, sfe = 56 mJ.m-2) and polytetrafluorethylene (PTFE, sfe = …