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Biomedical subjects

R E Baier

Publications and source records attributed to R E Baier.

At least 19 recordsLinked to original sources

Surface properties of mussel adhesive protein component films.

Mussel adhesive protein (MAP) is the adhesive agent used by the blue sea mussel (Mytilus edulis) to attach the animal to various underwater surfaces. It is composed of 75-->85 repeating decameric units with the reported primary sequence NH2-A(1)-K(2)-P(3)-S(4)-Y(5)-Hyp(6)-Hyp(7)-T(8)-DOPA(9)-K(10)-COOH. This study identifies and compares the surface properties of the decameric unit, selected fragments and individual amino acid constituents with the complete MAP preparation. These molecular systems were examined: (a) in the solid state as thin films formed on germanium substrata using multiple-attenuated-internal-reflectance infrared (MAIR-IR) spectroscopy, ellipsometry and contact angle analysis; and (b) in the solution state using circular dichroism (CD) spectroscopy. Extensive molecular modelling of the decamer was performed making integral use of the experimentally derived data. These cumulative semi-empirical and empirical results suggest a conformation for the decamer that closely associates the L-DOPA and tyrosine residues with the solid substratum. This model provides the first representation of MAP derived from a rational integration of theoretical and experimental data. On the basis of this model, a possible explanation for the bioadhesive properties of MAP is suggested.

Amino Acid Sequence

Reduced corrosion of CP Ti and Ti-6A1-4V alloy endosseous dental implants after glow-discharge treatment: a preliminary report.

Inductively coupled argon plasma spectroscopic analysis was used to quantify titanium, aluminum, and vanadium corrosion products released by superficial layers of identically shaped commercially pure titanium and Ti-6A1-4V alloy dental implants. Halves of each of two originally sterile groups were analyzed directly from the manufacturer's packaging and the other halves received glow-discharge treatment prior to their corrosion assay. The implants were incubated in pH 3 modified saline solution and were analyzed after 2- and 12-week corrosion periods. Implants treated with glow discharge showed statistically significant reduction in the amounts of corrosion products released, apparently as a result of glow-discharge-produced surface oxides of greater passivity than originally present.

Aluminum

Radiofrequency gas plasma (glow discharge) disinfection of dental operative instruments, including handpieces.

The radiofrequency-stimulated argon gas plasma (glow discharge) technique, already well-known for surface cleaning and activation of adhesion, was investigated for determination of its separate potential for rapid disinfection of dental operative instruments. Disinfection effectiveness was judged from diminished post-treatment recovery of viable organisms from the instruments agitated in saline. Streptococcus salivarius, Bacillus stearothermophilus, and Escherichia coli were used as primary contaminant organisms, dried from gelatin-thickened laboratory cultures onto the instruments and not subjected to any preliminary cleaning steps. Significant disinfection was obtained, with no sensible temperature increases, in under 10 minutes in laboratory apparatus consuming fewer than 5 Watts of power per cycle. Also, clinically used and deliberately-saliva-contaminated high-speed handpieces were gas-discharge-treated, with their resultant rapid disinfection noted by complete suppression of the viability of any transferred natural contaminant organisms within two minutes. With regard to preservation of instrument quality, it was also shown that this low-temperature gas-discharge method provides the noted substantial disinfection without deterioration of sharp edges. Work now in progress suggests that the method can provide cool, rapid, and complete sterilization when hydrogen peroxide vapors are present in the gas plasma used for treatment of instruments first given the normally recommended thorough pre-cleaning.

Argon

Improved integration potential for calcium-phosphate-coated implants after glow-discharge and water-storage.

Laboratory and clinical data support the conclusion that Radio-Frequency-Glow-Discharge-Treatment (RFGDT) of calcium-phosphate-coated implants can accelerate their functional integration with bone at host sites. In addition to the benefits of surface cleaning and activation associated with RFGDT, a period of water-storage prior to implantation also seems to be beneficial in eluting easily solubilized alkaline calcium components.

Dental Implantation, Endosseous

Principles of adhesion.

Understanding interfacial phenomena has been of direct relevance and practical benefit to extending the use of dental adhesives. Both surface physics, which describes properties of the inorganic materials' interfacial zones from their actual phase boundaries toward the bulk phases of the solids, and surface chemistry, which describes phenomena at the solid/biological interface and beyond it into the variable organic environment, have been important. High-energy materials include solids that are very hard, have high melting points, strong intermolecular forces, and basically crystalline structures, such as dental enamel. Low-energy materials, such as dentinal collagen, salivary films, and the organic resins of restorative materials, are softer, lower melting, and have weaker intermolecular forces, poorer crystallinity, and surface energies generally less than 100 ergs/cm. It has been a properly renewed emphasis on wetting of dental surfaces and their modification by primer coats, displacing or mixing with water and adsorbed proteinaceous films, that has promoted the success of many recently developed fourth-generation dentin adhesives. Their improved wettability for biological phases correlates directly with their better infiltration and anchoring of composites.

Acid Etching, Dental

Surface characterization of the cuticle of Dirofilaria immitis.

The surface reactivity of the dog heartworm (D. immitis) was evaluated by comprehensive contact angle measurements and a platelet retention test. Contact angle data yielded calculated surface energy terms very similar to those previously reported for intact vascular endothelium. The platelet test revealed the native worm surface to be nonreactive, retaining fewer platelets than glass or worms whose surfaces had been modified by extraction with acid and high salt solutions. The cuticular morphology of the heartworm was studied with both light and electron microscopy, the latter coupled with ferritin-conjugated double-layer immunolabeling to reveal adsorbed host protein on the cuticle surfaces. Multiple attenuated internal reflection (MAIR) IR spectroscopy confirmed the general composition of this surface layer to be glycoproteinaceous. Morphological and histochemical studies confirmed and extended previous descriptions of nematode cuticle, adding ultrastructural detail on cortical, medial, and basal layers. A trilaminar membrane, apparently corresponding to a mammalian cell membrane (plasmalemma), constituted the external cortical layer as observed in high magnifications. The existence of a glycocalyx of varying thickness was demonstrated in ruthenium red-stained sections. MAIR IR spectra showed this glycoproteinaceous film to appear, in fully hydrated samples, as a loose biological gel. Ferritin-antibody conjugate labeling confirmed the presence of adsorbed dog albumin, dog immunoglobulin class G (IgG) and dog complement fraction 3 (C3) in the cuticular surface layer. It is likely, therefore, that D. immitis heartworms demonstrate long-term thromboresistance at least in part due to their passive low-surface-energy overcoating with host proteins.

Animals

Effect of critical surface tension on retention of oral microorganisms.

The effect of critical surface tension on the initial retention of microorganisms from unstimulated human saliva was tested in a flow cell system. Prior to each experiment the total numbers and the morphotypes of microorganisms present in saliva were recorded. The test surfaces were prepared to display known increasing critical surface tensions, as verified and standardized by contact angle measurements. Surfaces of initially low (20-22 mN/m), medium (35-38 mN/m) and high (greater than 50 mN/m) critical surface tension were exposed to saliva at a flow rate of 1 ml/min. Microbiota and biofilm material associated with the test surface after 15 min of salivary exposure, were then subjected to standard detachment forces, by introducing a cell-free rinsing fluid at two different shear rates. Both the attachment and the detachment phases were executed at room temperature or 37 degrees C. The retained population was counted in three different zones of the test surfaces with a light microscope and statistically tested for correlation to the main variables (critical surface tension, flow rate and temperature) and interactions. Retention success was significantly dependent on the initial critical surface tension and the flow rate. Surfaces of medium critical surface tension, representative of human tooth surfaces and most restorative dental materials, retained the highest numbers of microorganisms in comparison with the other surfaces tested, with no statistically verified selectivity in proportions of retained coccoid and rodshaped microorganisms for any surface. A 30-fold increase of the flow rate resulted in a 70-80% reduction of the retention success, with a higher relative number of cocci present on all the test surfaces. These results demonstrate that initial retention of microorganisms to surfaces is non-specific with regard to morphotypes, but is strongly related both to the mechanical removal forces and the surface energetic state of the solid surface exposed. Retention of microbial populations at interfaces might, therefore, be controllable by advance selection of the critical surface tensions and predicted if shear forces at given sites are known.

Bacterial Adhesion

Electron microscopic studies of human mixed saliva.

With foam components removed, mixed saliva from three donors were solidified in liquid nitrogen and sectioned, mounted, and fixed. Examination by transmission (TEM) and scanning (SEM) electron microscopy and energy-dispersive X-ray (EDAX) analysis were performed for paraformaldehyde-fixed sections, some of which were OsO4-postfixed. The TEM and certain SEM examinations showed the presence of fine and dense salivary network structures, seemingly originating from the major fibrous components. In OsO4-treated sections, TEM pictures showed reticulated arrangements with open cellular diameters down to 0.2 microns. The EDAX analyses particularly showed the presence of Ca, Fe, K, P, and S, with increased Ca readings in major components. Untreated sections showed that strands, with diameters of more than 1-2 microns, had more electron-dense central portions than peripheries and sometimes had interior, very electron-dense, granules. The observed features indicate that saliva has internal structures consistent with its colloid chemical characteristics.

Adult

Structural studies of human saliva.

Samples of mixed saliva and of parotid and sublingual/submandibular saliva fractions from four donors were subjected to instantaneous solidification in liquid nitrogen followed by sectioning in a microtome/cryostat. The sections were stained with hematoxylin-eosin, periodic acid-Schiff, Alcian blue, Oil-red-O, or Sudan Black B and then examined at the light-microscopic level. In all the sectioned samples several previously never described features were observed, the most pronounced of which were a loose overall network structure and collections of lipoid droplets often in a loose arrangement. In the mixed saliva sample sections many of the microorganism-like structures were observed in large bunches associated with epithelial cells and densely staining saliva components. The present method was tested in a series of experiments for possible errors.

Adult

Thin-section transmission electron microscopy of human saliva.

Thin sections (90-100 nm) of flash-frozen human saliva fractions and whole saliva were examined by transmission electron microscopy. Inside the major filamentous structural components numerous ultrastructural details were observed, especially for whole saliva and submandibular/sublingual fractions. An outer reticulated zone believed to contain the major salivary glycoproteins surrounded an inner core consisting of a continuous, more electron dense phase with multiple vacuoles and granules of different sizes, shapes, and electron densities. The observed structures suggest a complex microarchitectural model for whole saliva.

Actin Cytoskeleton

Silicone rubber temporomandibular joint (TMJ) meniscal replacements: postimplant histopathologic and material evaluation.

Medical grade silicone rubber has long been considered a suitable meniscal replacement, but there has been increasing concern about migration of this material into adjacent tissues. The objectives of this study were to determine the definitive composition of tissue-incorporated material which is presumed by light microscopy to be silicone and to identify long term histopathologic sequelae of meniscal replacements. Adult female patients underwent meniscectomy and replacement with silicone rubber (Silastic) implants. After 12 to 18 months, recurrence of symptoms in 8% of these cases led to implant removal with excision of peri-implant fibrous pseudocapsules. Excised tissues, including one preauricular lymph node and implants were submitted for light microscopy, SEM, and energy dispersive x-ray microanalysis (EDX) for the identification of elemental composition, critical surface tension measurement, and internal reflection infrared spectroscopy. EDX revealed prominent peaks for silicon in both pseudocapsular and nodal tissues. Morphologic findings surrounding the long-term implants included foreign body reaction, synovitis, dystrophic calcification, fibrocartilaginous metaplasia, hyalinization, and scarring. Particulate silicone debris induced a pathologic response in the tissues and migrated to nodes. These findings suggest that periodic evaluation be performed over the life of such implants to rule out breakdown under function. These findings should intensify the search for improvements or replacements for silicone rubber as an interpositional material in the temporomandibular joint.

Calcinosis

Role of temperature and shear forces on microbial detachment.

A flow cell system was used to assess the effects of shear stress and temperature on adhesion and retention of oral microorganisms from unstimulated whole human saliva. The saliva passed between two parallel mounted test plates, prepared and calibrated to present a surface energetic state similar to that of natural teeth and most restorative materials. The initial attachment of microorganisms occurred at a constant flow rate of 1 ml/min. Microbiota and biofilm material associated with the surface after 15 min of exposure were then challenged by introducing a cell-free rinsing fluid at increased flow rates. The remaining population was counted in a reflected light microscope and correlated to the calculated shear stress for each experiment. A reduction of 70-80% of attached microorganisms was seen after a 30-fold increase of the detaching force. No statistically significant differences could be detected in the proportions of initially attached or the remaining cocci and rods. The experiments were conducted at two temperature levels, both within a physiologic range representative of the oral environment. Temperature did not significantly affect the total numbers of attached or retained microorganisms, within the range of 22-37 degrees C. These findings demonstrate that non-specific attachment and detachment processes are important in the initial stage of microbial adhesion. Although biologically specific adhesive interactions were not addressed in this study, the data suggest that these may occur only after a minimum contact time of non-specifically surface associated cells.

Bacterial Adhesion

Advanced biomaterials development from "natural products".

Natural substances and structures can serve increasingly well as biomedical products, given recent advances in understanding of requirements for biocompatibility and of methods for their preservation and surface tailoring. A successful example is the derivation of limb salvaging vessels, used in arterial reconstructive surgery, from human umbilical cords. There are numerous opportunities for additional product development from the umbilical cords' main ingredient, Wharton's gel, ranging from biolubricants to wound-healing aids. Major problems yet to be overcome with natural starting materials are their propensity for calcification and eventual biodeterioration. Surface modification of biomaterials to exhibit desired degrees of interaction with contacting viable tissues promises the greatest beneficial results. General principles of bioadhesion have broad applicability, predicting material behavior in environments as diverse as blood, saliva, and seawater.

Adhesiveness

Future directions in surface preparation of dental implants.

Clean, intrinsically high-surface-energy dental implants are both safe and effective, but ambiguity remains with regard to the true surface qualities of many implant materials that have been sterilized and are about to be placed in properly instrumented host sites. Future dental implant requirements should include proper surface preparation and surface quality maintenance of the implants themselves. Recommended directions for research on dental implant materials include greater use of surface analytical techniques to identify, understand, and ultimately control common surface contaminants. Regulatory agencies should require specific reporting of critical surface parameters for all dental implant materials in their final, sterile states, including any changes from desirable initial values. Different implants demand different degrees of interaction with adjacent biological phases, for example, to promote or inhibit bioadhesion, and different surface preparations can assure these results.

Biocompatible Materials

Investigation of tissue/implant interactions during the first two hours of implantation.

The objective of this study was to determine whether detectable differences exist in the initial process of tissue incorporation of implant materials of different original surface energies. Our earlier work focused on materials implanted for days to months; the experiments reported here addressed the initial 2 hours of implantation. Surface-treated metal implants were placed in the fascial plane in the back of New Zealand white rabbits for 5 minutes to 2 hours. Prior to implantation, the metal plates were characterized by infrared spectroscopy, ellipsometry, and contact potential methods; the same analytical techniques, as well as scanning electron microscopy and contact angle measurements were applied to the explanted samples. No significant cellular adhesion was observed within the first 2 hours of implantation. Films rich in lipids and proteins were detected on the implants, however, within 5 minutes. There is good evidence for specific film differentiation as a function of differing initial substratum surface energies.

Animals