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Biomedical subjects

E Wintermantel

Publications and source records attributed to E Wintermantel.

At least 37 records · Page 2Linked to original sources

Titanium containing amorphous hydrogenated carbon films (a-C: H/Ti): surface analysis and evaluation of cellular reactions using bone marrow cell cultures in vitro.

Amorphous hydrogenated carbon (a-C : H) coatings, also called diamond-like carbon (DLC), have many properties required for a protective coating material in biomedical applications. The purpose of this study is to evaluate a new surface coating for bone-related implants by combining the hardness and inertness of a-C : H films with the biological acceptance of titanium. For this purpose, different amounts of titanium were incorporated into a-C : H films by a combined radio frequency (rf) and magnetron sputtering set-up. The X-ray photoelectron spectroscopy (XPS) of air-exposed a-C : H/titanium (a-C : H/Ti) films revealed that the films were composed of TiO2 and TiC embedded in and connected to an a-C : H matrix. Cell culture tests using primary adult rat bone marrow cell cultures (BMC) were performed to determine effects on cell number and on osteoblast and osteoclast differentiation. By adding titanium to the carbon matrix, cellular reactions such as increased proliferation and reduced osteoclast-like cell activity could be obtained, while these reactions were not seen on pure a-C : H films and on glass control samples. In summary, a-C : H/Ti could be a valuable coating for bone implants, by supporting bone cell proliferation while reducing osteoclast-like cell activation.

Analysis of Variance↗

Titanium dioxide ceramics control the differentiated phenotype of cardiac muscle cells in culture.

A new approach, the cultivation of heart muscle cells on biocompatible scaffolds made from titanium dioxide ceramics was established to provide a mechanism for in vitro engineering of a vital heart tissue. Terminally differentiated ventricular myocytes isolated from hearts of adult rats were kept in primary culture for long periods of time and used as an experimental model. The microenvironmental properties of titanium dioxide ceramics helped to maintain the tissue-like structural organisation of the cardiac cells in vitro. Coating of the cell substrata with fine-grained titanium dioxide ceramics imitating cell surface topography favoured the formation of focal adhesion complexes in the ventral plasma membrane of cardiomyocytes. It also promoted the cellular expression of vinculin, a protein that connects the ECM integrin receptors to the network of cytoplasmic filaments, which define cell shape. This topographical reinforcement of cell-material interactions led to stabilisation of the molecular linkage between the extracellular contacts and the intracellular cytoskeleton and thus assisted the preservation and maintenance of the heart muscle cell differentiated phenotype in long-term primary culture. The results of this work demonstrate a promising pathway for the regulation of cellular organisation in vitro by local geometric control.

Animals↗

Feasibility study of an online toxicological sensor based on the optical waveguide technique.

Morphological properties of the cells often change as an early response to the presence of a pharmacologically acting toxic substance [Etcheverry, S.B., Crans, D.C., Keramidas, A.D., Cortizo, A.M., Arch. Biochem. Biophys. 338 (1997) 7-14]. Recently it has been shown that living animal cell adhesion and spreading can be monitored online and quantitatively via the interaction of the cells with the evanescent electromagnetic field present at the surface of an optical waveguide [Ramsden, J.J., Li, S.Y., Heinzle, E., Prinosil, J.E. Cytometry 19 (1995) 97-102]. In the present study, optical waveguide lightmode spectroscopy (OWLS) and confocal laser scanning microscopy (CLSM), which provides information about the shape of the cells at the surface, were compared under identical experimental conditions. This allowed for the correlation between the cell-shape information from CLSM and the cell-surface interaction measurements from OWLS. The proposed design of the microsystem sensor involves the establishment of a cell layer on the surface of the waveguide and the subsequent online measurement of the morphological response of the cells to various toxic substances. In the present study, the setup was evaluated using cells from an osteoblastic MC 3T3-E1 cell line, and sodium hypochlorite was used as model toxic substance. Comparing the OWLS signal to the morphological response measured by CLSM reveals that OWLS is effective in monitoring not only cell attachment and spreading but also the cellular response to toxic compounds (i.e. by means of change in cell morphology). For the model toxin, the OWLS measurements indicate that, at concentrations above 0.01%, the cells exhibit a clearly discernable morphological effect (i.e. a decrease in average cell contact area). Thus, the potential of an on-line sensor based on OWLS to applications in toxicology, pharmacy and biocompatibility was demonstrated.

3T3 Cells↗

[Growth of human respiratory epithelium on collagen foil].

BACKGROUND: Clinical application of bioartificial tracheal prosthesis must still be regarded as an experimental concept because restoration of a functional respiratory epithelium outlining the prosthesis is still not possible. Tissue engineering as a relatively new biotechnological discipline may offer new methods in expanding differentiated respiratory epithelium in vitro. In this study we compare two different cell and tissue culture procedures for growing human nasal mucosa on commercially available collagen foil. MATERIAL AND METHODS: Harvested specimens of human nasal mucosa (n = 6, 4 x 4 cm) were placed on collagen foil and incubated as tissue cultures for 4, 6 and 8 weeks. A suspension of enzymatically dispersed nasal epithelium seeded on collagen foil (5 x 10(5) cells) served as control. Cell growth and ciliary beat were monitored through an inverted microscope with Hoffman's modulation contrast and video set-up. Histological examination was performed after 4, 6 and 8 weeks. RESULTS: In the tissue cultures, the collagen foil was initially covered with fibroblasts growing from the mucosa specimen before epithelial cells spread out. The epithelial layer showed mostly ciliated cells which developed metachronous ciliary beat after 4 weeks in vitro. Ciliary activity was observed until the end of the experiments in 8 weeks. New cells on the suspension cultures were mesenchymal and did not exhibit any ciliary activity. CONCLUSIONS: Mucosa specimens seem to be more appropriate for tissue engineering of respiratory epithelium than cell suspensions from nasal epithelium. Collagen foil as tissue scaffold initiates epithelial-mesenchymal interaction and may play an important role in epithelial differentiation of new respiratory epithelium.

Biomedical Engineering↗

[Biomaterials, human tolerance and integration].

Biomaterials and related process engineering in order to obtain optimal surface and structural biocompatibility of implants and devices are presented. Vital-avital composites for tissue engineering, cell culture models, porous ceramics and degradable polymers are introduced as examples. Emphasis is laid on the conversion of basic research results into clinical applications and on the exchange of technologies from the non-medical into the medical field and vice versa.

Animals↗

Partially degradable film/fabric composites: textile scaffolds for liver cell culture.

In this study, a composite scaffold combining textile superstructures and biomimetic glycopolymers is introduced, which may allow engineering of organotypic liver tissue in vitro. Woven poly(ethylene therephtalat) (PET) fabrics were coated on one side with a thin biodegradable polymer film (poly[D-L-lactic-co-glycolic acid] PLGA), in order to obtain a polar structure. The composite structure ensured the stability of the membrane during in vitro degradation, independently of mesh size. Matrix porosity increased when a polymer blend matrix was used. For hepatocyte culturing studies, the scaffolds were additionally coated with an artificial glycopolymer (poly[N-p-vinylbenzyl-D-lactoamide], PVLA) in order to improve cell attachment. It was observed that formation of aggregates depends on the scaffold geometry as well as on the pretreatment and medium conditions. After 4 days in culture, the pores of the fabric were filled with aggregates illustrating the possibility of immobilizing hepatocyte aggregates in well-defined spatial configurations on textile structures.

Biocompatible Materials↗

[Failure conditions of glass filament yarns: a contribution to the valuation of carcinogenic potentials of fiber fragments].

Failure of glass filament yarns results in the formation of many fragments. Through inhalation, these particles can intrude into the human body. If the fragments are sufficiently bioresistant and have a fiber dust geometry, according to the MAK-values (6), i.e. if they are longer than 5 microns and thinner than 3 microns and show an aspect ratio greater than 3, they have a carcinogenic potential. Since the glass filaments show a diameter greater than 3 microns, no fiber dust particles will be formed if transversal fiber failure occurs without crack-branching. In the present study the geometric distribution of fragments after failure of glass filament yarns under combined stress was investigated in order to estimate the carcinogenic potential of the fragments. The knot tension test was shown to be a suitable method for this investigation. A defined fraction of the total amount of fragments were analysed with scanning electron microscopy (SEM) by measuring their length and diameter. To investigate whether the analysed particles are fragments of the glass filament yarns, chemical analysis was performed with energy dispersive X-ray analysis (EDX). Morphologically, two different fragment types were observed: a) Fragments with their entire filament cross-section which were formed by transversal fiber fraction. b) Smaller fragments which were formed through crack-branching. These smaller fragments were observed to adhere on the bigger fragments due to high surface forces. During each knot tension test, 5-60 fragments per filament were formed. However, the fraction of fiber dust particles was very low and showed a maximum of 1.5%. Only in one of the four tested yarn types (high temperature yarn HT 75) the formation of fiber dust particles was observed. The other yarns showed fragments with dimensions close to fiber dust geometry. Therefore, it cannot be excluded that some fragments with fiber dust geometry may have been formed during mechanical testing. Fragment distribution of the studied E-glass yarns was shown to be dependent on the modification method. To date, it cannot be excluded that there are types of glass filament yarns forming a major quantity of fiber dust particles during failure. The fragments of type b such as fiber dust particles were observed to adhere on bigger fragments which are themselves too big to reach the alveoli. In the present study the stability of these agglomerates under various environmental conditions was not investigated. Moreover, fragment agglomerates should not be considered on their own but in connection with the application. The presented tests were carried out with simple yarns and, therefore, represent isolated observations.

Air Pollutants↗

Autosterilization of biodegradable implants by injection molding process.

Sterilization of degradable implants by standard procedures may damage the parts due to the labile chemical nature of the polymers. This study examined whether the injection molding process used for the production of polymeric parts may itself sterilize the implant due to high temperature, pressure, and shear forces applied. Poly-D,L-lactic acid (PDLLA) and poly-L-lactic acid (PLLA) granules were contaminated with thermoresistant spores of Bacillus stearothermophilus (>10(5) spores/g). Sterile and contaminated granules of both polymers were injection molded and tested for sterility. All 27 samples produced with sterile PDLLA and processed at 120 degrees C and all 18 samples produced with sterile PLLA at 200 degrees C remained sterile after injection molding and handling. However, in five out of 28 PDLLA samples and in one out of 26 PLLA samples produced with contaminated material, spores had survived the process. In conclusion, the injection molding process could not reliably sterilize parts produced with polylactic acid granules that were heavily contaminated with thermoresistant organisms. However, the number of viable spores was significantly reduced by more than 99.99%. Thus, the injection molding process might allow the autosterilization of parts produced with raw material that is not heavily contaminated.

Biocompatible Materials↗

Surface activation of polyetheretherketone (PEEK) and formation of calcium phosphate coatings by precipitation.

Plasma activation of polyetheretherketone (PEEK) surfaces and the influence on coating formation in a supersaturated calcium phosphate solution was investigated in this study. It was observed that plasma treatment in a N2/O2 plasma had a significant effect on the wettability of the PEEK surface. The contact angle decreased from 85 degrees to 25 degrees after plasma treatment. Cell culture testing with osteoblastic cell lines showed plasma activation not to be disadvantageous to cell viability. X-ray photoelectron spectroscopy (XPS) analysis was performed to characterize the chemical composition of the PEEK surfaces. It was observed that the O1s intensity increased with plasma activation time. At the C1s peak the appearance of a shoulder at higher binding energies was observed. Coating of PEEK was performed in a supersaturated calcium phosphate solution. Coating thicknesses of up to 50 microm were achieved after 24 days of immersion. Plasma activation followed by nucleation in a highly saturated hydroxyapatite solution had a positive effect on the growth rate of the layer on PEEK. Chemical analysis revealed that the coating consists of a carbonate-containing calcium phosphate.

Journal Article↗

NaOH treatment of vacuum-plasma-sprayed titanium on carbon fibre-reinforced poly(etheretherketone).

Carbon fibre-reinforced polyetheretherketone (CF-PEEK) substrates were coated with titanium by vacuum-plasma-spraying and chemically treated in 10 M sodium hydroxide (NaOH) solution. After NaOH treatment, the specimens were immersed in simulated body fluid (SBF) containing ions in concentrations similar to those of human blood plasma. Scanning electron microscopy, energy-dispersive X-ray analysis and diffuse reflectance Fourier transformed-infrared spectroscopy were used to analyse the NaOH-treated VPS-Ti surface and the calcium phosphate layer formed during immersion in SBF. It was observed that a carbonate-containing calcium phosphate layer was formed on the NaOH-treated VPS-Ti surface during immersion in SBF, whereas no calcium phosphate precipitation occurred on the untreated surfaces. It is therefore concluded that vacuum-plasma-spraying with titanium and subsequent chemical modification in 10 M NaOH solution at 60 degrees C for 2 h is a suitable method for the preparation of bioactive coatings for bone ongrowth on CF-PEEK.

Journal Article↗

Topographical characterization and microstructural interface analysis of vacuum-plasma-sprayed titanium and hydroxyapatite coatings on carbon fibre-reinforced poly(etheretherketone).

In the present study, topographical characterization and microstructural interface analysis of vacuum-plasma-sprayed titanium and hydroxyapatite (HA) coatings on carbon fibre-reinforced polyetheretherketone (CF/PEEK) was performed. VPS-Ti coatings with high roughness values (Ra=28.29+/-3.07 microm, Rz=145.35+/-9.88 microm) were obtained. On this titanium, intermediate layer HA coatings of various thicknesses were produced. With increasing coating thickness, roughness values of the HA coatings decreased. A high increase of profile length ratio, Lr, of the VPS-Ti coatings (Lr=1.45) compared to the grit-blasted CF/PEEK substrate (Lr=1.08) was observed. Increasing the HA coating thickness resulted in a reduction of the Lr values similar to the roughness values. Fractal analysis of the obtained roughness profiles revealed that the VPS-Ti coatings showed the highest fractal dimension of D=1.34+/-0.02. Fractal dimension dropped to a value of 1.23-1.25 for all HA coatings. No physical deterioration of the CF/PEEK substrate was observed, indicating that substrate drying and the used VPS process parameter led to the desired coatings on the composite material. Cross-section analysis revealed a good interlocking between the titanium intermediate layer and the PEEK substrate. It is therefore assumed that this interlocking results in suitable mechanical adhesive strength. From the results obtained in this study it is concluded that VPS is a suitable method for manufacturing HA coatings on carbon fibre-reinforced PEEK implant materials.

Journal Article↗

Tissue engineering scaffolds using superstructures.

Here, scaffolds as cell and tissue carriers are approached from an engineering point of view, emphasizing material superstructuring in the design of supports. Superstructure engineering provides optimal spatial and nutritional conditions for cell maintenance by the arrangement of structural elements (e.g. pores or fibres) so as to vary the order of cell to cell contact. This approach is illustrated in the design of several scaffolds: knitted fabrics as three-dimensional superstructures for optimized osteosynthesis implants, a new injectable open porous implant system, an angiopolar non-degradable ceramic cell carrier, and an injectable or microsurgically implantable entangled carrier system. The implications for tissue engineering are discussed.

Anisotropy↗

Angiopolarity of cell carriers: directional angiogenesis in resorbable liver cell transplantation devices.

The purpose of this study was to obtain directional angiogenesis of small blood vessels and capillaries to an implant made from a resorbable polymer for hepatocyte transplantation. It was intended to mimic the native acinar structure of the liver in order to facilitate replication of the cells and organ growth. The implant device structure was designed for injection to minimize surgical trauma. Hollow microspheres with an open porous wall structure and one large central opening were made from poly(d,l-lactic-co-glycolic acid) (85:15 lactic:glycolic). This polymeric scaffold was seeded with hepatocytes and implanted into the abdominal wall muscle of syngeneic Fisher rats. Specimens explanted up to 56 days p.o. showed hepatocyte survival and the development of a directional blood supply. This phenomenon is coined "angiopolarity". The study should help in addressing the issue as to whether avascular cell implants with post-transplantation organ growth should be attempted. Processing options in applying heat to the polymer solution allow manufacturing of larger microspheres with different diameters of central openings. This would allow the use of the scaffold for other cell transplantations than hepatocytes.

Animals↗

Future directions in biomaterials.

Biomaterials have made a great impact on medicine. However, numerous challenges remain. This paper discusses three representative areas involving important medical problems. First, drug delivery systems; major considerations include drug-polymer interactions, drug transformation, diffusion properties of drugs and, if degradation occurs, of polymer degradation products through polymer matrices developing a more complete understanding of matrix degradation in the case of erodible polymers and developing new engineered polymers designed for specific purposes such as vaccination or pulsatile release. Second, cell-polymer interactions, including the fate of inert polymers, the use of polymers as templates for tissue regeneration and the study of polymers which aid cell transplantation. Third, orthopaedic biomaterials, including basic research in the behaviour of chondrocytes, osteocytes and connective tissue-free interfaces and applied research involving computer-aided design of biomaterials and the creation of orthopaedic biomaterials.

Biocompatible Materials↗

Intradiscal collagenase for treatment of lumbar disc herniations. A comparison of clinical results and computed tomography follow-up.

In a series of 34 patients with herniated lumbar discs, treated by intradiscal injection of highly purified collagenase, the post-treatment course has been followed-up clinically and by repeated computed tomographies (CT). Good or excellent results have been achieved in 17 patients. An only slight improvement of pain was noted in 2 patients. Fifteen patients had to be operated on due to not improved or worsened clinical symptoms. The most striking result of our CT follow-up was a tendency of the disc herniation to increase initially after collagenase injection. About two thirds of the patients had such an increase at the one week after injection control. After 6 weeks this rate had decreased to only about one quarter, but in the meantime 13 patients had to be operated. Only after 6 months most hernias of the up till then not operated patients were smaller and none were larger than before treatment. There was also a transient density decrease of the treated disc, most pronounced one week after collagenase injection. At controls 6 months later density had reached again pre-treatment levels. It is likely that the volume increase tendency of the disc material after collagenase injection is responsible for a worsening of the clinical symptoms, which not seldomly occurs during the initial post-treatment period, and in some patients makes an operation necessary.

Back Pain↗

The thermic vascular anastomosis (TVA). IV. Analysis by scanning electron microscopy.

In the present study 48 thermic vascular anastomoses performed in Sprague Dawley Wistar rats were subjected to analysis by scanning electron microscopy. Depending on the period of postoperative survival (up to 18 months) normal smooth endothelium, dissected cuffs and aneurysms were seen. Endothelial damage through heat application following bipolar coagulation was only slight. In one 18 months old anastomosis an aneurysm with a delicate network of endothelialized filaments was seen leaving a free passage for the blood flow centrally. A comparison of the literature describing the healing processes of intima following various traumas is made.

Animals↗

The thermic vascular anastomosis (TVA). II. Microvascular auscultation applied to thermic vascular anastomoses.

The study shows that microvascular auscultation with a diplomicrophone is a useful method in recording and analyzing the blood flow in nonsuture thermic vascular anastomoses. In 320 anastomoses on the rat carotid artery microvascular auscultation was found to be applicable immediately after performing an anastomosis and up to 30 days postoperatively. Records of sound tracks made at various periods of postoperative survival of the animals are presented and analyzed. A comparison with auscultation results of sutured microvascular anastomoses and of ligated arteries is made.

Animals↗