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Biomedical subjects

H Bienert

Publications and source records attributed to H Bienert.

12 recordsLinked to original sources

Scale-independent shape analysis for quantitative cytology using mathematical morphology.

A system for automatic quantification of morphological changes of cell lines, proposed for cytotoxicity tests of biomaterials, is presented. Light-micrographs of cultured cells are segmented by adaptive thresholding within a local adaptive window. Connected cells in binarized micrographs are separated by a novel morphological multiscale method, treating cells in their size-specific scale and hence resulting in scale-independent separations. Significant shape descriptors correlating well with cell toxicity are extracted from single cells. Size and compactness distributions turned out to be reliable and useful parameters, providing an alternative to the common subjective grading of shape deformations by visual inspection. The system is evaluated for several standardized toxical reference substances and is now in use for clinical biocompatibility testing.

Cell Line↗

Suramin inhibits proliferation of human arterial smooth muscle cells in vitro: potential drug for prevention of restenosis by local drug delivery.

PURPOSE: Suramin is known to inhibit proliferation of various tumor cells. This study was performed to investigate the effect of suramin on proliferating human arterial smooth muscle cells (HASMC) and thus to examine its suitability for the prevention of restenosis. METHOD: Proliferation of HASMC was stimulated with human whole blood serum (HWBS), as well as with platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF). Proliferation was assessed by measuring DNA synthesis and cell viability was assessed by measuring mitochondrial activity. RESULTS: Suramin exerted a strong attenuating effect on the proliferation stimulators HWBS, PDGF, and bFGF. A suramin concentration of 0.5 mmol/L, which is approximately twice the dosage used for systemic application, was sufficient for complete neutralization of proliferation stimulation by 10% HWBS added to the cell culture medium. CONCLUSION: The authors' data demonstrate a strong proliferation inhibiting effect of suramin on HASMC in vitro. Because it is known to interact not only with one but with a multitude of relevant growth factors, these results make suramin a most interesting substance for local application for prevention of hyperplastic neointima formation.

Arteries↗

Functionally adapted surfaces on a silicone keratoprosthesis.

BACKGROUND: Silicone intraocular lenses as well as silicone sponges and encircling bands on the bulbar surface are widely used and are well tolerated. The aim of this project is a new one-piece silicone keratoprosthesis with enhanced cell adhesion in the haptic region to optimize the keratoprosthesis stability. These investigations show how enhanced profileration of conjunctival fibroblasts and, therefore, improved tissue compatibility can be achieved by hydrophilizing and by protein immobilisation on a hydrophobic silicone surface. This allows a combination of desired chemical and mechanical properties of the silicone bulk material with surfaces of improved tissue compatibility. METHODS: Silicone foils with surface modifications of different kinds were tested. Experiments were done using cell cultures with murine fibroblasts L-929 and human conjuctival fibroblasts. Cytotoxicity assays were carried out with cells grown on the material in direct contact, as well as in indirect contact, with extracts (EN 30993-5). Viability stains by means of fluoresceindiacetate and ethidiumbromide together with morphology analyses by hemalaun-staining were performed. RESULTS: For the unmodified and modified foils themselves and their extracts any negative influence on cell cultures of murine and human cells could be excluded. There was a gradual improvement of cell morphology, spreading and proliferation dependent on the degree of surface modification. Covalently immobilised fibronectin showed the best results in contrast to adsorptive binding. CONCLUSIONS: Silicone surfaces can be modified chemically with bioactive proteins. These modifications are cell compatible and do not result in toxic reactions. The degree and type of silicone hydrophilization results in improved development of cell morphology, spreading and proliferation. Even better results are obtained after covalent binding of bioactive proteins like fibronectin. Improved biocompatibility with enhanced cellular overgrowth has been demonstrated in vitro for the modified silicone of the haptic region. We believe that this type of modification will help in reducing extrusion problems observed with former keratoprostheses.

Animals↗

Development of a new biodegradable intravascular polymer stent with simultaneous incorporation of bioactive substances.

OBJECTIVE: Due to the thrombogenicity and permanent implant nature of metallic stents, bioresorable synthetic polymers have been proposed for stents and local drug delivery systems. Bioresorbable polyesters like poly(D,L-lactide) demonstrated excellent biocompatibility in various tissues. This paper describes a novel method for the molding of these polymers. The specific CESP-process (Controlled Expansion of Saturated Polymers) is characterised by the use of the plasticizer carbon dioxide and allows the incorporation of bioactive substances at physiologic temperatures into the polymer bulk and the production of complex designed implants. METHODS: The CESP-process is characterised by the exposure of an amorphous polymer to an inert gas at high pressure with a significant lower glass transition point. The plasticizing effect makes it possible to process polylactides at a temperature close to room temperature. The low process temperature constitutes a key advantage for thermally sensitive polymers and allows the incorporation of thermally sensitive pharmaceutical additives. To obtain some preliminary information on the biocompatibility, in vitro cell toxicity testing as well as drug release assessment was performed. RESULTS: Different polymer sheets were produced using the CESP-process. Cytotoxicity was not observed in any molded polymer material. According to the mechanical and biocompatibility results Poly(D,L-lactide) (P-DL-LA) was investigated in the CESP-process. Finite element analysis was used to test the possible geometry of an adequate stent. A helical design was chosen and a stent-prototype was produced using the CESP-process. Peroxidase activity as an incorporated marker enzyme could be measured over 6 weeks. Different drug release profiles were obtained due to various pore sizes of the polymer. CONCLUSIONS: The new CESP-process can be used to process biodegradable polymers and to mold different stent geometries without inducing cytotoxic effects to the material. Furthermore, this procedure permits the simultaneous incorporation of bioactive substances during the molding process. Drug release kinetics can be regulated by different pore sizes of the material.

Absorbable Implants↗

[Reduction of cataract by plasma etching of intraocular lenses. An animal experiment study].

BACKGROUND: We studied if a modification of the silicon intraocular lens (IOL) by plasma etching is able to promote a bonding of the IOL surface and the capsular bag which might inhibit proliferation and migration of lens epithelial cells. METHODS: Silicon-disc lenses (90D, Adatomed), as disposable for regular cataract surgery, were used. Their haptic surface was etched via the use of a SO2 plasma, leaving the optic unmodified. The experiments were done on dwarf rabbits to allow for tight apposition of IOL and bag. Nine rabbits underwent extracapsular lensectomy using propofol anaesthesia and phaco/clear cornea surgical technique. Six eyes each received either no, a regular or a modified IOL. After 11 weeks the eyes were enucleated. Capsular bag and IOL were digitized using a flatbed scanner with transparency adapter. The data obtained were calibrated against a densitometric standard. The densities of the various specimen were analyzed quantitatively using self designed software. RESULTS: In aphacic eyes no significant posterior capsule opacification (PCO) was detectable. In the same time-span the regular IOL had developed a dense, heterogenous PCO. The plasma-treated IOL showed, especially in the central areas, a significant reduction of PCO as compared to untreated IOL. CONCLUSION: The reduction of PCO could not be explained by adhesion of the IOL surface and the capsular bag, which would impair migration of lens epithelial cells and thereby PCO. Likewise, lower PCO may be related to improved hydrophilic properties of the surface-modified IOL.

Animals↗

Development of a biodegradable ureteric stent: surface modification and in vitro assessment.

The aim of the present study was to develop a short bioresorbable ureteric stent and to characterize the chosen polymers with respect to surface modification, biocompatibility, and loading of a biologically active compound. As materials for the stent, poly(D,L-lactide) and poly(D,L-lactide-co-glycolide) were chosen. Degradation experiments were carried out and analytical data were obtained by contact angle measurement, X-ray photoelectron spectroscopy (XPS), and infrared spectroscopy in the attenuated reflection mode (FTIR-ATR). Gas loading technology was used to incorporate biologically active compounds, and biocompatibility of the polymers was assessed by in vitro cellular assays, applying measures such as cell morphology, proliferative activity, and membrane integrity. Our results indicate that surface modification of bioresorbable polymers is a suitable and efficient approach to improve the surface properties. Incorporation of biologically active compounds was possible without loss of activity, and in vitro assessment of cellular responses demonstrated the biocompatibility of the chosen polymers and modifications.

Absorption↗

A novel method for quantifying shape deformation applied to biocompatibility testing.

Cytotoxicity tests are important for the screening and evaluation of biocompatibility of artificial organs. Morphologic changes of cells that were contacted biomaterials or biomaterial extracts indicate their toxicity. However, information on cytotoxic effects is still obtained by subjective visual inspection of microscopic samples. In this article, a novel computer assisted method is introduced. The automatic analysis of digitized micrographs is achieved in several stages: segmentation, separation, classification, and measurement. The segmentation of the image is provided by a new local adaptive thresholding technique, which adapts the threshold window sizes onto local gray level distribution and yields optimal window sizes. The actual threshold is obtained by maximizing interclass variances and minimizing intraclass variance. For the separation of connected cells, the binarized samples are cleaned from "false" markers by morphologic filtering. The subsequent separation is a two phase approach. Information levels are generated top-down by successively applying an enhanced erosion operator, which yields markers and filters noise usually evolving from multiple erosions. The converse bottom-up integration of the eroded markers is performed by successively applying an enhanced dilation operator, which reconstructs the cells and prevents merging of already separated objects. The subsequent measuring provides quantitative parameters of the distribution of size and compactness of the cells contained within the sample. The method was evaluated by L-929 fibroblasts that were in contact with 0%, 5%, and 10% concentrations of ethanol. For each concentration, 268 images of the cell populations were captured. The obtained quantitative parameters are highly correlated to the common verbal description of morphologic changes. Therefore, the proposed automatic method has several advantages compared with subjective examinations. The results allow an objective comparison of the quantification of phenomena; the subjective influence of the observer is eliminated; and the laboratory staff is relieved of time consuming routine work.

Animals↗