Biomedical subjects

D Bakker

Publications and source records attributed to D Bakker.

At least 55 records · Page 3Linked to original sources

PubMed1993

Cell-seeding and in vitro biocompatibility evaluation of polymeric matrices of PEO/PBT copolymers and PLLA.

A bilayered matrix has been evaluated in vitro as a carrier for autografts of cultured epidermal keratinocytes and dermal fibroblasts, to be used as a skin substitute in deep dermal skin defects. A poly-L-lactide (PLLA) and an elastomeric and biodegradable poly(ethyleneoxide)-poly(butyleneterephthalate)(PEO-PBT++ +)copolymer, called Polyactive, were chosen as the constituents of the matrix. The substrate properties of the bilayers for human and rat epidermal keratinocytes and dermal fibroblasts were assessed. Keratinocytes attached and expanded into confluent sheets on both the routine cell culture plastic (TCPS) and the experimental substrates. Morphology of the cells cultured on the biomaterials was found to be comparable with the morphology of those grown on TCPS. In contrast to dense films, porous PEO:PBT copolymer and PLLA appeared poor substrates for fibroblasts. Long-term (in vivo) degradation of the biomaterials was mimicked in vitro to screen the biomaterials for any release of toxic substances. Culturing keratinocytes and fibroblasts in media based on the artificially aged biomaterials did not result in any negative effects on proliferative activity or morphological appearance of the cells.

Animals↗

PubMed1993

A new biodegradable matrix as part of a cell seeded skin substitute for the treatment of deep skin defects: a physico-chemical characterisation.

A bilayered matrix, as an integral part of a cell seeded skin substitute for deep dermal skin injury, is described. The skin substitute has been given a dense top layer, to function as a substrate for keratinocyte culture, and a porous under layer for wound adhesion and to serve as a template for dermal regeneration. This porous under layer may be seeded with dermal fibroblasts for improvement of neodermis formation. The elastomeric hydrogel Polyactive¿, a biodegradable polyether/polyester block copolymer, has been selected as the top layer component and Polyactive or Poly-L-lactide (PLLA) as the constituents of the under layer; this to ensure that the matrix is degradable and fulfils certain mechanical requirements. Polyactive top layers were found to be highly permeable to water vapour (+/-30 g/m -2 h -1 kPa) -1 and to molecules up to 150 kD (human IgG). Tensile properties of the bilayered matrices were to a considerable extent dependent on top layer and under layer composition and thickness. Elasticity moduli were in the range of those previously reported for human skin. Varying the weight ratios of the soft/hard segments of the Polyactives and/or substituting PLLA for Polyactive as the under layer component, allows the modulation of the matrix to specific application site dependent demands.

Biodegradation, Environmental↗

PubMed1992

Discrepancies between single stimulus and multiple stimulus visual field examinations with the Peritest semi-automated perimeter in glaucoma patients.

In a total of 24 randomly selected eyes of glaucoma patients, tested with the automated (single stimulus) and the manual (multiple stimulus) modes of the semi-automated Peritest perimeter, the numbers and depths of defects were compared as estimated with these two different measurement strategies. It was found that with the manual technique considerably less defects were detected than with the automated strategy.

Glaucoma↗

PubMed1992

Characterization of the antigenic and adhesive properties of FaeG, the major subunit of K88 fimbriae.

The two K88 serotypes, K88ab and K88ac, differ in terms of antigenic and adhesive properties. The structural determinants of the serotype-specific epitopes and the identify of the amino acid residues involved in fimbriae-receptor interaction were studied by the construction and analysis of K88 hybrid proteins in which various parts of the K88ab and K88ac fimbrial subunit FaeG were exchanged, and by in vitro mutagenesis of non-conserved amino acid residues. Using a set of monoclonal antibodies, several regions or amino acid residues involved in the formation of serotype-specific antigenic determinants were located. The haemagglutinating activity of the hybrid and mutant proteins revealed several amino acid residues involved in the formation of the receptor binding site. A clear correlation was found between the receptor binding site and the serotype-specific antigenic determinants.

Amino Acid Sequence↗

PubMed1992

Identification of minor fimbrial subunits involved in biosynthesis of K88 fimbriae.

The nucleotide sequences of the genes faeF, faeH, faeI, and faeJ encoding K88 minor fimbrial subunits were determined. Analysis of the primary structure of the gene products revealed that all four proteins are synthesized with an amino-terminal signal sequence. The molecular masses of the mature FaeF, FaeH, FaeI, and FaeJ proteins were calculated to be 15,161, 25,461, 24,804, and 25,093 Da, respectively. FaeH, FaeI, and FaeJ showed significant homology with FaeG, the major fimbrial subunit of K88 fimbriae. Mutations in the respective genes were constructed. Analysis of the mutants showed that the minor fimbrial subunits FaeF and FaeH play an essential role in the biogenesis but not in the adhesive properties of the K88 fimbriae. Mutations in faeI or faeJ had no significant effect on K88 production or adhesive capacity. Specific antisera against FaeF and FaeH were raised by immunization with hybrid Cro-LacZ-FaeF and Cro-LacZ-FaeH proteins. Immunoblotting and immunoelectron microscopy revealed that FaeF and FaeH are located in or along the K88 fimbrial structure.

Amino Acid Sequence↗

PubMed1991

Reactions of cells at implant surfaces.

The surface of implants is an important parameter in host-implant integration. Several strategies can be used to obtain integration, such as the application of grooves or pores at the implant surface. Most of these surface alterations, however, will lead to an increase of total implant surface area which might influence the inflammatory response to an implant. As far as integration with bone is concerned several biomaterials have been successful in mimicking this material, by having similar crystals at their surface (calcium phosphate ceramics) or by containing a certain amount of calcium and phosphorus. Polyactive, a poly(ethylene oxide)-poly(butylene terephthalate) segmented copolymer, also possesses favourable integration properties with bone, but initially lacks calcium and phosphorus. It is proposed that the application of hydrogels as biomaterial may add a new dimension to integration capacity.

Adsorption↗

PubMed1991

Localization and function of FanH and FanG, minor components of K99 fimbriae of enterotoxigenic Escherichia coli.

Specific antisera against FanG and against FanH were prepared by immunization with hybrid Cro-LacZ-FanG and Cro-LacZ-FanH proteins, respectively. Immunoblotting with these antisera revealed the presence of FanG and FanH as minor components in purified K99 fimbriae. Mutations were constructed in fanG and fanH and cells defective in FanG or FanH were characterized by ELISA, immunoblotting, adhesion assays and electron microscopy. A minicell experiment showed that the mutations in fanG or fanH had no effect on the expression of the other K99-specific proteins. Cells defective in FanG produced no fimbriae and did not agglutinate horse erythrocytes, but cell-free heat-shock preparations of these cells still bound the K99 glycolipid receptor. Cells defective in FanH produced 1-2% of the K99 fimbriae as compared with wild-type K99 producing cells. These mutant fimbriae appeared to be shorter but were still capable of binding the K99 glycolipid receptor. Apparently, FanG and FanH are not required for binding the K99 receptor. These results and analysis of K99 mutants by immunoblotting using a specific antiserum against another K99 minor component, FanF, indicated that the combinations FanF/FanG and FanF/FanH are required for the initiation and elongation (length determination) of K99 fimbriae formation, respectively.

Adhesins, Escherichia coli↗

PubMed1991

Structure and function of periplasmic chaperone-like proteins involved in the biosynthesis of K88 and K99 fimbriae in enterotoxigenic Escherichia coli.

The nucleotide sequence of faeE and fanE, two genes involved in the biosynthesis of K88 and K99 fimbriae, respectively, was determined and the amino acid sequence of the FaeE and FanE proteins was deduced. Immunoblotting of subcellular fractions with an antiserum raised against purified FaeE confirmed that FaeE is located in the periplasm. Indications were obtained that FaeE functions as a chaperone-like protein. Its interaction with the fimbrial subunit (FaeG) in the periplasm stabilized this polypeptide and prevents its degradation by the cell-envelope protease DegP. Furthermore, FaeE prevents the formation of FaeG multimers which cannot be incorporated into fimbriae. The reactions of the FaeE/FaeG dimers with a set of monoclonal antibodies directed against the various epitopes present on K88 fimbriae revealed that the fimbrial subunits associated with FaeE were present in a conformation resembling their native configuration. Indications about the domains in FaeG involved in the interaction with FaeE are discussed.

Amino Acid Sequence↗

PubMed1991

New alloplastic tympanic membrane material.

For patients with a totally empty middle ear a total alloplastic middle ear prosthesis (TAM) has been developed consisting of a macroporous hydroxyapatite canal-wall segment as a foundation system from which a dense hydroxyapatite ossicular chain is suspended. To connect the ossicular chain, we developed an alloplastic tympanic membrane made from a polymer. Light microscopy, morphometry, and autoradiography as well as various electron microscopy techniques were used in this study to evaluate the biocompatibility of Polyactive, a polyether polyester copolymer, after implantation in the rat middle ear. After between 2 and 4 weeks, implants were completely covered by tympanic-membrane epidermis and epithelium. Polyactive provoked a mild foreign-body reaction, showed a degradation rate of 54 percent after 1 year, and was nontoxic. Growth of fibrous tissue and bone into Polyactive copolymer indicated appropriate implant fixation by mechanical interlocking. The fixation of Polyactive by ingrowth of bone is promising, not only in terms of the amount of bone but also in terms of the bone/polymer interface. The latter is indicative of bonding osteogenesis in a way similar to that reported for hydroxyapatite implants. The results of this study showed that Polyactive copolymer is suitable as a degradable alloplastic tympanic membrane, both as a temporary scaffolding for the repair of tympanic membrane perforations and as a tympanic membrane in the TAM.

Animals↗

PubMed1991

[The Dutch Reading Test for Adults: a measure of premorbid intelligence level].

The construction of the Dutch Adult Reading Test (DART) is described. The DART is the Dutch version of the National Adult Reading Test. Both tests consist of a series of words with an irregular pronunciation. The score on the test is a predictor of premorbid intelligence of brain damaged patients. Furthermore, results of reliability and validation studies with the DART are reported. The main findings, which were obtained with the NART, were replicated by the DART. These findings consist of a high correlation (.85) with verbal intelligence in healthy controls (n = 22) and insensitivity to cerebral deterioration in brain damaged and demented patients (n = 53). The test also appeared to be insensitive to cognitive deterioration in a group of psychotic patients (n = 43).

Adult↗

PubMed1990

[Trends in incidence of cancer in southeast North Brabant and North Limburg during 1975-1986; report from the IKZ/SOOZ cancer registration (Integrated Cancer Center South/Cooperative Organization Oncology Hospitals)].

Trends in the incidence of cancer have been determined in the Southeast of the Netherlands from 1975 to 1986 inclusive by means of the regional SOOZ cancer registry. Age-adjusted registration rates of cancer in males rose rather steeply from more than 371 per 10(5) men-years to 424 in 1982-3 and fell thereafter to 407 per 10(5) in 1986. A similar changing pattern was seen in cancer of the lung, larynx, prostate and kidney and possibly also in cancer of the head and neck. The incidence of colorectal cancer had skin melanoma increased while that of cancer of the stomach fell. Age-adjusted incidence of cancer in females rose steadily from 266 to 290 per 10(5) woman-years. A steady increase was observed for cancer of the lung, breast, bladder and melanoma and temporarily for cancer of the kidney. The incidence of cancer of the stomach and gallbladder fell continuously. These changes in incidence generally corresponded with trends in cancer mortality as registered with the Central Bureau of Statistics. Bias from changes in data collection and coding appears to be negligible, but a certain influence on the registration rates from the markedly increased number of consultants managing cancer patients is likely. Contrasting trends in male and female rates of cancer of the airways illustrate the marked changes in smoking patterns since the sixties.

Female↗

PubMed1990

Tissue/biomaterial interface characteristics of four elastomers. A transmission electron microscopical study.

The tissue/biomaterial interface reactions of four elastomers--selected as candidates for scaffolding for tympanic membrane tissue in a total alloplastic middle ear prosthesis--were studied at the electron microscopical level after implantation in the rat middle ear. Time-dependent changes in the phagocyte/polymer interface suggested degradation of porous implants made of Estane polyether urethane, polypropylene oxide, and a poly(ethylene oxide hydantoin) and poly(tetramethylene terephthalate) segmented polyether polyester copolymer (HPOE/PBT copolymer), but not of dense Silastic silicone rubber implants. Silastic was always encapsulated in fibrous tissue. Contact between fibrous tissue and HPOE/PBT copolymer or Estane was established in the third month, but fibrous tissue was never seen close to polypropylene oxide. Bone made contact only with Estane and HPOE/PBT copolymer implants. The bone/copolymer interface showed an electron-dense layer morphologically similar to that seen between bone and hydroxyapatite ceramic, suggesting that with respect to bone HPOE/PBT copolymer behaves like a bioactive implant material. The electron-dense layer was absnet at the bone/Estane interface. Estane and especially HPOE/PBT copolymer seem to be suitable as alloplastic tympanic membrane because of their interface behavior with respect to fibrous tissue and bone.

Animals↗

PubMed1990

Biocompatibility of a polyether urethane, polypropylene oxide, and a polyether polyester copolymer. A qualitative and quantitative study of three alloplastic tympanic membrane materials in the rat middle ear.

The biocompatibility of porous implants made of Estane 5714 F1 polyether urethane, polypropylene oxide, and a poly(ethylene oxide hydantoin) and poly(tetramethylene terephthalate) segmented polyether polyester copolymer (HPOE/PBT copolymer), which were selected as candidates for an alloplastic tympanic membrane, was assessed after implantation in rat middle ears for periods of up to 1 year. Implantation of the materials led to tissue reactions initially associated with the wound-healing process, whereas after 1 month not only the presence of macrophages and foreign-body giant cells surrounding the implant materials but also implant degradation were characteristic for a foreign-body reaction. Macrophages and foreign-body giant cells dominated the picture of the tissue surrounding polypropylene oxide. The altered morphology of these cells, the persistent infiltration of the implantation sites by exudate cells, and the premature death of five rats in the 1-year group suggest that polypropylene oxide degradation was accompanied by the release of toxic substances. Estane and copolymer degradation did not induce tissue responses reflecting implant toxicity, and tympanic membranes given these alloplasts showed a normal healing pattern. Inclusions in the cytoplasm of macrophages associated with degradation and phagocytosis of all of the polymers under study were found to contain iron, silicon, titanium, and aluminum. Growth of fibrous tissue and bone, the latter into Estane and HPOE/PBT copolymer implants, indicated appropriate implant fixation by tissue, although macrophages and foreign-body giant cells were present as well. Especially the fixation of copolymer by ingrowth of bone seems promising in terms of the amount of bone in the pores and the electron-dense bone/copolymer interface. The latter is indicative for bonding osteogenesis. The HPOE/PBT copolymer is a better candidate for alloplastic tympanic membrane than Estane, and the use of polypropylene oxide cannot be recommended.

Animals↗

PubMed1990

The behavior of alloplastic tympanic membranes in Staphylococcus aureus-induced middle ear infection. I. Quantitative biocompatibility evaluation.

The biocompatibility of dense Silastic implants and porous implants made of Estane 5714 F1 polyether urethane, polypropylene oxide, and an HPOE/PBT segmented polyether polyester copolymer was evaluated during an induced Staphylococcus aureus middle ear infection. The middle ear response to infection seemed not to be affected by the presence of implants made of either of the polymers. Light microscopical morphometry and transmission electron microscopy showed degradation of the porous implants under study, but not of Silastic implants, which were invariably surrounded by a fibrous capsule. This finding, combined with the degree of porous implant degradation, the composition of the tissues surrounding the implants, and the tissue/biomaterial interface reactions are consistent with the results obtained in the noninfected middle ear. Round-cell infiltrates however, were predominantly associated with implants made of polypropylene oxide and HPOE/PBT copolymer; while the presence of (phagocytosed) microbial debris was associated with copolymer. The present findings indicate that with respect to implant behavior in infected surroundings Estane is the best porous material, whereas the behavior of Silastic implants did not deviate from that in non-infected ears.

Animals↗

PubMed1990

The behavior of alloplastic tympanic membranes in Staphylococcus aureus-induced middle ear infection. II. Morphological study of epithelial reactions.

Epithelial reactions to Silastic, Estane polyether urethane, polypropylene oxide, and a poly(ethylene oxide hydantoin) and poly(tetramethylene terephthalate) segmented polyether polyester copolymer were investigated after implantation in tympanic membranes and submucosa of noninfected and Staphylococcus aureus-infected rat middle ears. Porous implants made of Estane and polypropylene oxide were completely covered by tympanic-membrane connective tissue, epidermis, and epithelium in 2 weeks and those made of copolymer in between 2 and 4 weeks postoperatively. Silastic implants, which were dense, were not enveloped by tympanic-membrane tissue but rejected. Starting in the 6th postoperative month the proliferative activity and structure of both the tympanic membrane epithelium and epidermis became normal except for the presence of iron-containing secretory epithelium near polypropylene oxide. After initial swelling caused by the surgical trauma, neither the proliferative activity nor the composition of the epithelium covering submucosal implants was affected by the presence of any of the biomaterials. Infection of middle ears bearing implants induced epithelial reactions similar to those associated with infected middle ears without an implant.

Animals↗

PubMed1990

K88 fimbriae as carriers of heterologous antigenic determinants.

The K88 fimbriae of enterotoxigenic Escherichia coli are strongly immunogenic antigens that can be used to evoke protective immunity. To find out whether these fimbriae can be used as carriers for foreign epitopes, a highly variable region present in the primary structure of the different K88 variants was replaced with five different heterologous epitopes to investigate to what extent these insertions affected the expression, assembly (biogenesis), stability and immunogenic properties of the resulting hybrid fimbriae. Amino acid residues 163-173, were replaced using site-directed in vitro mutagenesis and the hybrid fimbriae were tested for these aspects using ELISA, immunoelectronmicroscopy and immunoblotting. Replacement of this highly variable region did not affect the biosynthesis of fimbriae, although all mutations tested resulted in a reduced expression depending on the epitope inserted. Testing of the different hybrid fimbriae with a panel of monoclonal antibodies raised against the various K88 serotypes K88ab, K88ac and K88ad indicated that replacement of amino acid sequence 163-173 did not affect conserved or K88ab specific epitopes but the K88ac and K88ad specific conformation was lost. Immunization with hybrid fimbriae raises antibodies specific for the inserted heterologous epitopes.

Amino Acid Sequence↗

PubMed1990

Growth-rate-dependent synthesis of K99 fimbrial subunits is regulated at the level of transcription.

Increase in the production of the fimbrial adhesion K99 by enterotoxigenic Escherichia coli in continuous cultures at specific growth rates above 0.25 h-1 was shown to be independent of the nature of the growth-limiting nutrient. The correlation between specific growth rate and K99 production was also found to be independent of the copy number of the K99 operon. Introduction of additional copies of the K99 regulatory region did not affect growth-rate-dependent K99 production in wild-type strains, indicating that no hypothetical regulatory host factor is titrated by the K99 regulatory region. Regulation at the transcriptional level was measured with galactokinase gene fusions. The transcription of the fimbrial subunit gene increased with an increase in specific growth rate. This growth-rate-dependent transcription was found to originate from the strong promoter PA. Transcription originating from the weaker promoter PB was independent of growth rate. The results indicated that transcriptional regulation at PA is involved in the growth-rate-dependent regulation of K99 production.

Adhesins, Escherichia coli↗

PubMed1990

Structure, localization and function of FanF, a minor component of K99 fibrillae of enterotoxigenic Escherichia coli.

The DNA sequence of the K99 fanF gene, encoding FanF, was determined. An open reading frame of 999 bp was found. The primary structure of FanF was deduced and analysis revealed the presence of a signal sequence of 22 amino acid residues. The mature protein contains 311 amino acid residues (Mr 33,905 D). The amino acid sequence of FanF showed similarity with the K88ab major subunit FaeG. A specific mouse antiserum against FanF was prepared by constructing and purifying a hybrid Cro-LacZ-FanF protein. Minicell analysis, immunoblotting and immunoelectronmicroscopy revealed a pool of FanF in the periplasm of K99-producing cells and showed, furthermore, that FanF is a minor component of K99 fibrillae, present at the top and in or along the shaft of the K99 fibrillar structures. A fanF mutant plasmid was constructed. Cells harbouring this plasmid produced all K99-specific proteins, except FanF, but produced 0.1% of the K99 fibrillae relative to 'normal' K99-producing cells. Electron microscopic observations showed that cells defective in fanF produce only a few (apparently short) K99 fibrillae. FanF, therefore, was supposed to play a role in initiation and elongation of K99 fibrillae formation. Thin-layer chromatography experiments involving purified receptor material showed that FanF is not required for binding of K99 fibrillae to the ganglioside receptor. Fibrillae produced by an adhesion-negative strain carrying a mutation in the K99 major fibrillar subunit were shown to contain a normal amount of FanF.

Amino Acid Sequence↗