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Biomedical subjects

K Schindhelm

Publications and source records attributed to K Schindhelm.

At least 19 recordsLinked to original sources

Phagocytosis of carbon particles by macrophages in vitro.

Particles of known size ranges of carbon fibre-reinforced carbon were presented to in vitro cultures of murine macrophages. Particles of up to 20 microns diameter were phagocytosed. Larger particles were not phagocytosed but became surrounded by aggregations of macrophages, some of which migrated on to the particle surfaces. Mean rates of phagocytosis up to 2.5 particles per hour were observed. Cells presented with a large excess of particles became rounded, detached from the substrate and some underwent lysis. The implications of these findings for the fate of particulates released from implanted medical devices is discussed. It is argued that a mechanism exists where particles in the size range 8-20 microns, released from medical devices, are small enough to be phagocytosed by macrophages and transported to the lymphatics and subsequently to the vascular circulation but large enough to lodge in capillary beds of tissues remote from the implant site.

Animals

Treated xenografts as gliding tendon prostheses in an ovine model.

A model for testing the properties of gliding tendon grafts has been developed that allows anastomoses to be evaluated separately from the mid-portion of the graft. In addition, two different graft materials may be implanted in one sheep foreleg whilst maintaining control (not operated) tendons in both the operated leg and contralateral foreleg. The model has been used to evaluate the response of xenografts made from chemically treated kangaroo tail tendon (KTT) compared with autografts. At 3 month the mid-sections of the glutaraldehyde-fixed xenografts maintained between 57 and 82% of their initial ultimate tensile strength whereas lyophilized KTT dropped to 10% and autografts retained 91% of initial strength. Sterilization by gamma-radiation of wet xenografts did not affect the material and implant properties significantly. Longer term studies are necessary to determine the resorption behaviour of the xenografts. Anastomosis strengths were found to be about the same for all grafts, at about 25% of the strength of the original tendon. Alternatives need to be investigated to improve this strength.

Anastomosis, Surgical

Comparative evaluation of treated bovine pericardium as a xenograft for hernia repair.

Two forms of bovine pericardium (BPC) were assessed as hernia repair materials: non-cross-linked (lyophilized) and cross-linked through treatment with glutaraldehyde (GA). These were compared with polypropylene mesh (Marlex) in a rabbit model. Over 52 wk implantation, the GA BPC grafts developed a strong, stable, fibrous tissue replacement with good incorporation into the abdominal muscle wall. The lyophilized BPC grafts were substantially resorbed within 12 wk of implantation, however the thin, fibrous replacement tissue was inadequate for abdominal wall support. Marlex grafts provided sufficient abdominal support, however these grafts were associated with extensive adhesion formation and, in this model, fat deposition around the perimeter of the graft. Control (ungrafted) rabbit abdominal muscle in the transverse orientation had an ultimate tensile load (UTL) of 11.4 +/- 5.1 N (x +/- s.d.) and a strain at UTL of 35 +/- 12% (n = 169). At 52 weeks the UTL of the repair sites was 7.3 +/- 4.5 N (n = 6), 5.1 +/- 3.5 N (n = 6) and 5.6 +/- 2.7 N (n = 6) for GA BPC, lypophilized BPC and Marlex grafts, respectively.

Animals

Beta 2-microglobulin kinetics in end-stage renal failure.

The kinetics of beta 2-microglobulin (beta 2m) were studied in five anephric or anuric hemodialysis patients. Human beta 2m was isolated from peritoneal dialysate using ion-exchange and gel chromatography and radiolabeled with 125I. Patients were injected with 10 microCi labeled beta 2m. In one study (N = 4), plasma activity was measured over 72 hours. In a second study (N = 4), patients received low-flux dialysis 24 hours after injection and high-clearance dialysis (Bellco BL655) at 48 hours. Plasma activities were fitted to a three-compartment, variable volume model. Endogenous beta 2m levels (radioimmunoassay) were 56 +/- 6 mg/liter. The beta 2m distribution volume was 12.7 +/- 2.0 liter (0.20 +/- 0.03 liter/kg) and the non-renal clearance was 3.0 +/- 0.4 ml/min. The generation rate, 9.9 +/- 1.7 mg/hr (0.16 +/- 0.04 mg/kg/hr), was similar to that measured in subjects with normal renal function. The three compartment model derived from the turnover data gave an adequate fit of the arterial concentrations of endogenous and exogenous beta 2m during low-flux (nil beta 2m clearance) and high-clearance (beta 2m clearance of 19 ml/min) dialysis. Simulations based on this model indicate that extracorporeal treatment can at best remove about 50% of weekly production. These results suggest that beta 2m production is not increased in dialysis patients, that there is substantial non-renal beta 2m clearance, and that the amount of beta 2m that can be removed by extracorporeal therapy is therefore limited.

Adult

Autograft and Leeds-Keio reconstructions of the ovine anterior cruciate ligament.

This study compared the Leeds-Keio prosthesis with grafting of autogeneic patellar tendon for the reconstruction of the ovine anterior cruciate ligament under controlled conditions. Reconstructed knees from six sheep of each group were evaluated at 12, 26, and 52 weeks postreconstruction with respect to clinical assessment, gross pathology, mechanical properties, and histology. Although no difference in clinical assessment (anteroposterior draw, range of motion, and function) was noted between the prosthesis reconstruction and the autograft reconstruction, the prosthesis provided a higher strength initially, which remained relatively constant over the one-year study. However, prosthesis wear was observed, with up to 50% of Dacron fibers ruptured in some cases. Histologic sections indicated that in the ovine model, the Leeds-Keio prosthesis should be considered an artificial device and not a scaffold or stent that supports aligned collagenous growth. The autograft had low strength at 12 weeks, which increased over the study period. Despite acceptable clinical performance and adequate mechanical properties up to one year postimplantation, neither reconstruction approached the clinical or mechanical performance of the normal anterior cruciate ligament in the ovine model.

Animals

Measurement of the mechanical properties of the ovine anterior cruciate ligament bone-ligament-bone complex: a basis for prosthetic evaluation.

The reported ultimate tensile stress of the anterior cruciate ligament varies greatly, ranging from 13 to 147 MPa. This study shows that the orientation and degree of flexion of the bone-ligament-bone complex significantly alter the apparent ultimate tensile properties (ultimate tensile stress ranging from 60 +/- 3 to 123 +/- 15 MPa, ultimate specific extension from 37 +/- 7 to 93 +/- 20%), whilst the method chosen for measuring extension also affects the calculated specific extension of the bone-ligament-bone complex. It is suggested that, for considerations of prosthesis design and evaluation, the mechanical properties of the bone-ligament-bone complex should be measured in anterior draw and extension measured using points as close as possible to the positions of the ligamentous attachment sites.

Animals

Endothelial cell seeding of small diameter vascular grafts.

This study examines, under flow conditions, the adhesion of endothelial cells to 3 mm diameter fibronectin (Fn)-coated expanded polytetrafluoroethylene (PTFE) vascular grafts. Cultured ovine carotid artery endothelial cells were labelled with 35S-methionine. The grafts were seeded with endothelial cells (1.5 x 10(6)/ml) by rolling for 1 h at 37 degrees C and then either cultured to confluence for 48 h or flow tested immediately. Cell attachment to grafts (n = 5) was evaluated in an in vitro flow circuit, using flows of up to 330 ml/min. Ex vivo studies (n = 5 grafts) were conducted without anticoagulant using autologous cells in a sheep model. Grafts were inserted into an externalized carotid-jugular shunt and exposed to blood flows of approximately 150 ml/min for 3 h. One hour seeded and 48 h cultured grafts demonstrated greater than 95% cell retention following in vitro flow studies. Ex vivo studies of 48 h cultured grafts gave endothelial cell retention of 81% with no sign of thrombogenicity. Furthermore, a preliminary 24 h ex vivo study has shown greater than 95% retention. This study demonstrates the firm attachment of seeded endothelial cells to Fn-coated PTFE grafts in the sheep model.

Animals

Collagen cross-linking and resorption: effect of glutaraldehyde concentration.

Cross-linked collagen bioprostheses usually are designed to be inert and nonresorbable, resulting in fatigue and wear failure in high-stress environments. Eventual replacement of the implant, although minimizing strength loss during resorption, would result in a graft with reparative ability. Kangaroo tail tendon (KTT) partially cross-linked with glutaraldehyde (GA) was evaluated in vitro for resistance to bacterial collagenase digestion and in vivo for biocompatibility and resorbability in an intramuscular implant assay. Cross-linking was quantified by thermal denaturation studies. Incomplete cross-linking was achieved with concentrations of GA less than 0.1% (w/v). KTT cross-linked in greater than or equal to 0.05% GA were collagenase resistant being incompletely digested after 240 h. Cross-linking of KTT with low concentrations of GA resulted in partial collagenase resistance and slowed resorption.

Animals

Transport and kinetics in synthetic and immunospecific adsorption columns.

Immunological or physicochemical adsorption can be used to remove specific proteins from plasma. The most common adsorption systems are based on the principle of affinity or adsorption chromatography. An adsorption column can be considered to be analogous to a packed bed, and transport is modeled by accounting for solute convection, accumulation, uptake, and axial dispersion within the bed. Adsorption kinetics and transport within particles of the packed bed need to be described. The solution of model equations is solute specific and generally requires numerical methods. The application of column adsorption systems to on-line therapeutic adsorption is constrained by flow considerations, which influence processing time; adsorbent capacity, which dictates column volume and hence design; and adsorbent stability, which may impact on patient safety.

Adsorption

IgG immunoadsorption in experimental allergic neuritis: effect on antibody levels and clinical course.

The effect of IgG immunoadsorption upon the course of chronic experimental allergic neuritis (EAN) is described. Miniature membrane plasma separators coupled with a Protein A (PA)-Sepharose immunoadsorbent column were used to perform upon conscious rabbits 5 IgG immunoadsorption treatments over 6 days. Quantitation of anti-myelin IgG and IgM by ELISA revealed that 55-65% of plasma IgG was removed per treatment. Rapid post-treatment antibody rebound was observed for anti-myelin IgG although no antibody overshoot above control levels could be observed. Anti-myelin IgM levels remained relatively unaffected by PA immunoadsorption. Comparisons of clinical scores between control and treatment animals showed that IgG immunoadsorption was significantly beneficial (day 1 post-treatment p less than 0.001; day 2 post-treatment p less than 0.05). However, rapid relapse was observed in all treatment animals such that by day 3 post-treatment no significant clinical difference between control and treatment groups could be observed. IgG immunoadsorption suppresses the clinical progression of chronic EAN in a manner similar to that seen with plasma exchange. This finding suggests that antibody modulates early disease pathogenesis.

Animals

Mixed-mode therapy: kinetic analysis and acute clinical evaluation.

A mixed therapeutic modality was devised in which patients with chronic renal failure were treated with a combination of continuous ambulatory peritoneal dialysis (CAPD; two daily 4-hour exchanges per day; 16 h dry belly) and hemodialysis (1 session per week). Kinetic modeling analysis indicated that a time-averaged urea concentration equivalent to CAPD could be obtained with a Kt/V value of 1.2-1.6, depending on patient parameters, for the single-weekly hemodialysis. The therapy format was acutely evaluated in a 2-week clinical trial on 4 patients. Excursions in small-solute concentration were virtually equivalent to those predicted from theory. Adequate fluid removal was obtained in the 2 CAPD exchanges and blood pressure was well controlled. As a result of the success of the acute trials, and since this format may offer potential lifestyle advantages to patients who possess dual access, a chronic trial of mixed-mode therapy seems advised.

Aged

Experimental allergic neuritis: effect of plasma infusions.

The effect of intravenous fresh frozen plasma (FFP) and artificial plasma infusions upon the clinical course of chronic experimental allergic neuritis (EAN) in the rabbit was investigated. A total of 12 animals allocated to treatment groups received rabbit FFP or a gelatin plasma expander Haemaccel (Hoechst) and were compared to 13 control non-treated animals. Animals receiving Haemaccel at a rate of 15 ml/kg/day for 7 days showed no significant clinical benefit at any stage. However, animals receiving 15 ml/kg/day FFP for 8 days showed significant clinical benefit during treatment initiated at the onset of definite neurological symptoms of EAN (Mann-Whitney U test, day 4 post-allocation P less than 0.05; day 6 post-allocation P less than 0.01; day 8 post-allocation P less than 0.05). Relapse was observed after cessation of treatment such that comparisons of clinical scores at day 14 and 22 post-allocation revealed no significant differences. Analysis of plasma anti-myelin IgG levels by ELISA showed that non-immunogenic plasma volume expansion decreased anti-myelin IgG concentrations immediately by an average of 34% but had no long-term effect. In contrast, anti-myelin IgG concentrations in FFP infused animals were significantly decreased, compared to controls, when measured 24 h after the last infusion (Student's t-test P less than 0.05). Identical percentage weight losses for both control and treatment groups post-allocation indicated that this decrease was immunologically mediated and not due to plasma dilution. Similar plasma cortisol concentrations measured in both groups showed no significant artifactual induction of endogenous steroid production. Infusions of FFP during early disease progression are able to mediate clinical remission in animals with chronic EAN.

Animals

Induction of endothelial cell migration by proline analogs and its relevance to angiogenesis.

The proline analogs cis-4-hydroxy-L-proline, 3,4-dehydro-L-proline, and 2-azetidinecarboxylic acid induced increases in the migration rate of retinal capillary endothelial cells, aortal endothelial cells, corneal endothelial cells, aortal smooth muscle cells, and retinal pericytes. cis-4-hydroxy-D-proline did not. The optimal concentration for migration induction by any of the active agents was approximately 10(-5) M. At higher concentrations (5 x 10(-4) M) migration was not induced or was inhibited. When tested by subcutaneous implant assays in rabbits, cis-4-hydroxy-L-proline and 2-azetidinecarboxylic acid consistently elicited a marked angiogenic response. Whereas these compounds are known to modulate collagen synthesis and secretion, the concentrations at which they are effective inducers of migration suggest that they may have a more specific target than general collagen synthesis.

Animals

Membrane plasma exchange in experimental allergic neuritis: effect on antibody levels and clinical course.

The effect of membrane plasma exchange on the course of chronic experimental allergic neuritis in rabbits is described. Using miniature membrane plasma separators conscious animals were treated with 4 exchanges over 5 days removing one plasma volume per procedure and using a non-immunogenic gelatin plasma solution as replacement. Comparisons of clinical scores between control and treated groups indicated that treated animals received significant benefit from plasma exchange (day 1 post-treatment P less than 0.002; day 3 post-treatment P less than 0.01). However, relapse was observed in all plasma exchanged animals such that by day 8 post-treatment there was no significant difference in clinical scores between the two groups. Quantitation of anti-myelin IgG and IgM by ELISA assay showed that 55-60% of circulating antibody was removed per exchange. Rapid post-exchange antibody rebound was observed for both IgG and IgM so that pre-exchange levels were re-established within 24-48 h. However, no significant overshoot in circulating levels of anti-myelin IgG nor IgM could be observed. It is probable that long-term remission as a result of therapeutic plasma exchange is a function of effective circulating plasma removal and if present, the suppression of ongoing antigenic stimuli.

Animals

A comparison of the effects of plasma exchange and immunoadsorption on anti-insulin antibody synthesis in rabbits.

Plasma exchange (PE) and ex vivo immunoadsorption (IA) may be applicable to the removal of anti-insulin antibodies (AI-Ab) from diabetic patients. However, the removal of antibodies may prompt an increase in their rate of synthesis and an overshoot of antibody levels which may be deleterious to the patient. The effects of both PE and IA on AI-Ab synthesis were studied in a rabbit model. Rabbits were immunized with insulin and the resulting AI-Abs removed by both plasma exchange and specific immunoadsorption. Following AI-Ab removal by PE no increase in AI-Ab synthesis or antibody overshoot occurred. However a large increase in AI-Ab synthesis and overshoot occurred following specific AI-Ab removal by immunoadsorption. Despite similar reductions in AI-Ab levels by PE and IA, no increase in antibody synthesis occurred due solely to antibody removal. It is likely that antigen released from the immunoadsorbent stimulated the increase in antibody synthesis following immunoadsorption. These findings are relevant to the clinical application of both PE and IA.

Animals

Chronic experimental allergic neuritis. An electrophysiological and histological study in the rabbit.

Ten adult outbred New Zealand white rabbits were inoculated with a single multiportal dose of purified bovine peripheral nerve myelin and Freund's adjuvant containing 500 mg of nerve antigen. Seven animals developed chronic relapsing or progressive disease which was followed by clinical examination for 14 months. Electrophysiological studies showed marked slowing of motor conduction velocity, dispersion of the evoked muscle action potential (MAP) and reduction in amplitude of the MAP derived from distal stimulation. Histological examination of the peripheral nervous system showed at 12 months a marked hypertrophic neuropathy in the nerve roots with well developed onion bulbs, active demyelination and a moderate nerve fibre loss. It is suggested that these animals provide a reliable and predictable model for human chronic inflammatory demyelinating polyneuropathy (CIDP) which should prove valuable for therapeutic trials and studies of pathogenetic mechanisms.

Animals

Linear response curves from an ELISA assay: measurement of anti-myelin IgG and IgM during experimental allergic neuritis.

An ELISA method for generating linear dose-response curves for measuring anti-myelin IgG and IgM is described. Linearity is achieved without logarithmic transformations. This test is used to measure plasma levels of anti-myelin antibodies in rabbits with EAN. Results are expressed as relative concentrations of antibody in arbitary mass units with 95% confidence limits fitted by inverse prediction. In the 8 animals studied, appearance of anti-myelin antibody always preceded onset of clinical signs and neither anti-myelin IgG nor IgM could be detected in any animal pre-inoculation. Five out of 8 animals displayed steady and progressive increases in anti-myelin IgG with the remaining 3 animals showing plateaus in IgG levels 21-30 days post-inoculation. Increases in anti-myelin IgG were generally parallelled by increases in disease severity. However, 2 animals showed recovery and relapse from EAN with no change in plasma levels of anti-myelin IgG. Patterns of production of anti-myelin IgM varied with some animals displaying early peaks while others showed gradual increases.

Animals