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Biomedical subjects

B K Milthorpe

Publications and source records attributed to B K Milthorpe.

14 recordsLinked to original sources

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

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

The polymerization pattern of zinc(II)-insulin at pH 7.0.

Sedimentation equilibrium experiments were conducted at pH 7.0 using solutions of bovine insulin containing 2 mol of zinc(II) ions per six base-mol of insulin. A detailed analysis of these results revealed the existence of a stable zinc-insulin hexamer together with linked polymerization reactions. Specifically these are a background polymerization of zinc-free insulin as previously described by Jeffrey et al. ((1976) Biochemistry 15, 4660--4665) and a slight tendency for the zinc-insulin hexamer to undergo indefinite self-association. Equilibrium constants governing these reactions are reported together with equations which permit calculation of the composition of the solution at any given total concentration. Comment is made on the possible biological significance of this linked polymerization pattern, and on the likely identity of the structure of the stable zinc-insulin hexamer with that previously reported from X-ray crystallographic studies.

Animals

Polymerization pattern of insulin at pH 7.0.

Sedimentation equilibrium results, obtained with bovine zinc-free insulin (with and without a component of proinsulin) at pH 7.0, I o.2, 25 degrees C, and up to a total concentration of 0.8 g/l., are shown to be consistent with three different polymerization patterns, all involving an isodesmic indefinite self-association of specified oligomeric species. The analysis procedure, based on closed solutions formed by summing infinite series, yields for each pattern a set of equilibrium constants, It is shown that a distinction between the possible patterns can be made by analyzing sedimentation equilibrium results obtained in a higher total concentration range (up to 4 g/1.) with insulin freed of zinc and proinsulin, account being taken of the composition dependence of activity coefficients. The favored pattern, which differs from that previously reported in the literature, involves the dimerization of monomeric insulin (mol wt 5734), governed by a dimerization constant of 11 X 10(4) M-1 and the isodesmic indefinite self-association of the dimer, described by an association constant of 1.7 X 10(4) M-1. This polymerization pattern is also shown to be consistent with the reaction boundary observed in sedimentation velocity experiments.

Hydrogen-Ion Concentration

The sedimentation equilibrium of heterogeneously associating systems and mixtures of non-interacting solutes: analysis without determination of molecular weight averages.

Sedimentation equilibrium is first considered of a system in which a ligand of any size binds to an acceptor at p sites, the experimental result, obtained with either interference or absorption optics, being a distribution of total solute concentration as a function of radial distance. Theory illustrated by a numerical example, is presented which shows that this distribution may be analysed to give the activity of the unbound ligand as a function of total weight concentration. It is shown that this information may be used together with conservation of mass equations written in terms of the initial mixing composition to evaluate the equilibrium constant(s) relevant to the system. Correlation with composition evaluation by use of absorption optics (when possible) is also discussed. The procedure does not involve solution of simultaneous equations which are sums of exponentials nor differentiation of experimental results to obtain apparent weight-average molecular weights. It is general in that it leads to the evaluation of the activity of the species characterized by the smallest M(1-vp) product and, accordingly, is shown to be useful in the analysis of non-interacting as well as of interacting systems.

Binding Sites

The direct analysis of sedimentation equilibrium results obtained with polymerizing systems.

Theory is presented in relation to sedimentation equilibrium results obtained with polymerizing systems, which permits evaluation of the activity of the monomer as a function of total weight concentration. In contrast to established methods, the suggested procedure does not involve the solution of simultaneous equations which are sums of exponentials or the determination of weight-average molecular weights. A major advantage of the method is that it avoids errors inherent in differentiation and integration steps. An extrapolation to infinite filution is involved, but this is to a defined limit and is uncomplicated by the existence of critical points in the relevant plot. The method is capable of detecting possible volume changes inherent on polymer formation, of treating systems where activity coefficients of solute species are functions of total concentration and of describing the system in terms of relevant equilibrium constants. These points and comparisons with existing methods of analysis are illustrated with numerical examples and with results obtained with lysozyme at pH 6.7. The lysozyme results are interpretable in terms of either a non-ideal monomer-dimer system or a monomer-dimer-trimer system.

Binding Sites