Use of tissue transglutaminase and fibronectin to influence osteoblast responses to tricalcium phosphate scaffolds.

To explore the possibility of controlling cell interaction with biomaterials, tricalcium phosphate scaffolds were modified using the enzyme tissue transglutaminase (tTgase) in conjunction with fibronectin. Previous reports in the literature have highlighted a number of favourable responses that this protein-enzyme complex can stimulate, including enhancing both cell adhesion, and mineralisation. Fibronectin and tTgase alone were used as controls, and a series of different concentrations of tTgase and fibronectin in combination were assessed. Cell metabolic activity, alkaline phosphatase production, and collagen content were all measured in cultures up to 28 days. Using tetracycline labelling, calcium containing multilayered regions were imaged and quantified. Addition of 6 microg fibronectin resulted in increased alkaline phosphatase activity in all combinations, while increased transglutaminase resulted in more collagen in the cell lysates. Samples treated with fibronectin produced many small mineralised areas, those with 6 microg fibronectin and transglutaminase produced numerous large mineralised areas. The mixture of fibronectin and transglutaminase may prove to be a useful treatment for producing increased osteoblast differentiation on scaffolds.

Cell interactions with laser-modified polymer surfaces.

The performance of a polymeric biomaterial depends on the bulk and surface properties. Often, however, the suitability of the surface properties is compromised in favour of the bulk properties. Altering the surface properties of these materials will have a profound effect on how cells and proteins interact with them. Here, we have used an excimer laser to modify the surface wettability of nylon 12. The surface treatment is rapid, cost-effective and can cause reproducible changes in the surface structure of the polymers. Polymers were treated with short wavelength ( < 200 nm) UV light. These wavelengths have sufficient photon energy (6.4eV) to cause bond scission at the material surface. This results in a surface reorganisation with incorporation of oxygen. Surface wettability changes were confirmed using contact angle measurements. Cell interactions with the surfaces were examined using 3T3 fibroblast and HUVEC cells. Cells morphology was examined using a confocal laser scanning microscope (CLSM). Cell activity and cell number on the treated nylon were assessed using biochemical assays for up to seven days. Both fibroblasts and endothelial cells initially proliferated better on treated compared with untreated samples. However, over seven days activity decreased for both cell types on the control samples and endothelial cell activity and cell number also decreased on the treated polymer.

Properties of calcium phosphate coatings deposited and modified with lasers.

Physical, chemical and biological properties of calcium phosphate coatings fabricated by a pulse laser deposition method at room temperature (RT PLD) have been studied. In vitro evaluation of RT PLD coatings on bioresorbable polymers (Poly-epsilon-caprolactone and Poly-L-lactide) have been carried out. It was shown that both polymers support osteoblast growth, with increased cell activity, alkaline phosphatase activity and total protein content on those surfaces that have been coated. The advantages of RT PLD coatings in biomaterials surface optimization are discussed.

Effects of acetabular resurfacing component material and fixation on the strain distribution in the pelvis.

A 3D finite element (FE) model of an implanted pelvis was developed as part of a project investigating an all-polymer hip resurfacing design. The model was used to compare this novel design with a metal-on-metal design in current use and a metal-on-polymer design typical of early resurfacing implants. The model included forces representing the actions of 22 muscles as well as variable cancellous bone stiffness and variable cortical shell thickness. The hip joint reaction force was applied via contact modelled between the femoral and acetabular components of the resurfacing prosthesis. Five load cases representing time points through the gait cycle were analysed. The effect of varying fixation conditions was also investigated. The highest cancellous bone strain levels were found at mid-stance, not heel-strike. Remote from the acetabulum there was little effect of prosthesis material and fixation upon the von Mises stresses and maximum principal strains. Implant material appeared to have little effect upon cancellous bone strain failure with both bended and unbonded bone-implant interfaces. The unbonded implants increased stresses in the subchondral bone at the centre of the acetabulum and increased cancellous bone loading, resembling behaviour obtained previously for the intact acetabulum.

Activities and conformational fitting of 1,4-naphthoquinone derivatives and other cyclic 1,4-diones tested in vitro against Pneumocystis carinii.

Atovaquone is a chemotherapeutic agent used to treat pneumonia caused by Pneumocystis carinii in some immunocompromised patients. A set of cyclic 1,4-diones were tested in vitro for ability to inhibit growth of P. carinii, including 22 variously substituted 1,4-naphthoquinones, one bis-1,4-naphthoquinone, and three other quinones. For comparison, the antipneumocystic primaquine and its 5-hydroxy-6-desmethyl metabolite were also tested. At 1.0 microg/ml, seven compounds inhibited growth by at least 39%, with atovaquone at 92%; of these seven, five are 2-hydroxy-1,4-naphthoquinones, while one is a 2-chloro- and another is a 2-methyl-1,4-naphthoquinone. At 0.1 microg/ml, however, the most active compound tested was the primaquine metabolite, which inhibited growth by more than 42% at this concentration. To ascertain a structure-activity relationship, all 1,4-naphthoquinones were compared conformationally by means of computer-based molecular modeling (Spartan) incorporating the Sybyl force field. Without exception, for all 21 monomers tested, the substituent at position 3 of the 1,4-naphthoquinone favored activity most strongly when it simultaneously occupied (i) space centered at about 3 A from position 3, without projecting steric bulk from the area encompassed by atovaquone's cyclohexyl ring, and (ii) roughly planar space at about 7.3 A from position 3, without projecting steric bulk perpendicularly. This structure-activity relationship may prove useful in the rational design of better antipneumocystis agents.

Osteoblast growth on titanium foils coated with hydroxyapatite by pulsed laser ablation.

Pulsed laser ablation is a new method for deposition of thin layers of hydroxyapatite (HA) on to biomaterial surfaces. In this paper, we report activity and morphology of osteoblasts grown on HA surfaces fabricated using different laser conditions. Two sets of films were deposited from dense HA targets, at three different laser fluences: 3, 6 and 9 Jcm(-2). One set of the surfaces was annealed at 575 degrees C to increase the crystallinity of the deposited films. Primary human osteoblasts were seeded onto the material surfaces and cytoskeletal actin organisation was examined using confocal laser scanning microscopy. The annealed surfaces supported greater cell attachment and more defined cytoskeletal actin organisation. Cell activity, measured using the alamar Blue assay, was also found to be significantly higher on the annealed samples. In addition, our results show distinct trends that correlate with the laser fluence used for deposition. The cell activity increases with increasing fluence. This pattern was repeated for alkaline phosphatase production by the cells. Differences in cell spreading were apparent which were correlated with the fluence used to deposit the HA. The optimum surface for initial attachment and spreading of osteoblasts was one of the HA films deposited using 9 J cm(-2) laser fluence and subsequently annealed at 575 degrees C.

Tensile mechanical properties of polyacetal after one and six months' immersion in Ringer's solution.

The engineering polymer polyacetal (also known as polyoxymethylene) has a long history of clinical use including in the femoral component of the Freeman all-polymer knee replacement. The polymer is a promising material for novel applications including the femoral component of hip resurfacing prostheses. However, no work following the effect on mechanical properties of extended immersion in the physiological environment has been reported. Samples of five grades of polyacetal were obtained and tested on an MTS Bionix 858 at 37 degrees C in physiological saline. Three groups of specimens were tested: "as received"; immersed in Ringer's solution at 37 degrees C for one month; immersed in Ringer's solution at 37 degrees C for six months'. While there were negligible reductions in the ultimate tensile strengths, the Young's moduli of all polymer grades showed significant reductions (p < 0.01) of up to 19.5% following one months' immersion. There were negligible changes in ultimate tensile strength and stiffness between one and six months' immersion, however some polymers showed a small, but significant (p < 0.05) increase in stiffness. The percentage elongation and true stress for plastic instability showed negligible changes following immersion for one and six months'. These results encourage the use of polyacetal in load-bearing orthopaedic applications.

Acetabular morphology and resurfacing design.

The bony surfaces of 18 archaeological hemipelves were scanned using a 3D laser surface scanner and CyDir software on a Silicon Graphics workstation. The acetabular area was selected and point data from the approximately spherical bone surface saved. These data were input to a MATLAB routine that calculated the radius and centre of the best-fit sphere. The goodness of fit was estimated using the mean and standard deviation of the distance of the bone surface points from the sphere surface. Eight points, at approximately equal distances around the acetabular rim, were selected with reference to bony landmarks. A plane containing three of these points served as an orientation reference plane. The vectors joining the eight rim points to the centre of the best-fit sphere were found. The angles between these vectors and the normal to the reference plane were calculated. Paired angles were summed to give the angle subtended by the acetabular rim in four directions. The overall mean angle was 158 degrees (range of mean angles 145 degrees -173 degrees ). The largest individual angles, some exceeding 180 degrees, were in the superior-inferior direction, while the mean angle in the anterior-posterior direction, i.e. that controlling flexion-extension, was 152 degrees. Males had larger subtended angles than females, although the difference was not statistically significant. Simulated reaming increased all angles by approximately 10 degrees. The subtended angles are important parameters in the design of the acetabular component of a hip replacement and particularly important in resurfacing hip replacement when the volume available is tightly constrained.

Physical, chemical, and biological characterization of pulsed laser deposited and plasma sputtered hydroxyapatite thin films on titanium alloy.

The physical, chemical, and biological properties of pulsed laser deposited (PLD) and plasma sputtered (PS) hydroxyapatite (HA) coatings were compared. Human osteoblast-like cell responses to these coatings in vitro were assayed for proliferation and phenotypic expression. PS coatings formed smooth and continuous thin films that followed the contours of the substrate surface. PLD coatings consisted of numerous spheroidal micro- and macroparticles. The crystallinity of all coatings was quantified by comparison with the HA target used for both the PS and PLD processes. The XRD and FTIR results indicated that unannealed PLD coatings deposited at room temperature had X-ray spectra consistent with an amorphous structure and were found to dissolve after only a few hours in saline solution. Annealing at 400 degrees C increased the crystallinity (87-98%), which resulted in improved stability and cell activity. The PS coatings showed greater chemical stability than the unannealed PLD coatings and contained an approximate 15% crystalline phase, increasing to 65% postannealing. Cell proliferation and alkaline phosphatase production were significantly higher on unannealed PS specimens than the other coating treatments. There may be benefits in engineering the presence of a minor percentage of a microcrystalline phase in an amorphous or nanometer scale polycrystalline HA structure.

In vivo measurement of acetabular cement pressurization using a simple new design of cement pressurizer.

Aseptic loosening of the acetabular component remains a limiting factor in the long-term success of total hip replacement. An instrumented pressurizer has been designed to allow the intraoperative measurement of acetabular cement pressurization, which is known to contribute to implant fixation. Average intraoperative cement pressures in 16 operations performed by 2 surgeons were 49 +/- 17 kPa (6.4 +/- 2.3 psi) and 47 +/- 17 kPa (6.2 +/- 2.2 psi), and peak pressures were 76 +/- 5 kPa (10.0 +/- 0.6 psi) and 93 +/- 15.kPa (12.2 +/- 1.9 psi), comparable to previous work in vitro. The pressurization required for optimal cement penetration into cleaned low-density cancellous bone is reported to be of the order of 35 to 50 kPa (4.6-6.6 psi) for 30 to 60 seconds, and the present data show that this is attainable in vivo using a simple device.

Proximal femoral osteotomy as the primary operation for young adults who have osteoarthrosis of the hip.

We report the results of proximal femoral osteotomy that was performed to treat osteoarthrosis in twenty-three consecutive young adults (twenty-five hips) who had a mean age of thirty-eight years (range, eighteen to fifty-three years). The mean duration of follow-up was seven years (range, two to twelve years). With conversion to a total hip replacement as the end point, the rate of survival at twelve years was 67 per cent (95 per cent confidence interval, 37 to 88 per cent). Four hips (16 per cent) were converted to a total hip replacement at a mean of eight years after the osteotomy. For the patients who did not have conversion to a total hip replacement, the mean score for pain, according to the system of Merle d'Aubigné and Postel as modified by Charnley, improved from 3.4 points preoperatively to 5.1 points postoperatively, the mean score for walking ability improved from 3.9 to 4.7 points, and the mean score for range of motion improved from 3.2 to 4.2 points. These results compare favorably with those following other forms of operative treatment of osteoarthrosis of the hip in young adults. In addition, the osteotomy does not preclude subsequent replacement arthroplasty if one is necessary.

An in vitro study of a new design of acetabular cement pressurizer.

Aseptic loosening of the acetabular component remains one of the limiting factors in the long-term success of total hip arthroplasty. Cement pressurization has been shown to improve fixation. A new pressurizer has been designed that seals around the rim of the acetabulum and covers the transverse ligament notch with a flap. The results of in vitro testing of this device are presented and compared with those of pressure generated by insertion of an acetabular cup. The pressurizer allowed sustained, uniform cement pressurization. Peak pressures with the new pressurizer were 180 kPa at both the iliac region of the rim and the pole of an instrumented model acetabulum, compared with 55 kPa at the rim and 120 kPa at the pole on cup insertion. Pressures were maintained in the range of 80-90 kPa. The flap was effective in preventing cement leakage from the notch, and pressures were higher than when the flap was absent. Cup insertion alone gave only transient pressurization, substantially less near the rim of the acetabulum than at the pole. Peripheral pressurization may be significant in producing secure local fixation at the rim of the acetabulum, in particular in the region of the ilium (Charnley zone 1), where radiolucencies are most commonly observed and where stresses in the implanted acetabulum are highest. Improved rim fixation may also play a role in preventing the ingress of wear debris.

Correction of deficiencies in flexible fiberoptic sigmoidoscope cleaning and disinfection technique in family practice and internal medicine offices.

To assess whether deficiencies exist in the processing of contaminated flexible fiberoptic sigmoidoscopes in family practice and internal medicine offices and whether training of office personnel results in a correction of identified deficiencies, we conducted a prospective review of sigmoidoscope processing in family practice and internal medicine offices before and after a training course. Participants were questioned on their current endoscope processing for 17 standards before and 2 months after receiving training. The 19 offices had between 4 and 11 deficiencies per office before training, with an average of 6.8 deficiencies per office. After training, deficiencies ranged from 0 to 8, with an average of 0.9 deficiencies per office (P < or = .001; Student t test). Personnel responsible for processing flexible sigmoidoscopes in family practice and internal medicine offices are insufficiently trained for this function. Endoscopes are not being processed according to current standards. After a 2-hour training period, these persons maintain their equipment close to or according to standards.

The detection of lice (Bovicola ovis) in mobs of sheep: a comparison of fleece parting, the lamp test and the table locks test.

Knowledge of the presence or absence of lice in a flock of sheep enables wool growers to make informed decisions as to the need for insecticidal treatments. However, with inapparent infestations, traditional methods of detection are not sufficiently sensitive and, as a consequence, flocks may be left untreated. Conversely, the routine application of insecticide to sheep with no sign of infestation is an unnecessary cost. The sensitivity of 3 procedures for detecting lice was evaluated in 68 mobs of sheep from 50 farms. In 24 mobs of sheep known to be lightly infested, lice were detected in 17% (71%) [corrected] by either parting the fleece of 10 sheep or by the lamp test in which 8g samples of shorn wool from 30 randomly selected fleeces were placed under lamps for 10 min to repel the lice. Twenty of 23 mobs (87%) were found to be infested by the table locks test in which a 30 g sample of locks wool was dissolved in 10% sodium hydroxide and the filtered residue examined with x 40 magnification. A screening test, in which either fleeces on 5 sheep were examined by fleece parting or lice were repelled from 30 shorn fleeces for 5 minutes, detected about 60% of lightly infested mobs. When this was followed by the table locks test 91% of lightly infested mobs were detected. Conducting any one of the tests on more than one mob, and in large mobs testing more frequently, increases the sensitivity of detection of lice within the whole flock.

Long-term results of revision hip arthroplasty. Survival analysis with special reference to the femoral component.

We report a clinical and radiological survival analysis of 148 consecutive revisions of hip arthroplasties. All patients referred were offered revision if it was indicated and they were medically fit. About one-third (32%) had had at least one previous revision, and about one-third had an established deep infection. The revisions were performed in ultra-clean air with body-exhaust suits. The usual method of fixation was by contained and pressurised cement of standard viscosity, to which appropriate antibiotics had been added in infected cases. Some patients had cementless revision. Clinical failure meant that one or both of the implants had been removed; radiological failure was assessed from serial radiographs. The clinical survival at ten years was 95%. The Merle D'Aubigné and Postel rating for pain improved from a mean of 2.9 to 5.2; and in Charnley group A and B cases, walking ability improved from 2.3 to 4.3. In contrast to some reports we also found good radiological survival; this was 90.5% for cemented femoral stems. Isoelastic revision stems inserted without cement gave poor results.