The cbsA gene of Lactobacillus crispatus strain JCM 5810, encoding a protein that mediates adhesiveness to collagens, was characterized and expressed in Escherichia coli. The cbsA open reading frame encoded a signal sequence of 30 amino acids and a mature polypeptide of 410 amino acids with typical features of a bacterial S-layer protein. The cbsA gene product was expressed as a His tag fusion protein, purified by affinity chromatography, and shown to bind solubilized as well as immobilized type I and IV collagens. Three other Lactobacillus S-layer proteins, SlpA, CbsB, and SlpnB, bound collagens only weakly, and sequence comparisons of CbsA with these S-layer proteins were used to select sites in cbsA where deletions and mutations were introduced. In addition, hybrid S-layer proteins that contained the N or the C terminus from CbsA, SlpA, or SlpnB as well as N-and C-terminally truncated peptides from CbsA were constructed by gene fusion. Analysis of these molecules revealed the major collagen-binding region within the N-terminal 287 residues and a weaker type I collagen-binding region in the C terminus of the CbsA molecule. The mutated or hybrid CbsA molecules and peptides that failed to polymerize into a periodic S-layer did not bind collagens, suggesting that the crystal structure with a regular array is optimal for expression of collagen binding by CbsA. Strain JCM 5810 was found to contain another S-layer gene termed cbsB that was 44% identical in sequence to cbsA. RNA analysis showed that cbsA, but not cbsB, was transcribed under laboratory conditions. S-layer-protein-expressing cells of strain JCM 5810 adhered to collagen-containing regions in the chicken colon, suggesting that CbsAmediated collagen binding represents a true tissue adherence property of L. crispatus.

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The wear of acetabular cups made from conventional gamma-sterilized, and electron-beam cross-linked ultra-high-molecular-weight polyethylene was studied with a biaxial hip wear simulator. The femoral heads were either polished or roughened so that they represented the type of roughening and the value of surface roughness (R(a) = 0.14-0.18 micro m) observed in the roughest explanted femoral heads. The lubricant was diluted calf serum, and the test length 3 million cycles. The mean wear rate and standard deviation of conventional polyethylene cups against polished and against roughened heads was 11.6 +/- 0.07 and 64.4 +/- 10.1 mg per 1 million cycles, respectively. The latter value closely corresponds to that measured from explanted Charnley prostheses. Against polished heads, cross-linked polyethylene cups showed net weight gain, whereas against roughened heads, their mean wear rate was 2.4 +/- 0.3 mg per 1 million cycles. The mean equivalent circle diameters of polyethylene wear particles produced in the above four categories were: conventional/roughened 0.32 micro m, cross-linked/roughened 0.29 micro m, conventional/polished 0.28 micro m, cross-linked/polished 0.23 micro m. The size ranges and shapes were close to those seen in particles isolated from tissue samples. In conclusion, the tests indicated that electron-beam irradiation effectively reduces the harmful polyethylene wear particle production in total hip arthroplasty.

The effect of counterface roughness on the wear of conventional gamma-sterilized, and electron-beam-crosslinked ultrahigh molecular weight polyethylene was studied with a circularly translating pin-on-disk device. The counterfaces, CoCr disks, were either polished, or roughened so that they represented the type of roughening and the range of surface roughness values (R(a) = 0.014-0.24 microm) observed in explanted femoral heads of total hip prostheses. The lubricant was diluted calf serum, and the test length 3 million cycles. A total of 24 tests were done. With both types of polyethylene, there was a strong correlation between R(a) and wear factor k. The power equations were k = 5.87 x 10(-5)(R(a))(0.91) for conventional polyethylene (R(2) = 0.94), and k = 7.87 x 10(-5)(R(a))(2.49) for crosslinked polyethylene (R(2) = 0.82). Crosslinking improved wear resistance significantly. The wear of crosslinked polyethylene against the roughest counterfaces was lower than the wear of conventional polyethylene against the polished counterfaces. Against rough counterfaces, the wear of crosslinked polyethylene was an order of magnitude lower than that of conventional polyethylene. On the crosslinked polyethylene pins that were tested against polished counterfaces, remains of original machining marks were still visible after the test. The average size of wear particles produced by both types of polyethylene against rough counterfaces was similar, 0.4 microm, whereas that produced by conventional and crosslinked polyethylene against polished counterfaces was significantly smaller, 0.2 and 0.1 microm, respectively.

Prosthetic joints appear to show a strong relationship between the type of relative motion and wear, requiring careful consideration in the design of wear simulators. This relationship was studied with a 12-station pin-on-disk device, specifically adapted for the wear simulation of prosthetic hip joints. Each station had a unique motion, characterized by the so-called slide track, the track of the pin on the disk. The slide track shapes included 10 ellipses, their aspect ratio (AR) varying from 1.1 to 11.0, and a circle and a straight line as extreme cases. Hence for the first time in hip wear simulation, the motion was systematically varied over a wide range. Conventional UHMWPE pins were tested against polished CoCr disks in diluted calf serum three times for 3 million cycles. Below the AR value of 5.5, the polyethylene wear factor and wear mechanisms agreed with clinical observations. Above this value, the wear factor decreased to unrealistically low values, and the wear surface topography differed from that of retrieved acetabular cups. The wear particles, however, were similar to those isolated from periprosthetic tissues, irrespective of the AR value. In conclusion, it is recommended that the AR value be kept well below the critical point of 5.5.