Intra-articular depletion of macrophages increases acute synovitis and alters macrophage polarity in the injured mouse knee.

OBJECTIVE: Acute synovial inflammation following joint trauma is associated with posttraumatic arthritis. Synovial macrophages have been implicated in degenerative changes. In this study, we sought to elucidate the role of intra-articular macrophages in the acute inflammatory response to fracture in the mouse knee. METHOD: A closed articular fracture was induced in two models of synovial macrophage depletion: genetically-modified MaFIA mice administered AP20187 to induce programmed macrophage apoptosis, and wild-type C57BL/6 mice administered clodronate liposomes, both via intra-articular injection. Synovial inflammation, bone morphology, and levels of F4/80+ macrophages, NOS2+ M1 macrophages, and CD206+ M2 macrophages were quantified 7 days after fracture using histology and micro-computed tomography. RESULTS: Intra-articular macrophage depletion with joint injury did not reduce acute synovitis or the number of synovial macrophages 7 days after fracture in either macrophage-depleted MaFIA mice or in clodronate-treated C57BL/6 mice. In macrophage-depleted MaFIA mice, macrophage polarity shifted to a dominance of M1 macrophages and a reduction of M2 macrophages in the synovial stroma, indicating a shift in M1/M2 macrophage ratio in the joint following injury. Interestingly, MaFIA mice depleted 2 days prior to fracture demonstrated increased synovitis (P = 0.003), reduced bone mineral density (P = 0.0004), higher levels of M1 macrophages (P = 0.013), and lower levels of M2 macrophages (not statistically significant, P=0.084) compared to control-treated MaFIA mice. CONCLUSION: Our findings indicate that macrophages play a critical immunomodulatory role in the acute inflammatory response surrounding joint injury and suggest that inhibition of macrophage function can have prominent effects on joint inflammation and bone homeostasis after joint trauma.

Loss-of-function mutations in the cathepsin C gene result in periodontal disease and palmoplantar keratosis.

Papillon-Lefèvre syndrome, or keratosis palmoplantaris with periodontopathia (PLS, MIM 245000), is an autosomal recessive disorder that is mainly ascertained by dentists because of the severe periodontitis that afflicts patients. Both the deciduous and permanent dentitions are affected, resulting in premature tooth loss. Palmoplantar keratosis, varying from mild psoriasiform scaly skin to overt hyperkeratosis, typically develops within the first three years of life. Keratosis also affects other sites such as elbows and knees. Most PLS patients display both periodontitis and hyperkeratosis. Some patients have only palmoplantar keratosis or periodontitis, and in rare individuals the periodontitis is mild and of late onset. The PLS locus has been mapped to chromosome 11q14-q21 (refs 7, 8, 9). Using homozygosity mapping in eight small consanguineous families, we have narrowed the candidate region to a 1.2-cM interval between D11S4082 and D11S931. The gene (CTSC) encoding the lysosomal protease cathepsin C (or dipeptidyl aminopeptidase I) lies within this interval. We defined the genomic structure of CTSC and found mutations in all eight families. In two of these families we used a functional assay to demonstrate an almost total loss of cathepsin C activity in PLS patients and reduced activity in obligate carriers.

Recycling of spent filter backwash water using coagulation-assisted membrane filtration: effects of submicrometre particles on membrane flux.

Membrane separation technology has been widely used for recycling of spent filter backwash water (SFBW) in water treatment plant. Membrane filtration performance is subject to characteristics of the particles in the SFBW. A bench-scale microfiltration (MF) coupled with pre-coagulation was set up to evaluate the recovery efficiency of SFBW. Effect of particle size distribution and zeta potential of the coagulated SFBW on the membrane filtration as well as the coagulation strategies were investigated. Pore clogging was more severe on the membrane with 1.0 mum pore size than on the membrane with 0.5 mum pore size due to the fact that submicrometre particles are dominant and their diameters are exactly closed to the pore size of the MF membrane. Pre-settling induced more severe irreversible fouling because only the submicrometre particles in the water become predominant after settling, resulting in the occurrence of more acute pore blocking of membrane. By contrast, pre-coagulation mitigates membrane fouling and improves membrane flux via enlarging particle size on membrane surface. The variations of zeta potential in response to coagulant dosing as well as fractal dimension were also compared with the performance of the subsequent filtration. The result showed that pre-coagulation induced by charge neutralization at the optimum dosage where the zeta potential is around zero leads to the optimal performance of the subsequent membrane filtration for SFBW recycling. At such condition, the fractal dimension of coagulated flocs reached minimum.

Effects of implant surface roughness and stiffness of grafted bone on an immediately loaded maxillary implant: a 3D numerical analysis.

This study investigated the effects of the stiffness of a maxillary sinus graft and the surface roughness of an immediately loaded implant using a non-linear three-dimensional finite element (FE) analysis (3D). Six FE models were created, including two stiffness values of grafted bone (345 and 3450 MPa of elastic modulus) and three conditions of implant-bone interfaces (Frictional coefficient of 0.3 for machined surface, 0.45 for rough implant surface and a bonded implant-bone interface for an osseointegrated implant). Computer tomographic images of a human skull were used to construct a posterior maxillary model. All implants were designed via the computer aided design software with a spiral threaded configuration. Three loading scenarios were investigated for each of the six models; axial loading and lateral loadings at 30 degrees and 60 degrees . The results showed that a 60 degrees lateral loading has scored the highest level of bone stresses among the three loading conditions. Immediately loaded implants with 0.3 frictional coefficient have suffered the highest bone stresses which were higher than those with bonded interface by about 57%. Increasing the frictional coeffecient to 0.45, however, did not show any benefits in reducing the peak bone stress. Raising the stiffness of grafted bone diminished the bone stress by about 10% in both the immediately loaded and the osseointegrated implants. It was also noted that increasing graft stiffness and implant surface roughness reduced the sliding at the implant-bone interface which may improve the implant stability and long-term survival.

Purification and biochemical characterization of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate-sensitive L-glutamate receptors of pig brain.

Two preparations of glutamate receptors were purified from the synaptic junctions of pig brain by a combination of detergent solubilization, anion-exchange chromatography, wheat-germ agglutinin affinity chromatography and sedimentation through sucrose gradients. These preparations were enriched in specific L-[3H]glutamate binding activity (> 5000 pmol of glutamate binding sites/mg of protein), and the rank order of ligand affinity for binding to these preparations was: quisqualate > 6-cyano-7- nitroquinoxaline-2,3-dione > alpha-amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid (AMPA) > L-glutamate > kainate > > N-methyl-D-aspartate approximately L-2-amino-4-phosphonobutyrate. SDS/PAGE analysis revealed that more than 80% of the protein in either of these preparations appeared as a single protein band of 106 kDa. Two-dimensional gel electrophoresis further revealed that these 106 kDa proteins consisted of a series of acidic proteins which were recognized by antibodies against rat AMPA receptor subunits. These 106 kDa proteins were also recognized by wheatgerm agglutinin and concanavalin A; in addition, peptide N-glycosidase F treatment of these preparations decreased their size to 99 kDa. Our results suggest that the putative glutamate receptors isolated here are likely to belong to the AMPA subtype of glutamate receptors in pig brain. Using the purification procedure reported here, 5 micrograms of AMPA receptor proteins can be isolated from 250 g of pig brain tissue.