Faculty Bibliography

OBJECTIVE: The migration of cells of chondrocyte lineage is believed to play a role in cartilage growth and repair. The present study examined 1) whether chondrocytes are capable of migration in vitro; and 2) the effects of nitric oxide (NO) on chondrocyte migration, adhesion, and cytoskeletal assembly. METHODS: Chondrocyte migration was evaluated by 2 assays: 1) 'centrifugal' migration within a 3-dimensional collagen matrix (dot culture); and 2) directed migration under agarose in response to bone morphogenetic protein. To assess the effects of NO, chondrocytes were treated with either exogenous NO (S-nitrosoglutathione [SNO-GSH]) or a mixture of cytokines known to induce endogenous NO production. The effects of NO on chondrocyte adhesion to fibronectin-coated surfaces, as well as on actin polymerization (determined by indirect immunofluorescence), were also examined. RESULTS: The capacity of chondrocytes to migrate was demonstrated both by the dot culture and by agarose methods. Both SNO-GSH and endogenous NO induced by cytokines inhibited this migration. Exposure to NO also inhibited attachment of chondrocytes to fibronectin and disrupted assembly of actin filaments. These effects of SNO-GSH and cytokine-induced NO production were reversed in the presence of hemoglobin and the NO synthase inhibitor NG-monomethyl arginine, respectively. CONCLUSION: NO interferes with chondrocyte migration and attachment to fibronectin, an extracellular matrix protein, probably via effects on the actin cytoskeleton. These effects of NO may result in impairment of cartilage repair, by interfering with the extracellular matrix regulation of chondrocyte function

The interfacial shear strength and bone tissue response was investigated for an arc deposited (AD) commercially pure titanium implant surface, with (AD/HA) and without (AD) plasma-sprayed hydroxyapatite (HA) coating. Ten purpose bred coonhounds received bilateral femoral stem implantation (AD and AD/HA) in the proximal femurs (hemiarthroplasty). The femoral prosthesis consisted of a modular CoCr alloy head, modular Ti-6Al-4V neck, and a 10-mm diameter cylindrical Ti-6Al-4V femoral stem. The AD surface had 30-35% greater surface roughness than the AD/HA surface. The HA coating had a purity greater than 90% and a crystallinity greater than 65%. After 6, 12, and 24 weeks, the implants were retrieved and analyzed with mechanical testing, qualitative and quantitative histology, and electron microscopy. The AD/HA implants had equivalent interfacial shear strengths to the AD implants at all time periods. The AD/HA implants had significantly greater linear bone contact than the AD implants. The 6-week implants had significantly thicker cortical bone than the 12- and 24-week implants. The HA coating was very stable in vivo, evidenced by no thickness reduction at any time period. Qualitatively, the AD/HA implants primarily had bone contacting the implant surface with little fibrous tissue present, and the AD implants had bone and fibrous tissue contacting the implant surface. The electron microscopy analysis showed that the mechanically tested implants exhibited a mixed failure mode at the bone, HA coating, and titanium interfaces

Neonatal rat calvarial osteoblasts were cultured on Ti-6Al-4V, Co-Cr-Mo alloy, 316L stainless steel and polystyrene (reference substrate) in the presence of ascorbic acid and 10 mM beta-glycerophosphate for 16, 17, 18, 19, 20, 21, 24 and 28 d. Scanning electron microscopy examination revealed that osteoblasts cultured on these orthopaedic/dental implant metals synthesized and deposited an extracellular matrix containing collagenous and non-collagenous components, as well as mineral nodules of various morphologies. Energy dispersive spectrometry revealed that the mineral deposits consisted of three distinct chemical compositions: calcium phosphate, calcium-sulphur-phosphorus, and calcium only. Backscattered electron imaging demonstrated that both the calcium phosphate and calcium-only deposits were electron dense, while the calcium-sulphur-phosphorus deposits were electron translucent. X-ray diffraction analysis indicated that the bulk of the osteoblast mineral deposits was amorphous hydroxyapatite; in addition, electron diffraction analysis revealed small regions of crystalline hydroxyapatite

The disadvantages of autogenous bone grafts has prompted a search for a dependable onlay bone graft substitute. A combination of tricalcium phosphate, a resorbable ceramic, and osteogenin, an osteoinductive protein, was evaluated as an onlay bone graft substitute in a rabbit calvarial model. Twenty-eight tricalcium phosphate implants (15 mm diameter x 5 mm; pore size, 100-200 microns) were divided into experimental and control groups and placed on the frontal bone of 14 adult New Zealand White rabbits. In the experimental animals, 185 micrograms of osteogenin was added to each implant. In the control animals, the implants were placed untreated. Implants were harvested at intervals of 1, 3, and 6 months, and evaluated using hematoxylin and eosin histology, microradiography, and histomorphometric scanning electron microscope backscatter image analysis. At 1 month there was minimal bone ingrowth and little tricalcium phosphate resorption in both the osteogenin-treated and control implants. At 3 months, both the osteogenin-treated and control implants showed a modest increase in bone ingrowth (8.85 percent versus 5.87 percent) and decrease in tricalcium phosphate (32.86 percent versus 37.08 percent). At 6 months, however, the osteogenin-treated implants showed a statistically significant increase in bone ingrowth (22.33 percent versus 6.96 percent; p = 0.000) and decrease in tricalcium phosphate (27.25 percent versus 37.80 percent; p = 0.004) compared with the control implants. The bone within the control implants was mostly woven at 6 months, whereas the osteogenin-treated implants contained predominantly mature lamellar bone with well-differentiated marrow. All implants maintained their original volume at each time interval studied. The tricalcium phosphate/osteogenin composite, having the advantage of maintaining its volume and being replaced by new bone as the tricalcium phosphate resorbs, may be applicable clinically as an onlay bone graft substitute

Light and electron microscopy was used for a histologic examination of normal heel fat pads and atrophic heel fat pads from patients with peripheral neuropathies. Histomorphometric analysis revealed an average 30% smaller mean cell area and 16% smaller mean cell diameters in the atrophic pads compared with the normal heel fat pads. Septal walls in the atrophic fat pads were often fragmented and approximately 75% wider than normal. Perineural fibrosis was also found in the atrophic heel fat pads. The Verhoeff elastic staining technique was used to determine the relative percentage of collagen to elastic tissue within the septae. No significant differences were noted between the normal and atrophic heels. The ultrastructure of the adipocytes from the normal and atrophic heel pads was similar to those found in abdominal subcutaneous fat. Lipid droplets of variable size and density thin the center of the adipocyte were surrounded by a thin border of cytoplasm. The interphase between adipocytes contained fine collagen and elastic fibers