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Objectives: Platelet Rich Plasma (PRP) is a biological compound rich of growth factors able to support healing processes, and it represents a fascinating tool for clinicians. However results on its potentiality as intra-articular treatment for cartilage degenerative lesions in osteoarthritis (OA) are still controversial, due to the lack of well designed studies and the differences in the preparation procedures. The aim of this study was to analyze the modifications induced by two different PRP preparations on chondrocytes and sinoviocytes in culture, by documenting changes in proliferations, gene expression, and extracellular matrix components deposition. Methods: Blood collected from 10 volunteers was used to obtain two PRP preparations derived respectively from a ‘‘one-spinning procedure’’, to obtain a pure platelet concentrate without leukocytes but with a limited number of platelets (P-PRP), and by a ‘‘two-spinning approach’’, producing a product characterized by a higher platelet concentration and the presence of leukocytes (L-PRP). Results: Our data showed that L-PRP contains higher levels of growth factors and cytokines compared to P-PRP. Both PRPs stimulate chondrocytes proliferation during all the culture period evaluated; a significant enhancement in cell growth was induced by P-PRP respect to L-PRP only at day 7. Moreover, P-PRP stimulates chondrocyte anabolism as demonstrated by the expression of collagen type II and aggrecan, while L-PRP promotes catabolic pathways in which some different cytokines are involved. Interesting, L-PRP induces a higher expression of hyaluronic acid synthase-2 and hyaluronan compared to P-PRP. On sinoviocytes, L-PRP is able to sustain the up-regulation of proinflammatory factors, such as IL-1beta, IL-8 and FGF-2, together with a down modulation of HGF and TIMP-4 expression, two factors that have been recognized as anti-catabolic mediators in cartilage, whereas P-PRP was not able to exert a differential pattern of biological effect compared to PPP. Conclusions: Our study highlights that the two PRPs induce distinct effects on cultured human chondrocytes and sinoviocytes. This is due to the different content in PRP of platelets, leukocytes, growth factors and molecules that act synergically to regulate several biological processes which could influence the regeneration of a functional tissue.

Glucocorticoids (GC) are the mainstay of treatment of large-vessel vasculitis (LVV), but a sizeable number of patients relapse upon tapering the GC dose or after discontinuation of GC therapy. In addition, GC cause numerous adverse events. Therefore, in patients with longstanding disease and in those at risk for GC-related adverse events, the use of alternative therapeutic agents should be considered. Interleukin-6 (IL-6) is a key player in the pathogenesis of LVV. Preliminary data suggest the efficacy of the IL-6 receptor inhibitor tocilizumab (TCZ) in patients with LVV. We report 2 treatment-naïve patients with a recent diagnosis of LVV who received monthly TCZ infusions (8 mg/kg body weight) for 6 consecutive months as monotherapy because of relative contraindications and patients' reluctance to take GC. In both cases we observed a complete clinical response and normalisation of inflammatory markers as well as a decrease in vascular FDG uptake and SUV ratio on fluorodeoxyglucose positron emission/computerised tomography. Serum IL-6 and soluble IL-6 receptor (sIL-6R) levels rose in both patients after TCZ therapy. TCZ may be an effective alternative to GC treatment for LVV patients at risk for GC-related adverse events. Larger studies are required to confirm our findings.

Anti-TNF-α therapy has successfully been used to treat Takayasu arteritis (TA) refractory to conventional immunosuppressive treatment. However, some patients fail to respond even to TNF-α blockers. Interleukin-6 (IL-6) is a key player in the pathogenesis of TA. Preliminary data also suggest efficacy of the IL-6 receptor inhibitor tocilizumab in patients with large-vessel vasculitis. We report a patient with TA refractory to multiple conventional immunosuppressive agents and two TNF-α blockers successfully treated with monthly tocilizumab infusions (8 mg/kg body weight) for 6 consecutive months. Clinical indices of disease activity, inflammatory markers, and 18Ffluorodeoxyglucose positron emission/computerised tomography findings normalised, while the prednisone dosage could be tapered. Serum IL-6 and soluble IL-6 receptor (sIL-6R) levels raised during tocilizumab treatment consistent with the mode of action of tocilizumab. Tocilizumab holds promise for patients with refractory TA. Larger studies are required to confirm our findings.

To evaluate PTX3 feasibility to provide a prognostic tool in PMR clinical practice.Circulating PTX3 levels were measured in 93 PMR patients at disease onset and during corticosteroid therapy and in 46 normal controls (NC) by ELISA.No difference in PTX3 concentrations was observed between NC and PMR either at disease onset and during follow-up or between groups of patients defined according to the presence of recurrence/relapse.PTX3 serum levels do not increase significantly in active PMR. Further studies on patients with giant-cell arteritis could evaluate whether large vessel involvement may be associated to increased PTX3 levels.

Evaluation of the role of VEGF in cartilage pathophysiology.VEGF release from chondrocytes in the presence of IL-1beta, TGFbeta and IL-10 was detected by immunoassay. VEGF receptor -1 and -2 expression and VEGF ability to modulate caspase -3 and cathepsin B expression were detected by immunohistochemistry on cartilage biopsies and cartilage explants. VEGF effects on chondrocyte proliferation was analysed by a fluorescent dye that binds nucleic acids.VEGF production by osteoartritis (OA) chondrocytes was significantly reduced by IL-1beta while it was increased in the presence of TGFbeta. Cartilage VEGFR-1 immunostaining was significantly downregulated in 'early' OA patients compared to normal controls (NC). VEGFR-2 expression was negligible both in OA and in NC. VEGF decreased the expression of caspase-3 and cathepsin B, whereas it did not affect proliferation.VEGF is able to down-modulate chondrocyte activities related to catabolic events involved in OA cartilage degradation.

Objective. To evaluate potential associations of vascular endothelial growth factor (VEGF) gene polymorphisms with Behçet's disease (BD) and disease expression. Methods. Case patients were 122 consecutive Italian patients with BD followed at the Rheumatology, Ophthalmology, and Neurology Units in Bologna, Ferrara, Milano, Palermo, Potenza, Prato, Reggio Emilia, and Trento over a 3-year period (1997-99) and who satisfied the International Study Group criteria for BD. Also selected as a control group were 200 healthy age and sex matched blood donors. All patients with BD and controls were genotyped by polymerase chain reaction and allele-specific oligonucleotide techniques for +936 C/T (rs3025039) and -634 C/G (rs2010963) mutations and for an 18 base pair (bp) insertion/deletion (I/D) polymorphism at -2549 of the the VEGF promoter region. In vitro release of VEGF by peripheral blood mononuclear cells (PBMC) was investigated by ELISA in healthy controls homozygous for the polymorphisms studied. Results. The carriage rates of the alleles I and -634C were significantly more frequent in patients with BD than in healthy controls [p corr = 0.036, OR 1.8 (95% CI 1.1-2.9) and p corr = 0.05, OR 1.8 (95% CI 1.1-3.0), respectively]. While the distribution of allele +936T was similar in patients with BD and healthy controls, its frequency was significantly higher in BD patients with posterior uveitis/retinal vasculitis than in those without (p = 0.022, OR 2.4, 95% CI 1.1-5.0). Lipopolysaccharide-stimulated VEGF production from PBMC of healthy subjects was higher in II homozygous than in DD homozygous. Conclusion. Our data indicate that carriers of -634C and I alleles are associated with susceptibility to developing BD.

Elevated RANTES serum levels are present in polymyalgia rheumatica (PMR) patients with active disease. Chemokines may contribute to the inflammatory PMR process through their binding to CC chemokine receptor 5 (CCR5). The aim of this study was to examine if the 32 base pair deletion allele in CCR5 (CCR5 delta 32 allele) might be associated with PMR susceptibility and influence the disease outcome.We enrolled 88 consecutive patients with PMR residing in the Reggio Emilia area (Italy) who had a follow-up duration of at least one year. As a control group we used 86 healthy blood donors from the same geographic area. The CCR5 genotype of all PMR patients and controls was studied by polymerase chain reaction amplification of the region which includes the 32 deletion (CCR5 delta 32). RANTES serum levels were measured by commercial ELISA kits in CCR5 delta 32 heterozygous and CCR5 homozygous PMR patients at diagnosis before starting corticosteroid therapy and again after 6 months of therapy, as well as in 28 healthy subjects over 50 years of age.Frequencies of the CCR5 and CCR5 delta 32 alleles in patients and controls did not differ significantly. Homozygosity for CCR5 delta 32 was not detected in PMR patients and was detected in only one of the controls. No significant differences were observed between the patients carrying the CCR5 delta 32 allele and those homozygous for the normal CCR5 allele when we compared sex, presence of distal synovitis and systemic signs and/or symptoms, initial and cumulative prednisone dose, duration of therapy, ESR at diagnosis, frequency of relapse/recurrence and RANTES serum levels at diagnosis and after 6 months of corticosteroids.These results indicate that the frequency of the 32 deletion of the CCR5 receptor was not significantly different between PMR patients and healthy controls, and this genotype does not appear to be associated with the susceptibility to or severity of PMR.