Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis
DOI:
https://doi.org/10.31053/1853.0605.v57.n1.37207Keywords:
synoviocytes incuhated, mmps, collagenase, synovial fluid, rheumatoid arthnitis, osteoanthnitisAbstract
Cartilage is a specializcd conneetive tissue. It contains few cells into ari extracell matrix. The matrix mainly constituents are
col lageri and proteoglycans. lts degradation depends on synoviocytes activity, that secrete nictalloproteases. agents tu proteoglycans cataholisrn. The.re are two types ofsynoviocytes: macrophagics (type "A:') and fihrohlastics (type "B"). The proteoglvean destruction can he LTdependent or LT-independent. The aun of this work is synoviocytes function ex i'(vo study, free immune systeni iniluence. In order tu do it, heparinized synovial fluid samples were obtained from 6 osteoarthritic (OA) and 6 arthritic (RA) both sex unireated patients, diLignosed accordiiig ACR entena, which discase duration was longer than 6 months. Patients average age was 70 ± 2 years. Control samples were synovial fluid from traumatic arthritis or non inflammatony honemuscle pathology. Synovial fluid was ce.ntifugated al 1500 g for 30 flhiiiUtCS ti) isolate synoViocyles. Sedimentcontalnin9 cells vas 6 hs incubed with Du!becco-Eagles media, thai has HEPES Gihco (26mM) NaHCO3 (O.Sg/I); glutamine (21-nM), streptornicine (100mg/1). G-peniciiline (1 U/ mi); anphotericine B (2.5mgl1). Cells cal ification a n d vi ah jI it y were cvtopathologically determined.
Bel'orc and after incubation, col lagenase activily was measured by ELISA-double- sandwich, using 10flg/m 1 monoclonal anti-MMPs
i n phosphate-hut'fer-sal inc. The antigenantibodv complex production with inespecific prolclns vas hlocked by hovine serie albumine. Streptavidin peroxidase vas added and washed w it h 2,2 ,az i n,di (3-el hy 1- benztazoi Ii suiphonic) acid iii develop color. The link of laheled aiitibody by absorbance al 410 nm vas determined in ELISA-spectrophothometcr. RA patient.s earlicr MMPs snovioctes production was 1 373±11 5ng/m 1. Then 611s incubation 2143±132 n<,/ml was reached. The menease (56) had high significance (p< 0.0001.). OA eanlier MMPs cclis production was 276±23 ng/mk hut after incubation it reached 542±47 ng/ml. (96Y incrcased with highly significativa diffenence too: p<0.000 1 Microscopic study was carnied out hefore and after incubation, and shows a lot of synoviocytes with plenty of cytopiasme when the collagenase leveis werc. highest. Qn the contrary. when Tow MMPs production by synovial fluid, as no incubatcd ostcoarthritic material, a few cells containingpicnotics nucleous were observed..
Significani quantitative differences in AR and OA enzvmatic secretion were observed. Although in rheumatoid arthritic MMPs leveis
synoviocytes production were 4.6 times ihan OA levels, after 6 lis incubation percentage of increase in OA cells secrction was highesl.
Descnihed results confirm MMP- 1 ss'nthesis by synoviocytes, and these leveis conelate with inflamrnation, more pronunced in acute (RA) ihan chronic pathology (OA). Synoviocyte incubation ]el us lo test desease changes in synovial fluid according lo cells
nuniher and phagocytic activity. Authors agnee lo assert that synovial fluid may rellect what is happening in an articular cartilago. hecaLise SE provides markers of joini desease. MMPs are giving information about pathways involved in OA and RA cartilage degradation
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