Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis
Cartilage is a specializcd conneetive tissue. It contains few cells into ari extracell matrix. The matrix mainly constituents arecol lageri and proteoglycans. lts degradation depends on synoviocytes activity, that secrete nictalloproteases. agents tu proteoglycans cataholisrn. The.re are two types o...
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Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2000
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/37207 |
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I10-R327-article-37207 |
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| record_format |
ojs |
| institution |
Universidad Nacional de Córdoba |
| institution_str |
I-10 |
| repository_str |
R-327 |
| container_title_str |
Revista de la Facultad de Ciencias Médicas de Córdoba |
| language |
Español |
| format |
Artículo revista |
| topic |
synoviocytes incuhated mmps collagenase synovial fluid rheumatoid arthnitis osteoanthnitis sinoviocitos incubados MMPsr colagenasa líquido sinoval artritis reumatoidea osteoartrosis |
| spellingShingle |
synoviocytes incuhated mmps collagenase synovial fluid rheumatoid arthnitis osteoanthnitis sinoviocitos incubados MMPsr colagenasa líquido sinoval artritis reumatoidea osteoartrosis Montrull, Hilda Brizuela , Nilda Demurtas, Silvia Strusberg, Alberto Spitale, Luis Meirovich, Carlos Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| topic_facet |
synoviocytes incuhated mmps collagenase synovial fluid rheumatoid arthnitis osteoanthnitis sinoviocitos incubados MMPsr colagenasa líquido sinoval artritis reumatoidea osteoartrosis |
| author |
Montrull, Hilda Brizuela , Nilda Demurtas, Silvia Strusberg, Alberto Spitale, Luis Meirovich, Carlos |
| author_facet |
Montrull, Hilda Brizuela , Nilda Demurtas, Silvia Strusberg, Alberto Spitale, Luis Meirovich, Carlos |
| author_sort |
Montrull, Hilda |
| title |
Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| title_short |
Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| title_full |
Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| title_fullStr |
Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| title_full_unstemmed |
Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| title_sort |
incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis |
| description |
Cartilage is a specializcd conneetive tissue. It contains few cells into ari extracell matrix. The matrix mainly constituents arecol 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-MMPsi 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 leveissynoviocytes 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 cellsnuniher 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 |
| publisher |
Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología |
| publishDate |
2000 |
| url |
https://revistas.unc.edu.ar/index.php/med/article/view/37207 |
| work_keys_str_mv |
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2024-09-03T21:03:33Z |
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I10-R327-article-372072022-09-23T19:37:50Z Incremento em la producción de colagenasa en incubado de sino viocitos de pacientes con artritis reumatoidea y osteoartrosis Montrull, Hilda Brizuela , Nilda Demurtas, Silvia Strusberg, Alberto Spitale, Luis Meirovich, Carlos synoviocytes incuhated mmps collagenase synovial fluid rheumatoid arthnitis osteoanthnitis sinoviocitos incubados MMPsr colagenasa líquido sinoval artritis reumatoidea osteoartrosis Cartilage is a specializcd conneetive tissue. It contains few cells into ari extracell matrix. The matrix mainly constituents arecol 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-MMPsi 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 leveissynoviocytes 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 cellsnuniher 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 El cartílago articular ar es un tejido paucicelular. con colágeno y proteoglicanos en la matriz extracelular. Su degradación es función de los sinoviocitos, que segregan metal oproteasas que catabolizan a los proteoglicanos. Se distinguen los sinoviocitos A o macrofágicos y los sinoviocitos B o fibrohiásticos. La destrucción de proteoglicanos puede ser UI'- dependiente o independiente.Nuestro objetivo fue estudiar ex u/ro el rol de los sinoviocitos, sin la influencia del sistema inmune. Líquido sinovial de pacientes de ambos sexos, 70±2años, Con OA(6) y AR(6), vírgenes de tratamiento, se centrifugó 30 minutos a 1500 g. para aislar sinoviocitos.El sedimento se incubó 6 hs en medio Dulbecco-Eaglcs, con 26 mM de HEPES Gibco, NaHCO3 ( 0.5g/1), gl ut.am i na (2 mM),estreptomicina l OOmg/l). penicilina (1 U/mI) y anfotericina 13 (2.5mg/l ). Identidad y viabilidad celulares se determinaron con técnicas citopatológicas. Las muestras control provinieron de artritis traumática o patología osleoarticular no-inflamatoria.Con an' icuerpos monoclonales antiMMPs( loftg/ml), previo bloqueo de producción de proteínas incspecíficas con albúminasérica bovina, se midió actividad colagenasa MM P-1) antes y después de incubar con ELISA-doble-sandwich. Con streptavidinperox idasa se desarrolló color y por absorbancia a 410 nm. se leyó la complejación de los anticuerpos marcados.La secreción de MMP- 1 por sino viocitos AR fue 1373±115 ng/ml. Con 6 hs de incubado aumentó hasta 2 143± 132ng/l Rey. I'ac. Cienc. 1Ied Córdoba 57 (1) 51-57- 2000 (-5617c)(p<0.000 1). probablemente por lah i perce 1 uI ari dad. Los sinoviocitos OA secretaron 276 ± 23 no/ml . y 542 ± 47 ng/ml tras la incubación (9611;)(p<0.001). Hay paralelismo entre la producción deMMP-1 y la observación microscópica. Sinoviocitos con abundante citoplasma corresponden a altos niveles de enzima. La baja secreción de MMPs responde a escasa población celular y núcleos picnóticos. Aunque en AR la producción de MMPs fue 4.6 veces mayor que en OA, en cambio el incremento porcentual tras la incubación fue casi el doble en OA ClUC en AR.Esos resultados confirman que la producción enzi mát ca varía con la in tianiación, que es mayor en los procesos agudos, y que laincubación de sinoviocitos permite detectar cambios patológicos locales Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2000-11-30 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion texto texto texto application/pdf https://revistas.unc.edu.ar/index.php/med/article/view/37207 10.31053/1853.0605.v57.n1.37207 Revista de la Facultad de Ciencias Médicas de Córdoba.; Vol. 57 No. 1 (2000); 51-58 Revista de la Facultad de Ciencias Médicas de Córdoba; Vol. 57 Núm. 1 (2000); 51-58 Revista da Faculdade de Ciências Médicas de Córdoba; v. 57 n. 1 (2000); 51-58 1853-0605 0014-6722 10.31053/1853.0605.v57.n1 spa https://revistas.unc.edu.ar/index.php/med/article/view/37207/37423 Derechos de autor 2022 Universidad Nacional de Córdoba http://creativecommons.org/licenses/by-nc/4.0 |