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Estudo imuno-histoqu?mico da presen?a de miofibroblastos e da express?o do fator transformador de crescimento-beta1, interferon gama, metaloproteinase de matriz 13 e indutor de metaloproteinases de matriz em les?es odontog?nicas epiteliaisSantos, Pedro Paulo de Andrade 28 February 2012 (has links)
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Previous issue date: 2012-02-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Myofibroblasts are cells that exhibit a hybrid phenotype, sharing the morphological
characteristics of fibroblasts and smooth muscle cells, which is acquired during a process
called differentiation. These cells then start to express -SMA, a marker that can be used for
their identification. Studies suggest that myofibroblasts are related to the aggressiveness of
different tumors and that TGF-1 and IFN- play a role in myofibroblast differentiation,
stimulating or inhibiting this differentiation, respectively. The objective of this study was to
investigate the role of myofibroblasts in epithelial odontogenic tumors, correlating the
presence of these cells with the aggressiveness of the tumor. Immunohistochemistry was used
to evaluate the expression of TGF-1 and IFN- in myofibroblast differentiation, as well as
the expression of MMP-13, which is activated by myofibroblasts, and of EMMPRIN
(extracellular matrix metalloproteinase inducer) as a precursor of this MMP. The sample
consisted of 20 solid ameloblastomas, 10 unicystic ameloblastomas, 20 odontogenic
keratocysts, and 20 adenomatoid odontogenic tumors. For evaluation of myofibroblasts, anti-
-SMA-immunoreactive cells were quantified in connective tissue close to the epithelium.
Immunoexpression of TGF-1, IFN-, MMP-13 and EMMPRIN was evaluated in the
epithelial and connective tissue components, attributing scores of 0 to 4. The results showed a
higher concentration of myofibroblasts in solid ameloblastomas (mean of 30.55), followed by
odontogenic keratocysts (22.50), unicystic ameloblastomas (20.80), and adenomatoid
odontogenic tumors (19.15) (p=0.001). No significant correlation between TGF-1 and IFN-
was observed during the process of myofibroblast differentiation. There was also no
correlation between the quantity of myofibroblasts and MMP-13 expression. Significant
correlations were found between MMP-13 and TGF-1 (r=0.087; p=0.011), between MMP-
13 and IFN- (r=0.348; p=0.003), as well as between EMMPRIN and MMP-13 (r=0.474;
p<0.001) and between EMMPRIN and IFN- (r=0.393; p=0.001). The higher quantity of
myofibroblasts observed in solid ameloblastomas, odontogenic keratocysts and unicystic
ameloblastomas suggests that these cells are one of the factors responsible for the more
aggressive biological behavior of these tumors, although the myofibroblast population was
not correlated with TGF-1, IFN-, MMP-13 or EMMPRIN. The correlation between MMP-
13 and TGF-1 suggests that the latter induces the expression of this metalloproteinase. The
present results also support the well-established role of EMMPRIN as an inducer of MMP-13.
Furthermore, the relationship between EMMPRIN and IFN- and between MMP-13 and IFN-
suggests synergism in the antifibrotic effect of these markers / Os miofibroblastos s?o c?lulas que apresentam um fen?tipo h?brido exibindo caracter?sticas morfol?gicas de fibroblastos e de c?lulas musculares lisas, sendo a aquisi??o de tal fen?tipo denominada diferencia??o, passando ent?o a expressar a -SMA, a qual ?
importante na identifica??o dessas c?lulas. Estudos t?m sugerido que os miofibroblastos
apresentam rela??o com a agressividade de diversas les?es e que o seu processo de
diferencia??o estaria relacionado ? express?o do TGF- 1 e do IFN- atuando,
respectivamente, no est?mulo e na inibi??o dessa diferencia??o. O objetivo deste trabalho foi
investigar o papel dos miofibroblastos em les?es odontog?nicas epiteliais, relacionando-os ?
agressividade das les?es e analisar por meio da imuno-histoqu?mica, a express?o do TGF- 1 e
IFN- no processo de diferencia??o, al?m da an?lise da MMP-13 que ? ativada por
miofibroblastos e do indutor de metaloproteinases de matriz (EMMPRIN) como precursor
desta MMP. A amostra foi constitu?da por 20 ameloblastomas s?lidos, 10 ameloblastomas
unic?sticos, 20 ceratocistos odontog?nicos e 20 tumores odontog?nicos adenomat?ides. Para a
avalia??o dos miofibroblastos, foram quantificadas as c?lulas imunorreativas ao anticorpo -
SMA presentes no tecido conjuntivo, pr?ximo ao tecido epitelial. As express?es de TGF- 1,
IFN- , MMP-13 e EMMPRIN, foram avaliadas no componente epitelial e no conjuntivo,
estabelecendo-se o percentual de imunorreatividade e atribuindo-se escores de 0 a 4. A an?lise
dos miofibroblastos evidenciou maior concentra??o nos ameloblastomas s?lidos (m?dia de
30,55), seguido pelos ceratocistos odontog?nicos (22,50), ameloblastomas unic?sticos (20,80)
e tumores odontog?nicos adenomat?ides (19,15) com valor de p= 0,001. N?o foi encontrada
correla??o significativa entre TGF- 1 e IFN- no processo de diferencia??o dos
miofibroblastos, bem como na rela??o entre a quantidade de miofibroblastos e a express?o da
MMP-13. Constatou-se, correla??o estat?stica entre MMP-13 e TGF- 1 (r= 0,087; p= 0,011)
al?m de significante correla??o entre MMP-13 e IFN- (r=0,348; p=0,003). Entre EMMPRIN
e MMP-13 verificou-se signific?ncia (r= 0,474; p<0,001) assim como entre EMMPRIN e
IFN- (r=0,393; p=0,001). A maior quantidade de miofibroblastos evidenciada nos
ameloblastomas s?lidos, ceratocistos odontog?nicos e ameloblastomas unic?sticos sugere que
estas c?lulas podem ser um dos fatores respons?veis para um comportamento biol?gico mais
agressivo destas les?es, embora a popula??o de miofibroblastos n?o tenha apresentado
correla??o com TGF- - 1, IFN- ,MMP-13 e EMMPRIN. Quanto a correla??o evidenciada
entre MMP-13 e TGF- 1, isto pode sugerir um papel indutor do TGF- 1 para a express?o da
MMP-13, assim como os resultados deste estudo refor?am a rela??o bem estabelecida do
EMMPRIN como indutor da MMP-13. Constatou-se tamb?m rela??o entre EMMPRIN e
IFN- assim como entre MMP-13 e IFN- sugerindo, dessa forma, um sinergismo na a??o
anti-fibr?tica desses marcadores
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Estudo da imunoexpress?o de RANKL e OPG, do ?ndice angiog?nico (CD34) e da presen?a de miofibroblastos (?-SMA) em ceratocistos odontog?nicos isolados e associados ? s?ndrome de GorlinNonaka, Cassiano Francisco Weege 23 September 2010 (has links)
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Previous issue date: 2010-09-23 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The odontogenic keratocysts are distinguished from other odontogenic cystic lesions by their potentially aggressive clinical behavior and association, in some cases, with Gorlin syndrome. Studies have suggested that syndrome keratocysts, in comparison with sporadic lesions, have higher growth and infiltration capacity and higher recurrence tendency. The aim of this study was to analyze, by means of immunohistochemistry, the expressions of receptor activator of nuclear factor ?B ligand (RANKL) and osteoprotegerin (OPG), the angiogenic index (CD34) and the presence of myofibroblasts (?-SMA) in primary and recurrent sporadic keratocysts and in keratocysts associated with Gorlin syndrome. The sample was composed by 30 sporadic keratocysts (22 primary and 8 recurrent) and 22 syndrome keratocysts. In the epithelium and in the fibrous capsule of the lesions, the immunoexpression of RANKL and OPG was evaluated by determination of the percentage of positive cells, according to the following scores: 0 (less than 10% of positive cells), 1 (11% - 50% of positive cells), 2 (51% - 75% of positive cells) and 3 (more than 76% of positive cells). In addition, cases were classified according to the RANKL score/ OPG score ratio, as follows: RANKL > OPG, RANKL < OPG, and RANKL = OPG. The angiogenic index was analyzed by counting the microvessels immunoreactive to anti-CD34 antibody in 5 fields (200?). The analysis of myofibroblasts was performed by counting the cells immunoreactive to anti-?-SMA antibody in 10 fields (400?). The analysis of the expressions of RANKL and OPG in the epithelial lining and in the fibrous capsule did not reveal significant differences between groups (p > 0.05). Regarding the RANKL/ OPG ratio in the epithelial lining, most sporadic primary (54.5%) and syndrome lesions (59.1%) showed RANKL < OPG ratio and RANKL = OPG ratio, respectively (p > 0.05). With respect to the RANKL/ OPG ratio in the fibrous capsule, the majority of sporadic primary (81.8%) and sporadic recurrent lesions (75.0%) and most syndrome lesions (45.5%) showed RANKL = OPG ratio (p > 0.05). The mean number of microvessels was 69.2 in sporadic primary lesions, 67.6 in recurrent lesions, and 71.6 in syndrome lesions, with no significant differences between groups (p > 0.05). The mean number of myofibroblasts was 34.4 in sporadic primary lesions, 29.3 in recurrent lesions, and 33.7 in syndrome lesions, with no significant differences between groups (p > 0.05). In conclusion, the results of the present study suggest that the differences in the biological behavior between sporadic keratocysts and keratocysts associated with Gorlin syndrome may not be related to the expressions of RANKL and OPG, the RANKL/ OPG ratio, the angiogenic index or the number of myofibroblasts in these lesions / Os ceratocistos odontog?nicos se destacam em rela??o a outras les?es c?sticas odontog?nicas pelo comportamento cl?nico potencialmente agressivo e por se apresentarem associados, em alguns casos, ? s?ndrome de Gorlin. Estudos t?m sugerido que os ceratocistos sindr?micos, em compara??o ?s les?es isoladas, possuem maior capacidade de crescimento e infiltra??o e maior tend?ncia ? recorr?ncia. O objetivo do presente trabalho consistiu em analisar, por meio de imuno-histoqu?mica, as express?es do ligante do receptor ativador do fator nuclear ?B (RANKL) e da osteoprotegerina (OPG), o ?ndice angiog?nico (CD34) e a presen?a de miofibroblastos (?-SMA), em ceratocistos isolados prim?rios e recorrentes e ceratocistos associados ? s?ndrome de Gorlin. A amostra foi composta por 30 ceratocistos isolados (22 prim?rios e 8 recorrentes) e 22 ceratocistos sindr?micos. A express?o de RANKL e OPG foi avaliada no epit?lio e na c?psula fibrosa das les?es, estabelecendo-se o percentual de c?lulas imunopositivas, de acordo com os escores: 0 (? 10% das c?lulas positivas), 1 (11% - 50% das c?lulas positivas), 2 (51% - 75% das c?lulas positivas) e 3 (? 76% das c?lulas positivas). Al?m disso, os casos foram categorizados, segundo a propor??o RANKL/ OPG, em: RANKL > OPG, RANKL < OPG e RANKL = OPG. O ?ndice angiog?nico foi analisado por meio da contagem dos microvasos imunomarcados pelo anticorpo anti-CD34, em 5 campos (200?). Para a avalia??o dos miofibroblastos, foram quantificadas as c?lulas imunorreativas ao anticorpo anti-?-SMA, em 10 campos (400?). A an?lise das express?es de RANKL e OPG, no revestimento epitelial e na c?psula fibrosa, n?o revelou diferen?as significativas entre os grupos (p > 0,05). Em rela??o ? propor??o RANKL/ OPG no revestimento epitelial, grande parte das les?es isoladas prim?rias (54,5%) e sindr?micas (59,1%) exibiu propor??o RANKL < OPG e propor??o RANKL = OPG, respectivamente (p > 0,05). Em rela??o ? propor??o RANKL/ OPG na c?psula fibrosa, a maioria das les?es isoladas prim?rias (81,8%) e isoladas recorrentes (75,0%) e grande parte das les?es associadas ? s?ndrome de Gorlin (45,5%) revelaram propor??o RANKL = OPG (p > 0,05). O n?mero m?dio de microvasos foi de 69,2 nas les?es isoladas prim?rias, 67,6 nas les?es recorrentes e 71,6 nas les?es sindr?micas, sem diferen?as significativas entre os grupos (p > 0,05). A an?lise dos miofibroblastos revelou valores m?dios de 34,4 nas les?es isoladas prim?rias, 29,3 nas les?es recorrentes e 33,7 nas les?es sindr?micas, sem diferen?as significativas entre os grupos (p > 0,05). Em conclus?o, os resultados do presente estudo sugerem que as diferen?as no comportamento biol?gico entre ceratocistos isolados e associados ? s?ndrome de Gorlin podem n?o estar relacionadas ?s express?es de RANKL e OPG, ? propor??o RANKL/ OPG, ao ?ndice angiog?nico ou ? quantidade de miofibroblastos presentes nas les?es
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