Tenascin and fibronectin expression after pulp capping with different hemostatic agents: a preliminary study.

This study investigated the expression of extracellular matrix glycoproteins tenascin (TN) and fibronectin (FN) in pulp repair after capping with calcium hydroxide (CH), following different hemostasis protocols. Class I cavities with a pulp exposure were prepared in 42 human third molars scheduled for extraction. Different hemostatic agents (0.9% saline solution, 5.25% sodium hypochlorite and 2% chlorhexidine digluconate) were used and pulps were capped with CH cement. After 7, 30 or 90 days, teeth were extracted, formalin-fixed, and prepared for immunohistochemical technique. Hemostatic agents did not influence the expression of TN and FN. Both glycoproteins were found in the entire the pulp tissue and around collagen fibers, but were absent in the mineralized tissues. In the predentin, TN showed positive immunostaining and FN had a variable expression. Within 7 days post-treatment, a slightly more pronounced immunostaining on the pulp exposure site was observed. Within 30 days, TN and FN demonstrated a positive expression around the dentin barrier and at 90 days, a thin and linear expression of TN and FN was delimitating the reparative dentin. In conclusion, hemostatic agents did not influence TN and FN expression. Immunostaining for TN and FN was seen in different regions and periods, demonstrating their role in pulp repair.

Tenascin and Fibronectin Expression after Pulp Capping with Different Hemostatic Agents: A Preliminary Study

This study investigated the expression of extracellular matrix glycoproteins tenascin (TN) and fibronectin (FN) in pulp repair after capping with calcium hydroxide (CH), following different hemostasis protocols. Class I cavities with a pulp exposure were prepared in 42 human third molars scheduled for extraction. Different hemostatic agents (0.9% saline solution, 5.25% sodium hypochlorite and 2% chlorhexidine digluconate) were used and pulps were capped with CH cement. After 7, 30 or 90 days, teeth were extracted, formalin-fixed, and prepared for immunohistochemical technique. Hemostatic agents did not influence the expression of TN and FN. Both glycoproteins were found in the entire the pulp tissue and around collagen fibers, but were absent in the mineralized tissues. In the predentin, TN showed positive immunostaining and FN had a variable expression. Within 7 days post-treatment, a slightly more pronounced immunostaining on the pulp exposure site was observed. Within 30 days, TN and FN demonstrated a positive expression around the dentin barrier and at 90 days, a thin and linear expression of TN and FN was delimitating the reparative dentin. In conclusion, hemostatic agents did not influence TN and FN expression. Immunostaining for TN and FN was seen in different regions and periods, demonstrating their role in pulp repair.

Este estudo investigou a expressão das glicoproteínas Tenascina (TN) e Fibronectina (FN) da matriz extracelular no reparo pulpar após capeamento com hidróxido de cálcio (HC), seguindo diferentes protocolos de hemostasia. Cavidades de classe I com exposição pulpar foram preparadas em 42 terceiros molares humanos indicados para extração. Diferentes agentes hemostáticos (solução salina a 0,9%, hipoclorito de sódio a 5,25% e clorexidina a 2%) foram usados e as polpas foram capeadas com cimento de HC. Após 7, 30 ou 90 dias, os dentes foram extraídos, fixados em formalina e preparados para análise imunoistoquímica. Os agentes hemostáticos não influenciaram a expressão de TN e FN. Ambas glicoproteínas foram encontradas em todo tecido pulpar, ao redor das fibras colágenas e estiveram ausentes nos tecidos mineralizados. Na pré-dentina, a TN mostrou forte imunoexpressão e a FN teve uma expressão variável. Após 7 dias, foi observada uma expressão levemente mais pronunciada no lugar da exposição pulpar. Aos 30 dias, a TN e a FN demonstraram uma expressão mais forte sob a barreira dentinária e aos 90 dias, uma expressão fina e linear da TN e FN apresentava-se delimitando a dentina reparativa. Em conclusão, os agentes hemostáticos não influenciaram e expressão da TN e da FN. A imunoexpressão da TN e FN foi observada em diferentes regiões e períodos, demonstrando o seu papel no reparo pulpar.

Imatinib atenua a fibrose miocárdica em associação com a inibição da atividade do PDGFRα / Imatinib attenuates myocardial fibrosis in association with inhibition of the PDGFRα activity

FUNDAMENTO: O Imatinib é um inibidor do receptor tirosina-quinase que foi confirmada como exercendo um efeito inibidor sobre a atividade do receptor do PDGF, fator de crescimento plaquetário (PDGFRα e PDGFRβ). OBJETIVO: Investigar o efeito protetor do Imatinib na fibrose miocárdica em acetato de deoxicorticosterona (DOCA)/ratos com hipertensão induzida por sal. MÉTODOS: Sessenta ratos Sprague-Dawley machos, uninefrectomizados foram distribuídos em três grupos: ratos controles (grupo CON): grupo deoxicorticosterona (grupo DOCA); grupo deoxicorticosterona e Imatinib (grupo DOCA IMA). A Pressão Arterial Sistólica (PAS) foi medida quinzenalmente. Foi estudada a porção apical do ventrículo esquerdo. Foram empregados: coloração vermelho sirius, coloração de hematoxilina-eosina, imuno-histoquímica e ensaio de western blot. RESULTADOS: A PAS nos grupos DOCA e IMA+DOCA foi maior que no grupo CON nos dias 14 e 28. Os animais do grupo DOCA apresentaram fibrose intersticial e perivascular grave no dia 28, e as expressões de PI, PIII, tenascina-C e fibronectina foram significativamente maiores que nos grupos DOCA+IMA e CON. Quando comparados com o grupo CON, os grupos DOCA e DOCA+IMA apresentaram resposta inflamatória de tecido miocárdico e infiltração de monócitos/macrófagos de diferentes graus. As expressões proteicas do PDGF-A, PDGF-C e PDGFRα foram significativamente maiores nos grupos DOCA e DOCA+IMA que no grupo CON, mas a expressão proteica do p-PDGFRα no grupo DOCA+IMA foi menor que no DOCA. CONCLUSÃO: O Imatinib pode exercer efeitos inibitórios sobre a fibrose miocárdica em ratos com hipertensão induzida por DOCA/sal, os quais podem ser atribuídos à inibição da atividade do PDGFR-α.

BACKGROUND: Imatinib is a tyrosine kinase receptor inhibitor that has been confirmed to exert inhibitory effect on the platelet derived growth factor PDGF receptor (PDGFRα and PDGFRβ) activity. OBJECTIVE: To investigate the protective effect of imatinib on the myocardial fibrosis in deoxycorticosterone-acetate (DOCA)/salt induced hypertensive rats. METHODS: Sixty male uninephrectomized Sprague-Dawley rats were assigned to three groups: control rats (CON group); deoxycorticosterone group (DOCA group); deoxycorticosterone and imatinib group (DOCA+IMA group). Systolic blood pressure (SBP) was measured biweekly. The apical portion of the left ventricle was studied. Sirius-Red staining, Hematoxylin-Eosin staining, immunohistochemistry and Western blot assay were employed. RESULTS: SBP in the DOCA group and DOCA+IMA group was higher than that in the CON group on day 14 and 28. Animals in the DOCA group showed severe interstitial and perivascular fibrosis on day 28, and the expressions of PI, PIII, tenascin-C and fibronectin were significantly higher than those in the DOCA+IMA group and CON group. When compared with the CON group, myocardial tissue inflammatory response and monocyte/macrophage infiltration of different degrees were observed in the DOCA group and DOCA+IMA group. Protein expressions of PDGF-A, PDGF-C and PDGFRα were signiflcantly higher in the DOCA and DOCA+IMA groups than those in the CON group, but the p-PDGFRα protein expression in the DOCA+IMA group was lower than that in the DOCA group. CONCLUSION: Imatinib can exert inhibitory effects on myocardial fibrosis in DOCA/salt induced hypertensive rats, which may be attributed to the inhibition of PDGFR-α activity.

Imatinib attenuates myocardial fibrosis in association with inhibition of the PDGFRα activity.

BACKGROUND: Imatinib is a tyrosine kinase receptor inhibitor that has been confirmed to exert inhibitory effect on the platelet derived growth factor PDGF receptor (PDGFRα and PDGFRß) activity. OBJECTIVE: To investigate the protective effect of imatinib on the myocardial fibrosis in deoxycorticosterone-acetate (DOCA)/salt induced hypertensive rats. METHODS: Sixty male uninephrectomized Sprague-Dawley rats were assigned to three groups: control rats (CON group); deoxycorticosterone group (DOCA group); deoxycorticosterone and imatinib group (DOCA+IMA group). Systolic blood pressure (SBP) was measured biweekly. The apical portion of the left ventricle was studied. Sirius-Red staining, Hematoxylin-Eosin staining, immunohistochemistry and Western blot assay were employed. RESULTS: SBP in the DOCA group and DOCA+IMA group was higher than that in the CON group on day 14 and 28. Animals in the DOCA group showed severe interstitial and perivascular fibrosis on day 28, and the expressions of PI, PIII, tenascin-C and fibronectin were significantly higher than those in the DOCA+IMA group and CON group. When compared with the CON group, myocardial tissue inflammatory response and monocyte/macrophage infiltration of different degrees were observed in the DOCA group and DOCA+IMA group. Protein expressions of PDGF-A, PDGF-C and PDGFRα were significantly higher in the DOCA and DOCA+IMA groups than those in the CON group, but the p-PDGFRα protein expression in the DOCA+IMA group was lower than that in the DOCA group. CONCLUSION: Imatinib can exert inhibitory effects on myocardial fibrosis in DOCA/salt induced hypertensive rats, which may be attributed to the inhibition of PDGFR-α activity.

Malignant lesions in the ventral prostate of alloxan-induced diabetic rats.

The aim of this study was to evaluate the changes caused by chronic diabetes in the rat ventral prostate and to establish a correlation between diabetes and the development of prostatic lesions. Male rats received alloxan (42 mg/kg b.w.) to induce diabetes. Ninety days after diabetes diagnosis, animals were sacrificed and the ventral prostate was removed and prepared for general and immunohistochemical analyses. The total area showing different types of lesions was estimated. Diabetes led to a decrease in the body and prostatic weights, as well as in testosterone levels. The prostate morphology and stereology showed high variation in the diabetic group. Some animals had light changes; the great majority had an intense epithelial atrophy; and other rats showed premalignant and malignant lesions in the prostate. Such epithelial atrophy was, in some samples, combined with chronic inflammation, similar to proliferative inflammatory atrophy (PIA). The diabetic group also presented high incidence of prostatitis, adenocarcinoma and prostatic intra-epithelial neoplasia (PIN). Samples with adenocarcinoma had poorly differentiated acini with high levels of cellular proliferation and nuclear atypia. These lesions exhibited an invasive feature showing Bcl-2-positive cells and interruptions in the basement membrane. An association of PIA, PIN and adenocarcinoma was detected in one sample. Reduced androgen levels have a synergic effect to insulin dysfunction promoting negative effects in the rat prostate. Diabetic individuals had a high incidence of prostatitis, and this inflammation could stimulate the incidence of other forms of prostatic pathology.

Tenascin and fibronectin expression in carcinoma ex pleomorphic adenoma.

This study was conducted to analyze the participation of tenascin and fibronectin, components of the extracellular matrix, in different types of carcinoma ex pleomorphic adenoma (CXPA). Seventeen cases of CXPA, classified according to the presence of epithelial and myoepithelial cells and the degree of invasion-intracapsular, minimally, and frankly invasive carcinoma-were immunohistochemically labeled for tenascin and fibronectin. Normal salivary gland included in the specimens showed tenascin only around the excretory duct, and fibronectin slightly expressed all over the stroma of the gland. In reminiscent pleomorphic adenoma, tenascin and fibronectin were observed around tubular structures and in the stroma. Both tenascin and fibronectin were expressed in all the CXPA studied. In areas of in situ carcinoma of the intracapsular type, the expression of these extracellular matrix proteins was enhanced compared with areas of residual pleomorphic adenoma. In intracapsular and minimally invasive types of CXPA, some areas of the tumor border presented tenascin and no fibronectin, pattern that may represent the real invasive front. In frankly invasive CXPA type with only epithelial component, fibronectin was strongly observed in a fibrillar network pattern, and tenascin was only focal. In frankly invasive type with myoepithelial component, tenascin staining was very strong and diffuse. This study showed different patterns of expression of tenascin and fibronectin along the process of tumorigenesis and tumor progression in CXPA, a fact that might play a role in invasion properties of these tumors.

Caracterização imuno-histoquímica de lesão híbrida de ameloblastoma desmoplásico e convencional / Hybrid lesion of desmoplastic and conventional ameloblastoma: immunohistochemical aspects

O ameloblastoma é uma neoplasia odontogênica benigna comumente encontrada nos ossos maxilares. Histologicamente, mostra diversos padrões, incluindo a ameloblastoma plexiforme e folicular. Quando estes padrões histológicos coexistem com um ameloblastoma que exibe abundante desmoplasia, são então denominados de lesão æhíbrida" de ameloblastoma desmoplásico e convencional. No presente trabalho, nos propomos a relatar um caso de lesão híbrida de ameloblastoma desmoplásico e convencional destacando os aspectos imuno-histoquímicos relativos a expressão das proteínas da matriz extracelular (tenascina, fibronectina e colágeno I).

Ameloblastoma is a benign epithelial odontogenic tumor and is the most commonly encountered odontogenic tumor in the jaws. Histologically, ameloblastomas occur in different patterns, including plexiform pattern and follicular pattern. "Hybrid " lesion of ameloblastoma is a tumor variant in which histologically, areas of follicular or plexiform ameloblastoma coexist with characteristic areas of ameloblastoma exhibiting pronounced stromal desmoplasia (desmoplastic ameloblastoma). The purpose of this article is to present a case of "hybrid" lesion of desmoplastic ameloblastoma (AD) and conventional, and investigate extracellular matrix proteins such as tenascin, fibronectin, and type I collagen.

"Hybrid" lesion of desmoplastic and conventional ameloblastoma: immunohistochemical aspects.

Ameloblastoma is a benign epithelial odontogenic tumor and is the most commonly encountered odontogenic tumor in the jaws. Histologically, ameloblastomas occur in different patterns, including plexiform pattern and follicular pattern. "Hybrid" lesion of ameloblastoma is a tumor variant in which histologically, areas of follicular or plexiform ameloblastoma coexist with characteristic areas of ameloblastoma exhibiting pronounced stromal desmoplasia (desmoplastic ameloblastoma). The purpose of this article is to present a case of "hybrid" lesion of desmoplastic ameloblastoma (AD) and conventional, and investigate extracellular matrix proteins such as tenascin, fibronectin, and type I collagen.

Immunoexpression of extracellular matrix proteins in human salivary gland development.

Immunoexpression of the extracellular matrix (ECM) proteins laminin, fibronectin, tenascin and types I, III and IV collagen was analyzed in the major and minor salivary glands of seven human fetuses at different gestational ages. The results showed the presence and localization of laminin, collagen IV and fibronectin around glandular structures at all stages of development. Tenascin was only detectable around excretory ducts. In the earliest stages of development, type I and type III collagen were presented as fine fibers delineating the glandular structures and delimiting the extension of the future lobule. As glandular development proceeded, the lobule was gradually filled with collagens and glandular tissue.

Immunohistochemical assessment of extracellular matrix components in syndrome and non-syndrome odontogenic keratocysts.

OBJECTIVE: Investigate the immunohistochemical distribution of fibronectin, tenascin, laminin and collagen IV in syndrome (SOKC) and non-syndrome odontogenic keratocysts (NSOKC). MATERIALS AND METHODS: Ten cases of SOKC and five of NSOKC were selected and streptoavidin-biotin technique was applied. The specimens were analyzed taking into account the following evaluation parameters: presence, continuity and thickness in the basement membrane and intensity, distribution and association with inflammatory cells in the cyst wall. RESULTS: Differences could be detected regarding tenascin, fibronectin and collagen IV between the SOKC and NSOKC. Tenascin was present in all cases along the basement membrane in SOKC and in five cases of NSOKC predominated negative areas. Furthermore, tenascin distribution was focal in the cyst wall in SOKC whereas in NSOKC it was diffuse. Concerning fibronectin, it was detected as a discontinuous band when present in SOCK and as a continuous band in NSOKC. Collagen IV was not present in the majority of the cases in SOKC. Negative areas for laminin predominated in the basement membrane in both groups. CONCLUSIONS: These findings show differences between the immunohistochemical expression of tenascin, fibronectin and collagen IV which might indicate a more aggressive biological behavior of SOKC as compared with NSOKC.

Tenascin and fibronectin expression in odontogenic cysts.

BACKGROUND: Odontogenic cysts (OCs) present distinct evolution and clinical behavior. This study was performed in order to investigate if some components of the extracellular matrix (ECM) may drive these differences. METHODS: Thirty OCs were selected: 10 radicular cysts (RCs), 10 dentigerous cysts (DCs), 10 non-syndrome odontogenic keratocysts (OKCs) and they were immunohistochemically analyzed to verify the expression pattern of tenascin and fibronectin. RESULTS: Tenascin immunostaining was mainly intense as a thick band deep to the epithelial-mesenchymal interface in both RCs and OKCs. The intense tenascin immunoexpression observed in the RCs was usually associated with inflammation. An intense fibronectin reactivity was observed in the basement membrane region and at the cystic wall of OKCs. CONCLUSIONS: Our results demonstrate differences between the expression of ECM proteins in the OCs studied. The higher tenascin and fibronectin expression in the capsule of OKCs suggests marked instability in the cystic structure and may contribute to its aggressive behavior.

Immunohistochemical study of the orthokeratinized odontogenic cyst: a comparison with the odontogenic keratocyst.

OBJECTIVE: Orthokeratinized odontogenic cyst (OOC) is a developmental cyst that occurs in the maxilla and the mandible and is defined by the World Health Organization as the uncommon orthokeratinized type of odontogenic keratocyst (OKC). However, studies have shown that OOC has peculiar clinicopathologic aspects and biologic behavior when compared with other developmental odontogenic cysts, especially OKCs. Therefore, in this study, the immunohistochemical profile of the OOC was delineated and compared with that of the OKC. STUDY DESIGN: Twelve cases of OOC were submitted to a panel of antibodies composed of cytokeratins (10, 13, and 14) and extracellular matrix proteins: fibronectin, types I and III collagen, and tenascin. For comparative means, 12 cases of OKC also were submitted to the same panel of antibodies. RESULTS: The results obtained showed that OOCs expressed cytokeratin 10 and showed variable expression of cytokeratins 13 and 14. Fibronectin and collagen types I and III also were expressed in OOC in a fibrillar aspect. OKC showed only the superficial keratin layer positive to cytokeratin 10 and the basal and suprabasal layers with variable expression of cytokeratin 14, and cytokeratin 13 was present in the upper epithelial layers. The extracellular matrix proteins showed a nonfibrillar expression. Tenascin was immunoexpressed only in OKC. CONCLUSION: The immunohistochemical profile of the studied cysts clearly showed that OOC presents a well-formed cystic enveloping, whereas the OKC profile is compatible with a more aggressive biologic behavior.

A novel cell line that retains the morphological characteristics of the cells and matrix of odontogenic myxoma.

Little is known about the histogenesis of the human odontogenic myxoma or the relation between tumour cells and the matrix. In order to attempt to remedy this situation, we established and investigated a cell line derived from a human odontogenic myxoma. To our knowledge this is the first cell line derived from this tumour. The cell line, named Mix 1, preserved features of the tumour cells. Mix 1 cells expressed vimentin, type I collagen, fibronectin, tenascin and hyaluronic acid. Ultrastructural analysis of cells of the tumour and cell line demonstrated similarities, both containing Golgi apparatus, rough endoplasmic reticulum and mitochondria indicative of secretory cells. Ultrastructural analysis showed the matrix to be represented by bundles of collagen fibrils in the tumour, and by irregular filaments in cultures more than 60 days old. The Mix 1 cell line promises to be an excellent model for investigating the biology of the odontogenic myxoma.

Extracellular matrix proteins increase the expression of pro-TRH and pro-protein convertase PC1 in fetal hypothalamic neurons in vitro.

External clues for neuron development include extracellular matrix (ECM) molecules. To explore ECM influence on the early development of peptide phenotype in the CNS, we have compared pro-TRH levels in primary cultures of rat hypothalamic cells plated either on poly-lysine (PL) (control) or on PL plus one of various ECM molecules at 10 microgram/ml. Fetal day 17 cells plated at a density of 1250/mm(2) were grown in a serum free medium made of Neurobasal medium supplemented with B27 (GIBCO). Cultures, consisting mainly of neurons, were analyzed at DIV 2. ECM proteins induced morphological effects in agreement with previously published studies. The amount of pro-TRH per dish, quantified by Western blotting, was increased to 275% for laminin, 191% for fibronectin and 173% for tenascin-C (control=100%); there was no effect of vitronectin. Laminin or fibronectin did not change pro-TRH mRNA or TRH levels but enhanced levels of the pro-protein convertase PC1 suggesting that the ECM molecules did regulate the translational status of pro-TRH. In conclusion, we have shown that some ECM proteins increased pro-TRH level in vitro; this may contribute to the enhancement of pro-TRH levels observed early in vivo in the hypothalamus.

Immunohistochemical localization of tenascin, fibronectin, and type III collagen in human dental pulp.

The distribution of tenascin (TN), fibronectin (FN), and type III collagen (col III) in the extracellular matrix of the connective tissue of normal, inflamed, and hyalinized human dental pulp was studied by immunohistochemical staining with monoclonal antibodies against these molecules. TN, FN, and col III were present in all normal tissues studied. In areas of hyalinization only col III was observed. None of the molecules studied were seen in areas of inflammatory exudate. Strong staining for TN and FN was found in the periphery of all specimens analyzed next to the odontoblastic layer. We therefore conclude that TN, FN, and col III are present in the extracellular matrix of normal human dental pulp. TN, FN, and col III distribution in inflammatory and degenerative processes is different from that observed in normal human dental pulp.

Molecular tunnels and boundaries for growing axons in the anterior commissure of hamster embryos.

We have analyzed the immunohistochemical expression of chondroitin sulfate proteoglycan (CSPG), fibronectin (FN), laminin (LN), tenascin (TN), and glial fibrillary acidic protein (GFAP) along the anterior commissure (AC) of hamster embryos (n=175; from embryonic day (E)12 to E16). Frozen sections were cut at different planes from embryonic brains between E12 and E16, treated for immunohistochemistry, and observed under epifluorescence microscopy. During the pre-crossing stage (E12-E13), CSPG was expressed as a sagittal stratum between the interhemispheric fissure and the prospective AC region. TN appeared rostral to the third ventricle and along the medial subventricular zone of the lateral ventricles. LN and FN both presented a faint expression, and GFAP was not detected. Although AC axons started crossing the midline region (E13.5-E14), CSPG, FN, LN, and, much less intensely, GFAP circumscribed the AC bundle, forming a tunnel through which AC fibers elongate. TN was no longer seen at the midplane but remained visible laterally. During the post-crossing stage (E14.5-E16), CSPG and TN were no longer seen at the midline, although both could be observed between the AC limbs, seeming to form boundaries for AC lateral growth. LN and FN were then absent near the AC bundle. During this late stage, GFAP expression became most intense, forming a distinct tunnel around the AC. We have shown that the expression of extracellular matrix molecules and GFAP follow a time- and space-regulated course related to AC development, plausibly representing influential factors for growth and guidance of commissural fibers.