espanolIntroduccion: la displasia cemento osea periapical es una lesion osea benigna de etiologia... more espanolIntroduccion: la displasia cemento osea periapical es una lesion osea benigna de etiologia desconocida. Presenta mayor prevalencia en mujeres, melanodermas, de descendencia africana o del suroeste asiatico. Es comunmente descubierta en examenes radiograficos de rutina, una vez que no presenta signos y sintomas clinicos. Debido a sus caracteristicas radiograficas, puede ser confundida con lesiones periapicales inflamatorias (quistes y granulomas periapicales); por tanto su diagnostico debe ser realizado de forma que el odontologo obedezca la conducta clinica adecuada al caso, con el fin de evitar iatrogenias. Objetivo: El objetivo de este trabajo es disertar sobre displasia cemento osea periapical (etiologia, tratamiento y pronostico), destacando las dificultades del diagnostico de esta lesion y su impacto en la terapia endodontica. Relato de Caso: paciente femenino, 37 anos de edad, con diagnostico de displasia cemento osea periapical con 4 anos de seguimiento en el area de e...
BACKGROUND Despite advances in cancer diagnosis and therapeutics, the overall 5-year survival rat... more BACKGROUND Despite advances in cancer diagnosis and therapeutics, the overall 5-year survival rate of oral squamous cell carcinoma (OSCC) remains low. Tumor formation, progression, recurrence, and chemo-resistance are associated with the presence of cancer stem cells (CSC) that show phenotypic heterogeneity, but how they influence tumor behavior remains poorly understood. We aimed to describe how two CSC phenotypes from an OSCC cell line, CD44High ESAHigh (Epi-CSC) and CD44High ESALow (EMT-CSC), behave in vitro and in vivo. METHODS In vitro behavior of FACS-sorted Epi-CSC and EMT-CSC from OSCC cells was characterized by their ability to form colonies, migrate, proliferate, and to invade a solid matrix. In vivo experiments were conducted in immunodeficient (NOD/SCID) mice by orthotopic xenografting of FACS-sorted OSCC subpopulations. RESULTS In vitro, the Epi-CSC phenotype was more proliferative and generated more holoclones than the EMT-phenotype. On the other hand, EMT-CSC cells migrate and invaded more than Epi-CSC cells in 3D culture, suggesting the CSC phenotype affects tumor cell behavior. When inoculated orthotopically into the tongues of immunodeficient mice, both subpopulations generated OSCC, but EMT-CSC formed fewer and smaller tumors. CONCLUSIONS Our results suggest that while cells in the Epi-CSC form the subpopulation that enables tumor growth, the EMT-CSC are related to migration and invasion. Clinically, this may reflect the importance of Epi-CSC for tumorigenesis and of the EMT-CSC for metastasis and highlights that variation in the proportion of CSC phenotypes from patient to patient may be relevant to the design of individual treatment protocols.
This study evaluated the viability, proliferation, and protein expression after photobiomodulatio... more This study evaluated the viability, proliferation, and protein expression after photobiomodulation (PBM) of stem cell from human exfoliated deciduous teeth (SHED). The groups were the following: G1 (2.5 J/cm2), G2 (3.7 J/cm2), and control (not irradiated). According to the groups, cells were irradiated with InGaAlP diode laser at 660 nm wavelength, continuous mode, and single time application. After 6 h, 12 h, and 24 h from irradiation, the cell viability and proliferation, and the protein expression were analyzed by MTT, crystal violet, and ELISA multiplex assay, respectively. Twenty-four hours after PBM, SHED showed better proliferation. Over time in the supernatant, all groups had an increase at the levels of VEGF-C, VEGF-A, and PLGF. In the lysate, the control and G2 exhibited a decrease of the VEGF-A, PECAM-1, and PLGF expression, while control and G3 decreased VEGF-C, VEGF-A, and PDGF expression. The dosimetries of 2.5 J/cm2 and 3.7 J/cm2 maintained viability, improved proliferation, and synthesis of the angiogenic proteins in the supernatant in the studied periods on SHED.
The initiation and progression of periodontitis might involve a local renin-angiotensin system (R... more The initiation and progression of periodontitis might involve a local renin-angiotensin system (RAS) in periodontal tissue. This study hypothesized that Losartan treatment could promote protection to rats submitted to experimental periodontitis (EP) by attenuating alveolar bone loss due to reduction in inflammatory cytokines, better ROS regulation and maintenance of the balance between bone formation and resorption factors. One hundred and thirty rats were submitted to EP with a silk suture thread (4.0) placed around the lower right first molar for 1, 3, 7 and 14 consecutive days. The study comprised 4 groups: G1 - control without EP; G2 - animals with EP treated with water; G3 - Losartan-treated animals (treatment started at the same day of EP induction) and G4 - animals previously treated with Losartan for 30 days followed by induction of EP and continuity of treatment. G2 rats had greater bone loss volume, increased number and thickness and decreased separation of trabeculae. On the other hand, G4 animals showed significant improvements in these parameters. Histological analysis revealed that EP favors inflammatory cell infiltration and junctional epithelium, cementum with alveolar bone crest destruction, but animals pretreated with Losartan (G4) did not show these features. Although the G3 animals did not demonstrate the improvements detected in G4, mRNA expression results were similar. In mandibular tissue, EP promoted mRNA increases for ACE, AT1 receptor and inflammatory mediators as well as decreases for antioxidant enzymes. However, Losartan treatments attenuated these responses in addition to promoting an increase in bone formation markers and transcription factors. This article is protected by copyright. All rights reserved.
The biological process of epithelial-to-mesenchymal transition (EMT) has been studied in oral squ... more The biological process of epithelial-to-mesenchymal transition (EMT) has been studied in oral squamous cell carcinoma (OSCC) metastasis, but it is rarely evaluated at several stages of oral carcinogenesis. This study aimed to analyze the presence of SNAIL and E-cadherin proteins, markers of EMT, in the development and progression of OSCC, evaluating excised specimens of potentially malignant lesions (oral leukoplakia with and without dysplasia-OL and OLD, respectively), tumor tissues (OSCC), metastatic lymph nodes (LN), and normal oral mucosa (NOM) by immunohistochemistry, considering subcellular localization. Additionally, SNAIL and E-cadherin transcripts were evaluated in vitro by qPCR, using SCC-9 cell line in comparison to human keratinocytes (HPEC). There was a significant increase in nuclear expression of SNAIL from NOM to OLD followed by a noticeable decrease in nuclear expression accompanied by increased cytoplasmic expression in OSCC (p<0.05). The E-cadherin cytoplasmic ...
Oral squamous cell carcinoma (OSCC) is an extremely aggressive disease associated with a poor pro... more Oral squamous cell carcinoma (OSCC) is an extremely aggressive disease associated with a poor prognosis. Previous studies have established that cancer stem cells (CSCs) actively participate in OSCC development, progression and resistance to conventional treatments. Furthermore, CSCs frequently exhibit a deregulated expression of normal stem cell signalling pathways, thereby acquiring their distinctive abilities, of which self-renewal is an example. In this study, we examined the effects of GLI3 knockdown in OSCC, as well as the differentially expressed genes in CSC-like cells (CSCLCs) expressing high (CD44high) or low (CD44low) levels of CD44. The prognostic value of GLI3 in OSCC was also evaluated. The OSCC cell lines were sorted based on CD44 expression; gene expression was evaluated using a PCR array. Following this, we examined the effects of GLI3 knockdown on CD44 and ESA expression, colony and sphere formation capability, stem-related gene expression, proliferation and invasio...
Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology, Jan 23, 2018
Tumour metastasis has been associated with cancer stem cells, a small population with stem-like c... more Tumour metastasis has been associated with cancer stem cells, a small population with stem-like cells properties, higher rate of migration and metastatic potential compared to cells from the tumour bulk. Our aim was to evaluate the immunoexpression of the putative cancer stem cell biomarkers ALDH1 and CD44 in primary tumour and corresponding metastatic lymph nodes. Tumour tissue specimens (n = 50) and corresponding metastatic lymph nodes (n = 25) were surgically obtained from 50 patients with oral squamous cell carcinoma and submitted to immunohistochemistry. CD44 and ALDH1 were semi-quantitatively scored according to the proportion and intensity of positive cells within the invasive front and metastatic lymph nodes as a whole. A combined score was obtained by multiplying both parameters and later dichotomized into a final score classified as low (≤2) or high (>2) immunoexpression. ALDH1 immunoexpression and CD44 immunoexpression were detected in both tumour sites, although the m...
Research on cancer stem cells (CSCs) has greatly increased in the field of medicine and pathology... more Research on cancer stem cells (CSCs) has greatly increased in the field of medicine and pathology; however, some conceptual misunderstandings are still present among the public as well as within the general scientific community that is not yet familiar with the subject. The very first problem is the misinterpretation of CSCs as a synonym of their normal counterparts, the well-known stem cells (SCs). Particularly in Dentistry, another common mistake is the misinterpretation of oral CSCs as normal tooth-derived SCs. The present review aims to clarify important concepts related to normal SCs and CSCs, as well as discuss the relevance of CSCs to the development, metastasis and therapy resistance of oral squamous cell carcinoma.
ABSTRACT The search for molecular markers to improve diagnosis, individualize treatment and predi... more ABSTRACT The search for molecular markers to improve diagnosis, individualize treatment and predict prognosis has been the focus of several groups studying oral squamous cell carcinoma (OSCC). Objectives: The purpose of this study was to evaluate the expression of IRX4 homeobox gene and its respective protein in OSCC. Methods: After microarray analysis of tumor samples and their tumor-free margins, two-fold differentially expressed homeobox genes were selected for further validation by real time RT-PCR, and the expression levels were then correlated to clinical-pathological parameters. Immunohistochemical staining and western blot analyses were also performed. Results: Among other homeobox genes presenting altered expression, IRX4 showed the highest and statistically significant (p= 0.003) median expression levels in tumors (2169.4) when compared to their non-tumoral margins (653.5). IRX4 up-regulation was also associated with a decreased overall survival rate. IRX4 protein localization and quantitation were assed by immunohistochemistry and western blot, respectively. Tumor samples revealed strong and widely distributed nuclear immunostaining in atypical keratinocytes within squamous cell carcinoma islands. In accordance, immunoblotting showed an increased protein level in tumor samples. Subsequent analysis of homeobox gene expression in OSCC cell lines (SCC-4, -9, 15 and -25) revealed IRX4 overexpression in most neoplastic cell lines compared to normal human oral keratinocytes, which was also confirmed by western blot analysis. Conclusion: These results suggest that IRX4 participate in the maintenance of the malignant phenotype of OSCC and is associated with prognosis. Additional functional studies are being conducted to better understand IRX4 participation in oral carcinogenesis. Financial support: FAPESP 06/04736-8 and 09/07359-9.
espanolIntroduccion: la displasia cemento osea periapical es una lesion osea benigna de etiologia... more espanolIntroduccion: la displasia cemento osea periapical es una lesion osea benigna de etiologia desconocida. Presenta mayor prevalencia en mujeres, melanodermas, de descendencia africana o del suroeste asiatico. Es comunmente descubierta en examenes radiograficos de rutina, una vez que no presenta signos y sintomas clinicos. Debido a sus caracteristicas radiograficas, puede ser confundida con lesiones periapicales inflamatorias (quistes y granulomas periapicales); por tanto su diagnostico debe ser realizado de forma que el odontologo obedezca la conducta clinica adecuada al caso, con el fin de evitar iatrogenias. Objetivo: El objetivo de este trabajo es disertar sobre displasia cemento osea periapical (etiologia, tratamiento y pronostico), destacando las dificultades del diagnostico de esta lesion y su impacto en la terapia endodontica. Relato de Caso: paciente femenino, 37 anos de edad, con diagnostico de displasia cemento osea periapical con 4 anos de seguimiento en el area de e...
BACKGROUND Despite advances in cancer diagnosis and therapeutics, the overall 5-year survival rat... more BACKGROUND Despite advances in cancer diagnosis and therapeutics, the overall 5-year survival rate of oral squamous cell carcinoma (OSCC) remains low. Tumor formation, progression, recurrence, and chemo-resistance are associated with the presence of cancer stem cells (CSC) that show phenotypic heterogeneity, but how they influence tumor behavior remains poorly understood. We aimed to describe how two CSC phenotypes from an OSCC cell line, CD44High ESAHigh (Epi-CSC) and CD44High ESALow (EMT-CSC), behave in vitro and in vivo. METHODS In vitro behavior of FACS-sorted Epi-CSC and EMT-CSC from OSCC cells was characterized by their ability to form colonies, migrate, proliferate, and to invade a solid matrix. In vivo experiments were conducted in immunodeficient (NOD/SCID) mice by orthotopic xenografting of FACS-sorted OSCC subpopulations. RESULTS In vitro, the Epi-CSC phenotype was more proliferative and generated more holoclones than the EMT-phenotype. On the other hand, EMT-CSC cells migrate and invaded more than Epi-CSC cells in 3D culture, suggesting the CSC phenotype affects tumor cell behavior. When inoculated orthotopically into the tongues of immunodeficient mice, both subpopulations generated OSCC, but EMT-CSC formed fewer and smaller tumors. CONCLUSIONS Our results suggest that while cells in the Epi-CSC form the subpopulation that enables tumor growth, the EMT-CSC are related to migration and invasion. Clinically, this may reflect the importance of Epi-CSC for tumorigenesis and of the EMT-CSC for metastasis and highlights that variation in the proportion of CSC phenotypes from patient to patient may be relevant to the design of individual treatment protocols.
This study evaluated the viability, proliferation, and protein expression after photobiomodulatio... more This study evaluated the viability, proliferation, and protein expression after photobiomodulation (PBM) of stem cell from human exfoliated deciduous teeth (SHED). The groups were the following: G1 (2.5 J/cm2), G2 (3.7 J/cm2), and control (not irradiated). According to the groups, cells were irradiated with InGaAlP diode laser at 660 nm wavelength, continuous mode, and single time application. After 6 h, 12 h, and 24 h from irradiation, the cell viability and proliferation, and the protein expression were analyzed by MTT, crystal violet, and ELISA multiplex assay, respectively. Twenty-four hours after PBM, SHED showed better proliferation. Over time in the supernatant, all groups had an increase at the levels of VEGF-C, VEGF-A, and PLGF. In the lysate, the control and G2 exhibited a decrease of the VEGF-A, PECAM-1, and PLGF expression, while control and G3 decreased VEGF-C, VEGF-A, and PDGF expression. The dosimetries of 2.5 J/cm2 and 3.7 J/cm2 maintained viability, improved proliferation, and synthesis of the angiogenic proteins in the supernatant in the studied periods on SHED.
The initiation and progression of periodontitis might involve a local renin-angiotensin system (R... more The initiation and progression of periodontitis might involve a local renin-angiotensin system (RAS) in periodontal tissue. This study hypothesized that Losartan treatment could promote protection to rats submitted to experimental periodontitis (EP) by attenuating alveolar bone loss due to reduction in inflammatory cytokines, better ROS regulation and maintenance of the balance between bone formation and resorption factors. One hundred and thirty rats were submitted to EP with a silk suture thread (4.0) placed around the lower right first molar for 1, 3, 7 and 14 consecutive days. The study comprised 4 groups: G1 - control without EP; G2 - animals with EP treated with water; G3 - Losartan-treated animals (treatment started at the same day of EP induction) and G4 - animals previously treated with Losartan for 30 days followed by induction of EP and continuity of treatment. G2 rats had greater bone loss volume, increased number and thickness and decreased separation of trabeculae. On the other hand, G4 animals showed significant improvements in these parameters. Histological analysis revealed that EP favors inflammatory cell infiltration and junctional epithelium, cementum with alveolar bone crest destruction, but animals pretreated with Losartan (G4) did not show these features. Although the G3 animals did not demonstrate the improvements detected in G4, mRNA expression results were similar. In mandibular tissue, EP promoted mRNA increases for ACE, AT1 receptor and inflammatory mediators as well as decreases for antioxidant enzymes. However, Losartan treatments attenuated these responses in addition to promoting an increase in bone formation markers and transcription factors. This article is protected by copyright. All rights reserved.
The biological process of epithelial-to-mesenchymal transition (EMT) has been studied in oral squ... more The biological process of epithelial-to-mesenchymal transition (EMT) has been studied in oral squamous cell carcinoma (OSCC) metastasis, but it is rarely evaluated at several stages of oral carcinogenesis. This study aimed to analyze the presence of SNAIL and E-cadherin proteins, markers of EMT, in the development and progression of OSCC, evaluating excised specimens of potentially malignant lesions (oral leukoplakia with and without dysplasia-OL and OLD, respectively), tumor tissues (OSCC), metastatic lymph nodes (LN), and normal oral mucosa (NOM) by immunohistochemistry, considering subcellular localization. Additionally, SNAIL and E-cadherin transcripts were evaluated in vitro by qPCR, using SCC-9 cell line in comparison to human keratinocytes (HPEC). There was a significant increase in nuclear expression of SNAIL from NOM to OLD followed by a noticeable decrease in nuclear expression accompanied by increased cytoplasmic expression in OSCC (p<0.05). The E-cadherin cytoplasmic ...
Oral squamous cell carcinoma (OSCC) is an extremely aggressive disease associated with a poor pro... more Oral squamous cell carcinoma (OSCC) is an extremely aggressive disease associated with a poor prognosis. Previous studies have established that cancer stem cells (CSCs) actively participate in OSCC development, progression and resistance to conventional treatments. Furthermore, CSCs frequently exhibit a deregulated expression of normal stem cell signalling pathways, thereby acquiring their distinctive abilities, of which self-renewal is an example. In this study, we examined the effects of GLI3 knockdown in OSCC, as well as the differentially expressed genes in CSC-like cells (CSCLCs) expressing high (CD44high) or low (CD44low) levels of CD44. The prognostic value of GLI3 in OSCC was also evaluated. The OSCC cell lines were sorted based on CD44 expression; gene expression was evaluated using a PCR array. Following this, we examined the effects of GLI3 knockdown on CD44 and ESA expression, colony and sphere formation capability, stem-related gene expression, proliferation and invasio...
Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology, Jan 23, 2018
Tumour metastasis has been associated with cancer stem cells, a small population with stem-like c... more Tumour metastasis has been associated with cancer stem cells, a small population with stem-like cells properties, higher rate of migration and metastatic potential compared to cells from the tumour bulk. Our aim was to evaluate the immunoexpression of the putative cancer stem cell biomarkers ALDH1 and CD44 in primary tumour and corresponding metastatic lymph nodes. Tumour tissue specimens (n = 50) and corresponding metastatic lymph nodes (n = 25) were surgically obtained from 50 patients with oral squamous cell carcinoma and submitted to immunohistochemistry. CD44 and ALDH1 were semi-quantitatively scored according to the proportion and intensity of positive cells within the invasive front and metastatic lymph nodes as a whole. A combined score was obtained by multiplying both parameters and later dichotomized into a final score classified as low (≤2) or high (>2) immunoexpression. ALDH1 immunoexpression and CD44 immunoexpression were detected in both tumour sites, although the m...
Research on cancer stem cells (CSCs) has greatly increased in the field of medicine and pathology... more Research on cancer stem cells (CSCs) has greatly increased in the field of medicine and pathology; however, some conceptual misunderstandings are still present among the public as well as within the general scientific community that is not yet familiar with the subject. The very first problem is the misinterpretation of CSCs as a synonym of their normal counterparts, the well-known stem cells (SCs). Particularly in Dentistry, another common mistake is the misinterpretation of oral CSCs as normal tooth-derived SCs. The present review aims to clarify important concepts related to normal SCs and CSCs, as well as discuss the relevance of CSCs to the development, metastasis and therapy resistance of oral squamous cell carcinoma.
ABSTRACT The search for molecular markers to improve diagnosis, individualize treatment and predi... more ABSTRACT The search for molecular markers to improve diagnosis, individualize treatment and predict prognosis has been the focus of several groups studying oral squamous cell carcinoma (OSCC). Objectives: The purpose of this study was to evaluate the expression of IRX4 homeobox gene and its respective protein in OSCC. Methods: After microarray analysis of tumor samples and their tumor-free margins, two-fold differentially expressed homeobox genes were selected for further validation by real time RT-PCR, and the expression levels were then correlated to clinical-pathological parameters. Immunohistochemical staining and western blot analyses were also performed. Results: Among other homeobox genes presenting altered expression, IRX4 showed the highest and statistically significant (p= 0.003) median expression levels in tumors (2169.4) when compared to their non-tumoral margins (653.5). IRX4 up-regulation was also associated with a decreased overall survival rate. IRX4 protein localization and quantitation were assed by immunohistochemistry and western blot, respectively. Tumor samples revealed strong and widely distributed nuclear immunostaining in atypical keratinocytes within squamous cell carcinoma islands. In accordance, immunoblotting showed an increased protein level in tumor samples. Subsequent analysis of homeobox gene expression in OSCC cell lines (SCC-4, -9, 15 and -25) revealed IRX4 overexpression in most neoplastic cell lines compared to normal human oral keratinocytes, which was also confirmed by western blot analysis. Conclusion: These results suggest that IRX4 participate in the maintenance of the malignant phenotype of OSCC and is associated with prognosis. Additional functional studies are being conducted to better understand IRX4 participation in oral carcinogenesis. Financial support: FAPESP 06/04736-8 and 09/07359-9.
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