ABSTRACT
The aim of this study was to investigate the effect of acid challenge on the activation of matrix metalloproteinases (MMPs) in the Dentinoenamel junction of primary and permanent teeth submitted to radiotherapy. For this purpose, a total of 178 dental fragments obtained from molars were used, and randomly divided into 2 groups (primary and permanent teeth) / 4 experimental subgroups (irradiated and non-irradiated, demineralized and non-demineralized). The fragments were exposed to radiation, with a dose fraction of 2 Gy, for 5 consecutive days, until a total dose of 60 Gy was reached, with a total of 30 cycles, for 6 weeks. To determine the activity of MMPs on the dentinoenamel junction (DEJ), in situ zymography assays on 0.6mm dental fragments were performed. To assess whether MMP activity would be impacted by an acidic environment, the fragments were placed in a demineralizing solution (pH of 4.8). The finding was that irradiation activated MMPs in DEJ and these effects were more evident in permanent when compared with primary teeth. When the effect of an acid challenge on MMPs activity was investigated, demineralization was observed not to increase MMPs activity in non-irradiated teeth, but it did increase MMPs activity in irradiated teeth. In conclusion, an acid challenge was found to exacerbate activation of MMPs in DEJ of permanent teeth submitted to irradiation, but not in primary teeth.
Subject(s)
Matrix Metalloproteinases , Matrix Metalloproteinases/metabolism , Matrix Metalloproteinases/radiation effects , Matrix Metalloproteinases/analysis , Humans , Time Factors , Tooth, Deciduous/radiation effects , Tooth, Deciduous/drug effects , Dentin/radiation effects , Dentin/drug effects , Dentin/enzymology , Dentition, Permanent , Random Allocation , Hydrogen-Ion Concentration , Tooth Demineralization , Statistics, Nonparametric , Analysis of Variance , Reference Values , Enzyme Activation/radiation effects , Enzyme Activation/drug effectsABSTRACT
Small ruminant lentiviruses (SRLV) are difficult to diagnose due to their escape mechanisms. Therefore, proteomics is an alternative in the search for biomarkers through extracellular matrix metalloproteinases (MMPs), enzymes related to the immune response. In this sense, this study aimed to analyze the profile of MMPs in healthy and infected Toggenburg goats with chronic SRLV infection in Southeast Brazil. Five positive and five negative goats for SRLV were selected using the agar gel immunodiffusion (AGID) microtechnique, western blot (WB), and nested polymerase chain reaction (nPCR). All animals were submitted to blood collection by puncture of the jugular vein, followed by centrifugation to obtain blood plasma, protein quantification by the Bradford method, one-dimensional electrophoretic separation (1D), and identification of protease activity by zymography and confirmation via reverse zymography in the presence of MMP-2 through the action of tissue inhibitors (TIMP-2). The analysis of protein bands was performed using descriptive statistics and densitometry values for zymography were subjected to the Shapiro-Wilk test to determine normality. Little difference was observed in the occurrence of protein bands between groups. Regarding MMPs, no differences were observed in the expression of proMMP-9, MMP-9, and MMP-2 in animals affected by SRLV. TIMP-2 inhibited proMMP-2 and MMP-2 in all animals. Thus, the profile of protein bands does not change in healthy goats with chronic SRLV infection. The TIMP-2 expression allowed proving the existence of MMP-2 in animals chronically infected by SRLV via reverse zymography
Lentivírus de pequenos ruminantes (LVPR) demonstram diagnóstico complexo devido seus mecanismos de escape. Desse modo, a proteômica apresenta-se como alternativa na busca por biomarcadores através das metaloproteinases da matriz extracelular (MMPs), enzimas ligadas a resposta imunológica. Assim, objetivou-se analisar o perfil das MMPs em cabras Toggenburg sadias e com infecção crônica por LVPR no Sudeste brasileiro. Selecionou-se cinco cabras positivas e cinco negativas para LVPR utilizando: microtécnica de imunodifusão em gel de agarose (MIDGA), Western Blot (WB) e reação em cadeia da polimerase nested (nPCR). Todas foram submetidas à coleta de sangue por punção da veia jugular, seguido de centrifugação para obtenção do plasma sanguíneo, quantificação proteica pelo método Bradford, separação via eletroforese unidimensional (1D), e identificação da atividade das proteases por zimografia e confirmação via zimografia reversa na presença da MMP-2 por meio da ação de inibidores teciduais (TIMP-2). A análise das bandas proteicas ocorreu através de estatística descritiva e para a zimografia os valores de densitometria foram submetidos ao teste de Shapiro-Wilk para determinar a normalidade. Observou-se pouca distinção na ocorrência das bandas proteicas entre os grupos. Em relação as MMPs, não houve diferenças na expressão da proMMP-9, MMP-9 e MMP-2 nos acometidos por LVPR. Observou-se que a TIMP-2 inibiu a proMMP-2 e MMP-2 em todos os animais. Dessa forma, o perfil de bandas proteicas não se altera em cabras sadias e com infecção crônica por LVPR. Através da expressão da TIMP-2 foi possível comprovar a existência da MMP-2 em animais cronicamente infectados por LVPR via zimografia reversa
Subject(s)
Animals , Female , Goats/virology , Biomarkers/analysis , Lentiviruses, Ovine-Caprine/isolation & purification , Matrix Metalloproteinases/analysis , Polymerase Chain Reaction/veterinary , Immunodiffusion/methodsABSTRACT
OBJECTIVE: The aim of the present study was to investigate the effects of a calcium hydroxide-based intracanal medication (ICM) on periodontal and endodontic infectious/inflammatory contents and on periodontal clinical parameters in teeth with primary periodontal lesion and secondary endodontic involvement. MATERIALS AND METHODS: Ten patients with abnormal pulp test results and deep probing depth derived from primary periodontal disease with secondary endodontic involvement were included. Samples were collected from root canals (RC) and periodontal pockets (PP) in order to investigate the microbiological status, levels of endotoxin (LPS), cytokines, and matrix metalloproteinases (MMP), before and after ICM. PCR was used for microbiological assessment. The kinetic-chromogenic LAL assay was used for LPS quantification. Quantikine ELISA kits were used for measurement of IL-1 α, IL-1 ß, TNF-α, PGE2, MMP-2, MMP-3, MMP-8, MMP-9, and MMP-13 levels. The statistical analyses were made using the Friedman and Wilcoxon tests (p < 0.05). T test was used to compare data on periodontal characteristics. RESULTS: ICM did not reduce the number of microorganisms in PP and RC, except for Fusobacterium nucleatum in RC. There was a significant reduction in LPS, MMPs, IL-1 ß, and TNF-α levels in PP after ICM. In RC, LPS, MMP13, PGE2, and IL-1ß levels remained unaltered (p > 0.05); however, the levels of the other MMPs and cytokines were reduced (p < 0.05). After 1 year of the root canal treatment, tooth mobility was significantly reduced (p ≤ 0.05). CONCLUSIONS: The use of a calcium hydroxide-based ICM showed positive effects for periodontal treatment prognosis, as it reduced LPS, cytokine, and MMP levels in periodontal pockets. CLINICAL SIGNIFICANCE: Patients presenting deep probing depth and undergoing periodontal treatment for at least 6 months, with no positive response to periodontal therapy, might benefit with the endodontic treatment.
Subject(s)
Calcium Hydroxide/pharmacology , Dental Pulp Cavity/drug effects , Dental Pulp Cavity/microbiology , Periapical Periodontitis/drug therapy , Periapical Periodontitis/microbiology , Root Canal Irrigants/pharmacology , Root Canal Therapy , Adult , Cytokines/analysis , Endotoxins/analysis , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Matrix Metalloproteinases/analysis , Middle Aged , Polymerase Chain ReactionABSTRACT
PURPOSE: To investigate the effect of oxymatrine on periodontitis in rats and related mechanism. Methods: Ninety SD rats were divided into control, model, 10, 20 and 40 mg/kg oxymatrine and tinidazole groups. The periodontitis model was established in later 5 groups. The 10, 20 and 40 mg/kg oxymatrine groups were intragastrically administrated with 10, 20 and 40 mg/kg oxymatrine, respectively. The tinidazole group was intragastrically administrated with 100 mg/kg tinidazole. The treatment duration was 4 weeks. The tooth mobility, gingival and plaque indexes, serum inflammatory factor levels and gingival tissue matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase (TIMP) protein levels were detected. Results: After treatment, compared with model group, in 40 mg/kg oxymatrine group the rat general conditions were obviously improved, the tooth mobility, gingival index and plaque index were significantly decreased (P<0.05), the serum tumor necrosis factor-α, interleukin-1ß and prostaglandin E2 levels were significantly decreased (P<0.05), the MMP-2 and MMP-9 protein levels were significantly decreased (P<0.05), and the TIMP-2 protein level was significantly increased (P<0.05). Conclusions: Oxymatrine can alleviate the experimental periodontitis in rats. The mechanism may be related to its inhibiting inflammatory factor secretion and regulating MMPs/TIMP protein expression.
Subject(s)
Alkaloids/pharmacology , Anti-Inflammatory Agents/pharmacology , Matrix Metalloproteinases/drug effects , Periodontitis/drug therapy , Quinolizines/pharmacology , Tissue Inhibitor of Metalloproteinases/drug effects , Animals , Dental Plaque Index , Dinoprostone/blood , Female , Gingiva/pathology , Interleukin-1beta/blood , Male , Matrix Metalloproteinases/analysis , Periodontitis/metabolism , Random Allocation , Rats, Sprague-Dawley , Reference Values , Reproducibility of Results , Tinidazole , Tissue Inhibitor of Metalloproteinases/analysis , Treatment Outcome , Tumor Necrosis Factor-alpha/bloodABSTRACT
Abstract Purpose: To investigate the effect of oxymatrine on periodontitis in rats and related mechanism. Methods: Ninety SD rats were divided into control, model, 10, 20 and 40 mg/kg oxymatrine and tinidazole groups. The periodontitis model was established in later 5 groups. The 10, 20 and 40 mg/kg oxymatrine groups were intragastrically administrated with 10, 20 and 40 mg/kg oxymatrine, respectively. The tinidazole group was intragastrically administrated with 100 mg/kg tinidazole. The treatment duration was 4 weeks. The tooth mobility, gingival and plaque indexes, serum inflammatory factor levels and gingival tissue matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase (TIMP) protein levels were detected. Results: After treatment, compared with model group, in 40 mg/kg oxymatrine group the rat general conditions were obviously improved, the tooth mobility, gingival index and plaque index were significantly decreased (P<0.05), the serum tumor necrosis factor-α, interleukin-1β and prostaglandin E2 levels were significantly decreased (P<0.05), the MMP-2 and MMP-9 protein levels were significantly decreased (P<0.05), and the TIMP-2 protein level was significantly increased (P<0.05). Conclusions: Oxymatrine can alleviate the experimental periodontitis in rats. The mechanism may be related to its inhibiting inflammatory factor secretion and regulating MMPs/TIMP protein expression.
Subject(s)
Animals , Male , Female , Periodontitis/drug therapy , Quinolizines/pharmacology , Tissue Inhibitor of Metalloproteinases/drug effects , Matrix Metalloproteinases/drug effects , Alkaloids/pharmacology , Anti-Inflammatory Agents/pharmacology , Periodontitis/metabolism , Reference Values , Tinidazole , Dinoprostone/blood , Random Allocation , Dental Plaque Index , Reproducibility of Results , Tumor Necrosis Factor-alpha/blood , Treatment Outcome , Rats, Sprague-Dawley , Tissue Inhibitor of Metalloproteinases/analysis , Matrix Metalloproteinases/analysis , Interleukin-1beta/blood , Gingiva/pathologyABSTRACT
Evidence shows the polymicrobial etiology of endodontic infections, in which bacteria and their products are the main agents for the development, progression, and dissemination of apical periodontitis. Microbial factors in necrotic root canals (e.g., endotoxin) may spread into apical tissue, evoking and supporting a chronic inflammatory load. Thus, apical periodontitis is the result of the complex interplay between microbial factors and host defense against invasion of periradicular tissues. This review of the literature aims to discuss the complex network between endodontic infectious content and host immune response in apical periodontitis. A better understanding of the relationship of microbial factors with clinical symptomatology is important to establish appropriate therapeutic procedures for a more predictable outcome of endodontic treatment.
Subject(s)
Dental Pulp Cavity/microbiology , Dental Pulp Diseases/complications , Dental Pulp Diseases/microbiology , Periapical Periodontitis/microbiology , Bacterial Infections/complications , Bacterial Infections/microbiology , Cytokines/analysis , Cytokines/physiology , Dental Pulp Cavity/pathology , Dental Pulp Diseases/pathology , Endotoxins/physiology , Humans , Lipopolysaccharides/physiology , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases/physiology , Periapical Periodontitis/pathologyABSTRACT
Abstract: Evidence shows the polymicrobial etiology of endodontic infections, in which bacteria and their products are the main agents for the development, progression, and dissemination of apical periodontitis. Microbial factors in necrotic root canals (e.g., endotoxin) may spread into apical tissue, evoking and supporting a chronic inflammatory load. Thus, apical periodontitis is the result of the complex interplay between microbial factors and host defense against invasion of periradicular tissues. This review of the literature aims to discuss the complex network between endodontic infectious content and host immune response in apical periodontitis. A better understanding of the relationship of microbial factors with clinical symptomatology is important to establish appropriate therapeutic procedures for a more predictable outcome of endodontic treatment.
Subject(s)
Humans , Periapical Periodontitis/microbiology , Dental Pulp Cavity/microbiology , Dental Pulp Diseases/complications , Dental Pulp Diseases/microbiology , Periapical Periodontitis/pathology , Bacterial Infections/complications , Bacterial Infections/microbiology , Lipopolysaccharides/physiology , Cytokines/analysis , Cytokines/physiology , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases/physiology , Dental Pulp Cavity/pathology , Dental Pulp Diseases/pathology , Endotoxins/physiologyABSTRACT
O objetivo deste estudo foi avaliar a influência do plasma rico (PRP) e pobre (PPP) em plaquetas na proliferação celular e expressão de metaloproteinases de matriz (MMPs), durante a reparação de úlceras corneais profundas. Foram utilizadas 45 coelhas, distribuídas em 3 grupos (G) experimentais (n=15), designados como grupos PRP (GR), PPP (GP) e Controle (GC), de acordo com o tratamento. Todos os animais foram submetidos à indução cirúrgica unilateral de úlcera corneal. No GR e GP, o sangue autólogo foi centrifugado, utilizando-se protocolo padronizado, e foram confeccionados os colírios de PRP e PPP, e instilados cinco vezes ao dia. No GC, foi utilizado colírio lubrificante. Cada grupo foi subdividido (n=5), segundo o momento final de avaliação, sendo 4 (M4), 7 (M7) e 30 dias (M30). As córneas dos animais foram processadas para avaliação morfológica e imuno-histoquímica para PCNA, MMP1, MMP2, MMP9, MT1-MMP e TIMP1. No M4, os níveis de MMP2 foram maiores no GP e GR, sendo que, no M7, esse comportamento foi observado apenas no GP. No M30, no GR, verificou-se maior número de células epiteliais e marcação para MMP1 que o GP. No GR, a proliferação celular foi maior no M4 que nos demais momentos, e a marcação para MMP2 foi maior no M4 que no M30. O PRP estimula a proliferação celular na fase inicial (M4) do tratamento quando comparado aos demais momentos, diferentemente dos demais tratamentos. O uso de colírios de plasma rico e pobre em plaquetas influencia a expressão de metaloproteinases de matriz envolvidas no processo de reparação corneal(AU)
The aim of this study was to evaluate the influence of platelet-rich (PRP) and poor (PPP) plasma in cell proliferation and matrix metalloproteinases (MMPs) expression during the repair of deep corneal ulcers. Forty-five female rabbits were distributed in 3 experimental groups (G) (n = 15), referred to as PRP (GR), PPP (GP) and Control (GC) groups, in accordance with the treatment. All animals underwent surgical induction of unilateral corneal ulcer. PRP and PPP eye drops were made by using centrifuged blood through standardized protocol, and instilled five times a day. In GC, lubricant eye drops were used. Each group was subdivided (n = 5) according to the final time point, 4 (M4), 7 (M7) and 30 days (M30). The animals' corneas were processed for morphological and immunohistochemical analysis for PCNA, MMP1, MMP2, MMP9, MT1-MMP and TIMP1. In M4, the levels of MMP2 were higher in GP and GR, and in M7, this behavior was only observed in the GP. In M30, more epithelial cells and MMP1 expression were found in GR than GP. In GR, cell proliferation was higher in M4 than at other time points and MMP2 expression was higher in M4 than M30. The PRP stimulates cell proliferation in the early phase (M4) of treatment when compared to other time points, different from other treatments. The use of eye drops of platelet-rich and poor plasma influences the expression of matrix metalloproteinases involved in the corneal repair process(AU)
Subject(s)
Animals , Female , Rabbits , Proliferating Cell Nuclear Antigen/analysis , Platelet-Rich Plasma/physiology , Matrix Metalloproteinases/analysis , Corneal Ulcer/surgery , Immunohistochemistry/veterinary , Wound Healing/physiology , Corneal Injuries/veterinary , Cell Proliferation/physiologyABSTRACT
O objetivo deste estudo foi avaliar a influência do plasma rico (PRP) e pobre (PPP) em plaquetas na proliferação celular e expressão de metaloproteinases de matriz (MMPs), durante a reparação de úlceras corneais profundas. Foram utilizadas 45 coelhas, distribuídas em 3 grupos (G) experimentais (n=15), designados como grupos PRP (GR), PPP (GP) e Controle (GC), de acordo com o tratamento. Todos os animais foram submetidos à indução cirúrgica unilateral de úlcera corneal. No GR e GP, o sangue autólogo foi centrifugado, utilizando-se protocolo padronizado, e foram confeccionados os colírios de PRP e PPP, e instilados cinco vezes ao dia. No GC, foi utilizado colírio lubrificante. Cada grupo foi subdividido (n=5), segundo o momento final de avaliação, sendo 4 (M4), 7 (M7) e 30 dias (M30). As córneas dos animais foram processadas para avaliação morfológica e imuno-histoquímica para PCNA, MMP1, MMP2, MMP9, MT1-MMP e TIMP1. No M4, os níveis de MMP2 foram maiores no GP e GR, sendo que, no M7, esse comportamento foi observado apenas no GP. No M30, no GR, verificou-se maior número de células epiteliais e marcação para MMP1 que o GP. No GR, a proliferação celular foi maior no M4 que nos demais momentos, e a marcação para MMP2 foi maior no M4 que no M30. O PRP estimula a proliferação celular na fase inicial (M4) do tratamento quando comparado aos demais momentos, diferentemente dos demais tratamentos. O uso de colírios de plasma rico e pobre em plaquetas influencia a expressão de metaloproteinases de matriz envolvidas no processo de reparação corneal.
The aim of this study was to evaluate the influence of platelet-rich (PRP) and poor (PPP) plasma in cell proliferation and matrix metalloproteinases (MMPs) expression during the repair of deep corneal ulcers. Forty-five female rabbits were distributed in 3 experimental groups (G) (n = 15), referred to as PRP (GR), PPP (GP) and Control (GC) groups, in accordance with the treatment. All animals underwent surgical induction of unilateral corneal ulcer. PRP and PPP eye drops were made by using centrifuged blood through standardized protocol, and instilled five times a day. In GC, lubricant eye drops were used. Each group was subdivided (n = 5) according to the final time point, 4 (M4), 7 (M7) and 30 days (M30). The animals' corneas were processed for morphological and immunohistochemical analysis for PCNA, MMP1, MMP2, MMP9, MT1-MMP and TIMP1. In M4, the levels of MMP2 were higher in GP and GR, and in M7, this behavior was only observed in the GP. In M30, more epithelial cells and MMP1 expression were found in GR than GP. In GR, cell proliferation was higher in M4 than at other time points and MMP2 expression was higher in M4 than M30. The PRP stimulates cell proliferation in the early phase (M4) of treatment when compared to other time points, different from other treatments. The use of eye drops of platelet-rich and poor plasma influences the expression of matrix metalloproteinases involved in the corneal repair process.
Subject(s)
Animals , Female , Rabbits , Proliferating Cell Nuclear Antigen/analysis , Matrix Metalloproteinases/analysis , Platelet-Rich Plasma/physiology , Corneal Ulcer/surgery , Wound Healing/physiology , Immunohistochemistry/veterinary , Corneal Injuries/veterinary , Cell Proliferation/physiologyABSTRACT
Introduction and objective: Overexpression of MMPs has been related to biochemical recurrence after radical prostatectomy. TIMP1 and TIMP2 are controllers of MMPs and the aim of this study is to evaluate the expression levels of MMPs and their regulators using immunohistochemistry in tissue microarray of localized prostate cancer (PC). Materials and Methods: Immune-expression of MMP-9, MMP-2, TIMP1, TIMP-2, MMP-14 and IL8, were analyzed by immunohistochemistry in radical prostatectomy specimens of 40 patients with localized PC who underwent surgery between September 1997 and February 2000. Protein expression was considered as categorical variables, negative or positive. The results of the immune-expression were correlated to Gleason score (GS), pathological stage (TNM), pre-operatory PSA serum levels and biochemical recurrence in a mean follow up period of 92.5 months. Results: The loss of TIMP1 immune-expression was related to biochemical recurrence. When TIMP1 was negative, 56.3% patients recurred versus 22.2% of those whose TIMP1 was positive (p=0.042). MMP-9, MMP-2, IL8 and MMP-14 were positive in the majority of PC. TIMP-2 was negative in all cases. Conclusion: Negative immune-expression of TIMP1 is correlated with biochemical recurrence in patients with PC possibly by failing to control MMP-9, an important MMP related to cancer progression.
Subject(s)
Adult , Aged , Humans , Male , Middle Aged , Matrix Metalloproteinases/analysis , Neoplasm Recurrence, Local/pathology , Prostatic Neoplasms/pathology , Tissue Inhibitor of Metalloproteinase-1/analysis , /analysis , Biomarkers, Tumor/analysis , Disease Progression , Immunohistochemistry , /analysis , Kaplan-Meier Estimate , Neoplasm Grading , Neoplasm Staging , Neoplasm Recurrence, Local/chemistry , Prostatectomy , Prostate-Specific Antigen/blood , Prostatic Neoplasms/chemistry , Prostatic Neoplasms/surgery , Statistics, NonparametricABSTRACT
Matrix metalloproteinases (MMPs), myofibroblasts (MFs) and epithelial proliferation have key roles in neoplastic progression. In this study immunoexpression of MMP-1, MMP-2 and MMP-9, presence of MFs and the epithelial proliferation index were investigated in actinic cheilitis (AC), lip squamous cell carcinoma (LSCC) and mucocele (MUC). Thirty cases of AC, thirty cases of LSCC and twenty cases of MUC were selected for immunohistochemical investigation of the proteins MMP-1, MMP-2, MMP-9, α-smooth muscle actin (α-SMA) and Ki-67. The MMP-1 expression in the epithelial component was higher in the AC than the MUC and LSCC. In the connective tissue, the expression was higher in the LSCC. MMP-2 showed lower epithelial and stromal immunostaining in the LSCC when compared to the AC and MUC. The epithelial staining for MMP-9 was higher in the AC when compared to the LSCC. However, in the connective tissue, the expression was lower in the AC compared to other lesions. The cell proliferation rate was increased in proportion to the severity of dysplasia in the AC, while in the LSCC it was higher in well-differentiated lesions compared to moderately differentiated. There were no statistically significant differences in number of MFs present in the lesions studied. The results suggest that MMPs could affect the biological behaviour of ACs and LSCCs inasmuch as they could participate in the development and progression from premalignant lesions to malignant lesions.
Subject(s)
Carcinoma, Squamous Cell/pathology , Cheilitis/pathology , Lip Neoplasms/pathology , Matrix Metalloproteinases/biosynthesis , Myofibroblasts/pathology , Precancerous Conditions/pathology , Carcinoma, Squamous Cell/metabolism , Cheilitis/metabolism , Humans , Immunohistochemistry , Ki-67 Antigen/biosynthesis , Lip Neoplasms/metabolism , Matrix Metalloproteinase 1/analysis , Matrix Metalloproteinase 1/biosynthesis , Matrix Metalloproteinase 2/analysis , Matrix Metalloproteinase 2/biosynthesis , Matrix Metalloproteinase 9/analysis , Matrix Metalloproteinase 9/biosynthesis , Matrix Metalloproteinases/analysis , Precancerous Conditions/metabolismABSTRACT
OBJECTIVE: To examine levels of matrix metalloproteinases (MMPs)-1, -2, -3, -7, -8, -12, and -13 in the gingival crevicular fluid (GCF) of periodontally compromised teeth at different time points during orthodontic movement. MATERIALS AND METHODS: Ten controlled periodontitis subjects were submitted to orthodontic treatment. One dental arch was subjected to orthodontic movement, and teeth in the opposite arch were used as controls. GCF samples were collected from the lingual sites of two movement and two control incisors 1 week before orthodontic activation (-7 d), immediately after orthodontic activation, and after 1 hour, 24 hours, and 7, 14, and 21 days. Multiplexed bead immunoassay was used to measure MMPs in GCF. Data were analyzed using Friedman and Wilcoxon statistical tests. RESULTS: The only significant change found over time was in the levels of MMP-1 in the movement group (P < .05). When the two groups were compared after activation, the only statistically significant difference found was in levels of MMP-12 24 hours after activation (P < .05). CONCLUSIONS: Our findings suggested that the orthodontic movement of periodontally compromised teeth without active pockets did not result in significant changes in the GCF levels of MMPs.
Subject(s)
Gingival Crevicular Fluid/enzymology , Matrix Metalloproteinases/analysis , Periodontal Diseases/pathology , Periodontal Diseases/therapy , Tooth Movement Techniques/methods , Adult , Female , Humans , Immunoassay , Male , Middle Aged , Time Factors , Treatment OutcomeABSTRACT
Background: Studies have attempted to understand the importance of metalloproteinase (MMPs) in the pathogenesis ofdiseases caused by lentiviruses, being the human immunodeficiency virus (HIV) the most investigated. Despite advancesin studies with MMPs in others diseases, research correlating the presence and activity of gelatinases in animals affectedby caprine arthritis encephalitis virus (CAEV), a lentiviruses, are incipient and there is a need for research aiming to understand the dynamic of these enzymes in animals infected and its relation to pathological condition. The aim of this studywas to evaluate the presence and activity of the MMPs in blood serum of chronically infected bucks by CAEV.Materials, Methods & Results: The experiment was constituted by two groups (n = 5 each group). The first one wascomposed of five naturally infected bucks (4-5 years) and second group constituted of five seronegative bucks (3-4 years)for CAE. Serology was performed using the Western Blotting (WB) and confirmed by Polymerase Chain Reaction (PCR).These bucks belong to the experimental flock at Embrapa Goats and Sheep and the seropositive bucks were confirmed forCAE in the first two years. Blood samples were collected by puncturing the jugular vein from animals and evaluated byzymography (SDS-PAGE) using gelatin as substratum. Clinical examination was performed by evaluating the temperature (T), cardiac frequency (FC), respiratory frequency (FR) and clinical articular index (IAC). The IAC was calculatedby measuring the circumference of carpal joint and metacarpal bone height (difference between greater extent carpal andmetacarpal lesser extent). In infected and control groups were found molecular mass bands of 66 kDa (MMP-2), 72 kDa(pro-MMP-2), 86 kDa (MMP-9) and 92 kDa (pro-MMP-9). The correlation...(AU)
Subject(s)
Animals , Ruminants/blood , Ruminants/virology , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases/blood , Arthritis-Encephalitis Virus, Caprine , Semi-Arid ZoneABSTRACT
INTRODUCTION AND OBJECTIVE: Overexpression of MMPs has been related to biochemical recurrence after radical prostatectomy. TIMP1 and TIMP2 are controllers of MMPs and the aim of this study is to evaluate the expression levels of MMPs and their regulators using immunohistochemistry in tissue microarray of localized prostate cancer (PC). MATERIALS AND METHODS: Immune-expression of MMP-9, MMP-2, TIMP1, TIMP-2, MMP-14 and IL8, were analyzed by immunohistochemistry in radical prostatectomy specimens of 40 patients with localized PC who underwent surgery between September 1997 and February 2000. Protein expression was considered as categorical variables, negative or positive. The results of the immune-expression were correlated to Gleason score (GS), pathological stage (TNM), pre-operatory PSA serum levels and biochemical recurrence in a mean follow up period of 92.5 months. RESULTS: The loss of TIMP1 immune-expression was related to biochemical recurrence. When TIMP1 was negative, 56.3% patients recurred versus 22.2% of those whose TIMP1 was positive (p=0.042). MMP-9, MMP-2, IL8 and MMP-14 were positive in the majority of PC. TIMP-2 was negative in all cases. CONCLUSION: Negative immune-expression of TIMP1 is correlated with biochemical recurrence in patients with PC possibly by failing to control MMP-9, an important MMP related to cancer progression.
Subject(s)
Matrix Metalloproteinases/analysis , Neoplasm Recurrence, Local/pathology , Prostatic Neoplasms/pathology , Tissue Inhibitor of Metalloproteinase-1/analysis , Tissue Inhibitor of Metalloproteinase-2/analysis , Adult , Aged , Biomarkers, Tumor/analysis , Disease Progression , Humans , Immunohistochemistry , Interleukin-8/analysis , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Grading , Neoplasm Recurrence, Local/chemistry , Neoplasm Staging , Prostate-Specific Antigen/blood , Prostatectomy , Prostatic Neoplasms/chemistry , Prostatic Neoplasms/surgery , Statistics, NonparametricABSTRACT
Background: Studies have attempted to understand the importance of metalloproteinase (MMPs) in the pathogenesis ofdiseases caused by lentiviruses, being the human immunodeficiency virus (HIV) the most investigated. Despite advancesin studies with MMPs in others diseases, research correlating the presence and activity of gelatinases in animals affectedby caprine arthritis encephalitis virus (CAEV), a lentiviruses, are incipient and there is a need for research aiming to understand the dynamic of these enzymes in animals infected and its relation to pathological condition. The aim of this studywas to evaluate the presence and activity of the MMPs in blood serum of chronically infected bucks by CAEV.Materials, Methods & Results: The experiment was constituted by two groups (n = 5 each group). The first one wascomposed of five naturally infected bucks (4-5 years) and second group constituted of five seronegative bucks (3-4 years)for CAE. Serology was performed using the Western Blotting (WB) and confirmed by Polymerase Chain Reaction (PCR).These bucks belong to the experimental flock at Embrapa Goats and Sheep and the seropositive bucks were confirmed forCAE in the first two years. Blood samples were collected by puncturing the jugular vein from animals and evaluated byzymography (SDS-PAGE) using gelatin as substratum. Clinical examination was performed by evaluating the temperature (T), cardiac frequency (FC), respiratory frequency (FR) and clinical articular index (IAC). The IAC was calculatedby measuring the circumference of carpal joint and metacarpal bone height (difference between greater extent carpal andmetacarpal lesser extent). In infected and control groups were found molecular mass bands of 66 kDa (MMP-2), 72 kDa(pro-MMP-2), 86 kDa (MMP-9) and 92 kDa (pro-MMP-9). The correlation...
Subject(s)
Animals , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases/blood , Ruminants/blood , Ruminants/virology , Arthritis-Encephalitis Virus, Caprine , Semi-Arid ZoneABSTRACT
O tecido ósseo possui alto grau de rigidez e resistência à pressão, característica relacionada às suas funções de proteção, sustentação e locomoção. Patologias como periodontite, artrite reumatóide e osteoporose decorrem do desequilíbrio da dinâmica óssea, que ocorre quando os osteoclastos estão em maior atividade em relação aos osteoblastos. O fluoreto (F-) é incorporado nos tecidos mineralizados após ingestão e a sua administração aumenta a formação óssea in vivo, de maneira dependente da dose e da linhagem dos animais. As linhagens A/J e 129P3/J possibilitam a análise dos fenômenos moleculares envolvidos na suscetibilidade e resistência de células ósseas aos efeitos do F- no osso. Neste estudo, avaliou-se a administração in vivo de F- na osteoclastogênese, através da análise da atividade da enzima fosfatase ácida resistente ao tartarato (TRAP), contagem de células TRAP- positivas e quantificação da atividade das metaloproteinases MMP-2 e -9. Para tal, células hematopoiéticas provenientes da medula óssea de camundongos das linhagens 129P3/J e A/J foram cultivadas com M-CSF e RANKL em associação ou não com F-, nas concentrações de 10- 9, 10-7, 10-5, 5x10-5, 5x10-5, 10-4 e 10-3 M. Os dados mostraram que a atividade da TRAP foi semelhante entre as células das duas linhagens de animais, independente do F-. Concomitante à diferenciação osteoclástica, o tratamento das células com F-, independente da concentração, aumentou significativamente a atividade da TRAP em células da linhagem A/J. O aumento da atividade de TRAP foi associada com o aumento do número de osteoclastos formados, na concentração de 10-3 M F-. Já em células dos animais 129P3/J, o F- não alterou a atividade da TRAP bem como a osteoclastogênese. A atividade de MMP-9 foi aumentada em relação à da MMP-2, porém ambas não foram moduladas pelo F-, independente da linhagem. Assim, concluiu-se que as células de ambas as linhagens não diferem com relação à diferenciação de osteoclastos,
The bone tissue has a high degree of rigidity and pressure resistance, characteristic related to its protection, support and locomotion functions. Diseases such as periodontitis, osteoporosis and rheumatoid arthritis arise from dynamic bone imbalance that occurs when osteoclasts present increased activity compared to osteoblasts. Fluoride (F-) is incorporated in mineralized tissues after ingestion and its administration increases in vivo bone formation in dose and strain-dependent manner. The strains A/J and 129P3/J make possible the analysis of molecular phenomena involved in susceptibility and resistance of bone cells to the effect of F- in bone. In this study, we evaluated the effect of F- on the in vitro osteoclastogenesis, by analyzing the fosfatase tartrate-resistant acid (TRAP) activity, TRAP positive cell counts and the activity of MMP-2 and -9 measurements in A/J and 129P3/J differentiated hematopoietic stem cells. Hematopoietic cells were differentiated with M-CSF and RANKL and treated or not with F- in the concentrations of 10-9, 10-7, 10-5, 5x10-5, 5x10-5, 10-4 and 10-3 M.The data show that TRAP activity, an osteoclast marker, was similar between cells from both strains of mice, regardless of F-. In addition, the treatment of A/J cells with F- significantly increased TRAP activity, in a dose-independent manner. This was accompained by the increased TRAP+-osteoclast counts, at 10-3 M F- dose. However, while the MMP-9 activity was aumented when compared to MMP-2, it was not altered by F-, regardless of the strain. Thus, we concluded that cells from both strains do not differ regarding the osteoclast differentiation potential, but they respond differently to F-, whereas A/J and 129P3/J cells are sensitive and resistant to F--induced osteoclastogenesis, respectively.
Subject(s)
Animals , Male , Female , Mice , Cariostatic Agents/pharmacology , Fluorine/pharmacology , Fluorides/pharmacology , Osteoclasts , Osteogenesis , Cell Count , Cells, Cultured , Cell Differentiation , Matrix Metalloproteinases/analysis , Matrix Metalloproteinases , Reproducibility of Results , Bone Resorption/prevention & control , Time FactorsABSTRACT
BACKGROUND: Calcifying cyst odontogenic tumour (CCOT) is a rare benign cystic neoplasm of odontogenic origin. MMPs are responsible for extracellular matrix remodelling and, together their inhibitors and inducer, determinate the level of its turnover in pathological processes, leading to an auspicious microenvironment for tumour development. Thus, our goal was to evaluate matrix metalloproteinases (MMPs-2, -7, -9 and -14), their inhibitors (TIMPs-2, -3, -4 and RECK) and its inductor (EMMPRIN) expression in CCOT. MATERIALS AND METHODS: We used 18 cases of CCOT submitted to immunolocalization of the target proteins and analysed in both neoplastic odontogenic epithelial and stromal compartments. RESULTS: All molecules evaluated were expressed in both compartments in CCOT. In epithelial layer, immunostaining for MMPs, TIMPs, RECK and EMMPRIN was found in basal, suprabasal spindle and stellate cells surrounding ghost cells and ghost cells themselves, except for MMP-9 and TIMP-2 which were only expressed by ghost cells. In stromal compartment, extracellular matrix, mesenchymal (MC) and endothelial cells (EC) were positive for MMP-2, -7, TIMP-3 and -4, while MMP-9, TIMP-2 and RECK were positive only in MC and MMP-14 only in EC. Statistical significance difference was found between both compartments for MMP-9 (P < 0.001), RECK (P = 0.004) and EMMPRIN (P < 0.001), being more expressed in epithelium than in stroma. Positive correlation between both stromal EMMPRIN and RECK expression was found (R = 0.661, P = 0.003). CONCLUSIONS: We concluded that these proteins/enzymes are differentially expressed in both epithelium and stroma of CCOT, suggesting an imbalance between MMPs and their inducer/inhibitors may contribute on the tumour behaviour.
Subject(s)
Basigin/analysis , GPI-Linked Proteins/analysis , Matrix Metalloproteinases/analysis , Odontogenic Tumors/chemistry , Tissue Inhibitor of Metalloproteinases/analysis , Adolescent , Adult , Endothelial Cells/chemistry , Endothelial Cells/enzymology , Epithelium/chemistry , Epithelium/enzymology , Extracellular Matrix/chemistry , Extracellular Matrix/enzymology , Female , Humans , Male , Matrix Metalloproteinase 14/analysis , Matrix Metalloproteinase 2/analysis , Matrix Metalloproteinase 7/analysis , Matrix Metalloproteinase 9/analysis , Mesoderm/chemistry , Mesoderm/enzymology , Middle Aged , Neoplasm Proteins/analysis , Odontogenic Tumors/enzymology , Tissue Inhibitor of Metalloproteinase-2/analysis , Tissue Inhibitor of Metalloproteinase-3/analysis , Tumor Microenvironment , Young Adult , Tissue Inhibitor of Metalloproteinase-4ABSTRACT
BACKGROUND: Matrix metalloproteinases (MMPs) are a family of host-derived proteinases reported to mediate multiple functions associated with periodontal breakdown and inflammation. High MMP levels in African-American children with localized aggressive periodontitis (LAgP) have been reported previously by the present authors. However, little is known about MMP reductions in gingival crevicular fluid (GCF) after therapy. This study aims to evaluate MMP levels in the GCF after treatment of LAgP and to correlate these levels with clinical response. METHODS: GCF samples were collected from 29 African-American individuals diagnosed with LAgP. GCF was collected from one diseased site (probing depth [PD] >4 mm, bleeding on probing [BOP], and clinical attachment level ≥ 2 mm) and one healthy site (PD ≤ 3 mm, no BOP) from each individual at baseline and 3 and 6 months after periodontal treatment, which consisted of full-mouth scaling and root planing (SRP) and systemic antibiotics. The volume of GCF was controlled using a calibrated gingival fluid meter, and levels of MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-12, and MMP-13 were assessed using fluorometric kits. RESULTS: MMP-1, MMP-8, MMP-9, MMP-12, and MMP-13 levels were reduced significantly up to 6 months, comparable to healthy sites at the same point. Significant correlations were noted between MMP-2, MMP-3, MMP-8, MMP-9, MMP-12, and MMP-13 levels and percentage of sites with PD >4 mm. MMP-3, MMP-12, and MMP-13 levels also correlated with mean PD of affected sites. CONCLUSION: Treatment of LAgP with SRP and systemic antibiotics was effective in reducing local levels of specific MMPs in African-American individuals, which correlated positively with some clinical parameters.
Subject(s)
Aggressive Periodontitis/therapy , Matrix Metalloproteinases/analysis , Adolescent , Black or African American , Aggressive Periodontitis/enzymology , Amoxicillin/therapeutic use , Anti-Bacterial Agents/therapeutic use , Child , Child, Preschool , Dental Scaling/methods , Female , Follow-Up Studies , Gingival Crevicular Fluid/enzymology , Humans , Male , Matrix Metalloproteinase 1/analysis , Matrix Metalloproteinase 12/analysis , Matrix Metalloproteinase 13/analysis , Matrix Metalloproteinase 2/analysis , Matrix Metalloproteinase 3/analysis , Matrix Metalloproteinase 8/analysis , Matrix Metalloproteinase 9/analysis , Metronidazole/therapeutic use , Periodontal Attachment Loss/enzymology , Periodontal Attachment Loss/therapy , Periodontal Index , Periodontal Pocket/enzymology , Periodontal Pocket/therapy , Root Planing/methods , Young AdultABSTRACT
This randomized split-mouth study aimed to examine the levels of matrix metalloproteinases (MMPs) -1, -2, -3, -7, -8, -12, and -13 in the gingival crevicular fluid (GCF) at different time points during orthodontic tooth movement. A total of 16 healthy orthodontic subjects (7 females, 9 males; mean age, 17.7 years) who needed their first upper premolars extracted were enrolled. One randomly chosen maxillary canine was subjected to a distalizing force and was considered to be the test side. The contralateral canine, which was not subjected to any force but was included in the orthodontic appliance, was used as a control side. GCF sampling was performed at both the mesial (tension) and distal (pressure) test and control sites at baseline, immediately before applying the orthodontic appliance, and after 1 and 24 hours and 7, 14, and 21 days. A multiplexed bead immunoassay was used to analyse the GCF samples. The mean levels of the MMP-1, -2, -3, -7, -8, -12, and -13 were not significantly different between the test and control groups in each time showed. The comparisons between the tension and pressure sites were also not significantly different at each individual time. A few variations focused on MMP-1 and -3, but the expression of MMP-8 was higher than that of the other MMPs. MMPs are released in sufficient quantities such that tooth movement occurs but with no significant increase in GCF levels.
Subject(s)
Gingival Crevicular Fluid/enzymology , Matrix Metalloproteinases/metabolism , Tooth Movement Techniques , Adolescent , Adult , Female , Humans , Longitudinal Studies , Male , Matrix Metalloproteinase 1/analysis , Matrix Metalloproteinase 12/analysis , Matrix Metalloproteinase 12/metabolism , Matrix Metalloproteinase 13/analysis , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 2/analysis , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 3/analysis , Matrix Metalloproteinase 3/metabolism , Matrix Metalloproteinase 7/analysis , Matrix Metalloproteinase 7/metabolism , Matrix Metalloproteinase 8/analysis , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinases/analysis , Maxilla/surgery , Orthodontic Appliances , Periodontium/surgery , Young AdultABSTRACT
Hydrogen peroxide is an oxidative agent commonly used for dental bleaching procedures. The structural and biochemical responses of enamel, dentin, and pulp tissues to the in vivo bleaching of human (n = 20) premolars were investigated in this study. Atomic force microscopy (AFM) was used to observe enamel nanostructure. The chemical composition of enamel and dentin was analyzed by infrared spectroscopy (FTIR). The enzymatic activities of dental cathepsin B and matrix metalloproteinases (MMPs) were monitored with fluorogenic substrates. The amount of collagen in dentin was measured by emission of collagen autofluorescence with confocal fluorescence microscopy. The presence of Reactive Oxygen Species (ROS) in the pulp was evaluated with a fluorogenic 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) probe. Vital bleaching of teeth significantly altered all tested parameters: AFM images revealed a corrosion of surface enamel nanostructure; FTIR analysis showed a loss of carbonate and proteins from enamel and dentin, along with an increase in the proteolytic activity of cathepsin-B and MMPs; and there was a reduction in the autofluorescence of collagen and an increase in both cathepsin-B activity and ROS in pulp tissues. Together, these results indicate that 35% hydrogen peroxide used in clinical bleaching protocols dramatically alters the structural and biochemical properties of dental hard and soft pulp tissue.