RESUMO
Abstract Specific proteases capable of degrading native triple helical or denatured collagen have been required for many years and have a large spectrum of applications. There are few complete reports that fully uncover production, characterization and purification of fungi collagenases. In this review, authors searched through four scientific on line data bases using the following keywords (collagenolytic OR collagenase) AND (fungi OR fungus OR fungal) AND (production OR synthesis OR synthesize) AND (characterization). Scientific criteria were adopted in this review to classify found articles by score (from 0 to 10). After exclusion criteria, 21 articles were selected. None obtained the maximum of 10 points defined by the methodology, which indicates a deficiency in studies dealing simultaneously with production, characterization and purification of collagenase by fungi. Among microorganisms studied the non-pathogenic fungi Penicillium aurantiogriseum and Rhizoctonia solani stood out in volumetric and specific collagenase activity. The only article found that made sequencing of a true collagenase showed 100% homology with several metalloproteinases fungi. A clear gap in literature about collagenase production by fungi was verified, which prevents further development in the area and increases the need for further studies, particularly full characterization of fungal collagenases with high specificity to collagen.
Assuntos
Colágeno/metabolismo , Colagenases/metabolismo , Fungos/metabolismo , Especificidade por Substrato , Colágeno/química , Colagenases/isolamento & purificação , Colagenases/biossíntese , Colagenases/química , Meios de Cultura , Ativação Enzimática , Proteólise , Fungos/classificaçãoRESUMO
Specific proteases capable of degrading native triple helical or denatured collagen have been required for many years and have a large spectrum of applications. There are few complete reports that fully uncover production, characterization and purification of fungi collagenases. In this review, authors searched through four scientific on line data bases using the following keywords (collagenolytic OR collagenase) AND (fungi OR fungus OR fungal) AND (production OR synthesis OR synthesize) AND (characterization). Scientific criteria were adopted in this review to classify found articles by score (from 0 to 10). After exclusion criteria, 21 articles were selected. None obtained the maximum of 10 points defined by the methodology, which indicates a deficiency in studies dealing simultaneously with production, characterization and purification of collagenase by fungi. Among microorganisms studied the non-pathogenic fungi Penicillium aurantiogriseum and Rhizoctonia solani stood out in volumetric and specific collagenase activity. The only article found that made sequencing of a true collagenase showed 100% homology with several metalloproteinases fungi. A clear gap in literature about collagenase production by fungi was verified, which prevents further development in the area and increases the need for further studies, particularly full characterization of fungal collagenases with high specificity to collagen.
Assuntos
Colágeno/metabolismo , Colagenases/metabolismo , Fungos/metabolismo , Colágeno/química , Colagenases/biossíntese , Colagenases/química , Colagenases/isolamento & purificação , Meios de Cultura , Ativação Enzimática , Fungos/classificação , Proteólise , Especificidade por SubstratoRESUMO
OBJECTIVES: To investigate whether low-level laser therapy (LLLT) alters the expression and activity of MMP-2 and MMP-9 in the trigeminal ganglion (TG) during different stages of temporomandibular joint (TMJ) inflammation in rats. It also evaluated whether LLLT modifies mechanical allodynia and orofacial hyperalgesia. MATERIALS AND METHODS: Wistar rats (±250 g) were divided into groups that received saline (SAL) or complete Freund's adjuvant (CFA, 50 µl) in the TMJ, and that later underwent LLLT (20 J cm(-2) ) at their TMJ or not (groups SAL, SAL + LLLT, CFA, and CFA + LLLT). LLLT was applied on days 3, 5, 7, and 9 after SAL or CFA. Mechanical allodynia was evaluated on days 1, 3, 5, 7, and 10; orofacial hyperalgesia was assessed on day 10. Gelatin zymography and in situ zymography aided quantification of MMPs in the TG. RESULTS: Low-level laser therapy abolished the reduction in the mechanical orofacial threshold and the increase in orofacial rubbing during the orofacial formalin test induced by CFA. LLLT also decreased the CFA-induced rise in the levels of MMP-9 and MMP-2 as well as the gelatinolytic activity in the TG. CONCLUSION: Low-level laser therapy could constitute an adjuvant therapy to treat temporomandibular disorders and prevent inflammation-induced alterations in the levels of MMP-2 and MMP-9 and in the gelatinolytic activity in TGs.
Assuntos
Artrite Experimental/terapia , Colagenases/biossíntese , Terapia com Luz de Baixa Intensidade/métodos , Transtornos da Articulação Temporomandibular/enzimologia , Transtornos da Articulação Temporomandibular/terapia , Articulação Temporomandibular/inervação , Gânglio Trigeminal/enzimologia , Animais , Colagenases/metabolismo , Dor Facial/terapia , Adjuvante de Freund/farmacologia , Gelatina/metabolismo , Hiperalgesia/terapia , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Ratos , Ratos Wistar , Cloreto de Sódio/administração & dosagem , Articulação Temporomandibular/efeitos dos fármacos , Articulação Temporomandibular/cirurgia , Transtornos da Articulação Temporomandibular/patologia , Gânglio Trigeminal/metabolismo , Gânglio Trigeminal/patologiaRESUMO
INTRODUÇÃO: Novas opções terapêuticas para o tratamento de lesões térmicas são constantemente buscadas, especialmente se reduzirem tempo de cicatrização e dor, sem aumentar as taxas de infecção das queimaduras. Estudos recentes sugerem que o uso tópico de heparina pode alcançar esses objetivos. Este estudo tem o objetivo de avaliar tempo de epitelização, dor e taxa de infecção, comparando o uso de heparina tópica ao uso de colagenase no tratamento de queimadura de segundo grau superficial de face e pescoço. MÉTODO: No total, 20 pacientes foram randomizados em dois grupos: grupo tratado com heparina sódica e grupo tratado com colagenase (controle). Os critérios de exclusão foram: história de sangramento, discrasia sanguínea, alergias ao produto, úlcera péptica ativa e queimadura há mais de 24 horas. O teste de Mann-Whitney foi utilizado para avaliar os resultados. A dor foi avaliada pela necessidade do uso de analgésicos opioides. RESULTADOS: A heparina não foi efetiva em diminuir o tempo de epitelização ou o uso de opioides, e a taxa de infecção não apresentou diferença estatística entre os grupos. CONCLUSÕES: A heparina pode ser usada com segurança no tratamento de queimadura de segundo grau superficial em face e pescoço, mas seus efeitos benéficos ainda precisam ser comprovados.
BACKGROUND: New treatment options for thermal injuries are very desirable, especially if they reduce healing time and pain without increase of infection rates. Recent studies suggest that heparin topical use can achieve those goals. This study has the objective to evaluate healing time, pain and infection rate comparing topical use of heparin and collagenase in the treatment of superficial second degree burns of face and neck. METHODS: Twenty patients were randomized into 2 groups: group treated with topical heparin and group treated with collagenase (control group). The exclusion criteria were: history of bleeding, blood discrasia, allergies to the product, active peptic ulcer and burns with more than 24 hours. Mann-Whitney test was applied to evaluate the results. The pain was measured by the use of opioid analgesics. RESULTS: The heparin was not effective in decrease of healing time nor the use of opioids, and the infection rate didn't present significant difference between the groups. CONCLUSIONS: The heparin can be used safely in treatment of superficial second degree burn of face and neck, but its beneficial effects need to be proven.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , História do Século XXI , Dor , Cicatrização , Ferimentos e Lesões , Queimaduras , Heparina , Ensaio Clínico Controlado Aleatório , Colagenases , Estudo de Avaliação , Cabeça , Pescoço , Dor/tratamento farmacológico , Cicatrização/efeitos dos fármacos , Ferimentos e Lesões/tratamento farmacológico , Queimaduras/tratamento farmacológico , Queimaduras/terapia , Heparina/uso terapêutico , Colagenases/biossíntese , Colagenases/efeitos dos fármacos , Colagenases/uso terapêutico , Cabeça/cirurgia , Pescoço/cirurgiaRESUMO
Medium composition and culture conditions for maximal collagenase production by Penicillium aurantiogriseum URM4622 were optimized using a response surface approach. A full two-level design on three factors (initial medium pH, soybean flour concentration, and temperature) was employed to identify the most significant fermentation parameters for collagenase production, and a subsequent central composite design (CCD) was used to find the optimal levels of the two most significant factors (initial medium pH and soybean flour concentration). The design results indicated that the initial medium pH and the temperature had significant negative main effects, whereas the substrate concentration had a positive effect on the collagenase production. The maximum collagenolytic activity predicted by the fitted response surface was expected to occur at pH 7.21, 1.645% soybean flour concentration and 24°C. Three replicate experiments were run at these conditions and yielded an activity response of 283.36 ± 1.33 U, which not only is the highest obtained in this study but also represents a 5-fold increase over the lowest response observed in the initial design. Since all experiments were carried out with an inexpensive substrate, the final results point out to a cost-effective medium for collagenase production with potential industrial-scale applications.
Assuntos
Colagenases/biossíntese , Fermentação , Penicillium/metabolismo , Biomassa , Meios de Cultura , Concentração de Íons de HidrogênioRESUMO
Parvimonas micra are gram positive anaerobic cocci isolated from the oral cavity and frequently related to polymicrobial infections in humans. Despite reports about phenotypic differences, the genotypic variation of P. micra and its role in virulence are still not elucidated. The aim of this study was to determine the genotypic diversity of P. micra isolates obtained from the subgingival biofilm of subjects with different periodontal conditions and to correlate these findings with phenotypic traits. Three reference strains and 35 isolates of P. micra were genotyped by 16S rRNA PCR-RFLP and phenotypic traits such as collagenase production, elastolytic and hemolytic activities were evaluated. 16S rRNA PCR-RFLP showed that P. micra could be grouped into two main clusters: C1 and C2; cluster C1 harbored three genotypes (HG1259-like, HG1467-like and ICBMO583-like) while cluster C2 harbored two genotypes (ATCC33270-like and ICBMO36). A wide variability in collagenolytic activity intensities was observed among all isolates, while elastolytic activity was detected in only two isolates. There was an association between hemolytic activity in rabbit erythrocytes and cluster C2. There was an association between hemolytic activity in rabbit erythrocytes and cluster C1. Although these data suggest a possible association between P. micra genetic diversity and their pathogenic potential, further investigations are needed to confirm this hypothesis.
Assuntos
Biodiversidade , Colagenases/biossíntese , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/metabolismo , RNA Ribossômico 16S/genética , Animais , Análise por Conglomerados , Impressões Digitais de DNA , DNA Bacteriano/genética , DNA Ribossômico/genética , Elastina/metabolismo , Eritrócitos/microbiologia , Genótipo , Gengiva/microbiologia , Bactérias Gram-Positivas/genética , Bactérias Gram-Positivas/isolamento & purificação , Hemólise , Humanos , Doenças Periodontais/microbiologia , Polimorfismo de Fragmento de Restrição , CoelhosRESUMO
As metaloproteinases da matriz (MMPs) foram relacionadas a diversas doenças inflamatórias como artrite e também ao câncer. O presente trabalho tem por objetivo estabelecer o papel da MMP-2, MMP-9 e MMP-8 no processo de inflamação pulpar. Foram adotadas as seguintes hipóteses nulas: (1) o padrão de expressão das MMP-2, MMP-9 e MMP-8 não sofre alteração nos diferentes estágios da polpa humana: normal, reversível, transição, irreversível ou necrose; (2) não há diferença de expressão das MMP-2, -9 e MMP-8, considerando-se um mesmo estágio de inflamação tecidual pulpar. Os métodos utilizados foram: (I) Obtenção dos espécimes, que foram divididos em grupos de acordo com critérios adotados de semiologia subjetiva e objetiva. Obtiveram-se os seguintes grupos: GI (Controle) dentes hígidos (n=7); GII (Pulpite Reversível n=4); GIII (Pulpite Transição n=4); GIV (Pulpite Irreversível/Necrose n=8). Logo após exodontia, os dentes obtidos foram cortados ligeiramente abaixo da junção amelodentinária e fixados em formol a 10% por 48h. Foram lavados em água corrente (24h) para então serem processados histologicamente. Foram obtidas secções de 4m, aderidas em lâminas silanizadas e submetidas à imunomarcação (Técnica da Peroxidase), utilizando os anticorpos anti MMP-2, MMP-9 e MMP-8 humanos. A presença de imunomarcação foi realizada através da análise semi-quantitativa por escores, sendo que a quantificação de marcação por corte seguiu o seguinte escore: 0= ausente; 1= leve; 2= moderada; 3= intensa. Realizou-se teste estatístico não paramétrico Kruskal-Wallis, p<0,05. As comparações intergrupos revelaram, para CO: (1)MMP-2 - GI=GII=GIII, GIII=GIV, GI>GIV (p<0,01) e GII>GIV (p<0,05); (2)MMP-9 GI=GII=GIV, GII=GIII e GIII>GI (p<0,01); (3)MMP-8 GI=GII=GIII=GIV. Na região central da polpa, obteve-se: (1)MMP-2 GI=GII=GIII, GIII=GIV, GI>GIV (p<0,001) e GII>GIV (p<0,01); (2)MMP-9 GI=GII=GIII, GIII=GIV, GIV>GI (p<0,001) e GIV>GII (p<0,01); (3)MMP-8 GI=GII, GIII=GIV, GIII>GI (p<0,05),...
The matrix metalloproteinases (MMPs) have been related to various inflammatory diseases, such as arthritis, as well as to cancer. The aim of the present study was to establish the role of MMP-2, MMP-9 and MMP-8 in the process of dental pulp inflammation. The following null hypotheses were adopted: (1) the pattern of MMP-2, MMP-9 and MMP-8 expression does not undergo alteration in the following different stages of human pulp: normal, reversible, transition, irreversible or necrosis; (2) there is no difference in the expression of MMP-2, -9 and MMP-8, when considering the same stage of pulp tissue inflammation. The methods used were: (I) Obtainment of specimens, which were divided into groups according to the subjective and objective criteria of semiology adopted. The following groups were obtained: GI (Control) healthy teeth (n=7); GII (Reversible Pulpitis n=4); GIII (Transition Pulpitis n=4); GIV (Irreversible Pulpitis/Necrosis n=8). Soon after extraction the teeth obtained were cut slightly below the amelodentinal junction and fixed in 10% formol for 48h. They were washed under running water (24h) and were histologically processed afterwards. Sections of 4m were obtained, adhered to silanized slides, and submitted to immunomarking (Peroxidase Technique), using human anti MMP-2, MMP-9 and MMP- 8 antibodies. The presence of immunomarking was determined through semi-quantitative analysis by scores, and marking by cut was quantified using the following score: 0= absent; 1= slight; 2= moderate; 3= intense. The Kruskal-Wallis non-parametric statistical test was performed, p<0.05. Intergroup comparisons revealed the following: for CO: (1)MMP-2 - GI=GII=GIII, GIII=GIV, GI>GIV (p<0.01) and GII>GIV (p<0.05); (2)MMP-9 GI=GII=GIV, GII=GIII and GIII>GI (p<0,01); (3)MMP-8 GI=GII=GIII=GIV. In the central region of the pulp, the following results were obtained: (1)MMP-2 GI=GII=GIII, GIII=GIV, GI>GIV (p<0.001) and GII>GIV (p<0.01); (2)MMP-9 GI=GII=GIII, GIII=GIV, GIV>GI...
Assuntos
Humanos , Colagenases/biossíntese , Gelatinases/biossíntese , Técnicas In Vitro , Polpa Dentária/química , Pulpite/patologia , Imuno-Histoquímica , Metaloproteinase 9 da Matriz/biossíntese , /biossíntese , /biossíntese , Estatísticas não ParamétricasRESUMO
The aquatic ecosystem is the natural habitat of microorganisms including Vibrio and Aeromonas genus which are pathogenic to human and animals. In the present investigation the frequency of these bacteria and the enzymatic characteristics of 34 Vibrio alginolyticus strains isolated from bivalves harvested in Venice Lagoon (Italy) and Guanabara Bay (Brazil) were carried out from November 2003 to February 2004. The mussels' samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1% of sodium chloride (NaCl) and APW plus 3% NaCl incubated at 37 degrees C for 18-24 h. Following the samples were streaked onto TCBS Agar (Thiossulfate Citrate Bile Sucrose Agar) and the suspected colonies were submitted to biochemical characterization. Also, the Vibrio alginolyticus strains were evaluated to collagenase, elastase and chondroitinase production. The results showed the isolation of 127 microorganisms distributed as follows: 105 Vibrio strains such as V. alginolyticus (32.4%), V. harveyi (19%) and V. parahaemolyticus (7.6%), 20 Aeromonas strains and two Plesiomonas shigelloides were the main pathogens isolated. We observed the production of the three enzymes from V. alginolyticus strains considered as the main virulence factors of the bacteria, especially in cases of human dermatological infection.
Assuntos
Aeromonas/classificação , Bivalves/microbiologia , Vibrio alginolyticus/enzimologia , Vibrio/classificação , Aeromonas/isolamento & purificação , Animais , Brasil , Condroitinases e Condroitina Liases/biossíntese , Colagenases/biossíntese , Itália , Elastase Pancreática/biossíntese , Vibrio/isolamento & purificação , Vibrio alginolyticus/isolamento & purificaçãoRESUMO
The aquatic ecosystem is the natural habitat of microorganisms including Vibrio and Aeromonas genus which are pathogenic to human and animals. In the present investigation the frequency of these bacteria and the enzymatic characteristics of 34 Vibrio alginolyticus strains isolated from bivalves harvested in Venice Lagoon (Italy) and Guanabara Bay (Brazil) were carried out from November 2003 to February 2004. The mussels' samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1 percent of sodium chloride (NaCl) and APW plus 3 percent NaCl incubated at 37 ºC for 18-24h. Following the samples were streaked onto TCBS Agar (Thiossulfate Citrate Bile Sucrose Agar) and the suspected colonies were submitted to biochemical characterization. Also, the Vibrio alginolyticus strains were evaluated to collagenase, elastase and chondroitinase production. The results showed the isolation of 127 microorganisms distributed as follows: 105 Vibrio strains such as V. alginolyticus (32.4 percent), V. harveyi (19 percent) and V. parahaemolyticus (7.6 percent), 20 Aeromonas strains and two Plesiomonas shigelloides were the main pathogens isolated. We observed the production of the three enzymes from V. alginolyticus strains considered as the main virulence factors of the bacteria, especially in cases of human dermatological infection.
O ecossistema aquático é o habitat natural de microrganismos incluindo aqueles dos gêneros Vibrio e Aeromonas os quais são patogênicos para o homem e animais. Na presente investigação foi avaliada a freqüência destas bactérias e a característica enzimática de 34 cepas de Vibrio alginolyticus isoladas de bivalves coletados na Lagoa de Venice (Itália) e Baía de Guanabara (Brasil) durante o período de Novembro-2003 a Fevereiro-2004. As amostras de mexilhões foram submetidas a enriquecimento em Água Peptonada Alcalina (APA) adicionada de 1 por cento de Cloreto de Sódio (NaCl) e APA com 3 por cento de NaCl (37 ºC/18-24h). Em seguida as amostras foram semeadas em Agar TCBS (Agar Tiossulfato Citrato Bile Sacarose) e as colônias suspeitas foram submetidas à caracterização bioquímica. As cepas de Vibrio alginolyticus foram avaliadas quanto à produção das enzimas colagenase, elastase e condroitinase. Os resultados demonstraram o isolamento de 127 microrganismos assim distribuídos: 105 cepas de Vibrio das quais V. alginolyticus (32,4 por cento), V. harveyi (19 por cento) e V. parahaemolyticus (7,6 por cento), 20 cepas de Aeromonas e 2 Plesiomonas shigelloides foram os principais patógenos isolados. Observou-se a produção das três enzimas a partir de V. alginolyticus, consideradas principais fatores de virulência da bactéria, em especial em casos de infecção dermatológica humana.
Assuntos
Animais , Aeromonas/classificação , Bivalves/microbiologia , Vibrio alginolyticus/enzimologia , Vibrio/classificação , Aeromonas/isolamento & purificação , Brasil , Condroitinases e Condroitina Liases/biossíntese , Colagenases/biossíntese , Itália , Elastase Pancreática/biossíntese , Vibrio alginolyticus/isolamento & purificação , Vibrio/isolamento & purificaçãoRESUMO
Overproduction of urokinase-type plasminogen activator (uPA) and metalloproteases (MMPs) is strongly correlated with tumorigenicity and with invasive and metastatic phenotypes of human and experimental tumors. We demonstrated previously that overproduction of uPA in tumor cells is mediated by a phospholipase D (PLD)- and protein kinase C-dependent mechanism. The oncogenic stimulus of v-Src and v-Ras results in the activation of PLD, which is dependent upon the monomeric GTPase RalA. We have therefore investigated whether RalA plays a role in uPA and MMP overproduction that is observed in response to oncogenic signals. We report here that NIH3T3 cells transformed by both v-Src and v-Ras, constitutively overproduce uPA and that expression of a dominant negative RalA mutant (S28N) blocks overproduction of uPA in both the v-Src-and v-Ras-transformed cells. v-Src and v-Ras also induced an upregulation of the activity of MMP-2 and MMP-9 as detected by zymograms, however only the v-Src induction correlated with MMP protein levels detected by Western blot analysis. The dominant negative RalA mutant blocked increased MMP-2 and 9 overproduction induced by v-Src, but not the increased activity of MMP-2 and 9 induced by v-Ras. And, consistent with a role for the RalA/PLD pathway in mitogenesis and tumor development, the dominant negative RalA mutant completely blocked tumor formation by v-Src- and v-Ras-transformed NIH3T3 cells injected subcutaneously in syngeneic mice. The data presented here implicate RalA and PLD as signaling mediators for tumor formation and protease production by transformed cells.
Assuntos
Transformação Celular Neoplásica/genética , GTP Fosfo-Hidrolases/genética , Genes ras , Genes src , Metaloendopeptidases/biossíntese , Ativador de Plasminogênio Tipo Uroquinase/biossíntese , Proteínas ral de Ligação ao GTP , Células 3T3 , Animais , Colagenases/biossíntese , Gelatinases/biossíntese , Regulação Neoplásica da Expressão Gênica , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da Matriz , Camundongos , Camundongos Endogâmicos BALB C , Mutação , Regulação para CimaRESUMO
The protein MMP-2 (type IV collagenase) belongs to the family of metalloproteinases. Its function is related to cellular matrix degradation including basement membrane type IV collagen. Its presence in the neoplastic cells might enhance its capacity for dissemination. To find out some of its clinico-pathological and immunohistochemical behavior, 98 adenocarcinomas of the stomach were immunohistochemically studied, in search for MMP-2 in neoplastic cells. The results showed a correlation between MMP-2 with parietal depth of infiltration (p = 0.03) and with metastases in regional lymph nodes (p = 0.05). On the other hand, no correlation was found with sex, gastric localization, size of the tumor, histological type or grade neither with expression of MIB-1, c-erbB-2 nor p53 proteins, recurrence nor 5 year survival or no recurrency.
Assuntos
Adenocarcinoma/enzimologia , Colagenases/biossíntese , Neoplasias Gástricas/enzimologia , Idoso , Feminino , Humanos , Masculino , Metaloproteinase 9 da Matriz , Proteínas Nucleares/análise , Proteínas Proto-Oncogênicas c-bcl-2/análise , Receptor ErbB-2/análise , Estudos Retrospectivos , Proteína Supressora de Tumor p53/análiseRESUMO
Type II pneumocytes are multifunctional alveolar epithelial cells that play a major role in the maintenance of lung structure and function. Recent evidence supports that these cells can synthesize a variety of extracellular matrix components in vitro, suggesting an active participation in connective tissue remodeling. However, their possible role in extracellular matrix degradation is unknown. In this study the production of matrix metalloproteinases (MMPs) was examined in primary cultures of rat alveolar type II pneumocytes after 2 and 7 days in culture. Under basal conditions, at both periods type II cells expressed interstitial collagenase mRNA. The immunoreactive protein was detected both in the cells and in conditioned media, and collagenolytic activity was revealed after trypsin activation. Gelatinolytic activity was detected by zymography showing a relative molecular mass of approximately 72 and 92 kDa (gelatinases A and B). Phorbol treatment increased collagenase and gelatinase activities. In addition, three alveolar epithelial cell lines were analysed for MMP production: MLE-12 (mice), L2 (rat), and A549 (human). The cell lines A549 and MLE-12 revealed collagenase and gelatinase A and B activities whereas the L2 cell line only exhibited gelatinase A activity, even after PMA induction. These findings demonstrate that alveolar epithelial cells synthesize in vitro several MMPs that confer on them the ability to degrade extracellular matrix and basement membrane components, a capacity of considerable importance for the remodeling of the stromal/epithelial interface.
Assuntos
Colagenases/biossíntese , Células Epiteliais/enzimologia , Gelatinases/biossíntese , Metaloendopeptidases/biossíntese , Alvéolos Pulmonares/enzimologia , Animais , Caseínas/análise , Linhagem Celular , Eletroforese em Gel de Poliacrilamida , Humanos , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da Matriz , Camundongos , Alvéolos Pulmonares/citologia , RatosRESUMO
Heparin present in tissue has been reported to be a potential locally active agent responsible for bone resorption by the stimulation of collagenase via osteoclast activation and increased collagenase synthesis by the osteoblast. The purpose of this study was to extract, identify and quantify heparin in the "clinically normal", mildly and severely inflamed gingiva of Beagle dogs, using a simple and reliable technique. The extraction was carried out by homogenization, fat elimination, proteolytic digestion and precipitation, with organic solvents. Identification was performed by microelectrophoresis conducted on an agarose coated microscope slide. Quantification was performed by measuring the optical density of the metachromatic toluidine blue stained spot an comparing with the standard reference curve of heparin run simultaneously. The results showed that the difference in concentration of heparin in units per gram of wet tissue, was not significant when the "clinically normal" (26 +/- 1.9) was compared with mildly inflamed (24.4 +/- 4.7) gingiva. However, the concentration of heparin in severe gingivitis (79 +/- 7) was significantly higher. Gingival heparin could play important role in the established periodontal lesions, acting as a local factor or co-factor in periodontal bone destruction.