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1.
Langmuir ; 37(49): 14345-14359, 2021 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-34855402

RESUMO

Molecular adsorption to the nanoparticle surface may switch the colloidal interactions from repulsive to attractive and promote nanoparticle agglomeration. If the nanoparticles are magnetic, then their agglomerates exhibit a much stronger response to external magnetic fields than individual nanoparticles. Coupling between adsorption, agglomeration, and magnetism allows a synergy between the high specific area of nanoparticles (∼100 m2/g) and their easy guidance or separation by magnetic fields. This yet poorly explored concept is believed to overcome severe restrictions for several biomedical applications of magnetic nanoparticles related to their poor magnetic remote control. In this paper, we test this concept using curcumin (CUR) binding (adsorption) to ß-cyclodextrin (ßCD)-coated iron oxide nanoparticles (IONP). CUR adsorption is governed by host-guest hydrophobic interactions with ßCD through the formation of 1:1 and, possibly, 2:1 ßCD:CUR inclusion complexes on the IONP surface. A 2:1 stoichiometry is supposed to promote IONP primary agglomeration, facilitating the formation of the secondary needle-like agglomerates under external magnetic fields and their magneto-microfluidic separation. The efficiency of these field-induced processes increases with CUR concentration and ßCD surface density, while their relatively short timescale (<5 min) is compatible with magnetic drug delivery application.


Assuntos
Curcumina , Nanopartículas , beta-Ciclodextrinas , Sistemas de Liberação de Medicamentos , Microfluídica
2.
Langmuir ; 37(35): 10612-10623, 2021 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34436906

RESUMO

This paper (part II) is devoted to the effect of molecular adsorption on the surface of magnetic iron oxide nanoparticles (IONP) on the enhancement of their (secondary) field-induced agglomeration and magnetic separation. Experimentally, we use Methylene Blue (MB) cationic dye adsorption on citrate-coated maghemite nanoparticles to provoke primary agglomeration of IONP in the absence of the field. The secondary agglomeration is manifested through the appearance of needlelike micron-sized agglomerates in the presence of an applied magnetic field. With the increasing amount of adsorbed MB molecules, the size of the field-induced agglomerates increases and the magnetic separation on a magnetized micropillar becomes more efficient. These effects are mainly governed by the ratio of magnetic-to-thermal energy α, suspension supersaturation Δ0, and Brownian diffusivity Deff of primary agglomerates. The three parameters (α, Δ0, and Deff) are implicitly related to the surface coverage θ of IONP by MB molecules through the hydrodynamic size of primary agglomerates exponentially increasing with θ. Experiments and developed theoretical models allow quantitative evaluation of the θ effect on the efficiency of the secondary agglomeration and magnetic separation.

3.
Photochem Photobiol Sci ; 17(8): 1023-1035, 2018 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-29850740

RESUMO

A blue luminescent and superhydrophobic coating based on an electropolymerized fluorinated-pyrene monomer and its planktonic bacteria and biofilm repellent properties are reported. Two different pathogenic bacterial strains (Gram-positive and Gram-negative) at two different incubation times (2 h planktonic bacterial and 24 h biofilm adhesion) were studied and monitored (analyzed) using multicolor scanning confocal fluorescence microscopy. The coating was proved to reduce bacterial adhesion by 65%. It is highly effective against biofilm attachment, with 90% reduction of bacteria surface coverage. This blue fluorescent surface provides a facile method to characterize the coating, observe the bacterial distribution and quantify the bacterial coverage rate by fluorescence imaging of different colors. Furthermore, the film does not show significant bacterial toxicity during the working incubation times.


Assuntos
Biofilmes/efeitos dos fármacos , Polímeros/farmacologia , Pseudomonas aeruginosa/fisiologia , Pirenos/química , Staphylococcus aureus/fisiologia , Aderência Bacteriana , Interações Hidrofóbicas e Hidrofílicas , Testes de Sensibilidade Microbiana , Microscopia de Fluorescência , Polímeros/química , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície
4.
J Periodontal Res ; 46(6): 756-62, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21793827

RESUMO

BACKGROUND AND OBJECTIVE: The aim of the present study was to analyse the expression of MMP-2, MMP-9 and MMP-13 in newly formed bone following maxillary sinus augmentation using inorganic bovine bone substitute, because these MMPs play a major role in bone remodeling and bone resorption. MATERIAL AND METHODS: Deproteinized bovine bone (Bio-Oss(®)) was used to fill cavities after elevating the sinus mucosa. Twenty patients with edentulous posterior maxilla were treated with 20 sinus-augmentation procedures using a two-stage technique. Forty-nine Straumann(®) endosseous implants were used to complete the implant-prosthetic rehabilitation. One cylinder-shaped bone biopsy from each patient was taken from the augmented maxillary region using trephine burs at the second stage of surgery, 8 months after grafting. A biopsy was also taken as a control from the upper molar region from six different patients who did not undergo the sinus procedure. All biopsies were subjected to biochemical analysis and staining for TRAP. RESULTS: No implant losses or failures occurred. The large number of TRAP-positive multinucleated osteoclasts in resorption lacunae indicated that the resorption was very active in all grafts, in contrast with the control group. Zymography and western blot analysis demonstrated a significantly increased expression of MMP-2, MMP-9 and MMP-13 in the newly formed bone compared with controls (p < 0.05). CONCLUSION: The quantity of osteoclastic cells and the increased expression of proteolytic enzymes suggest that 8 months after grafting, inorganic bovine bone is slowly resorbing and is the site of important remodeling of the newly formed bone by means of resorption and synthesis.


Assuntos
Regeneração Óssea/fisiologia , Metaloproteinase 13 da Matriz/biossíntese , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Levantamento do Assoalho do Seio Maxilar/métodos , Adulto , Idoso , Animais , Substitutos Ósseos , Bovinos , Implantação Dentária Endóssea , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Minerais , Osteoclastos
5.
Tissue Cell ; 41(2): 141-50, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19041995

RESUMO

The purpose of this in situ study is to quantify the inflammatory cell subsets and the area fraction (AA%) occupied by collagen fibers in human healthy and diseased (four different stages) gingival connective tissue in order to establish a possible correlation between periodontal disease resulting in collagen breakdown and specific inflammatory cell subsets. Paraffin gingival tissue sections from eight healthy controls (group 0), 10 patients with gingivitis (group 1), 10 patients with moderate periodontitis (group 2) and 10 patients with severe periodontitis (group 3) were immunohistochemically investigated using antibodies against CD-45+, CD-3+, CD-8+, CD-20+, CD-68+, and EMA+ (plasma cells). The AA% occupied by gingival collagen fibers significantly decreased from 54.12% in group (0) to 38.58% in group (1), to 31.87% in group (2), and to 25.46% in group (3). In progressive lesions of periodontal disease, CD-3(+) and CD-8+ cell numbers were increased in early stages within the connective tissue, while CD-20+ cell numbers were increased only in late stages. On the other hand, EMA+, CD-68+ and CD-45+ cell numbers were progressively increased from group (0) to group (3). We demonstrated that CD-68+ monocyte/macrophages, CD-45+ leukocyte common antigen and notably EMA+ plasma cells are pertinently correlated with the severity of periodontal disease and related collagen breakdown.


Assuntos
Colágeno/imunologia , Macrófagos , Monócitos , Doenças Periodontais/imunologia , Doenças Periodontais/patologia , Plasmócitos , Adolescente , Adulto , Antígenos CD/análise , Antígenos CD/imunologia , Antígenos CD20/análise , Antígenos CD20/imunologia , Complexo CD3/análise , Complexo CD3/imunologia , Antígenos CD8/análise , Antígenos CD8/imunologia , Estudos de Casos e Controles , Colágeno/metabolismo , Matriz Extracelular/imunologia , Matriz Extracelular/metabolismo , Matriz Extracelular/patologia , Feminino , Gengiva/química , Gengiva/imunologia , Gengiva/patologia , Gengivite/imunologia , Gengivite/metabolismo , Gengivite/patologia , Humanos , Imuno-Histoquímica , Antígenos Comuns de Leucócito/análise , Antígenos Comuns de Leucócito/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Monócitos/metabolismo , Monócitos/patologia , Periodontite/imunologia , Periodontite/metabolismo , Periodontite/patologia , Plasmócitos/imunologia , Plasmócitos/metabolismo , Plasmócitos/patologia , Adulto Jovem
6.
J Periodontal Res ; 44(5): 578-87, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18752566

RESUMO

BACKGROUND AND OBJECTIVE: The presence of lysozyme in human gingiva has not previously been demonstrated. In this study, we looked for evidence for the potential role of lysozyme as a protector of gingival elastic fibres. The objective of this study was also to determine the ex vivo susceptibility to hydrolysis of gingival elastic fibres from patients with or without periodontal disease by human leukocyte elastase and by human cathepsin G. MATERIALS AND METHODS: Using gingival tissue sections from eight control, 10 gingivitis and 10 periodontitis patients, we evaluated the area fraction occupied by gingival elastic fibres (after selective staining) by the use of automated image analysis. In the ex vivo experiments, serial tissue sections from four control, four gingivitis, four young periodontitis and four aged periodontitis patients were submitted to the action of human leukocyte elastase and cathepsin G, after which enzymatic activities were determined by image analysis. Indirect immunodetection of lysozyme was also done on tissue sections for all patients included in this study. RESULTS: Large variations of the area fraction occupied by elastic fibres were observed in human gingiva from young and aged patients with and without periodontal disease. In control and gingivitis patients, leukocyte elastase and cathepsin G had high comparable elastin solubilizing activities. With young and aged periodontitis patients, the two serine proteinases had weak elastin solubilizing activities. Lysozyme appeared to be present at the periphery of gingival elastic fibres in periodontitis patients. CONCLUSION: Lysozyme can be considered an important natural protector of elastic fibres in pathological gingiva.


Assuntos
Inibidores Enzimáticos/farmacologia , Gengiva/enzimologia , Gengivite/enzimologia , Muramidase/fisiologia , Periodontite/enzimologia , Adolescente , Adulto , Fatores Etários , Idoso , Catepsina G , Catepsinas/farmacologia , Proteínas Contráteis/análise , Tecido Elástico/efeitos dos fármacos , Tecido Elástico/enzimologia , Tecido Elástico/patologia , Elastina/análise , Proteínas da Matriz Extracelular/análise , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Gengiva/patologia , Hemorragia Gengival/enzimologia , Gengivite/patologia , Humanos , Hidrólise , Processamento de Imagem Assistida por Computador , Elastase de Leucócito/farmacologia , Masculino , Pessoa de Meia-Idade , Muramidase/análise , Perda da Inserção Periodontal/enzimologia , Bolsa Periodontal/enzimologia , Periodontite/patologia , Serina Endopeptidases/farmacologia , Adulto Jovem
7.
J Biomed Mater Res A ; 87(3): 666-75, 2008 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-18189302

RESUMO

In this work, we first tested the influence of low-molecular-weight (LMW) fucoidan extracted from pheophicae cell wall on bidimensional cultured normal human osteoblasts' behaviors. Second, by impregnation procedure with LMW fucoidan of bone biomaterial (Lubboc), we explored in this bone extracellular matrix context its capabilities to support human osteoblastic behavior in 3D culture. In bidimensionnal cultures, we evidenced that LMW fucoidan promotes human osteoblast proliferation and collagen type I expression and favors precocious alkaline phosphatase activity. Furthermore, with LMW fucoidan, von Kossa's staining was positive at 30 days and positive only at 45 days in the absence of LMW fucoidan. In our three-dimensional culture models with the biomaterial pretreated with LMW fucoidan, osteoblasts promptly overgrew the pretreated biomaterial. We also evidenced that osteoblasts increased proliferation with pretreated biomaterial when compared with untreated biomaterial. Osteoblasts secreted osteocalcin and expressed BMP2 receptor on control material as well as with LMW fucoidan impregnated biomaterial. In conclusion, in our experimental conditions, LMW fucoidan stimulated expression of osteoblastic markers differentiation such as alkaline phosphatase activity, collagen type I expression, and mineral deposition; furthermore, cell proliferation was favored. These findings suggest that fucoidan could be clinically useful for bone regeneration and bone substitute design.


Assuntos
Materiais Biocompatíveis/farmacologia , Osso e Ossos/metabolismo , Osteoblastos/efeitos dos fármacos , Polissacarídeos/farmacologia , Adulto , Fosfatase Alcalina/metabolismo , Substitutos Ósseos/metabolismo , Substitutos Ósseos/farmacologia , Proliferação de Células , Células Cultivadas , Colágeno Tipo I/metabolismo , Feminino , Humanos , Masculino , Microscopia Eletrônica de Varredura , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Phaeophyceae/química , Engenharia Tecidual
8.
J Eur Acad Dermatol Venereol ; 20(8): 980-7, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16922949

RESUMO

Cutaneous ageing, as a result of combined chronological and photo-ageing in sun-exposed areas, is accompanied by major modifications of the elastic fibres. We aimed to investigate qualitative and quantitative changes of dermal elastin fibres during cutaneous chronological and photo-ageing and the involvement of lysozyme in these processes. Morphological, age-related changes and variations in the relative elastin content in sun-protected (buttock) and sun-exposed (forearm and face) skin of healthy volunteers were studied (145 samples). The deposition of lysozyme in elastin fibres was studied using light and immuno-electron microscopy and taking into consideration the relative efficacy of different UV wavebands (UVA or SSR (solar simulated radiation)). Our studies also included the proteolytic degradation of elastin by human leucocyte elastase (HLE) in situ. Our results indicate a reduction of elastin content with age in sun-protected and sun-exposed skin, associated for the latter with high elastin content, resulting in elastosis. Total UVA (320-400 nm), and in particular long wave UVA (UVA-1, 340-400 nm), induces lysozyme deposition in elastin fibres to a higher extent than solar simulated radiation (SSR, 280-400 nm). Immuno-electron microscopy revealed lysozyme association with the electron-dense granular amorphous elastin structures, corresponding to a basophilic degeneration induced by sun exposure. Lysozyme has no elastolytic activity in situ; however, its binding to elastin limits elastin degradation by human leucocyte elastase (HLE). In addition, a direct inhibitory effect of lysozyme on HLE was observed. Our data suggest that lysozyme prevents elastin degradation by HLE after binding to the damaged parts of the elastin network and by direct lysozyme-HLE interaction, which reduces HLE proteolytic activity. These observations contribute to a better understanding of the chronological and photo-induced changes of the dermal elastic network.


Assuntos
Elastina/metabolismo , Muramidase/fisiologia , Envelhecimento da Pele/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Imunofluorescência , Humanos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Microscopia de Fluorescência , Pessoa de Meia-Idade , Pele/efeitos da radiação , Raios Ultravioleta
9.
Oral Microbiol Immunol ; 21(3): 197-200, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16626378

RESUMO

BACKGROUND: As antigen-presenting cells, Langerhans cells may play an important role in the initiation and maintenance of periodontal disease. This study is the first report that extends our knowledge of the expression of matrix metalloproteinases and their endogenous tissue inhibitors by Langerhans cells in healthy and diseased gingival tissues. METHODS: Single and double immunolabeling procedures were carried out using monoclonal antibodies against CD1a, matrix metalloproteinases 2 and 9, and tissue inhibitors of matrix metalloproteinases 1 and 2, and analyzed by conventional and confocal microscopes. RESULTS: Langerhans cells expressed matrix metalloproteinases 2 and 9, and tissue inhibitors of matrix metalloproteinases 1 and 2 in healthy and diseased gingival tissues. The tissue inhibitors of matrix metalloproteinase-positive Langerhans cells were mainly observed in the upper epithelial layers. Matrix metalloproteinase 9-positive Langerhans cells were observed especially during periodontitis and in the basal epithelial layer or crossing the basement membrane. CONCLUSION: During periodontal disease, changes in the expression of matrix metalloproteinases and their tissue inhibitors by gingival Langerhans cells could be implicated in the migration of the cells towards the connective tissue.


Assuntos
Gengiva/enzimologia , Células de Langerhans/enzimologia , Metaloproteinases da Matriz/biossíntese , Periodontite/enzimologia , Inibidores de Proteases/metabolismo , Inibidores Teciduais de Metaloproteinases/biossíntese , Movimento Celular , Gengiva/citologia , Humanos , Técnicas Imunoenzimáticas , Metaloproteinase 2 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Microscopia Confocal , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Inibidor Tecidual de Metaloproteinase-2/biossíntese
10.
Cytometry A ; 69(1): 20-6, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16342114

RESUMO

BACKGROUND: We characterize the application of multiphoton microscopy to the observation of the extracellular matrix of fresh unstained vessels. METHOD: Combined two-photon-excited fluorescence (2PEF) and second harmonic generation (SHG) imaging of large arteries reveals the architecture of elastin and collagen fibers in the vessel wall with remarkable specificity. RESULTS: We present elastin/collagen imaging in unstained rat vessels at both micrometer and whole vessel scales, and we characterize the optical properties of rat carotid artery and aorta walls. We apply this method to evidence deleterious effects of residual doses of a pesticide on the vessel wall. CONCLUSION: This study illustrates the potential of 2PEF/SHG microscopy for pharmacological studies in unlabeled arteries.


Assuntos
Vasos Coronários/ultraestrutura , Matriz Extracelular/metabolismo , Animais , Artérias Carótidas/metabolismo , Artérias Carótidas/ultraestrutura , Colágeno/metabolismo , Colágeno/ultraestrutura , Vasos Coronários/metabolismo , Elastina/metabolismo , Elastina/ultraestrutura , Matriz Extracelular/ultraestrutura , Feminino , Interpretação de Imagem Assistida por Computador , Técnicas In Vitro , Masculino , Microscopia de Fluorescência por Excitação Multifotônica , Ratos , Ratos Sprague-Dawley
11.
J Vasc Res ; 42(3): 190-201, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15832055

RESUMO

Supravalvular aortic stenosis (SVAS) and Williams Beuren syndrome (WBS) can be considered as inherited diseases affecting the whole arterial tree and causing narrowing of the vessels. It has been reported that abnormal deposition of elastin in arterial walls of patients with SVAS and WBS leads to increased proliferation of arterial smooth muscle cells (SMC), which result in the formation of hyperplastic intimal lesions. In this work, we conducted morphological and morphometrical analysis with stenotic aortas from patients suffering from SVAS and WBS and from healthy control subjects and demonstrated that the amount of elastic fibers and the loss of integrity of vascular elastic fibers in the aortas reflect similar changes in the skin of patients with SVAS or WBS, as reported in our previous work conducted on skin in these pathological states. On the other hand, we conducted investigations on metalloproteinases (MMP2, MMP9, MMP7) and their specific tissue inhibitors TIMP1 and TIMP2 to verify their possible involvement in the etiopathogeny of SVAS and WBS. We particularly evidenced an altered MMP9/TIMP1 balance in favor of matrix degradation which could facilitate SMC migration and neointimal hyperplasia. Our findings suggest that elastinolytic enzymes secreted by arterial SMC, possibly including matrilysin 1, are critical for the development of arterial lesions in SVAS and WBS and contribute to perpetuate arterial stenosis in either SVAS or WBS.


Assuntos
Aorta/fisiopatologia , Estenose Aórtica Supravalvular/fisiopatologia , Síndrome de Williams/fisiopatologia , Adulto , Aorta/patologia , Estenose Aórtica Supravalvular/patologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Humanos , Processamento de Imagem Assistida por Computador , Técnicas Imunológicas , Metaloproteases/metabolismo , Distribuição Tecidual , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Síndrome de Williams/patologia
12.
Opt Express ; 13(17): 6667, 2005 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-19498682

RESUMO

We present corrected versions of the list of authors and of Section 2.1.

13.
Arch Dermatol Res ; 296(5): 220-5, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15449075

RESUMO

Toxic epidermal necrolysis (TEN) and Stevens-Johnson syndrome (SJS) are considered to be drug-induced diseases, and are characterized by extensive mucocutaneous disorder and epidermal necrosis which result in the detachment of the epidermis. Inactive and active forms of metalloproteinases (MMP2 and MMP9) secreted by skin explants maintained in organ culture for 72 h and in blister fluid from two TEN and three SJS patients were investigated. Interestingly, lesional skin from both the TEN and the SJS patients cultured for 3 days in conditioned medium showed high levels of both 72 kDa progelatinase A and 66 kDa activated gelatinase A, and the 66 kDa activated form was not observed in cultures of skin from control individuals. Furthermore, indirect immunodetection showed the presence of MMP2 and MMP9 in TEN and SJS patients' skin. Increased gelatinase activity in the culture medium of TEN and SJS skin maintained in organ culture and in blister fluid indicates that these gelatinases may be responsible for the detachment of the epidermis in these drug-induced necrolyses.


Assuntos
Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Síndrome de Stevens-Johnson/enzimologia , Adulto , Idoso , Vesícula/enzimologia , Vesícula/etiologia , Vesícula/patologia , Western Blotting , Líquidos Corporais/enzimologia , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Técnicas Imunoenzimáticas , Técnicas Imunológicas , Masculino , Pessoa de Meia-Idade , Técnicas de Cultura de Órgãos , Pele/enzimologia , Pele/patologia , Coloração e Rotulagem , Síndrome de Stevens-Johnson/complicações , Síndrome de Stevens-Johnson/patologia
14.
Clin Oral Investig ; 7(4): 206-11, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14508666

RESUMO

The aim of this study was to determine if a vegetable extract from seeds of Lupinus albus (LU 105) has the capacity to inhibit human leukocyte elastase and/or protect gingival elastic fibers against proteolytic degradation. LU105 was extracted from seeds of L albus and is freely soluble in water. In this study the ex-vivo elastolytic activity of human leukocyte elastase and the potential inhibitory effect of LU 105 were determined using human gingival cryostat tissue sections and computerized morphometric analysis. The gingival tissue sections pre-treated or not with LU 105 were submitted to the action of human leukocyte elastase or submitted to the simultaneous action of human leukocyte elastase and LU 105, and then analyzed using automated image analysis. In such conditions, LU 105 at 0.1%, 0.01%, and 0.001% developed a dose dependent protection of gingival elastic fibers against enzymatic proteolysis due to human leukocyte elastase, and LU 105 at 0.1% or 0.01% was able to inhibit the elastolytic activity of leukocyte elastase itself. It is proposed that LU 105 is an option for the treatment of gingival inflammation in which leukocyte elastase is involved.


Assuntos
Tecido Elástico/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Elastase de Leucócito/antagonistas & inibidores , Lupinus , Oligopeptídeos/farmacologia , Extratos Vegetais/farmacologia , Inibidores de Proteases/farmacologia , Tecido Elástico/enzimologia , Humanos
15.
Exp Dermatol ; 12(4): 403-11, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12930296

RESUMO

The aim of this work was to validate an image analysis method, based on cell nuclei form factor determination, for counting fibroblasts within human dermis. We first used reconstructed dermal equivalents in which fibroblasts can also be counted directly after lysis of the collagen matrix. We found a good correlation between the results of direct counting and those of image analysis from day 10 to day 28 of culture. When applied to young normal donors' skin biopsies fixed in Bouin's solution and embedded in paraffin, the image analysis method yielded mid-dermis fibroblast counts of between 2100 and 4100 per mm3 of fresh tissue. A nuclear form factor (FF) comprised between 0.35 and 0.84 was found to be a biologic marker of fibroblasts. This was confirmed after fibroblast discrimination from other cell types, which had rounder nuclei (FF >/= 0.85) and were identified either by their location (e.g. endothelial cells) or by labeling with specific antibodies (e.g. lymphocytes and monocytes/macrophages). Similar results were obtained with seven healthy donors' skin biopsies that had been frozen in nitrogen liquid and cryostat-sectioned, showing that this counting method is independent of the histologic procedure. Finally, analysis of samples of hypertrophic scars from two patients revealed that fibroblast density in some parts of the dermis was more than twice the value found in other parts presenting a fibroblast density almost normal, showing that this cell counting method can also be used to assess fibroblast heterogeneity within a given tissue.


Assuntos
Contagem de Células/métodos , Derme/citologia , Derme/patologia , Fibroblastos/citologia , Fibroblastos/patologia , Núcleo Celular/ultraestrutura , Técnicas Histológicas , Humanos , Processamento de Imagem Assistida por Computador
16.
Clin Oral Investig ; 7(4): 198-205, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12802622

RESUMO

This study examined the effects of a vegetable extract from Lupinus albus (LU105) on MMPs and TIMPs secreted by human gingival fibroblasts in culture. LU105 was extracted from seeds of L. albus and is freely soluble in water. Gelatin zymography showed that control human gingival fibroblasts maintained in culture for 48 h express pro-MMP2 (progelatinase A) in the culture medium while the active form of MMP2 (gelatinase A), the active form of MMP9 (gelatinase B), and pro-MMP9 (progelatinase B) are not detected. Fibroblasts derived from inflamed gingiva expressed in the culture medium increased amounts of pro-MMP2 (progelatinase A) compared with controls and significant amounts of pro-MMP9 (progelatinase B). LU105 diminished the expression by gingival fibroblasts derived from inflamed tissue of both pro-MMP2 and pro-MMP9. Furthermore LU105 did not modify the amount of TIMP2 expressed in culture by controls or by gingival fibroblasts derived from inflamed tissue. TIMP1 and MMP1 significantly decreased when LU105 was added in the culture media of gingival fibroblasts derived from inflamed tissue compared with control fibroblasts. Thus LU105 seems to offer an opportunity to restore a correct balance between MMP2, MMP9, MMP1, and their natural inhibitors, i.e., TIMP1 and TIMP2 in human inflamed gingiva.


Assuntos
Gengiva/efeitos dos fármacos , Gengiva/enzimologia , Lupinus , Metaloproteinases da Matriz/efeitos dos fármacos , Oligopeptídeos/farmacologia , Periodontite/enzimologia , Extratos Vegetais/farmacologia , Inibidores de Proteases/farmacologia , Adolescente , Adulto , Análise de Variância , Western Blotting , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Gengiva/citologia , Humanos , Inibidores de Metaloproteinases de Matriz , Metaloproteinases da Matriz/análise , Metaloproteinases da Matriz/biossíntese , Sementes , Inibidores Teciduais de Metaloproteinases/análise , Inibidores Teciduais de Metaloproteinases/biossíntese , Inibidores Teciduais de Metaloproteinases/efeitos dos fármacos
17.
Magnes Res ; 16(1): 43-8, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12735482

RESUMO

Vascular alterations during magnesium deficiency have long been known but the implicated mechanisms have so far been poorly documented. In this preliminary assay, we compared the thoracic aortic histology in Swiss OF1 mice fed a severe magnesium-deficient diet (50 +/- 5 ppm) for 42 days to that of controls fed a standard diet (1700 +/- 100 ppm magnesium). It appeared (eosin-haematoxylin coloration) that, in magnesium-deficient mice, the aortic wall was thinner than in controls. Specific colorations of the two of main fibers vascular tissue (collagens and elastin) showed severe structural alterations of both components. These changes were consecutive to the expression of matrix metalloproteinases (MMP) -2 and -9 which were present as zymogens (inactive forms) in controls and supposed to be present in their active and inactive forms in magnesium-deficient mice (zymography). These changes which have not been reported so far would explain, at least in part, the sensitivity of magnesium-deficient mice to various stress or xenobiotics.


Assuntos
Endotélio Vascular/enzimologia , Endotélio Vascular/patologia , Deficiência de Magnésio/enzimologia , Metaloproteinase 2 da Matriz/fisiologia , Metaloproteinase 9 da Matriz/fisiologia , Animais , Aorta Torácica/enzimologia , Aorta Torácica/patologia , Bovinos , Células Cultivadas , Magnésio/sangue , Deficiência de Magnésio/patologia , Camundongos , Suínos
18.
Magnes Res ; 16(1): 59-64, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12735484

RESUMO

The medical management of Marfan Syndrome (MFS) mainly relies on early prevention of the aortic complications. Hemodynamic treatments try to diminish the forcefulness of cardiac contractions and to reduce blood pressure: for example long term administration of propranolol may significantly reduce the rate of increase in aortic ratio (aortic diameter/expected aortic diameter). Retardation of aortic dilatation may be most often observed by early treatment started when the baseline end-diastolic aortic root diameter is < 40 mm. It seems better to use beta-blockers without intrinsic sympathomimetic activity. Successful acceptance of beta-blockers may be limited by side-effects, but the efficiency of alternative hypotensive agents (calcium channel inhibitors, ACE inhibitors) is not yet validated. Gene therapy might constitute an etiologic specific treatment of MFS. FBN1-RZ1 hammerhead antisense ribozyme is able to suppress expression of the mutant FBN1 allele. The use of ribozymes as systemic therapeutic agents will depend on efficient delivery to its target, but the various proposed vectors raise yet unsolved problems. A hydrogel angioplasty balloon might be a possible vector for delivering an antisense ribozyme in the aortic wall specifically. Ribozymes--as deoxyribonucleotides--may be taken up by tissue upon local application. Further research should study ex vivo local application of antisense ribozyme on human aortic wall, before assessing in vivo efficiency and tolerance of this aortic local vectorisation. It is always necessary to maintain a balanced magnesium intake in patients with MFS. Firstly to prevent the multiple noxious effects of magnesium deficiency on cardiovascular targets. Secondly to ensure the best efficiency and the least toxicity of the hemodynamic drugs used as long term prophylactic treatment for cardiovascular complications and of the etiologic antisense magnesium-dependent gene therapy, in the future.


Assuntos
Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/prevenção & controle , Terapia Genética/métodos , Magnésio , Síndrome de Marfan/metabolismo , Síndrome de Marfan/terapia , RNA Antissenso/administração & dosagem , Doenças Cardiovasculares/etiologia , Hemodinâmica , Humanos , Magnésio/uso terapêutico , Deficiência de Magnésio/etiologia , Deficiência de Magnésio/prevenção & controle , Síndrome de Marfan/complicações , RNA Catalítico/administração & dosagem
19.
FASEB J ; 17(6): 644-51, 2003 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-12665477

RESUMO

Periodontitis are diseases of the supportive tissues of the teeth provoked by bacteria and characterized by gingival inflammation and bone destruction. We have developed a new strategy to repair tissues by administrating agents (RGTA) that mimic heparan sulfates by protecting selectively some of the growth factors naturally present within the injured tissue and interfering with inflammation. After periodontitis induction in hamsters, the animals were left untreated or received weekly i.m. injections of RGTA1507 at a dose of 100 microg/kg, 400 microg/kg, 1.5 mg/kg, or 15 mg/kg for 4 wk. RGTA treatment significantly reduced gingival tissue inflammation, thickened the pocket epithelium by increasing cell proliferation, and enhanced collagen accumulation in the gingiva. A marked reduction in bone loss was observed, resulting from depression of osteoclasia and robust stimulation of bone formation at the dose of 1.5 mg/kg. RGTA treatment for 8 wk at this dose reversed macroscopic bone loss, sharply contrasting with the extensive bone destruction in the untreated animals. RGTA treatment decreased gelatinase A (MMP-2) and B (MMP-9) pro-forms in gingival tissues. Our data indicate that a 4 wk treatment dose-dependently attenuated gingival and bone manifestations of the disease, whereas a longer treatment restored alveolar bone close to controls. By modulating and coordinating host responses, RGTA has unique therapeutic properties and is a promising candidate for the treatment of human periodontitis.


Assuntos
Dextranos/uso terapêutico , Periodontite/tratamento farmacológico , Animais , Regeneração Óssea/efeitos dos fármacos , Cricetinae , Dextranos/administração & dosagem , Dextranos/química , Relação Dose-Resposta a Droga , Precursores Enzimáticos/efeitos dos fármacos , Precursores Enzimáticos/metabolismo , Gengiva/efeitos dos fármacos , Gengiva/metabolismo , Gengiva/patologia , Masculino , Metaloproteinase 2 da Matriz/efeitos dos fármacos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/efeitos dos fármacos , Metaloproteinase 9 da Matriz/metabolismo , Mesocricetus , Periodontite/enzimologia , Periodontite/patologia
20.
Clin Oral Investig ; 6(1): 39-50, 2002 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11996162

RESUMO

Free-floating collagen lattice is considered a useful tool for assessing wound healing in vitro. This work compared extracellular matrix remodeling in collagen lattices populated by gingival or dermal fibroblasts. For 21 days we followed gel contraction and changes in cell number of collagen lattices seeded with l.5 x 10(5) fibroblasts of each tissue. We also used indirect immunodetection to study extracellular matrix components, metalloproteinases (MMPs), and their tissues inhibitors (TIMPs). In addition, the presence of MMPs and TIMPs in the culture media was analyzed by zymography and western blotting. No significant difference was found concerning gel contraction and changes in cell number. We observed the early expression of fibrillin I and collagen type III, apparently codistributed and at the end of the gel contraction their disappearance. Concomitantly we demonstrated the expression of MMPs and TIMPs, initially localized in cellular cytoplasm, then spreading in the extracellular compartment, and even found in the culture medium. This remodeling was more rapid and intense with gingival fibroblasts than dermal fibroblasts. In conclusion, gingival fibroblasts seem more efficient at remodeling the connective tissue than dermal fibroblasts and could lead to the better wound healing observed in vivo.


Assuntos
Derme/citologia , Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Gengiva/citologia , Adolescente , Adulto , Western Blotting , Contagem de Células , Células Cultivadas , Criança , Técnicas de Cocultura , Colágeno/metabolismo , Colágeno Tipo I/análise , Colágeno Tipo III/análise , Células do Tecido Conjuntivo/metabolismo , Citoplasma/enzimologia , Elastina/análise , Proteínas da Matriz Extracelular/análise , Fibrilinas , Fluoresceína-5-Isotiocianato , Técnica Indireta de Fluorescência para Anticorpo , Corantes Fluorescentes , Humanos , Metaloproteinases da Matriz/análise , Proteínas dos Microfilamentos/análise , Microscopia Eletrônica , Inibidores Teciduais de Metaloproteinases/análise , Cicatrização
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