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1.
Biomaterials ; 32(28): 6850-9, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21741701

RESUMO

Bioactive implants are promising tools in regenerative medicine. Here we describe a versatile procedure for preparing a gene-activated matrix on titanium. Lyophilized copolymer-protected gene vectors (COPROGs) suspended in poly(d,l-lactide) (PDLLA) solutions in ethyl acetate were used to varnish solid surfaces. The gene-activated PDLLA surfaces were first established on polypropylene 96-well plates. Vector release from these surfaces in aqueous buffer, cell viability and gene transfer efficiency to NIH 3T3 fibroblasts was strongly dependent on the vector dose and its ratio to PDLLA film thickness. A detailed analysis of these relationships allowed establishing correlations which can be used to calculate suitable combinations of COPROGs and PDLLA yielding optimal gene transfer efficiency. This was verified with COPROG-activated PDLLA coatings on titanium foils. HEK 293 and mesenchymal stem cells expressed the BMP-2 gene comprised in the gene-activated surface in a manner that was consistent with the predicted dose-response and toxicity profiles found in NIH 3T3 cells. The systematic procedure presented here for identifying optimal coating compositions can be applied to any combination of vector type and coating material.


Assuntos
Materiais Revestidos Biocompatíveis/química , Vetores Genéticos/química , Vetores Genéticos/genética , Poliésteres/química , Titânio/química , Animais , Materiais Revestidos Biocompatíveis/metabolismo , DNA/química , DNA/metabolismo , Técnicas de Transferência de Genes , Genes Reporter , Vetores Genéticos/metabolismo , Células HEK293 , Humanos , Teste de Materiais , Camundongos , Microscopia Eletrônica de Varredura , Células NIH 3T3 , Próteses e Implantes
2.
Int J Cancer ; 129(5): 1265-76, 2011 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-21710499

RESUMO

The human Y-box binding protein 1 (YB-1) is known to be a promising target for cancer therapy. We have demonstrated that YB-1 plays an important role in the adenoviral life cycle by regulating the adenoviral E2-gene expression. Thus, we studied the oncolytic effect of the recombinant adenovirus Ad-Delo3-RGD, in which the transactivation domain CR3 of the E1A protein is ablated to enable viral replication only in YB-1 positive cancer cells. In vitro Southern Blot analysis and cytopathic effect assays demonstrate high anti-glioma potency, which was significantly increased in combination with temozolomide (TMZ), daunorubicin and cisplatin. Since vascular endothelial growth factor (VEGF) is thought to promote the hypervascular phenotype of primary, malignant brain tumors, we also tested Ad-Delo3-RGD in regard to the inhibition of VEGF expression. Indeed, we found that Ad-Delo3-RGD induced VEGF down regulation, which was even amplified under hypoxic conditions. Tumor-bearing nudemice treated with the YB-1 dependent oncolytic adenovirus showed significantly smaller tumors than untreated controls. Furthermore, combination therapy with TMZ led to a regression in all treated animals with complete tumor regression in 33 % of analyzed mice, which was verified by bioluminescence imaging and histological studies. In addition, histopathological evaluation revealed enhanced apoptosis and a reduction in tumor vessel formation, indicating that Ad-Delo3-RGD has an anti-angiogenic effect in addition to its oncolytic capacity in vivo. Hence, our results demonstrate that the combination therapy of YB-1 dependent virotherapy and TMZ is effective in a xenograft glioma mouse model and might be useful in a YB-1 based clinical setting.


Assuntos
Adenoviridae/genética , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Encefálicas/terapia , Terapia Genética , Glioma/terapia , Terapia Viral Oncolítica , Proteína 1 de Ligação a Y-Box/genética , Animais , Southern Blotting , Western Blotting , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Cisplatino/administração & dosagem , Terapia Combinada , Dacarbazina/administração & dosagem , Dacarbazina/análogos & derivados , Daunorrubicina/administração & dosagem , Vetores Genéticos/uso terapêutico , Glioma/genética , Glioma/secundário , Humanos , Técnicas Imunoenzimáticas , Camundongos , Camundongos Nus , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Temozolomida , Células Tumorais Cultivadas , Replicação Viral
3.
J Anat ; 217(1): 76-82, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20456523

RESUMO

Polychrome sequential labeling with fluorochromes is a standard technique for the investigation of bone formation and regeneration processes in vivo. However, for human application, only tetracycline and its derivates are approved as fluorochromes. Therefore, the aim of this study was to determine the fluorescence characteristics of the different tetracycline derivates to assess the feasibility of sequential in vivo bone labeling using distinguishable fluorochromes. Eight different tetracycline derivates were injected subcutaneously into growing rats as a single dose or sequentially in different combinations. After preparation of resin-embedded undecalcified bone sections, the fluorescence properties of the tetracycline derivates in bone were analyzed using conventional fluorescence microscopy, spectral image analysis and confocal laser scanning microscopy. Each tetracycline derivate exhibited a characteristic fluorescence spectrum, but the differences between them were small. Chlortetracycline could be discriminated reliably from all other derivates and could therefore be combined with any other tetracycline derivate for reliably distinguishable double labeling. Tetracycline itself exhibited the brightest fluorescence of all the investigated derivates. Interestingly, in conventional microscopy the same tetracycline derivative can appear in different colours to the human eye, even if spectral analysis confirmed identical emission peaks. In conclusion, the data suggest that fluorescence double labeling of bone is feasible using appropriate tetracycline derivates in combination with spectral imaging modalities.


Assuntos
Osso e Ossos/metabolismo , Tetraciclinas , Animais , Estudos de Viabilidade , Corantes Fluorescentes , Processamento de Imagem Assistida por Computador/métodos , Masculino , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Ratos , Ratos Wistar
4.
Lasers Med Sci ; 23(1): 55-8, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17468897

RESUMO

Concerning dental implant systems, a main problem is the adhesion of peri-implant mucosa in the cervical region. The aim of the present study was to use a laser for modifying titanium implants to promote mucosal adhesion, which is indispensable as a biological barrier against bacterial infection. By the use of a KrF excimer laser, it was possible to induce a holey structure on the polished area of the implant surface, which was analysed by a scanning electron microscope. In addition, the attachment of fibroblast cells to the created structures was investigated with the aid of an environmental scanning electron microscope. It turned out that the cells preferentially attach to the holey structure. Thereby, the cells form bridges inside, leading to a complete covering of the hole. In this way, a more effective biological barrier against bacteria can be created.


Assuntos
Adesão Celular , Implantação Dentária Endóssea , Implantes Dentários , Lasers , Titânio , Humanos , Osseointegração , Desenho de Prótese , Doses de Radiação , Propriedades de Superfície
5.
Lasers Med Sci ; 22(4): 223-7, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17318664

RESUMO

Dental implants are becoming increasingly important in prosthodontic rehabilitation. Bacterial infections, however, can induce bone loss and jeopardize clinical success. Recent literature has demonstrated that infrared CO(2) laser light is suitable for the decontamination of exposed implant surfaces. The aim of the present study was to investigate the influence of 308-nm excimer laser irradiation on peri-implantitis-associated bacteria in vitro. In this study, a XeCl excimer laser (308 nm) was used (Summit Technology, Boston, USA). Both aerobe (Streptococcus mutans, S. sanguis, Actinomyces naeslundii) and anaerobe microorganisms (A. odontolyticus, Prevotella melaninogenica) were tested. According to previous studies, a constant energy of 0.8 J/cm(2) and a constant frequency of 20 Hz were used for all irradiations. Colony-forming units after laser irradiation were counted. Excimer laser irradiation showed significant influence on the growth of all microorganisms. As compared to S. mutans and S. sanguis, A. naeslundii demonstrated higher sensitivity to laser irradiation. Anaerobe microorganisms, in contrast, demonstrated that a total of 200 pulses were sufficient to reduce the replication of these germs for more than 99.9%. Excimer laser irradiation (lambda = 308 nm) can significantly reduce both aerobe and anaerobe microorganisms. Depending on the parameters chosen, 200 pulses are sufficient for sterilization. New studies are necessary to evaluate if this wavelength is more of value in the treatment of peri-implantitis than other wavelengths or conventional therapies.


Assuntos
Infecções Bacterianas/etiologia , Implantação Dentária Endóssea/efeitos adversos , Lasers de Excimer , Bactérias Aeróbias , Bactérias Anaeróbias , Infecções Bacterianas/microbiologia , Infecções Bacterianas/terapia , Humanos , Técnicas In Vitro , Projetos Piloto , Fatores de Risco
6.
J Anat ; 210(1): 117-21, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17229289

RESUMO

Fluorochrome sequential labelling of mineralizing tissues is commonly used in different fields of clinical and basic research. Recently we improved polychrome fluorescent sequential labelling of bone by applying spectral image analysis to discriminate seven different fluorochromes. Although basic mineralization processes of bone and teeth follow comparable principles, the respective tissues differ in terms of matrix composition and mineral assembly. The aim of this study therefore was to investigate the feasibility of this new technique for polychrome sequential labelling of teeth and to demonstrate the advantages in the field of dentistry. Furthermore, the exact labelled area of each fluorochrome could be measured, even in regions of overlapping fluorochromes. The technique presented may provide a basis for further investigations of mineralization processes of different anatomical dental structures.


Assuntos
Processamento de Imagem Assistida por Computador , Dente/química , Animais , Corantes Fluorescentes , Masculino , Microscopia de Fluorescência/métodos , Ratos , Ratos Wistar , Espectrofotometria
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