Yes / Infection is the major reason for guided tissue regeneration/guided bone regeneration (GTR/GBR) membrane failure in clinical application. In this work, we developed GTR/GBR membranes with localized drug delivery function to prevent infection by electrospinning of poly(ε-caprolactone) (PCL) and gelatin blended with metronidazole (MNA). Acetic acid (HAc) was introduced to improve the miscibility of PCL and gelatin to fabricate homogeneous hybrid nanofiber membranes. The effects of the addition of HAc and the MNA content (0, 1, 5, 10, 20, 30, and 40 wt.% of polymer) on the properties of the membranes were investigated. The membranes showed good mechanical properties, appropriate biodegradation rate and barrier function. The controlled and sustained release of MNA from the membranes significantly prevented the colonization of anaerobic bacteria. Cells could adhere to and proliferate on the membranes without cytotoxicity until the MNA content reached 30%. Subcutaneous implantation in rabbits for 8 months demonstrated that MNA-loaded membranes evoked a less severe inflammatory response depending on the dose of MNA than bare membranes. The biodegradation time of the membranes was appropriate for tissue regeneration. These results indicated the potential for using MNA-loaded PCL/gelatin electrospun membranes as anti-infective GTR/GBR membranes to optimize clinical application of GTR/GBR strategies.

Inspiriert durch natürliche Grenzflächenproteine in Knochen wurden bifunktionale Biokonjugate für verschiedene Grenzflächenstabilisierungen genutzt. In einem kombinatorischen Ansatz wurden materialspezifische Peptid-block-Polyethylenglycol-Konjugate eingesetzt, um die Grenzflächen von Nanopartikeln in Lösung sowie in Polymerkompositen zu stabilisieren. Dazu wurden biokombinatorisch ausgewählte Peptid-Sequenzen mit einer Affinität für MgF2-Oberflächen in Form eines MgF2-bindenden Konjugats (MBC) synthetisiert, welches die materialaffine Bindung der monodispersen Peptid-Domäne mit der zusätzlichen Funktion des synthetischen Polymer-Blocks verbindet. Aus detaillierten Untersuchungen der Bindungseigenschaften von MBC und davon abgeleiteten Konjugaten mit variierten Peptidarchitekturen bzw. Polymer-Blocklängen bei verschiedenen Inkubationsbedingungen ging hervor, dass das sequenzspezifisch bindende MBC das Potenzial zur Stabilisierung von MgF2-Nanopartikeln hat. Das Konjugat verhinderte die Agglomeration der Partikel und ermöglichte im Gegensatz zu etablierten Stabilisatoren die vollständige Redispergierbarkeit sogar nach Eintrocknung. Die Stabilisierung in Lösung wurde auf die Kompatibilisierung von Partikeln in Polycaprolacton (PCL)-Kompositen übertragen, in denen das grenzflächenaktive MBC die Materialeigenschaften von bioabbaubaren PCL/MgF2-Kompositen optimierte. Die Grenzflächenstabilisierung führte zur gleichzeitigen Erhöhung der Steifigkeit und der Zähigkeit der Materialien bis in den Bereich natürlicher Knochen. Durch die gemeinsame Zugabe von MBC-kompatibilisiertem MgF2 und Hyxdroxylapatit zu PCL wurde ein bioaktives Material geschaffen, das nachweislich die osteogene Differenzierung von mesenchymalen Stammzellen und die Mineralisierung von neuem Knochengewebe unterstützte. Damit stellt es ein vielversprechendes Komposit für die Regeneration von Knochengewebe und weitere Anwendungen dar. / Inspired by natural interface proteins in bone, bifunctional bioconjugates were exploited for different interface stabilization applications. The interfaces of nanoparticles both in solution and in polymeric composites were stabilized by combinatorially selected, material specific peptide-polymer conjugates. Peptide sequences showing affinity to MgF2 particle surfaces were selected from a phage display library and translated into a MgF2-binding peptide-block-poly(ethylene glycol) conjugate (MBC). The MBC combined the material-affine binding of the monodisperse peptide domain with an additional function of a synthetic polymer block. Detailed studies of the binding properties of MBC and congeneric conjugates with other peptide architectures or different polymer block lengths, as well as varied incubation conditions revealed the potential of the sequence-specific MBC to stabilize MgF2 nanoparticles in solution. The conjugate inhibited the agglomeration of the particles. In contrast to established stabilizers, it enabled fully redispersable nanoparticles even after complete drying. The stabilization approach in solution was expanded to the compatibilization of the particles in polycaprolactone (PCL) composites. Inspired by the structure of highly specific interface proteins, MBC optimized the material properties of biodegradable PCL/MgF2 composites. Additionally, the interface stabilization simultaneously increased both the stiffness and the toughness of the composites up to the range of natural bone. The addition of hydroxyapatite alongside MBC-compatibilized MgF2 to PCL created a bioactive material that showed enhanced osteogenic differentiation of mesenchymal stem cells and the mineralization of new bone tissue. Therefore, a mechanically reinforced, osteoinductive material was prepared showing high potential in extensive in vitro studies for biomedical applications such as guided bone regeneration, yet not limited to that.

No description available.

Made available in DSpace on 2014-07-29T15:22:00Z (GMT). No. of bitstreams: 1 Parte 1.pdf: 1874528 bytes, checksum: 7e46501ade5b782c1d4f3f9926c18a43 (MD5) Previous issue date: 2006-11-30 / Control of the process of repair or progression of periapical lesions after endodontic treatment is monitored by conventional or digital radiography. In this research digital subtraction radiography (DSR) was used that uses the subtraction of images longitudinally, in which the change in the alveolar bone is visualized against a uniform gray background. The objectives of this study were: (1) to evaluate the repair process of periapical lesions after endodontic treatment by using DSR; (2) to quantify by means of point/pixel (picture element), area (histogram) and linear measures (profile line), the gain or loss of mineral density in the area of the lesion, using the average of the pixel values; (3) to compare the diagnostic information, suggestive of the repair process, obtained through a subjective evaluation of DSR with a conventional radiographic evaluation and digitalized image and (4) to evaluate the contribution of DSR to an early identification of the repair of periapical lesions after endodontic treatment. The sample consisted of twelve patients with a total of seventeen periapical lesions. The x-rays were digitalized and submitted to DSR using DSR software. The pixel values of the subtracted images were determined by using Image Tool software. Both the conventional x-rays as well as the digitalized and subtracted images were qualitatively evaluated. The results showed a gain in mineral density with a meandp of 133.495.17, 130.275.77 and 129.414.46 for the points/pixel, histogram and profile line tools, respectively. In the evaluation of numerical gain Pearson s Coefficient of Correlation (r) presented these values: mean of points/histogram = 0.746; mean of points/profile line = 0.724 and histogram/profile line = 0.860. When the numerical values were transformed into percentile gain meandp of 0.674.01, 1.214.33 and 1.163.36 were obtained for the points/pixels, histogram and profile line tools, respectively. In the evaluation of the percentile gain Spearman s Coefficient of Correlation (rs) showed the following values: mean of points/histogram = 0.697; mean of the points/profile line = 0.646 and histogram/profile line = 0.844. In the qualitative analysis, the frequency of success in the ordering of the correct sequence of the repair process using conventional radiography, digitalized image and DSR was 37.3%, 31.4% and 31.4%, respectively. One concluded, therefore, that: (1) the process of repair of periapical lesions after endodontic treatment can be evaluated quantitatively by means of longitudinal analysis using DSR; (2) any one of the three tools can be used to quantify the repair, considering that correlation exists between the time of repair and the increase of the value of pixel; (3) the comparative evaluation between the subjective methods using conventional radiography, digitalized image and SDR, it showed that all had been capable to evidence the process of repair of periapical lesions from the first radiography (15 days), not having difference between them and (4) the quantitative evaluation by SDR obtained to after evidence the beginning of the repair with 15 days the beginning of the endodontic treatment, even so this repair was really effective from 105 days after the beginning of the endodontic treatment. / O controle do processo de reparo ou progressão de lesões periapicais pós-tratamento endodôntico é monitorado pelo exame radiográfico convencional ou digital. Nesta pesquisa foi utilizada a subtração digital radiográfica (SDR), que utiliza a subtração de imagens longitudinalmente, na qual a mudança no osso alveolar é visualizada contra um plano de fundo (background) cinza homogêneo. Os objetivos desse estudo foram: (1) avaliar o processo de reparo de lesões periapicais pós-tratamento endodôntico por meio de SDR; (2) quantificar por meio de ponto/pixel (picture element), área (histograma) e medida linear (perfil linha) na área da lesão, o ganho ou perda de densidade mineral por meio da média dos valores dos pixels; (3) comparar as informações diagnósticas, sugestivas do processo de reparo, obtidas por meio da avaliação subjetiva da SDR com a avaliação radiográfica convencional e imagem digitalizada; e (4) avaliar a contribuição da SDR na identificação precoce do reparo de lesões periapicais pós-tratamento endodôntico. A amostra constituiu-se de doze indivíduos totalizando dezessete lesões periapicais. As radiografias foram digitalizadas e submetidas à SDR utilizando o programa DSR. As imagens subtraídas tiveram os valores de pixel determinados utilizando o programa Image Tool. Tanto as radiografias convencionais quanto as imagens digitalizadas e subtraídas foram avaliadas qualitativamente. Os resultados evidenciaram ganho de densidade mineral com médiadp de 133,495,17; 130,275,77; 129,414,46 para as ferramentas ponto/pixel; histograma e perfil linha respectivamente. Na avaliação do ganho numérico o Coeficiente de Correlação de Pearson (r) mostrou valores de: média dos pontos/ histograma = 0,746; média dos pontos/ perfil linha = 0,724 e histograma/ perfil linha = 0,860. Quando os valores numéricos foram transformados em ganho percentual foram obtidas médiadp de 0,674,01; 1,214,33; 1,163,36 para as ferramentas ponto/pixel; histograma e perfil linha respectivamente. Na avaliação do ganho percentual o Coeficiente de Correlação de Spearman (rs) mostrou valores de: média dos pontos/ histograma = 0,697; média dos pontos/ perfil linha = 0,646 e histograma/ perfil linha = 0,844. Na análise qualitativa, a freqüência de acertos na ordenação da seqüência correta do processo de reparo usando radiografia convencional, imagem digitalizada e SDR foi de 37,3%; 31,4% e 31,4% respectivamente. Concluiu-se, portanto, que: (1) o processo de reparo de lesões periapicais pós-tratamento endodôntico pode ser avaliado quantitativamente por meio de análise longitudinal com SDR (2) qualquer uma das três ferramentas pode ser utilizada para quantificar o reparo, considerando que existe correlação entre o tempo de reparo e o aumento do valor de pixel; (3) a avaliação comparativa entre os métodos subjetivos, usando radiografia convencional, imagem digitalizada e a SDR, mostrou que todos foram capazes de evidenciar o processo de reparo de lesões periapicais desde a primeira radiografia (15 dias), não havendo diferença entre eles e (4) a avaliação quantitativa por meio de SDR conseguiu evidenciar o início do reparo com 15 dias após o início do tratamento endodôntico, embora esse reparo fosse realmente efetivo a partir de 105 dias após o início do tratamento endodôntico.

Indiana University-Purdue University Indianapolis (IUPUI) / Bone growth can be impaired due to disease, such as osteoporosis. Currently, intermittent parathyroid hormone (PTH) treatment is the only approved therapy in the United States for anabolic bone growth in osteoporosis patients. The anabolic effects of PTH treatment are due, at least in part, to modulation of the Wnt/β-catenin pathway. Activation of the Wnt/ β-catenin pathway using a small molecule inhibitor of GSK3β was previously shown to increase markers of bone formation in vitro. Our study utilized a zebrafish model system to study Wnt activated fin regeneration and bone growth. Wnt signaling is the first genetically identified step in fin regeneration, and bony rays are the main structure in zebrafish fins. Thus, zebrafish fin regeneration may be a useful model to study Wnt signaling mediated bone growth. Fin regeneration experiments were conducted using various concentrations of a GSK3β inhibitor compound, LSN 2105786, for different treatment periods and regenerative outgrowth was measured at 4 and 7 days post amputation. Experiments revealed continuous low concentration (4-5 nM) treatment to be most effective at increasing regeneration. Higher concentrations inhibited fin growth, perhaps by excessive stimulation of differentiation programs. In situ hybridization experiments were performed to examine effects of GSK3β inhibitor on Wnt responsive gene expression. Experiments showed temporal and spatial changes on individual gene markers following GSK3β inhibitor treatment. Additionally, confocal microscopy and immunofluorescence labeling data indicated that the Wnt signaling intracellular signal transducer, β-catenin, accumulates throughout GSK3β inhibitor treated tissues. Finally, experiments revealed increased cell proliferation in fin regenerates following LSN 2105786 treatment. Together, these data indicate that bone growth in zebrafish fin regeneration is improved by activating Wnt signaling. Zebrafish Wnt signaling experiments provide a good model to study bone growth and bone repair mechanisms, and may provide an efficient drug discovery platform.

Indiana University-Purdue University Indianapolis (IUPUI) / An excess of bone resorption over bone formation leads to osteoporosis, resulting in a reduction of bone mass and an increase in the risk of bone fracture. Anabolic and anti-resorptive drugs are currently available for treatment, however, none of these drugs are able to both promote osteoblastogenesis and reduce osteoclastogenesis. This thesis focused on the role of eukaryotic translation initiation factor 2 alpha (eIF2alpha), which regulates efficiency of translational initiation. The elevation of phosphorylated eIF2alpha was reported to stimulate osteoblastogenesis, but its effects on osteoclastogenesis have not been well understood. Using synthetic chemical agents such as salubrinal and guanabenz that are known to inhibit the de-phosphorylation of eIF2alpha, the role of phosphorylation of eIF2alpha in osteoclastogenesis was investigated in this thesis. The questions addressed herein were: Does the elevation of phosphorylated eIF2alpha (p-eIF2alpha) by salubrinal and guanabenz alter osteoclastogenesis? If so, what regulatory mechanism mediates the process? It was hypothesized that p-eIF2alpha could attenuate the development of osteoclast by regulating the transcription factor(s) amd microRNA(s) involved in osteoclastogenesis. To test this hypothesis, we conducted in vitro and in silico analysis of the responses of RAW 264.7 pre-osteoclast cells to salubrinal and guanabenz. First, the in vitro results revealed that the elevated level of phosphorylated eIF2alpha inhibited the proliferation, differentiation, and maturation of RAW264.7 cells and downregulated the expression of NFATc1, a master transcription factor of osteoclastogenesis. Silencing eIF2alpha by RNA interference suppressed the downregulation of NFATc1, suggesting the involvement of eIF2alpha in regulation of NFATc1. Second, the in silico results using genome-wide expression data and custom-made Matlab programs predicted a set of stimulatory and inhibitory regulator genes as well as microRNAs, which were potentially involved in the regulation of NFATc1. RNA interference experiments indicated that the genes such as Zfyve21 and Ddit4 were primary candidates as an inhibitor of NFATc1. In summary, the results showed that the elevation of p-eIF2alpha by salubrinal and guanabenz leads to attenuation of osteoclastogenesis through the downregulation of NFATc1. The regulatory mechanism is mediated by eIF2alpha signaling, but other signaling pathways are likely to be involved. Together with the previous data showing the stimulatory role of p-eIF2alpha in osteoblastogenesis, the results herein suggest that eIF2alpha-mediated signaling could provide a novel therapeutic target for treatment of osteoporosis by promoting bone formation and reducing bone resorption.