Made available in DSpace on 2017-07-24T19:22:04Z (GMT). No. of bitstreams: 1 Micheleelias.pdf: 598764 bytes, checksum: 189fdf6d0d1c8583d61adb9e336c15cf (MD5) Previous issue date: 2008-02-19 / Similar to natural teeth, implants are liable to destruction of support structures. This process of destruction is often the result of opportunistic infections by bacteria that colonizes restorations on the implants and adjacent theeth. A. actinomycetemcomitans is often isolated from periodontal and periimplantar sites but its hole in disease is already to be determined. The aim of this study was to compare the presence of A. actinomycetemcomitans in samples of crevicular fluid and saliva from 14 patients partially and 8 totally edentulous, aging from 42 to 78 years old, rehabilitated with osseointegrated titanium implants, Samples of crevicular fluid were obtained with paper points introduced in periimplantar pockets after removal of supragingival plaque with sterile gauze and left inside for 1 minute. The poits were transferred to tubes with triptic soy solution, homogeneized and plated, in duplicate, on TSBV agar and the plates incubated for 5 days in 37C. After this period, the number of cfu/mL of the microorganism. The presence of A. actinomycetemcomitans was also verified in samples of saliva. The samples were collected with sterile swabs and processed the same way. The results demonstrated that A. actinomycetemcomitans is isolated from gingival fluid and saliva in a small number of patients and in small quantity, both in partially edentulous and totally ones and the differences were not significant between groups (Test Mann-Whitney-gingival fluid: 0.7280; saliva: 0.3503). the results of this study suggest that the colonization of periimplantar sulcus and oral cavity of and the number of mL of A. actinomycetemcomitans in theses sites are not modulated by the presence or not natural theeth. / Semelhante aos dentes, os implantes são susceptíveis à destruição de estruturas de suportes. Este processo de destruição é com freqüência o resultado de infecções por bactérias oportunistas que colonizaram a restauração sobre o implante e a dentição adjacente. Aggregatibacter actinomycetemcomitans é frequentemente isolado de sítios periodontais e periimplantares, entretanto os fatores que modulam a colonização da mucosa bucal e o papel deste microrganismo na doença são controversos. Assim, o objetivo deste estudo foi comparar a presença de A. actinomycetemcomitans em amostras de fluido gengival periimplantar e saliva de 14 pacientes parcialmente edêntulos e 8 totalmente edêntulos, com idade entre 42 e 78 anos, reabilitados com implantes osseointegrados de titânio. Amostras do fluido gengival foram obtidas com cones de papel que foram introduzidos nos sulcos periimplantares após remoção do biofilme supragengival com gaze esterilizada e deixados por um minuto. Os cones foram transferidos para solução de caldo tripticaseína de soja, homegeneizados e as amostras foram semeados em duplicata em agar TSBV - ágar de soja tripticaseína acrescido de bacitracina e vancomicina, e incubadas a 37C por cinco dias em anaerobiose (Anaerobac). Após incubação, determinou-se o número de mL do microrganismo pesquisado. A. actinomycetemcomitans também foi pesquisado na saliva, que foi coletada com auxilio de suabes esterilizados e processada microbiologicamente do mesmo modo que as amostras obtidas da região periimplantar. Os resultados mostraram que não houve diferença estatística, demonstrada através do teste Mann-Whitney, no número de mL de A. actinomycetemcomitans, tanto na saliva quanto no fluido do sulco periimplantar, quando comparou-se os pacientes totalmente desdentados e parcialmente desdentados (sulco: 0.7280; saliva 0.3503). Os resultados obtidos sugerem que a colonização do sulco periimplantar e cavidade bucal e o número de A. actinomycetemcomitans nesses sítios não são modulados pela presença ou não de dentes naturais.

Osseointegration, or the direct integration of an implant into bone tissue, is necessary for implant success. Titanium is commonly used clinically in dental and orthopaedic implants because of its passivating oxide layer, which facilitates osseointegration, and its mechanical properties such as a modulus of elasticity similar to bone. Diverse studies have shown that surface microtopography, chemistry, and surface energy affect osteoblast behavior. The problem with these studies is that they access the average behavior of a culture in response to a substrate and not the behavior of individual cells. The objective of this study was to develop a method for correlating the behavior of individual cells with the characteristics of the surface underneath them. More specifically, this work developed a method to correlate integrin beta-1 (β1) expression with the surface characteristics under individual cells. Integrins are cell surface receptors that bind to specific proteins in the extracellular matrix adsorbed on the implant surface. Previous work has shown that expression of certain integrins is increased when osteoblasts on titanium substrates develop a more differentiated phenotype, and that integrin β1 is necessary for osteoblast response to roughness on titanium substrates. This study used molecular beacons specific to integrin β1 to quantify integrin β1 expression of MG63 cells cultured on titanium disks. A template was designed to coordinate the location of cells using fluorescence microscopy and scanning electron microscopy (SEM) in reference to laser etchings on the disks. After live cell imaging, cells were fixed, dried, and critical point dried for focused ion beam (FIB) milling. Transmission electron microscopy (TEM) sections of cells identified with high and low integrin β1 molecular beacon intensity were milled, and cells with high and low integrin β1 molecular beacon intensity were also serial sectioned. While our TEM results were inconclusive, SEM images from serial sectioning showed contact points between the cell body and the substrate, consistent with previous results. Cells cultured on pretreatment (PT) or sandblasted acid etched (SLA) titanium surfaces were also serial sectioned, showing that cells on SLA surfaces have more regions of contact between the cells and the substrate than cells on PT surfaces. This work is significant as it is the first study to develop a method to correlate individual cell behavior with the substrate surface characteristics under the individual cells. Previous studies have reported the average cell behavior in response to their substrates, while this work allows for the study of substrate surface characteristics that positively affect integrin β1 expression in individual cells. Further optimization of the fluorescence imaging process and FIB milling process could be done, and the method developed in this study could be used in future studies to investigate surface characteristics after using other fluorescent analyses of cell behavior, such as immunocytochemistry.

Biomaterial surface properties such as microtopography and energy can change cellular responses at the cell-implant interface. Phospholipase D (PLD) is required for differentiation of osteoblast-like MG63 cells on machined and grit-blasted titanium surfaces. Here, we determined if PLD is also required on microstructured/high-energy substrates and the mechanism involved. shRNAs for human PLD1 and PLD2 were used to silence MG63 cells. Wild-type and PLD1 or PLD1/2 silenced cells were cultured on smooth-pretreatment surfaces (PT); grit-blasted, acid-etched surfaces (SLA); and SLA surfaces modified to have higher surface energy (modSLA). PLD was inhibited with ethanol or activated with 24,25-dihydroxyvitamin-D₃ [24R,25(OH)₂D₃]. As surface roughness/energy increased, PLD mRNA and activity increased, cell number decreased, osteocalcin and osteoprotegerin increased, and protein kinase C (PKC) and alkaline phosphatase specific activities increased. Ethanol inhibited PLD and reduced surface effects on these parameters. There was no effect on these parameters after knockdown of PLD1, but PLD1/2 double knockdown had effects comparable to PLD inhibition. 24R,25(OH)₂D₃increased PLD activity and production of osteocalcin and osteoprotegerin, but decreased cell number on the rough/high-energy surfaces. These results confirm that surface roughness/energy-induced PLD activity is required for osteoblast differentiation and that PLD2 is the main isoform involved in this pathway. Here we showed that PLD is activated by 24R,25(OH)₂D₃ in a surface-dependent manner and inhibition of PLD reduced the effects of surface microstructure/energy on PKC, suggesting that PLD mediates the stimulatory effect of microstructured/high-energy surfaces via PKC-dependent signaling.

Thesis (Ph. D.)--University of Sydney, 2006. / Title from title screen (viewed 19 Dec. 2006). Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy to the School of Communication Sciences and Disorders, Faculty of Health Sciences. Includes bibliographical references. Also issued in print.

Thesis (Ph. D.)--University of Alabama at Birmingham, 1999. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M.S.)--University of Michigan, 1999. / Includes bibliographical references.

Thesis (M.S.)--University of Michigan, 2003. / Includes bibliographical references.

Thesis (doctoral)--Lund University, 1995. / Added t.p. with thesis statement inserted.

Thesis (Ph. D.)--University of Alabama at Birmingham, 1999. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M.S.)--University of Michigan, 1992.