O objetivo foi evidenciar a presença do espaço intercelular dilatado do epitélio esofágico após a infusão de ácido clorídrico (HCl) à 0,1N comparando com a infusão de soro fisiológico (SF) em pacientes sem sintomas típicos da DRGE com mucosa esofágica normal e compará-los com os de sintomas típicos e esofagite erosiva. Foram entrevistados e realizaram o exame de endoscopia digestiva alta 60 pacientes destes, 29 foram incluídos no estudo sendo 18 com esôfago normal (9 foram infundidos SF e 9 HCl) e 11 com esofagite erosiva (6 foram infundidos SF e 5 HCl) e foram realizados 4 biópsias da mucosa esofágica (2 antes e 2 depois das infusões). Não foi encontrado diferença estatisticamente significante no espaço intercelular da mucosa esofágica dos pacientes com e sem esofagite erosiva com ácido clorídrico ou soro fisiológico não sendo um marcador da DRGE / The purpose was to prove the presence of extended intercellular space of the esophagic epithelium after chloridric acid infusion (HCI) to 0,1N comparing to the physiologic serum infusion (PS) in patients without typical symptoms of DGER with normal esophagic mucous membrane and compare them to ones with typical symptoms and erosive esophagitis. 60 patients were interviewed and took the high digestive endoscopy; 29 were included in the research, among them 18 with normal esophagus (9 were infused PS, and 9 HCI) and 11 with erosive esophagitis (6 were infused PS and 5 HCI); 4 biopsies of esophagic mucous membrane were made (2 before and 2 after infusions). It was not found any statistically meaningful difference in intercellular space of esophagic mucous membrane in patients with or without erosive esophagitis with chloridric acid or physiologic serum, and thus it is not a DGER

<p>Ugljenične nanocevi (UNC) imaju sve veću primenu u elektronici, kompjuterskoj i optičkoj industriji, kao i u biomedicini. Dok proizvodnja jednoslojnih UNC nanocevi beleži sve veći rast poslednjih godina, rizik koji nosi izlaganje ovom nanomaterijalu ostaje nerazja&scaron;njen. Oskudni i često kontradiktorni podaci o toksičnim efektima jednoslojnih UNC ukazuju na potrebu za daljim ispitivanjima. U na&scaron;im istraživanjima ispitivane su promene u ćelijskom odgovoru kao i morfolo&scaron;ke promene nakon delovanja jednoslojnih ugljeničnih nanocevi na ćelijskoj liniji humanih fetalnih fibroblasta pluća MRC-5 i ćelijskoj liniji humanog adenokarcinoma pluća A549. U ovoj studiji kori&scaron;ćene su jednoslojne ugljenične nanocevi koje su sadržale rezidualne nečistoće poput gvožđa. Citotoksičnost jednoslojnih UNC (engl. single-walled carbon nanotubes &ndash; SWCNT) je ispitivana kolorimetrijskim MTT testom. Tokom 24 i 48h niske koncentracije jednoslojnih ugljeničnih nanocevi (&lt;250 &mu;g/mL) pokazale su nisku toksičnost na proliferaciju i vijabilnost u obe ispitivane ćelijske linije. Ipak, pri visokim koncentracijama UNC (250-750 &mu;g/mL) antiproliferativni efekat je bio blizu IC50 vrednostima. Na osnovu rezultata dobijenih MTT testom može se zaključiti da su maligne A549 ćelije osetljivije na delovanje jednoslojnih UNC u odnosu na normalne MRC-5 ćelije. Kombinacija ugljeničnih nanocevi sa prirodnim polifenolima (resveratrolom i proantocijanidolima) nije značajno uticala na citotoksičnost u MRC-5 ćelijama, za razliku od A549 ćelija gde je tretman kombinacijama umanjio toksičnost ugljeničnih nanocevi. Transmisionom elektronskom mikroskopijom ispitivan je efekat jednoslojnih ugljeničnih nanocevi na ćelijsku morfologiju i preživljavanje. Intracelularni agregati ugljeničnih nanocevi primećeni su u obe ćelijske linije, čime je potvrđeno da ugljenične nanocevi ulaze u ćelije. Imajući u vidu da nanomaterijali poput ugljeničnih nanocevi indukuju oksidativni stres i njime posredovanu apoptozu, na protočnom citometru je određivano prisustvo ćelija u apoptozi i nekrozi. Tretman ćelija sa jednoslojnim ugljeničnim nanocevima nije doveo do značajnog porasta broja apoptotskih ili nekrotičnih ćelija, &scaron;to ide u prilog niskoj toksičnosti ovog nanomaterijala, odnosno ukazuje na alternativne mehanizme toksičnosti. Međutim kombinacija jednoslojnih ugljeničnih nanocevi sa antioksidantima, resveratrolom i proantocijanidolima indukuje veći procenat apoptoze i nekroze u odnosu na tretman samo sa nanocevima. Promene u ekspresiji gena praćene su lančanom reakcijom polimeraza (PCR). Komparativna analiza rezultata genske ekspresije MRC-5 i A549<br />ćelija nakon tretmana sa jednoslojnim ugljeničnim nanocevima pojedinačno i u kombinaciji sa antioksidantima ukazala je na kompleksnost i raznolikost biolo&scaron;kog odgovora ispitivanih ćelija. U na&scaron;em istraživanju ispitivana je i promena aktivnosti enzima antioksidativne za&scaron;tite i količine glutationa u ćeliji. Primena jednoslojnih ugljeničnih nanocevi u MRC-5 ćelijama dovodi po smanjenja specifične aktivnosti enzima SOD i GR, povećava specifičnu aktivnost GPx i ne utiče na promenu specifične aktivnosti GST i količine glutationa u ćeliji Primena jednoslojnih ugljeničnih nanocevi u A549 ćelijama dovodi po smanjenja specifične aktivnosti enzima SOD, ne utiče na promenu specifične aktivnost enzima GR, GST i GPx, i dovodi do povećanja količine glutationa u ćeliji. Ćelijska vijabilnost, morfolo&scaron;ke promene, redoks homeostaza i ekspresija ispitivanih gena bile su promenjene nakon tretmana sa jednoslojnim ugljeničnim nanocevima. Iako su dobijeni rezultati značajni za procenu toksičnosti ugljeničnih nanocevi, neophodna su dalja istraživanja koja treba da doprinesu boljem razumevanju toksičnih efekata ugljeničnih nanocevi.</p> / <p>Carbon nanotubes are being actively introduced in electronics, computer science, and optics as well as for various biomedical applications. While production of single-walled carbon nanotubes (SWCNT) has escalated in recent years, the knowledge on risk associated with exposure remains unclear. Contradictory data on the toxic effects of single-walled carbon nanotubes highlights the urgent need for further studies. In this study we investigated the alterations in cellular response along with morphological changes induced by single-walled carbon nanotubes in human lung fibroblast cell line MRC-5 and adenocarcinoma human alveolar basal epithelial cells A549. In this study we used SWCNT containing large amounts of residual metallic impurities such is iron, and the iron concentration increased in dose dependent manner in cells exposed to SWCNT. Cytotoxicity was evaluated by MTT assay and SWCNT showed little cytotoxic effect on the proliferation and viability of two cell lines tested at the concentrations used (&lt;250 &mu;g/mL) within 24 and 48h. However exposing both cell lines to high concentrations (250-750 &mu;g/mL) resulted in near IC50 values. Based on MTT test SWCNT were more cytotoxic to A549 cell line. Cytotoxicity of SWCNT in combination with natural polyphenols (resveratrol and proanthocyanidins) did not noticeably affect the cytotoxicity of SWCNT to MRC-5 cells. However introduction of polyphenols did reduce the cytotoxicity of SWCNT to A549 cells. Transmission electron microscopy was used to complement cytotoxicity assays and to investigate the pathological effect of internalized SWCNT on cell morphology and survival. Intracellular bundles of CNTs, possibly aggregated/agglomerated were observed in both cell lines, confirming internalization after 24h exposure. Since nanoparticles like carbon nanotubes are toxic mainly because they cause oxidative stress, often associated with an increased apoptosis we checked for apoptotic and necrotic cells using flow cytometry. Incubation with SWCNT did not result in pronounced apoptosis or necrosis supporting its low toxicity and possibly alternative mechanism of cell damage. However incubation with SWCNT in combination with resveratrol and proanthocyanidins induced higher levels of both apoptosis and necrosis than SWCNT alone. Changes in gene expression following exposure to SWCNT were evaluated by polymerase chain reaction PCR array which indicated complex and diverse change in expression of genes involved in apoptosis, cell proliferation and oxidative stress. Finally we investigated the modulation of the antioxidant enzyme system and the changes in the cytosolic levels of GSH. SWCNT reduced the specific activity of SOD and GR enzymes, increased GPx activity. No changes in intracellular levels of GSH were observed in MRC-5 cell line. Same treatment in A549 cell reduced the specific activity of SOD, had no effect on GR, GST and GPx activity, but increased intracellular levels of GSH. Cell viability, morphologic changes, redox homeostasis and gene expression were affected by the presence of SWCNT. Although our findings are useful in predicting human response against SWCNT exposure, further study is needed for better understanding of the effects of SWCNT.</p>

Orientador: Hélio Amante Miot / Resumo: O melasma é hipermelanose crônica e adquirida decorrente de um complexo processo que envolve hipertrofia melanocítica e disfunção melanogênica. Acomete preferencialmente o sexo feminino e as lesões ocorrem nas áreas fotoexpostas, especialmente a face. Sua patogênese não é bem compreendida e os estudos clássicos avaliam apenas pele acometida e perilesional, mas pouco se sabe do comportamento da pele fotoprotegida, submetida aos mesmos fatores sistêmicos e genéticos. Neste estudo, objetivamos avaliar características histológicas, vias epidérmicas que influem na melanogênese (Wnt e ASIP) e características ultraestruturais da pele com melasma em comparação com a pele sã adjacente e retroauricular. Para a execução deste estudo transversal com controle intra-sujeito, foram coletadas três biópsias cutâneas (punch 3 mm) de onze mulheres com melasma facial. As áreas de coleta foram a pele com melasma, pele sã adjacente (distando no máximo 2 cm do limite da lesão) e pele retroauricular ipsilateral. Os fragmentos provenientes de dez participantes foram corados por hematoxilina-eosina, ácido periódico de Schiff, Fontana-Masson, picrosirius red, azul de toluidina e Verhöff; imunomarcados para CD34 e submetidos à imunofluorescência direta (IFD) de dupla marcação para proteínas Wnt1, WIF-1 e ASIP. Já os três fragmentos de uma das participantes foram processados para Microscopia Eletrônica de Transmissão (MET). Os dados obtidos foram comparados entre as topografias por modelo linear generali... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Submitted by Ana Cláudia Cavalcante Espósito null (anaclaudiaesposito@gmail.com) on 2017-07-27T20:34:29Z No. of bitstreams: 1 Ana Cláudia mestrado para depositar.pdf: 2933638 bytes, checksum: 3c192abd021482675fa186f4c4bba9e4 (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-07-31T19:29:06Z (GMT) No. of bitstreams: 1 lemos_acce_me_bot.pdf: 2933638 bytes, checksum: 3c192abd021482675fa186f4c4bba9e4 (MD5) / Made available in DSpace on 2017-07-31T19:29:06Z (GMT). No. of bitstreams: 1 lemos_acce_me_bot.pdf: 2933638 bytes, checksum: 3c192abd021482675fa186f4c4bba9e4 (MD5) Previous issue date: 2017-07-20 / Fundo de Apoio à Dermatologia de São Paulo (FUNADERSP) / O melasma é hipermelanose crônica e adquirida decorrente de um complexo processo que envolve hipertrofia melanocítica e disfunção melanogênica. Acomete preferencialmente o sexo feminino e as lesões ocorrem nas áreas fotoexpostas, especialmente a face. Sua patogênese não é bem compreendida e os estudos clássicos avaliam apenas pele acometida e perilesional, mas pouco se sabe do comportamento da pele fotoprotegida, submetida aos mesmos fatores sistêmicos e genéticos. Neste estudo, objetivamos avaliar características histológicas, vias epidérmicas que influem na melanogênese (Wnt e ASIP) e características ultraestruturais da pele com melasma em comparação com a pele sã adjacente e retroauricular. Para a execução deste estudo transversal com controle intra-sujeito, foram coletadas três biópsias cutâneas (punch 3 mm) de onze mulheres com melasma facial. As áreas de coleta foram a pele com melasma, pele sã adjacente (distando no máximo 2 cm do limite da lesão) e pele retroauricular ipsilateral. Os fragmentos provenientes de dez participantes foram corados por hematoxilina-eosina, ácido periódico de Schiff, Fontana-Masson, picrosirius red, azul de toluidina e Verhöff; imunomarcados para CD34 e submetidos à imunofluorescência direta (IFD) de dupla marcação para proteínas Wnt1, WIF-1 e ASIP. Já os três fragmentos de uma das participantes foram processados para Microscopia Eletrônica de Transmissão (MET). Os dados obtidos foram comparados entre as topografias por modelo linear generalizado de efeitos mistos. As participantes eram fototipo III ou IV de Fitzpatrick, com idade média (desvio-padrão) de 42,9 (8,9) anos e apresentavam lesões há 16,7 (7,9) anos. Houve adelgaçamento da camada córnea na pele com melasma e na pele adjacente. Na pele com melasma houve maior compactação da córnea, maior pigmentação melânica epidérmica, maior heterogeneidade do colágeno, elastose solar, maior número de mastócitos, falhas da integridade da zona da membrana basal, melanócitos em pêndulo, bem como maior celularidade e vasos na derme superficial. IFD evidenciou maior intensidade de marcação de Wnt1 na pele com melasma em relação à pele adjacente e maior intensidade na pele retroauricular em relação à pele sã adjacente. Não houve diferença estatística significativa na intensidade de marcação de WIF-1 e ASIP entre as topografias. À MET, houve maior dano estrutural na lâmina lúcida no melasma, bem como maior número de melanossomas maduros e organelas citoplasmáticas nos melanócitos e queratinócitos basais. Tais resultados evidenciam que a pele com melasma apresenta, além da hipertrofia melanocítica, alterações na barreira epidérmica, na derme superior, zona de membrana basal e maior ativação da via Wnt, que diferem da pele fotoexposta adjacente e da retroauricular, configurando um fenótipo individualizado e não somente uma extensão do fotoenvelhecimento ou do envelhecimento intrínseco. / Melasma is a chronic and acquired hypermelanosis resulting from a complex process which involves melanocytic hypertrophy and melanogenic dysfunction. Melasma mainly affects females and lesions occur in the photoexposed areas, especially the face. Its pathogenesis is not well understood, and classical studies evaluate only the affected and perilesional skin, but little is known about the behavior of the non-sun-exposed skin, subjected to the same systemic and genetic factors. In this study, we aimed to evaluate histological features, epidermal pathways that influence melanogenesis (Wnt and ASIP) and ultrastructural characteristics of the skin with melasma in comparison to healthy adjacent and retroauricular skin. For the execution of this cross-sectional study with intrasubject control, three skin biopsies (punch 3 mm) were collected from eleven women with facial melasma. The areas of collection were the skin with melasma, adjacent healthy skin and retroauricular skin. Fragments from ten participants were stained with hematoxylin-eosin, periodic acid from Schiff, Fontana-Masson, picrosirius red, toluidine blue and Verhöff; immunomarked for CD34 and subjected to double-labeled direct immunofluorescence (DIF) for Wnt1, WIF-1 and ASIP proteins. The three fragments of one of the participants were processed for Transmission Electron Microscopy (TEM). The data obtained were compared between topographies by generalized linear model of mixed effects. Participants were Fitzpatrick's phototype III or IV; the mean age (standard deviation) was 42.9 (8.9) years and they had lesions for 16.7 (7.9) years. There was thinning of the corneal layer on the skin with melasma and adjacent skin. In the skin with melasma, there was more corneal compaction, greater epidermal melanic pigmentation, greater collagen heterogeneity, solar elastosis, more mast cells, defects of the basement membrane area, pendulum melanocytes, as well as greater cellularity and vessels in the superficial dermis. DIF showed a greater intensity of Wnt1 marking in the skin with melasma in relation to the adjacent skin, and greater intensity in the retroauricular skin in relation to the adjacent healthy skin. There was no significant statistical difference in the intensity of WIF-1 and ASIP marking between topographies. At TEM, there was more structural damage to the lamina lucida in melasma, as well as more mature melanosomes and cytoplasmic organelles in melanocytes and basal keratinocytes. These results show that melasma skin presents, in addition to melanocytic hypertrophy, alterations in the epidermal barrier, upper dermis, basement membrane zone and greater activation of the Wnt pathway, which differ from adjacent and retroauricular photoexposed skin, forming an individualized phenotype and not only an extension of photoaging or intrinsic aging.

O objetivo foi evidenciar a presença do espaço intercelular dilatado do epitélio esofágico após a infusão de ácido clorídrico (HCl) à 0,1N comparando com a infusão de soro fisiológico (SF) em pacientes sem sintomas típicos da DRGE com mucosa esofágica normal e compará-los com os de sintomas típicos e esofagite erosiva. Foram entrevistados e realizaram o exame de endoscopia digestiva alta 60 pacientes destes, 29 foram incluídos no estudo sendo 18 com esôfago normal (9 foram infundidos SF e 9 HCl) e 11 com esofagite erosiva (6 foram infundidos SF e 5 HCl) e foram realizados 4 biópsias da mucosa esofágica (2 antes e 2 depois das infusões). Não foi encontrado diferença estatisticamente significante no espaço intercelular da mucosa esofágica dos pacientes com e sem esofagite erosiva com ácido clorídrico ou soro fisiológico não sendo um marcador da DRGE / The purpose was to prove the presence of extended intercellular space of the esophagic epithelium after chloridric acid infusion (HCI) to 0,1N comparing to the physiologic serum infusion (PS) in patients without typical symptoms of DGER with normal esophagic mucous membrane and compare them to ones with typical symptoms and erosive esophagitis. 60 patients were interviewed and took the high digestive endoscopy; 29 were included in the research, among them 18 with normal esophagus (9 were infused PS, and 9 HCI) and 11 with erosive esophagitis (6 were infused PS and 5 HCI); 4 biopsies of esophagic mucous membrane were made (2 before and 2 after infusions). It was not found any statistically meaningful difference in intercellular space of esophagic mucous membrane in patients with or without erosive esophagitis with chloridric acid or physiologic serum, and thus it is not a DGER

El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / PURPOSE: To assess the influence of simulated oral erosive/abrasive challenges on the bond strength of an etch-and-rinse two-step bonding system to enamel using an in situ/ex vivo protocol. METHODS: Bovine enamel blocks were prepared and randomly assigned to four groups: CONT - control (no challenge), ABR - 3x/day-1 minute toothbrushing; ERO - 3x/day - 5 minutes extraoral immersion into regular Coca Cola; and ERO+ABR - erosive protocol followed by a 1-minute toothbrushing. Eight blocks were placed into an acrylic palatal appliance for each volunteer (n = 13), who wore the appliance for 5 days. Two blocks were subjected to each of the four challenges. Subsequently, all the blocks were washed with tap water and Adper Single Bond 2/Filtek Z350 were placed. After 24 hours, 1 mm2 beams were obtained from each block to be tested with the microtensile bond strength test (50 N load at 0.5 mm/minute). The data were statistically analyzed by one-way RM-ANOVA and Tukey's tests (alpha = 0.05). RESULTS: No difference was detected among the ABR, ERO, and CONT groups (P > 0.05). ERO+ABR group yielded lower bond strengths than either the ABR and ERO groups (P < 0.0113). / Revisión por pares

<p>Već izvesno vreme u stručnoj literaturi vodi se rasprava o biokompatibilnosti materijala koji se koriste u periapikalnoj hirurgiji. Pored biokompatibilnosti, od materijala za retrogradnu opturaciju kanala korena zuba se očekuje da spreči prodor bakterija iz kanalnog sistema u okolna tkiva. Kvalitetno rubno zaptivanje, odnosno adekvatna marginalna adaptacija su među najznačajnijim faktorima za dugoročan uspeh tretmana. Cilj ovog istraživanja je bio ispitati biokompatibilnost i utvrditi da li postoji razlika u biokompatibilnosti između mineral-trioksid agregata, trikalcijum-silikatnog cementa i amalgama na osnovu tri standadna testa citotoksičnosti, kao i utvrditi marginalnu adaptaciju ispitivanih materijala za retrogradno zatvaranje kanala korena zuba. Materijali i metode: Eksperimentalni deo istraživanja je podeljen na dva dela. U prvom delu istraživanja je vr&scaron;eno ispitivanje biokompatibilnosti materijala, dok je u drugom delu vr&scaron;eno ispitivanje marginalne adaptacije na osnovu mikrofotografija dobijenih skening elektronskim mikroskopom. Ispitivanje biokompatibilnosti je izvr&scaron;eno na dve ćelijske linije, liniji mi&scaron;jih fibroblasta (L929) i liniji humanih fibroblasta (MRC-5). U eksperimentima su kori&scaron;ćene samo žive (vijabilne) ćelije. Broj ćelija i njihova vijabilnost je određena testom odbacivanja boje sa 0,1% tripan plavim. Vijabilnost ćelija kori&scaron;ćenih u eksperimentu je bila veća od 90%. Biokompatibilnost sve tri vrste materijala je ispitana na osnovu standardnih testova biokompatibilnosti: DET test, MTT test, Agar difuzioni test. Ispitivanje marginalne adaptacije je sprovedeno na 90 ekstrahovanih jednokorenih zuba interkaninog sektora gornje vilice sa intaktnom pulpom, zavr&scaron;enim rastom korena, bez frakture i resorpcije korena zuba. Izvr&scaron;ena je endodontska obrada svih zuba i nakon toga su zubi ostavljeni u vlažnoj sredini na 48h, da bi se sprečile frakture prilikom sečenja. Nakon mehaničko medikamentozne obrade i opturacije kanala korena zuba je vr&scaron;ena resekcija vrha korena zuba 3mm, a nakon toga su svi zubi preparisani do dubine od 3 mm unutar kanala, ultrazvučnim nastavcima (EMS, miniMaster Piezon scaler). Zubi su naizmenično podeljeni u 3 grupe (30 zuba po grupi). Prvu grupu čine zubi kojima je apikalni kavitet biti ispunjen amalgamom, u drugoj grupi, apikalni kavitet je ispunjen MTA, a u trećoj trikalcijum-silikatnim cementom. Nakon retrogradne opturacije, zubi su ostavljeni u vlažnoj sredini 48h, do potpunog vezivanja ispitivanih materijala. Nakon vezivanja ispitivanih materijala, zubi su sečeni longitudinalno, finom dijamantskom &scaron;ajbnom. Marginalna adaptacija ispitivanih materijala je procenjena skening elektronskim mikroskopom (SEM). Pripremljeni uzorci su posmatrani pod uvećanjima 30x, 40x, 80x i 100x. Uvećanje 30x je rađeno radi prikaza celokoupnog retrogradnog punjenja na jednom snimku, tj. spoja materijal-zub. Nakon toga je napravljeno vi&scaron;e uzastopnih snimaka sauvećanjem 80x do pune dužine materijala. Na taj način je utvrđeno postojanje i izvr&scaron;eno merenje marginalne pukotine u mikrometrima. Merenja ukupne dužine marginalne pukotine u mikrometrima u 5 tačaka sa obe strane preparata su vr&scaron;ena u softverskoj aplikaciji Image J software (National Institutes of Health, Bethesda, USA). Tačke su izabrane tako da tačka 1a i tačka 5a predstavljaju gornju i donju ivicu preparata. Tačka 3a predstavlja sredinu rastojanja između tačke 1a i tačke 5a. Tačka 2a i tačka 4a predstavljaju sredinu razmaka između tačke 1a i 3a, odnosno 3a i a5. Tačke 1b-5b su naspramne tačke. Rezultati biokompatibilnosti ukazuju na visok stepen ćelijske kompatibilnosti svih ispitivanih materijala. Međutim, DET testomnije utvrđena statistički značajna razlika u citotoksičnosti između istovetnih ispitivanih materijala u obe ćelijske linije, niti između sva tri ispitivana materijala u obe ćelijske kulture.Poređenjem rezultata MTT testa nakon 24h i 48h, kao i nakon 48h i 72h uočava se da su dobijene srednje vrednosti indeksa citotoksičnosti kod sva tri ispitivana materijala i na obe ćelijske linije manje nakon 48h, odnosno nakon 72h, &scaron;to ukazuje na oporavak ćelijskog metabolizma. Poređenjem rezultata nakon 24h i nakon 72h, uočava se znatno veći pad vrednosti indeksa citotoksičnosti nakon 72h kod sva tri ispitivana materijala i na obe ćelijske linije. U kulturi ćelija MRC5, kod sva tri ispitivana materijala postoji statistički značajna razlika između indeksa citotoksičnosti izmerenog nakon 24h i nakon 72h, kao i u kulturi ćelija L929. Agar difuzionim testom nije uočena dekolorizacija, niti liza ćelija ispod ispitivanih materijala. Ćelijski odgovor je 0/0 &scaron;to ukazuje da ovim testom nije utvrđeno postojanje citotoksičnog efekta ispitivanih materijala na ćelijske linije L929 i MRC-5. Ispitivanje marginalne adaptacije materijala je vr&scaron;eno na osnovu mikrofotografija dobijenih skening elektronskim mikroskopom.. Najpre su testirane razlike na prvoj tački merenja. Rezultati ovog testa pokazuju da postoje značajne razlike između materijala i da amalgam ima značajno vi&scaron;e vrednosti izmerenih pukotina u odnosu na preostala dva materijala, dok se vrednosti za MTA i biodentin međusobno značajno ne razlikuju. Kao jo&scaron; jedna referentna tačka uzeta je tačka merenja 5. I u odnosu na vrednosti u ovoj tački merenja zabeležene su značajne razlike između materijala. Post hoc Mann-Whitney test pokazuje da se amalgam značajno razlikuje od preostala dva materijala, dok nema značajnih razlika između MTA i biodentina. Na osnovu medijane može se videti da amalgam ima niže vrednosi u ovoj tački merenja u odnosu na preostala dva materijala. U tačkama 2-4, kao i u tačkama 1-5 (ukupno), ne postoji statistički značajna razlika u marginalnoj adaptaciji ispitivanih materijala. Rezultati biokompatibilnosti ukazuju na visok stepen ćelijske kompatibilnosti svih ispitivanih materijala. Rezultati sva tri testa pokazuju da ne postoji statistički značajna razlika u citotoksičnosti između ispitivanih materijala. MTT test pokazuje da u obe ćelijske kulture, kod istovetnih ispitivanih materijala postoji statistički značajna razlika između indeksa citotoksičnosti izmerenog nakon 24h i nakon 72h. Rezultati ispitivanja marginalne adaptacije pokazuju da u tački 1 najgore zaptiva amalgam, dok izmedju MTA i biodentina nema razlike. U tački 5 najbolje zaptiva amalgam.</p> / <p>Biocompatibility of materials, deployed in periapical surgery, has been a subject of debate in referential literature for some time now. Apart from biocompabillity, root end filling materials are expected to prevent bacteria from entering the surrounding tissue from canal system. The most important factors for successful long term treatment include marginal seal, i.e. adequate marginal adaptation. The aim of this research was to examine biocompatibility and establish the potential difference in biocompatibility between mineral trioxide aggregate, tricalcium silicate cement and dental amalgam according to three cytotoxicity tests, but also to corroborate marginal adaptation of the materials in question for retrograde seal of a root canal. Materials and methods: The experimental part of the research is divided in two parts. In the first part of the research biocompatibility of the materials was examined, while the examination of the marginal adaptation based on the micro images from scanning electron microscope was conducted in the second part. The examination of biocompatibility was executed on two cell lines, mouse fibroblast cell line (L929) and human cell line (MRC-5).Biocompatibility of all three types of material was examined based on three standard biocompatibility tests: DET test, MTT test, Agar diffusion test.The examination of marginal adaptation was carried out on 90 single-rooted tooth extractied human teeth of the intercanine sector of maxilla with intact pulp, mature apices, without root fractures or resorption. All teeth were endodontically treated. After the extirpation, irrigation and opturation, the resection of 3mm of root apex and retrograde preparation with ultrasonic instruments up to 3 mm depth inside the canal was done. Teeth were divided in three groups alternately. The First group include teeth which apical cavitation was filled with amalgam, in the Second group apical cavitation was filled with MTA, and in the Third group with tricalcium silicate cement. After the complete setting of the materials, teeth were cut in longitudinal manner, with fine, diamond tool. Marginal adaptation of the materials was assessed through scanning electron microscope (SEM). Software application Image J software was deployed to measure the total length of marginal fissure in micrometers in 5 pointson both sides of the preparation . The results of biocompatibility indicate high degree of cell compatibility of all tested materials. DET test did not assert any statistically significant difference in cytotoxicity between the same tested materials in both cell lines, nor between all three tested materials in both cell cultures. Comparing the results of MTT test after 24h and 48h, and 48h and 72h, it is noted that middle value of cytotoxicity index with all three tested materials and on both cell lines is lower after 48h, and after 72h, indicating the recovery of cell metabolism. In both cell cultures, with all three tested materials there is statistically significant difference between measured cytotoxicity indices after 24h and after 72h. Agar diffusion test did not show decolorization, nor cell lysis underneath the tested materials, which means that cytotoxic effect was not asserted on cell lines L929 i MRC-5. The examination of marginal adaptation was conducted according to micro images gained by scanning electron microscope. The results in the measure point 1 indicate there are significant differences between materials, and amalgam has significantly higher values of the measured fissures in relation to remaining two materials, whereas values for MTA and biodentine do not differ significantly. In measure point 5 significant differences were noted. Post hoc Mann-Whitney test shows that amalgam has lower values of the tested fissures in this measure point in relation to two other materials, while there were no significant differences between MTA i biodentine. In points 2-4, as well as points 1-5 (in total), there were no statistically significant differences in marginal adaptation of the examined materials. The results of all three tests show that there is no statistically significant difference in cytotoxicity between examined materials. MTT test shows that there is, in both cell cultures, with the same examined materials , statistically significant difference between cytotoxicity indices measured after 24h and after 72h. The results of the examination of marginal adaptation show that in point 1 amalgam has the worst seal, whereas between MTA and biodentine there is no difference. In point 5 amalgam has the best seal.</p>

Der Einsatz nanostrukturierter Magnetmaterialien als Speicherschichten in Festplatten stellt ein vielversprechendes Konzept zur weiteren Erhöhung der erreichbaren Speicherdichten im Vergleich zu den heute eingesetzten granularen Medien dar. Für die Realisierung dieses Konzeptes ist eine detaillierte Kenntnis der Struktureigenschaften und deren Einfluss auf das magnetische Verhalten der einzusetzenden Schichten erforderlich. Für die vorliegende Arbeit wurden drei verschiedene magnetische Materialien ausgewählt und insbesondere mit elektronenmikroskopischen Methoden in struktureller Hinsicht untersucht. Dazu zählen ferromagnetische (FePt)(100-x)Cu(x) -Schichten, ferromagnetische [Co/Pt]n -Multilagen sowie ferrimagnetische Fe(100-x)Tb(x) -Schichten. Der Schwerpunkt der Untersuchungen lag dabei auf der Korrelation zwischen strukturellen und magnetischen Eigenschaften sowie im Einfluss der Nanostrukturierung auf das magnetische Verhalten der Schichten. In dieser Hinsicht wurden Aspekte der durch die Struktur bedingten magnetischen Anisotropie in Form von magnetokristalliner und Grenzflächenanisotropie betrachtet. Zudem wurde das Kopplungsverhalten zwischen einzelnen Strukturelementen in nanostrukturierten Schichten untersucht. Aufbauend auf die Untersuchung der drei genannten Materialien wurden [Co/Pt]n und Fe(100-x)Tb(x) ausgewählt zum Aufbau eines Systems mit zwei magnetischen Komponenten: Fe(80)Tb(20) / [Co/Pt]10. Die Untersuchungen konzentrierten sich dabei auf die Morphologie der Grenzfläche zwischen den beiden Bestandteilen und deren Einfluss auf den Exchange Bias, der in diesem System vorliegt.

Die fortwährende Verkleinerung der Strukturbreiten in der Mikroelektronik erfordert es, herkömmliche SiO2 Dielektrika durch Materialien mit kleinerer Dielektrizitätskonstante zu ersetzen. Dafür sind verschiedene „low-k Materialien“ entwickelt worden. Unter diesen sind die Organosilikatgläser, die aus SiO2 Netzwerken mit eingelagerten Methylgruppen bestehen wegen ihrer ausgezeichneten Eigenschaften besonders interessant als Dielektrika zwischen metallischen Leiterbahnen. In dieser Arbeit sind fünf verschiedene dieser „low-k Materialien“ untersucht worden: ein dichtes und vier poröse Materialien, die alle durch plasmagestützte chemische Gasphasenabscheidung hergestellt wurden. Die strukturellen, chemischen und dielektrischen Eigenschaften der Materialien wurden mit Hilfe der analytischen Durchstrahlungselektronenmikroskopie unter Verwendung eines abbildenden GATAN-Energiespektrometers untersucht. Die Bestimmung der radialen Verteilungsfunktion (RDF) zur Charakterisierung der atomaren Nahordnung ermöglicht uns die Ermittlung mittlerer Bindungslängen und – winkel sowie der mikroskopischen Dichte des Materials. Gegenüber SiO2 wurden in den untersuchten „low-k Materialien“ stark veränderte mittlere Si-O, O-O und Si-Si Bindungslängen gefunden. Dieses wirkt sich natürlich auch auf die mittleren Si-O-Si bzw. O-Si-O Bindungswinkel aus, und wie erwartet war auch die mikroskopische Dichte der „low-k Materialien“ kleiner als die Dichte des SiO2. Elektronen Energieverlustspektroskopie (EELS) und Photoelektronenspektroskopie (XPS) wurden zur Charakterisierung der chemischen Umgebung der Atome in den „low-k Materialien“ herangezogen. Die Energien von Ionisationskanten und die Bindungsenergien der Silizium-2p und Sauerstoff-1s Elektronen waren in den „low-k Materialien“ größer als im SiO2. Die Kohlenstoffatome kamen in den „low-k Materialien“ sowohl sp2 als auch sp3 hybridisiert vor. sp2-Hybridisierung liegt vor, wenn Bindungen wie Si=CH2 und C=C im Netzwerk vorkommen, während sp3 Hybridisierung z.B. dann vorkommt, wenn freie Si-Bindungen durch –CH3 Gruppen abgesättigt werden. Die Anteile an sp2- bzw. sp3-hybridisierten Kohlenstoffatome wurden aus der Feinstruktur der K-Energieverlustkanten des Kohlenstoffs abgeschätzt. Das ergab, daß die meisten Kohlenstoffatome in den „low-k Materialien“ sp2-hybridisiert sind. Die dielektrischen Eigenschaften wurden durch Kramers-Kronig-Transformation einer Energieverlustfunktion ermittelt, die aus dem Niedrigverlust-EELS-Spektrum im Bereich der Plasmonenanregungen gewonnen wurde. Die Bandlücke des SiO2 beträgt ungefähr 9 eV während dichte „low-k Materialien“ aufgrund der Unregelmäßigkeiten in ihrem SiO2-Netzwerk zusätzliche Zustandsdichten innerhalb der Bandlücke aufweisen. Die Erzeugung von Poren im „low-k Material“ vermindert offenbar die Zustandsdichte im Bereich der Bandlücke und erweitert diese im Vergleich zum SiO2. Eine Modellrechnung mit der Dichtefunktionaltheorie für ein Strukturmodell, das den „low-k Materialien“ nahe kommt, ist zum Vergleich mit der experimentell gefundenen kombinierten Zustandsdichte herangezogen worden und zeigt eine gute Übereinstimmung. Die im Standard-Herstellungsprozeß vorkommenden Verfahren des Plasmaätzens und der Plasmaveraschung können die Struktur des „low-k Materials“ z.B. an den Seitenwänden von Ätzgräben verändern. Die gestörten Bereiche wurden mit der energiegefilterten Elektronenmikroskopie untersucht. Dabei wurde gefunden, daß sich die Strukturveränderungen der Seitenwände bis zu einer Tiefe in der Größenordnung von ungefähr 10 Nanometern erstrecken. Diese Bereiche sind verarmt an Kohlenstoff und ähneln folglich mehr einem SiO2-Dielektrikum. Die Kohlenstoffverarmung erstreckt sich in die „low-k Schicht“ in Form eines gaussartigen Profils mit maximaler Kohlenstoffkonzentration in der Mitte der Schicht. Die Sauerstoffkonzentration und die mikroskopische Dichte steigen in der Nähe der Seitenwände.:TABLES OF CONTENTS LIST OF FIGURES AND TABLES IX 1 INTRODUCTION 1 1.1 Motivation 1 1.2 Low-k dielectric material trends 2 1.3 Required properties of low-k dielectric materials 7 1.4 Technical issues of low-k dielectric materials 7 1.5 Research objectives 8 2 EXPERIMENTAL TECHNIQUES 10 2.1 Transmission electron microscopy 10 2.2 Interaction of fast electrons with a solid specimen 11 2.3 Electron energy-loss spectroscopy 16 2.4 Elemental quantification 19 3 RADIAL DISTRIBUTION FUNCTION 21 3.1 Introduction 21 3.2 Physical aspects of an electron diffraction experiment 22 3.3 Merits and demerits of electron diffraction analysis 24 3.4 Results and Discussion 25 3.5 Conclusion 33 4 CHEMICAL STATE ANALYSIS OF LOW-K DIELECTRIC MATERIALS: AN EELS AND XPS STUDY 34 4.1 EELS analysis 34 4.2 Spectra post-acquisition processing 35 4.3 XPS analysis 37 4.4 Results and Discussion 39 4.5 Conclusion 48 5 DIELECTRIC RESPONSE OF LOW-K DIELECTRIC MATERIALS 49 5.1 Introduction 49 5.2 Exact determination of the loss-function 49 5.3 Fourier-log deconvolution 50 5.4 Kramers-Kronig analysis 51 5.5 Model fitting of dielectric functions: Lorentz oscillator model 52 5.6 Data processing 54 5.7 Results and Discussion 54 5.8 Conclusion 65 6 SIDE-WALL DAMAGE ANALYSIS 66 6.1 Introduction 66 6.2 Energy-filtered transmission electron microscopy (EFTEM) 67 6.3 Merits of EFTEM 69 6.4 Results and Discussion 70 7 SUMMARY, CONCLUSION AND SUGGESTIONS FOR THE FUTURE WORK 88 7.1 Summary 88 7.2 Conclusion 89 7.3 Suggestions for the future work 90 8 REFERENCES 92 9 APPENDIX 100 9.1 Appendix A: Script for normalization of loss function. 100 9.2 Appendix B: Kramers-Kronig analysis script 101 9.3 Appendix C: Sum rule for verification of Kramers-Kronig relation 102 9.4 Appendix D: Lorentz oscillator model 103 9.5 Appendix E: EFTEM image spectrum script 104 10 SELBSTÄNDIGKEITSERKLÄRUNG 108

Cellulose is the most abundant biopolymer on Earth as it is found in every plant cell wall; therefore, it represents one of the most promising natural resources for the fabrication of sustainable materials. In plants, cellulose is mainly used for structural integrity, however, some species organise cellulose in helicoidal nano-architectures generating strong iridescent colours. Recent research has shown that cellulose nanocrystals, CNCs, isolated from natural fibres, can spontaneously self-assemble into architectures that resemble the one producing colouration in plants. Therefore, CNCs are an ideal candidate for the development of new photonic materials that can find use to substitute conventional pigments, which are often harmful to humans and to the environment. However, various obstacles still prevent a widespread use of cellulose-based photonic structures. For instance, while the CNC films can display a wide range of colours, a precise control of the optical appearance is still difficult to achieve. The intrinsic low thermal stability and brittleness of cellulose-based films strongly limit their use as photonic pigments at the industrial scale. Moreover, it is challenging to integrate them into composites to obtain further functionality while preserving their optical response. In this thesis, I present a series of research contributions that make progress towards addressing these challenges. First, I use an external magnetic field to tune the CNC films scattering response. Then, I demonstrate how it is possible to tailor the optical appearance and the mechanical properties of the films as well as to enhance their functionality, by combining CNCs with other polymers. Finally, I study the thermal properties of CNC films to improve the retention of the helicoidal arrangement at high temperatures and to explore the potential use of this material in industrial fabrication processes, such as hot-melt extrusion.