Electron microscopic detection of salivary <i>α</i>‐amylase in the pellicle formed <i>in situ</i>

Deimling, Daniela; Breschi, Lorenzo; Hoth-Hannig, Wiebke; Ruggeri, Alessandra; Hannig, Christian; Nekrashevych, Yuriy; Prati, Carlo; Hannig, Matthias

doi:10.1111/j.1600-0722.2004.00168.x

Cited by 47 publications

(49 citation statements)

References 26 publications

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“…So far, there are only studies which investigated enzyme activities in caries-free mature individuals [Deimling et al, 2004;Hannig et al, 2004Hannig et al, , 2008aHannig et al, , b, 2009. The aim of this study was therefore to investigate the process of initial bioadhesion in caries-active and -inactive individuals.…”

Section: Discussionmentioning

confidence: 99%

“…Host enzymes with protective properties are lysozyme and peroxidase; α-amylase and glycosyltransferase (GTF) facilitate the binding of microorganisms to the pellicle layer. They can all be found immobilized in the initial in situ pellicle in an active conformation [Deimling et al, 2004;Hannig et al, 2004Hannig et al, , 2008b. The enzyme activity of the pellicle enzymes might be related to the caries activity.…”

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confidence: 99%

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Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

et al. 2017

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Aim: The present study aimed to evaluate the impact of caries activity on the key enzymes and the ultrastructure of the in situ pellicle. Methods: Pellicle formation was performed on bovine enamel slabs. Intraoral exposure (3, 30, and 120 min) was accomplished by 14 caries-active (DMFS: 22.7 ± 12.1) and 13 caries-inactive (DMFS: 1.5 ± 1.8) individuals. The enzyme activities (lysozyme, peroxidase, α-amylase, glycosyltransferase [GTF]) in the in situ pellicle and resting saliva of all participants were analyzed directly after oral exposure. In addition, a simultaneous visualization of these enzymes, extracellular glucans, and adherent bacteria was carried out. Fluorescent patterns were analyzed with fluorescence labeling and 4′,6-diamidino-2-phenylindole/concanavalin A staining. In addition, the distribution of GTF B, C, and D and the ultrastructure of the pellicle were examined by gold immunolabeling and transmission electron microscopy with selected samples. Results: Enzyme activities of amylase, peroxidase, lysozyme, and GTF were detected on all enamel slabs in an active conformation. Neither exposure time nor caries activity had an impact on the enzyme activities. Gold immunolabeling indicated that the pellicle of caries-active subjects tends to more GTF D molecules. The pellicles of caries-inactive and -active individuals revealed a similar ultrastructural pattern. Conclusion: The enzyme activities as well as the pellicle's ultrastructure are of high similarity in caries-active and -inactive subjects. Thereby, oral exposure time has no significant influence. This reflects a high uniformity during the initial phase of bioadhesion (3-120 min) concerning enzymatic functions. However, there is a tendency towards more GTF D in caries-active individuals.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

et al. 2017

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“…The initial biofilm formation was carried out in situ on bovine enamel and dentin samples, an approach adopted in several previous studies on pellicle formation and initial bioadhesion [Deimling et al, 2004;Hannig et al, 2008a;Jung et al, 2010]. The prepared slabs allow the investigation of bioadhesion in a standardized manner and very like the situation in the oral cavity, although minor microscopic irregularities and residual organic material on the native tooth surface could modulate the interaction of the fluoride or chlorhexidine preparations with the enamel and dentin surface in vivo.…”

Section: Discussionmentioning

confidence: 99%

“…The slabs were washed twice for 5 min in double-distilled water. The enamel samples were treated by ultrasonication for 1 min in 3% sodium hypochlorite to remove the superficial smear layer [Deimling et al, 2004[Deimling et al, , 2007. Afterwards, 10-minute ultrasonication in 70% ethanol was performed for the purpose of disinfection, followed by 10-minute ultrasonication in double-distilled water.…”

Section: Specimens and Subjectsmentioning

confidence: 99%

Effect of Conventional Mouthrinses on Initial Bioadhesion to Enamel and Dentin in situ

et al. 2012

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Aim: The study aimed to investigate the effect of a customary fluoride solution, containing sodium fluoride and amine fluoride, on initial biofilm formation on enamel and dentin in situ compared directly to chlorhexidine. Methods: Bovine enamel and dentin specimens were mounted on maxillary splints carried by 9 subjects. After 1 min of pellicle formation, rinses with tap water (control), chlorhexidine (meridol med CHX 0.2%, GABA) and a fluoride mouthrinse (elmex, GABA) were performed for 1 min. Subsequently, the slabs were carried for another 8 h. The adherent bacteria were determined by DAPI staining, live-dead staining and determination of colony-forming units after desorption; glucan formation was visualized with concanavalin A. Additionally, energy-dispersive X-ray spectroscopy (EDX) analysis of the in situ biofilm layers was conducted, and contact angle measurements were performed. Statistical evaluation was performed by means of the Kruskal-Wallis test followed by the Mann-Whitney U test (p < 0.05). Results: In the control group, significantly higher amounts of adherent bacteria were detected on dentin (4.8 × 10⁶ ± 5.4 × 10⁶ bacteria/cm²) than on enamel (1.2 × 10⁶ ± 1.5 × 10⁶ bacteria/cm², DAPI). Chlorhexidine significantly reduced the amount of adherent bacteria (dentin: 2.8 × 10⁵ ± 3.4 × 10⁵ bacteria/cm²; enamel: 4.2 × 10⁵ ± 8.7 × 10⁵ bacteria/cm²). Rinses with the fluoride solution also significantly reduced bacterial adherence to dentin (8.1 × 10⁵ ± 1.5 × 10⁶ bacteria/cm²). Fluoride could not be detected by EDX analysis of the biofilms. Fluoride mouthrinsing did not influence the wettability of the pellicle-covered enamel surface. Conclusion: In addition to the reduction of demineralization and antibacterial effects, fluorides inhibit initial biofilm formation on dental hard tissues considerably, especially on dentin.

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“…As histatinas pertencem a uma família de peptídeos básicos ricos em histidinas que se encontram na saliva humana (OPPENHEIM et al, 1988 DEIMLING et al, 2004;HANNIG et al, 2001). …”

Section: Composição Da Paunclassified

Determinação da composição da película adquirida formada in situ sobre o esmalte e dentina humanos através de análise proteômica

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Determinação da composição da película adquirida formada in situ sobre esmalte e dentina humanos através de análise proteômica BAURU 2013 Nota: A versão original desta dissertação encontra-se disponível no Determinação da composição da película adquirida formada in situ sobre esmalte e dentina humanos através de análise proteômica / Melina Rodrigues Bellini-Bauru, 2013. 161p. : il. ; 31cm. Dissertação (Mestrado)-Autorizo, exclusivamente para fins acadêmicos e científicos, a reprodução total ou parcial desta dissertação/tese, por processos fotocopiadores e outros meios eletrônicos. Assinatura: Data: Comitê de Ética da FOB-USP Protocolo nº: 038/2011 Data: 28/04/2011 DEDICATÓRIA Dedico de todo meu coração este trabalho à Deus, que tem acompanhado sempre os meus passos, e aberto os caminhos, e direcionado sempre minha vida. Mostrando que a minha coragem deve ser maior que meu medo; e minha força deve ser tão grande quanto a minha fé. "Pois quando a sabedoria entrar no teu coração, e o conhecimento for agradável a tua alma, o bom siso te guardará e a inteligência te conservará". (Provérbios 2:10-11) (…) Staring up the road and pray to god I see headlights. (Darius Rucker). Dedico este trabalho aos meus pais, Rosângela e Vanderlei, os quais me deram o maior bem o qual eu possuo: a vida. Eu agradeço muito por tudo o que vocês sempre fazem por mim. Vocês são meus exemplos de vida, por mostrar que sem esforço jamais atingiremos o alto da montanha. Por não deixarem que eu desanime no meio da estrada, e mostrarem que só atingimos o cimo da montanha se estivermos decidido a enfrentar o esforço da caminhada. Obrigada por sempre me orientarem, e me auxiliarem quando eu mais preciso, sou muito feliz de ser filha de vocês! Eu amo MUITO vocês! "Vá firme na direção das suas metas. Porque o pensamento cria, o desejo atrai e a fé realiza". AGRADECIMENTOS ESPECIAIS Meus sinceros agradecimentos à minha querida orientadora Prof a. Dr a. Marília Afonso Rabelo Buzalaf. Professora, não teria palavras pra descrever a senhora, que sempre se disponibilizou de muita paciência para me auxiliar. Mas pode ter certeza que a senhora tem todo meu carinho, respeito e muita admiração. Meus agradecimentos provêm de todo esse tempo que trabalhamos juntas. A senhora sempre me recebeu de portas abertas na bioquímica. Tenho muito orgulho pela senhora ter sido minha orientadora. A senhora sabe que tem muito reconhecimento no mundo científico: e parabéns professora, por não perder a humildade e simplicidade. Pode ter certeza, que a senhora tem o maior merecimento do mundo, por estar onde esta. Sou muito feliz, por ter a senhora ao meu lado com uma postura séria, honesta e ética e acredito que todos que têm a oportunidade de conhecê-la sabem do que eu estou falando. Reconheço que apesar de estar nesse meio acadêmico que sempre exige muito, a senhora sempre está de coração aberto para os seus alunos. Agradeço pelo maravilhoso trabalho que fizemos juntas, e por me aceitar em ser sua orientada. Se tivesse que voltar ao passado escolher tudo de novo, escolheria...

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Electron microscopic detection of salivary α‐amylase in the pellicle formed in situ

Cited by 47 publications

References 26 publications

Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

Effect of Conventional Mouthrinses on Initial Bioadhesion to Enamel and Dentin in situ

Determinação da composição da película adquirida formada in situ sobre o esmalte e dentina humanos através de análise proteômica

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