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Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament of Healthy TeethLagerholm, Sara January 2019 (has links)
ABSTRAKT:Isolering och karaktärisering av mesenkymala stamceller från periodontalligamentet hos friskatänderSYFTE: Att isolera och odla celler från periodontalligamentet samt karaktärisera dem sommesenkymala stamceller.MATERIAL OCH METOD: Friska premolarer gjordes tillgängliga vid ortodontiskaextraktioner. Den mellersta 1/3 av periodontalligamentet skrapades varpå en enzymatiskmetod användes för isolering av individuella celler. Resulterande celler odlades understandardiserade metoder. Karaktärisering av celler skedde genom flödescymetri med 2 olikapaneler av cellyta markörer; en för etablerat positiva uttryck och en för kända negativauttryck hos mesenkymala stamceller. Möjlighet av celler att differentieras in vitro tilladipocyter och osteocyter testades genom tillförsel av specifika substanser till odlingsmediet.RESULTAT: Celler från 11 av 13 tänder isolerades och odlades framgångsrikt adherenta tillodlingsytan i upp till 8 generationer. Celluttryck av de positiva markörerna CD73, CD90 samtCD44 bekräftades genom flödescymetri. Inget uttryck observerades för den negativa panelenCD45, CD34, CD11b, CD19 eller HLA class II. Uttrycket av CD105 kunde inte fastställas pgaofullständigt data. Försök till differentiering av celler till adipocyter och osteocyter visade påfenotypiska förändringar efter 21 dagar.SLUTSATS: Den här studien har bidragit till framgångsrik isolering och delvis karaktäriseringav mesenkymala stamceller från periodontalligamentet hos friska tänder. En icke-invasivmetod av detta slag, resulterande i tillgång till denna cellpopulation utgör ett lovande verktygför framtida studier med goda möjligheter till ytterligare kunskap applicerbart till kliniskasituationer inom tandvården. / ABSTRACT:Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament ofHealthy TeethAIM: To isolate and culture viable cells from the periodontal ligament and confirming theiridentity as mesenchymal stem cells.METHODS AND MATERIALS: Healthy premolars were collected at the time oforthodontic extractions. The middle 1/3 of the periodontal ligament was scraped andsubsequent cell isolation was performed using an enzymatic method; yielding single cellisolates. Cells were cultured and maintained under standard culture conditions. Cellcharacterization was performed by flow cytometry using two sets of cell surface markers; oneknown to be present and one known to be absent in mesenchymal stem cells. Ability of thecells for in vitro differentiation into adipogenic and osteogenic lineages was tested usingspecifically formulated media supplements.RESULTS: Cells were successfully isolated from 11 of 13 teeth and were maintained asadherent cultures for up to 8 generations. Cellular expression of positive markers; CD73, CD90and CD44 were confirmed by flow cytometry. For the negative marker panel, expression ofCD45, CD34, CD11b, CD19 and HLA class II were not detectable. The expression of CD105was inconclusive. As determined by phenotypic changes, cells appeared to have undergoneadipogenic and osteocytic differentiation at 21 days.CONCLUSION: This study has resulted in successful isolation and partial characterization ofmesenchymal stem cells from the periodontal ligament of healthy teeth. Non-invasive accessto these cells, provides an excellent tool for future studies, potentially leading to beneficialknowledge transferable to the dental clinical situation.
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An Investigation of Surface Characteristics of Enamel Treated with Infiltrative Resin: A Scanning Electron Microscopy StudyEasterly, Danielle E 01 January 2017 (has links)
AN INVESTIGATION OF SURFACE CHARACTERISTICS OF ENAMEL TREATED WITH INFILTRATIVE RESIN: A SCANNING ELECTRON MICROSCOPY STUDY
Danielle E. Easterly, Doctorate of Dental Surgery.
A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Dentistry at Virginia Commonwealth University.
Virginia Commonwealth University. 2017.
Director: Dr. Eser Tüfekçi
D.D.S., M.S., Ph.D., M.S.H.A.
Objective: To evaluate the microstructural changes of a resin infiltrant (ICON®, DMG America LLC, Englewood, NJ) after six months of simulated toothbrushing.
Materials and Methods: Ten extracted third molars (n=10) were collected. Artificial white spot lesions were created and resin applied. Environmental SEM images at 250X and 500X were taken after application of Icon® (T1), and after six months of simulated toothbrushing (T2). Micrographs were evaluated for changes in surface characteristics.
Results: SEM showed some changes in the surface characteristics of the resin after simulated toothbrushing. However, changes in presence of enamel rods, microcracks, or fractures were not statistically significant (p>0.05). The effects of polymerization shrinkage were noted on most samples in the form of clefts and fissures.
Conclusions: Icon® resin seems to withstand challenge by toothbrush abrasion over a six-month period, with some evidence of microstructural wear.
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