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Differentiation and Activity of Murine Derived Stromal Osteoblasts After Electromagnetic Wave StimulationWu, Jennifer L. January 2022 (has links)
Indiana University-Purdue University Indianapolis (IUPUI) / Introduction: Elimination of bacteria and active infection within an infected root canal system is one of the primary objectives of nonsurgical root canal treatment. One of the measures of successful root canal treatment is subsequent bone healing of periapical lesions caused by previous infection. A previous study by Yumoto et al. showed that electromagnetic wave stimulation can increase proliferation of osteoblastic cells with no cytotoxicity, and it can also up-regulate growth factors such as vascular endothelial growth factor and platelet-derived growth factor.18 They also showed increased proliferation of an immortalized osteoblastic MC3T3-E1 cell line 3 days following electromagnetic stimulation (EMS).18 Previously, Pauly et al. found increased alkaline phosphatase (ALP) activity with 10 mA EMS application to primary murine calvaria-derived osteoblastic cells with 5 pulses at 1 second per pulse, but no significant differences were found for MTS proliferation nor mineral deposition compared to a negative control group.82 Optimization of the different variables including post-treatment incubation time, current delivery, and number of pulses per treatment may be necessary to improve osteogenic activity. The use of mesenchymal stem cells from murine bone marrow may also offer a physiologically relevant model for osteoblastic regeneration of periapical lesions.
Objectives: The goal of this study was to investigate and optimize the effects of electromagnetic wave stimulation (EMS) on murine bone marrow mesenchymal stem cells (MSCs) by evaluating the proliferation and differentiation of the cells after exposure to different EMS treatment regimens.
Materials and Methods: 5 x104 stromal osteoblasts (SOBs) were cultured in 24-well plates in α-MEM containing 10% fetal bovine serum. Cells were then subjected to pulsed EMS treatments of 1 mA, 10 mA, and 50 mA. EMS was generated using an electromagnetic apical treatment (EMAT) device created by J. Morita MFG Corp. Proliferation was assessed via MTS assay 1 days after treatment. For osteogenic differentiation, ascorbic acid and β-glycerol phosphate were added to the culture media, and SOBs were cultured for 14 days. Afterwards, alkaline phosphatase (ALP) activity and Alizarin-red S mineral deposition were quantified as measures of osteoblast activity. Cells grown in osteogenic media without EMS treatment served as the negative control.
Results: Although MSC proliferation was unaffected by different EMS treatment regimens, 50 mA EMS resulted in a decrease in ALP activity and mineral deposition by osteoblasts.
Conclusions: Our findings suggest bone healing by EMS may involve a different cellular mechanism, that is not reproduced in vitro in our studies. Utilizing different amperage and EMS regimens may improve osteogenic differentiation.
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Immunhistokemisk (IHC) analys av låggradigt inflammerade biopsier med apikal parodontit -en pilotstudie / Immunohistochemical (IHC) analysis of biopsies with apical periodontitis with assessed low-grade inflammation – a pilot studyPesonen, Izabell, Ismail, Midia January 2021 (has links)
Syfte: Att ta reda på hur relationen ser ut för B- respektive T-lymfocyter i biopsier av rotfyllda tänder med apikal parodontit med bedömd låggradig inflammation. Denna studie kommer även att analysera hur den inflammatoriska bilden ser ut i förhållande till beskrivna symtom i dessa biopsier. Material & metod: En pilotstudie utfördes på 10 biopsier från rotfyllda tänder med apikal parodontit med bedömd låggradig inflammation enligt Danesh et al 2019 klassificeringssystem. Biopsierna hämtades från Malmö universitets biobank. Immunhistokemisk infärgning av antigenerna CD20+ och CD3+ utfördes samt analyserades med hjälp av digitalmikroskop. Jämförelsen av symtomen från remisserna skedde efter att de histologiska resultaten sammanställts. Resultat: Det fanns fler T-lymfocyter än B-lymfocyter i 6 stycken av biopsierna. I de resterande 4 biopsierna var de lika många. I remisserna hade symtombilden inte angetts föralla biopsier. Slutsats: Enligt vår pilotstudie ser vi tendenser till att T-lymfocyter är fler än B-lymfocyter eller att de är lika många. Inga slutsatser kunde dras gällande symtombilden. Ett större material från olika remittenter krävs för definitiva slutsatser. En vidare infärgning av CD4+ samt CD8+ skulle vara intressant. / Aim: To investigate the relation between B- and T- lymphocytes in biopsies of root-filled teeth with apical periodontitis with assessed low-grade inflammation. This study will also analyse what the inflammation looks like in relation to the symptoms described in these biopsies. Study design: A pilot study was performed on 10 biopsies of root-filled teeth with apical periodontitis with assessed low-grade inflammation according to the classification system of Danesh et al 2019. The biopsies were collected from Malmö University's biobank. An immunohistochemical staining of antigens CD20+ and CD3+ were performed and analysed by a digital microscope. A comparison of the symptoms from the referrals were performed once the histological results were compiled. Results: There were more T-lymphocytes than B-lymphocytes in 6 of the biopsies. In the remaining 4 biopsies there were an equal amount of B- and T-lymphocytes. The symptoms were not stated for all biopsies in the referrals. Conclusion: According to our pilot study, we see tendencies that T lymphocytes are more than B lymphocytes or that they are equal. No conclusions could be drawn regarding the symptom picture. Larger material from different referrers is required for definitive conclusions. A further staining of CD4 + and CD8 + would be interesting.
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Avaliação do processo de raparo de lesões periopicais pós-tratamento endodôntico por meio de subtração digital radiográfica / Evaluation of the process of repair of periapical lesions after endodontic treatment by digital subtraction radiographySILVA, Janaína Benfica e 30 November 2006 (has links)
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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.
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