研究目的
To evaluate the effect of an MB experimental formulation (ethanol 20%) in antimicrobial photodynamic therapy (aPDT) used as an adjuvant to scaling and root planing (SRP) in the periodontal treatment of diabetic rats.
研究成果
Adjunctive aPDT with 0.01% MB solubilized in ultra-pure water/ethanol/carboxymethylcellulose (77:20:3) can have beneficial potential to periodontal treatment of diabetics, showing less bone loss, lower inflammatory infiltrate, and higher collagen content compared to SRP alone.
研究不足
The current study has some limitations: (i) diabetes and periodontitis were induced in a short period and the acute nature of these models might not fully reflect the chronic pathological conditions in humans; and (ii) short period (7 days) to analyze the periodontal healing.
1:Experimental Design and Method Selection:
A controlled, blinded, and randomized animal experiment using a ligature model to induce experimental periodontitis in diabetic rats. The study involved inducing diabetes with streptozotocin, periodontitis with silk ligature, and treatments including SRP and aPDT with different MB formulations.
2:Sample Selection and Data Sources:
Forty male Wistar rats (Rattus norvegicus albinus) aged 90 days, housed under controlled conditions. Diabetes was induced via intraperitoneal injection of streptozotocin. Periodontitis was induced by placing a silk ligature on the first right mandibular molar.
3:List of Experimental Equipment and Materials:
Streptozotocin (Sigma Chemical Company), glucose monitor (Accu-check Active, Roche Diagnosis), silk ligature (Ethicon 4-0, Johnson & Johnson), manual curettes (Hu-Friedy Co. Inc.), methylene blue (Sigma Aldrich), laser (InGaAlP, Thera Lase - DMC), syringes, insulin needles, formalin, EDTA, microtome, image analysis systems (Axiovision; Carl Zeiss MicroImaging, Zen lite 2012 blue edition, ImageJ v
4:50i), polarization microscopy (Leica, model DM2000). Experimental Procedures and Operational Workflow:
Diabetes induction, periodontitis induction, randomization into groups (NLG, LG, SRPG, aPDTW, aPDTEt), SRP procedure, aPDT application with laser irradiation, euthanasia after 7 days, tissue processing, histometric analysis of bone loss, inflammatory response, and collagen content.
5:Data Analysis Methods:
Statistical analysis using Shapiro-Wilk test for normality, one-way ANOVA, post hoc Bonferroni's test, repeated measures ANOVA, mixed models ANOVA, Tukey's test, intraclass correlation coefficient, and Kappa coefficient for reliability.
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image analysis system
Axiovision
Carl Zeiss MicroImaging
Used for histometric analysis of bone loss area.
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image-processing software
Zen lite 2012 blue edition
Carl Zeiss MicroImaging
Used for selecting regions of interest in histometric analysis.
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polarization microscopy
Leica DM2000
Leica
Used for digitalizing images of collagen fibers in picrosirius red stained sections.
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image-processing software
Leica DFC295
Leica
Used for selecting images in polarization microscopy.
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glucose monitor
Accu-check Active
Roche Diagnosis
Measuring blood glucose levels from tail veins of animals.
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silk ligature
Ethicon 4-0
Johnson & Johnson
Placed on the gingival margin to induce periodontitis.
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curettes
1-2 mini five
Hu-Friedy Co. Inc.
Used for scaling and root planing (SRP) procedure.
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laser
InGaAlP, Thera Lase
DMC
Applied for antimicrobial photodynamic therapy (aPDT) with specific wavelength and power settings.
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software
ImageJ v1.50i
National Institutes of Health
Used for binarizing images and calculating percentages of collagen fibers.
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