ABSTRACT
BACKGROUND: Previous studies have demonstrated that bone demineralization can improve consolidation in bone grafts. The biologic mechanisms underlying this phenomenon remain unclear. METHODS: Twelve adult male guinea pigs were used in this experiment. Forty-five bone samples removed from the calvaria of nine animals were divided in groups (n = 9) according to the time of demineralization with citric acid (50%, pH 1): 15, 30, 90, and 180 seconds and non-demineralized samples (control). Preosteoblasts (MC3T3-E1) were cultured on the bone samples for 24, 48, and 72 hours (n = 3). Fifteen samples removed from the remaining three animals were analyzed by scanning electron microscopy/energy dispersive spectrometry (SEM/EDS) after demineralization (n = 3). RESULTS: The number of preosteoblasts increased significantly with time in all groups. The bone surface area covered by these cells increased with time, except in the control group. Intragroup differences occurred between 24 and 72 hours (P < 0.05). Samples demineralized for 30 seconds showed greater area covered by preosteoblast cells than for the other times of demineralization in all periods of cell culture (P < 0.05) without a statistically significant difference compared with 15 seconds. SEM/EDS showed diminished content of calcium (Ca) after 15 seconds of demineralization, but the Ca content increased after 180 seconds of demineralization (P < 0.05). The phosphorus (P) amount increased significantly only after 30 seconds of demineralization (P < 0.5). The sulfur (S) content was increased in demineralized samples in relation to non-demineralized ones, reaching the highest level after 90 seconds, when the difference became significant in relation to all the other times of demineralization (P < 0.05). Magnesium (Mg) content did not differ significantly between demineralized and non-demineralized samples. CONCLUSIONS: Bone surfaces demineralized for 30 seconds increased the spreading of preosteoblasts as well as the surface area covered by these cells. Bone demineralization deserves to be studied in periodontal and maxillofacial regenerative procedures.
Subject(s)
Bone Demineralization Technique/methods , Bone and Bones/drug effects , Citric Acid/pharmacology , Osteoblasts/physiology , 3T3 Cells , Animals , Bone and Bones/chemistry , Calcium/analysis , Cell Adhesion/physiology , Cell Culture Techniques , Cell Movement/physiology , Cell Proliferation , Cell Shape , Cells, Cultured , Cytoplasm/ultrastructure , Guinea Pigs , Magnesium/analysis , Male , Mice , Microscopy, Electron, Scanning , Phosphorus/analysis , Spectrometry, X-Ray Emission , Sulfur/analysis , Time Factors , Tissue Scaffolds/chemistryABSTRACT
BACKGROUND: To the best of the authors' knowledge, a standard protocol for treating peri-implantitis is not yet established. METHODS: A total of 150 titanium disks with smooth or rough surfaces contaminated with microbial biofilm were implanted subcutaneously in rats after undergoing one of three treatments: 1) low-intensity laser (LIL); 2) antimicrobial photodynamic therapy (aPDT); or 3) toluidine blue O (TBO). Sterile and contaminated disks served as negative (NC) and positive (C) control groups, respectively. After days 7, 28, and 84, tissue inflammation was evaluated microscopically by measuring the density of collagen fibers (degree of fibrosis) and concentration of polymorphonuclear neutrophils. RESULTS: Surface texture did not affect the degree of inflammation, but the area of reactive tissue was significantly greater for rough implants (2.6 ± 3.7 × 10(6) µm(2)) than for smooth ones (1.9 ± 2.6 × 10(6) µm(2); P = 0.0377). Group C presented the lowest and group NC presented the highest degree of fibrosis with significance only after day 7; these groups had the highest and lowest scores, respectively, for degree of inflammation. Group C showed the largest area of reactive tissue (9.11 ± 2.10 × 10(6) µm(2)), but it was not significantly larger than group LIL (P = 0.3031) and group TBO (P = 0.1333). Group aPDT showed the smallest area (4.34 ± 1.49 × 10(6) µm(2)) of reactive tissue among the treatment groups. After day 28, groups LIL, aPDT, TBO, and C resembled group NC in all the studied parameters. CONCLUSION: Group aPDT showed more favorable results in parameter area of reactive tissue than the other methods after day 7, but over longer time periods all methods produced outcomes equivalent to sterile implants.
Subject(s)
Biofilms/radiation effects , Decontamination/methods , Dental Implants/microbiology , Low-Level Light Therapy , Peri-Implantitis/radiotherapy , Animals , Biofilms/drug effects , Coloring Agents/therapeutic use , Fibrosis/drug therapy , Fibrosis/radiotherapy , Male , Peri-Implantitis/drug therapy , Photochemotherapy , Rats , Rats, Wistar , Subcutaneous Tissue/pathology , Surface Properties , Titanium , Tolonium Chloride/therapeutic useABSTRACT
A peri-implantite acomete um número crescente de indivíduos e os protocolos de tratamento objetivam descontaminar as superfícies dos implantes e torná-las novamente osseointegráveis. Porém, ainda não foi estabelecido um padrão ouro de tratamento para peri-implantite. Este estudo testa o poder de descontaminação do laser em baixa intensidade (LBI), da terapia fotodinâmica (PDT) e do azul de toluidina O (TBO). Sobre discos de titânio de 1,5 mm de espessura e 4,0 mm de diâmetro de superfícies lisas ou rugosas foi permitida a deposição natural de biofilme microbiano por 7 dias após os quais os discos foram divididos em grupos experimentais (15 discos por grupo), de acordo com o método de descontaminação: grupo LBI (laser de InGaAlP de 660 nm, 30 mW, 45 J/cm2 no modo continuo por 30 s); grupo PDT; grupo TBO (aplicação de TBO por 60 s); grupo controle positivo (C) sem tratamento e grupo controle negativo (NC) estéril. Os discos foram implantados em tecido conjuntivo subcutâneo de ratos e, após 7, 28 e 84 dias, foram obtidas 5 biópsias de cada grupo para análise em microscopia óptica. Implantes lisos e rugosos não diferiram com relação ao grau de fibrosamento e severidade do infiltrado inflamatório, mas a área do tecido reacional perimaterial foi maior nos rugosos (2,6 ± 3,7 x 106 µm2) do que nos lisos (1,9 ± 2,6 x 106 µm2). O grupo C foi o que apresentou menor grau de fibrosamento do tecido reacional (1), mas somente houve diferença estatisticamente significante (p = 0,0230) entre os grupos C e NC. A severidade do infiltrado inflamatório somente diferiu significantemente entre os grupos aos 7 dias, quando o grupo NC apresentou o menor escore (2) em comparação aos demais grupos (3). Com relação à área do tecido reacional perimaterial, só houve diferenças entre os grupos aos 7 dias, quando o grupo C apresentou maior área que os demais grupos (9,11 ± 2,10 x 106 µm2), porém, sem diferença estatisticamente significante em comparação aos grupos LBI e TBO...
The peri-implantitis is a disease that is increasing in a number of individuals and the protocols of treatment has intended to decontaminate the implant surfaces and makes this biocompatible again. However, until this moment, a gold standard for peri-implantitis treatment was not established. The present study tests the power of decontamination of low intensity laser (LBI), photodynamic therapy (PDT) and toluidine blue O (TBO). The natural deposition of microbian biofilm was allowed on titanium discs with 1,5 mm of thickness and 4,0 mm of diameter and smooth and grooved surfaces during 7 days when the discs were separated in experimental groups (15 discs each group), in accordance with the decontamination method: LBI group (decontamination with InGaAlP laser, 660 nm, 30 mW, 45 J/cm2 in continuous way during 30 s); PDT group; TBO group (application of TBO during 60 s); controlled positive group (C) without any kind of treatment and controlled negative group (NC), sterile. The discs had been implanted in rats subcutaneous connective tissue and, after 7, 28 and 84 days, had been gotten 5 biopsies from each group for histology processing. Smooth and grooved implants did not differ on fibrosis grade and severity of the inflammatory infiltrate, however the perimaterial reaction tissue area was bigger on grooved implants (2,6 ± 3,7 x 106 µm2) that on smooth implants (1,9 ± 2,6 x 106 µm2). The C group was the one that showed the smallest reaction tissue fibrosis grade (1), but only was significant statistical difference (p = 0,0230) between C and NC groups. The severity of inflammatory infiltrate only significant differed between the groups on 7 days, when the NC group showed the smallest score (2) in comparison to the others groups (3). In relation to the perimaterial reaction tissue area, only were differences between the groups on 7 days, when C group showed bigger area than the others (9,11 ± 2,10 x 106 µm2), nevertheless, without significant statistical...