Laser and LED photobiomodulation effects in osteogenic or regular medium on rat calvaria osteoblasts obtained by newly forming bone technique.

The purposes of this study are to evaluate the effects of photobiomodulation (PBM) with laser and LED on rat calvaria osteoblasts (rGO lineage), cultured in osteogenic (OST) or regular (REG) medium, after induction of a quiescent state and to test if PBM is capable of osteogenic induction and if there is a sum of effects when combining OST medium with PBM. Before irradiation, the cells were put in a quiescent state (1% FBS) 24 h, when red (AlGaInP-660 nm) and infrared laser (GaAlAs-808 nm) and LED (637 ± 15 nm) were applied. The groups were as follows: red laser (RL3-5 J/cm2, 3 s and RL5-8.3 J/cm2, 5 s, 1.66 W/cm2); infrared laser (IrL3-5 J/cm2, 3 s and IrL5-8.3 J/cm2, 5 s); LED (LED3-3 s and LED5-5 s, 0.02 J/cm2, 0.885 W/cm2); positive (C+, 10% FBS) and negative control (C-, 1% FBS). For alkaline phosphatase (ALP) and mineralization assays, the cells were cultured in REG (DMEM 10% FBS) and OST medium (DMEM 10% FBS, 50 µg/mL ascorbic acid, 10 mM ß-glycerophosphate). Statistical analysis was performed using ANOVA and Tukey's tests (p < 0.05). RL5 and LED5 increased proliferation, in vitro wound closure, ALP, and mineralization in rGO cells (p < 0.05). PBM with red laser and LED induced mineralization by itself, without osteogenic medium, not observed for infrared laser (p < 0.05). A sum of effects was observed in osteogenic medium and PBM by infrared, red laser, and LED (5 s). Red laser and LED increased proliferation, migration, and secretory phases in rGO cells in a dose-dependent manner. PBM with red laser and LED promotes osteogenic induction by itself. PBM with infrared laser and osteogenic medium potentializes mineralization.

Free gingival graft and acellular dermal matrix for gingival augmentation: a 15-year clinical study.

OBJECTIVES: This study evaluated clinical outcomes of acellular dermal matrix (ADM) allograft compared with autogenous free gingival graft (FGG) for gingival augmentation after 15 years. MATERIAL AND METHODS: Twenty-two patients were originally included and evaluated by de Resende et al. (Clin Oral Investig 23:539-550, 2019), and 12 accepted to participate in this longitudinal evaluation. Clinical parameters evaluated were recession depth (RD), probing depth (PD), clinical attachment level (CAL), keratinized tissue width (KTW), and soft tissue thickness (TT). In addition, esthetic perception was evaluated by patients and by a calibrated periodontist. Data were evaluated by ANOVA complemented by Tukey tests (p < 0.05). RESULTS: After 15 years, both treatments provided a significant increase in KTW and TT but with superior results for the FGG group (p < 0.05). No differences were observed between groups for PD and CAL. In the ADM group, RD significantly increased in long term, as well as the rate of tissue contraction. The percentage of shrinkage for the ADM group was 59.6%. Conversely, the FGG group presented a creeping attachment of 17.6% and RD significantly decreased in long term. The ADM group presented superior results considering professional esthetic perception. CONCLUSIONS: Both treatments longitudinally promoted significant gain of keratinized tissue width and thickness with superior outcomes for the FGG group. The ADM group demonstrated more tissue contraction and gingival recession whereas the FGG group presented creeping attachment. Professional esthetic perception was superior for the ADM group. CLINICAL RELEVANCE: This study added important clinical data with long-term evaluation of ADM compared with FGG.

Acellular dermal matrix allograft versus free gingival graft: a histological evaluation and split-mouth randomized clinical trial.

OBJECTIVES: This split-mouth controlled randomized clinical trial evaluated clinical and histological results of acellular dermal matrix allograft (ADM) compared to autogenous free gingival graft (FGG) for keratinized tissue augmentation. MATERIAL AND METHODS: Twenty-five patients with the absence or deficiency of keratinized tissue (50 sites) were treated with FGG (control group) and ADM (test group). Clinical parameters included keratinized tissue width (KTW) (primary outcome), soft tissue thickness (TT), recession depth (RD), probing depth (PD), and clinical attachment level (CAL). Esthetic perception was evaluated by patients and by a calibrated periodontist using visual analog scale (VAS). Histological analysis included biopsies of five different patients from both test and control sites for each evaluation period (n = 25). The analysis included percentage of connective tissue components, epithelial luminal to basal surface ratio, tissue maturation, and presence of elastic fibers. Data were evaluated by ANOVA complemented by Tukey's tests (p < 0.05). RESULTS: After 6 months, PD and CAL demonstrated no differences between groups. ADM presented higher RD compared to FGG in all periods. Mean tissue shrinkage for control and test groups was 12.41 versus 55.7%. TT was inferior for ADM group compared to FGG. Esthetics perception by professional evaluation showed superior results for ADM. Histomorphometric analysis demonstrated higher percentage of cellularity, blood vessels, and epithelial luminal to basal surface ratio for FGG group. ADM group presented higher percentage of collagen fibers and inflammatory infiltrate. CONCLUSIONS: Both treatments resulted in improvement of clinical parameters, except for RD. ADM group presented more tissue shrinkage and delayed healing, confirmed histologically, but superior professional esthetic perception. CLINICAL RELEVANCE: This study added important clinical and histological data to contribute in the decision-making process between indication of FGG or ADM.

Comparison of the effect of root surface modification with citric acid, EDTA, and aPDT on adhesion and proliferation of human gingival fibroblasts and osteoblasts: an in vitro study.

Root demineralization is used in Periodontics as an adjuvant for mechanical treatment. The aim of this study was to evaluate the effects of root surface modification with mechanic, chemical, and photodynamic treatments on adhesion and proliferation of human gingival fibroblasts and osteoblasts. Root fragments were treated by scaling and root planing (C-control group), EDTA (pH 7), citric acid plus tetracycline (CA-pH 1), and antimicrobial photodynamic therapy (aPDT) with toluidine blue O and red laser (pH 4). Cells were seeded (104 cells/well, 6th passage) on root fragments of each experimental group and cultured for 24, 48, and 72 h. Cells were counted in scanning electron microscopy images by a calibrated examiner. For fibroblasts, the highest number of cells were present at 72-h period (p < 0.05). EDTA group showed a very low number of cells in relation to CA group (p < 0.05). CA and aPDT group presented higher number of cells in all periods, but without differences between other treatment groups (p > 0.05). For osteoblasts, there was a significant increase in cell numbers for aPDT group at 72 h (p < 0.05). In conclusion, aPDT treatment provided a positive stimulus to osteoblast growth, while for fibroblasts, aPDT and CA had a tendency for higher cell growth.

Laser and light-emitting diode effects on pre-osteoblast growth and differentiation.

The acceleration of bone regeneration by low-intensity laser irradiation may hold potential benefits in clinical therapy in orthopedics and dentistry. The purpose of this study is to compare the effects of light-emitting diode (LED) and laser on pre-osteoblast MC3T3 proliferation and differentiation. Cells were irradiated with red, infrared, and LED (3 and 5 J/cm(2)). Lasers had a power density of 1 W/cm(2) and irradiation time of 2 and 5 s. LED had a power density of 60 mW/cm(2) and irradiation time of 50 and 83 s. Control group did not receive irradiation. Cell growth was assessed by a colorimetric test (MTT) (24, 48, 72, and 96 h), and cell differentiation was evaluated by alkaline phosphatase (ALP) quantification after growth in osteogenic medium (72 and 96 h and 7 and 14 days). At 24 h, the cell growth was enhanced 3.6 times by LED (5 J/cm(2)), 6.8 times by red laser (3 J/cm(2)), and 10.1 times by red laser (5 J/cm(2)) in relation to control group (p < 0.05). At the other periods, there was no influence of irradiation on cell growth (p > 0.05). The production of ALP was not influenced by irradiation at any period of time (p > 0.05). Low-intensity laser and LED have similar effects on stimulation of cell growth, but no effect on cell differentiation.

Palatal mucosa derived fibroblasts present an adaptive behavior regarding cytokine secretion when grafted onto the gingival margin.

BACKGROUND: Considering that grafted gingival tissue might have to be adapted to the receptor area and that fibroblasts have the ability to respond to bacterial stimuli through the release of various cytokines, this study investigated whether fibroblasts from the palatal mucosa behave differently when grafted onto the gingival margin regarding cytokine secretion. METHODS: Biopsies from the palatal mucosa were collected at the time of free gingival graft surgery, and after four months re-collection was performed upon surgery for root coverage. Fibroblasts were isolated by the explant technique, cultured and stimulated with Porphyromonas gingivalis (Pg) and Escherichia coli (Ec) LPS for 24 or 48 h for comparative evaluation of the secretion of cytokines and chemokines, such as IL-6, IL-8/CXCL8, MIP-1α/CCL3, TGF-ß, VEGF and CXCL16. Unstimulated cells were used as the control group. Cells were tested for viability through MTT assay, and secretion of cytokines and chemokines was evaluated in the cell supernatants by Enzyme-Linked Immunosorbent Assay (ELISA). RESULTS: Fibroblasts from the palatal mucosa maintained the same secretion pattern of IL-6 when grafted onto the gingival margin. On the contrary, fibroblasts from the marginal gingival graft showed increased secretion of IL-8/CXCL8 even in the absence of stimulation. Interestingly, MIP-1α/CCL3 secretion by fibroblasts from the marginal gingival graft was significantly increased after 48 hours of stimulation with Pg LPS and after 24 h with Ec LPS. Only fibroblasts from the marginal gingival graft showed secretion of TGF-ß. VEGF and CXCL16 secretion were not detected by both subsets of fibroblasts. CONCLUSION: Fibroblasts from the palatal mucosa seem to be adapted to local conditions of the site microenvironment when grafted onto the gingival marginal area. This evidence supports the effective participation of fibroblasts in the homeostasis of the marginal periodontium through secretion modulation of important inflammatory mediators.

The influence of implant surface roughness on decontamination by antimicrobial photodynamic therapy and chemical agents: A preliminary study in vitro.

BACKGROUND: The aim of this preliminary study was to analyze the effectiveness of three different protocols of decontamination on five commercial moderate rough implants. MATERIAL AND METHODS: The types of implants investigated were: Neoporos Drive CM (CM; Neodent®), Drive CM Acqua (ACQ; Neodent®), SLActive (SLA; Straumann®), Osseotite (OT; Biomet 3i®) and Nanotite (NT; Biomet 3i®). Implant surface properties (n = 2/type of implant; control groups) were analyzed by scanning electron microscopy (SEM) images to determine surface roughness parameters (SRP) and energy disperse X-ray spectrometry to determine the chemical composition. Implants were then inoculated with Aggregatibacter actinomycetencomitans in vitro (n = 6/type of implant;experimental groups) and the contaminated areas were determined in SEM images (500x magnifications). Decontamination of implants was performed in duplicate by three protocols: antimicrobial photodynamic therapy (aPDT), EDTA associated with citric acid (EDTA + CA) and 0.12 % chlorhexidine (CHX). The remaining contaminated area (rCtA) was determined in SEM images (500x magnifications). All quantitative analysis through SEM images were analyzed in ImageJ® software for two-dimensional parameters. RESULTS: No significant differences were found in SRP among implants (control group), except for Rv (lowest valley) between SLA vs. OT (p=0.0031; Kruskal Wallis post hoc Dunn). NT implants showed highest contaminated area vs. ACQ implants (68.19 % ± 8.63 % and 57.32 % ± 5.38 %, respectively; p = 0.0016, Tukey's test). SRP after decontamination showed statistical difference for Ra (arithmetical mean deviation) for all decontamination groups when compared to control (p < 0.05; ANOVA with post-hoc Tukey's multiple comparisons test), only CM implants showed statistical difference when compared decontamination protocols to control with highest modification of SRP for EDTA + AC group. For decontamination analysis, for applicability of different protocols in the same type of implant, only SLA showed statistical significant difference for aPDT vs. EDTA + CA (p = 0.0114; ANOVA with post-hoc Tukey's multiple comparisons test) with lowest rCTA for aPDT, however for ACQ implants the aPDT showed lowest rCTA with no statistical difference (p > 0.05; ANOVA with post-hoc Tukey's multiple comparisons test). No statistical difference was observed between the decontamination protocols at other implant types. CONCLUSION: It can be suggested that the chemical-physical characteristics of dental implants can be effected by the process of contamination and decontamination by aPDT and chemical agents.

Clinical and patient-centered outcomes using two types of subepithelial connective tissue grafts: A split-mouth randomized clinical trial.

BACKGROUND: This split-mouth randomized clinical trial compared two different types of subepithelial connective tissue grafts (SCTG) considering clinical parameters and patient-centered outcomes in patients with bilateral recession type 1 multiple gingival recessions after 6 months postoperatively. METHODS: A total of 21 patients were surgically treated with coronally advanced flap (CAF) associated with SCTG harvested by: double blade scalpel (DBS) and de-epithelialized (DE) SCTG. Periodontal clinical parameters and esthetics were evaluated by a calibrated periodontist at baseline and after 6 months. Patient-centered outcomes related to pain/discomfort and esthetics were assessed with visual analogue scale after 7 days and 6 months, respectively. RESULTS: All clinical parameters, with the exception of probing depth, demonstrated differences in intragroup evaluation, comparing baseline to 6-month evaluation (P <0.05). Both groups presented reduction of recession depth and recession width and gain of keratinized tissue thickness, keratinized tissue width, and clinical attachment level (P <0.05). Intergroup comparison (DBS × DE) demonstrated no significant differences considering clinical parameters and periods. Both techniques improved esthetics evaluated by patients, without a difference between groups in patients and professional analysis. However, DBS group presented inferior pain/discomfort compared with DE (P <0.05). CONCLUSION: DBS and DE associated with CAF presented satisfactory clinical outcomes. However, DBS presented inferior morbidity, an important fact for decision-making process.

Deposition of Immune Complexes in Gingival Tissues in the Presence of Periodontitis and Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a complex chronic autoimmune disease characterized by tissue damage and widespread inflammation in response to environmental challenges. Deposition of immune complexes in kidneys glomeruli are associated with lupus nephritis, determining SLE diagnosis. Periodontitis is a chronic inflammatory disease characterized by clinical attachment and bone loss, caused by a microbial challenge - host response interaction. Deposition of immune complex at gingival tissues is a common finding in the course of the disease. Considering that, the primary aim of this study is to investigate the deposition of immune complexes at gingival tissues of SLE patients compared to systemically healthy ones, correlating it to periodontal and systemic parameters. Twenty-five women diagnosed with SLE (SLE+) and 25 age-matched systemically healthy (SLE-) women were included in the study. Detailed information on overall patient's health were obtained from file records. Participants were screened for probing depth (PD), clinical attachment loss (CAL), gingival recession (REC), full-mouth bleeding score (FMBS) and plaque scores (FMPS). Bone loss was determined at panoramic X-ray images as the distance from cementenamel junction to alveolar crest (CEJ-AC). Gingival biopsies were obtained from the first 15 patients submitted to surgical periodontal therapy of each group, and were analyzed by optical microscopy and direct immunofluorescence to investigate the deposition of antigen-antibody complexes. Eleven (44%) patients were diagnosed with active SLE (SLE-A) and 14 (56%) with inactive SLE (LES-I). Mean PD, CAL and FMBS were significantly lower in SLE+ than SLE-(p < 0.05; Mann Whitney). The chronic use of low doses of immunosuppressants was associated with lower prevalence of CAL >3 mm. Immunofluorescence staining of markers of lupus nephritis and/or proteinuria was significantly increased in SLE+ compared to SLE-, even in the presence of periodontitis. These findings suggest that immunomodulatory drugs in SLE improves periodontal parameters. The greater deposition of antigen-antibody complexes in the gingival tissues of patients diagnosed with SLE may be a marker of disease activity, possibly complementing their diagnosis.

Residual decontamination chemical agents negatively affect adhesion and proliferation of osteoblast-like cells on implant surface.

PURPOSE: To investigate the influence of implant surface decontaminated and uncontaminated on osteoblast-like cell adhesion and proliferation MATERIALS AND METHODS: Commercially available implants of different brands and surface characteristics were selected: Biomet 3i® Nanotite (NT) and Osseotite (OT), Straumann® SLActive (SLA), and Neodent® Acqua Drive (ACQ) and Neoporos Drive CM (CM). Physical and chemical properties of the implants were investigated by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), and wettability analysis (WETT). Implants were previously contaminated with Aggregatibacter actinomycetemcomitans strains; after that, samples were decontaminated by different chemical methods. Decontaminated (test group; n = 15/type of implant) and uncontaminated (control group; n = 5/type of implant) samples were analyzed according to the number of human osteoblastic osteosarcoma cells (Saos-2) adhered on the implant surface after 24 h and 72 h in SEM images. RESULTS: ACQ was found to be highly hydrophilic, and NT was the most hydrophobic implant. Increased variation of Saos-2 cell adhesion and proliferation were observed on all test and control groups. Controversially, at the proliferation analysis in 72 h, CM implant was the only implant that showed no significant difference between test and group (p = 0.2833; Tukey's multiple comparisons test). NT implants showed the greater value of cell proliferation when compared with all types of implant surface (p = 0.0002; Tukey's multiple comparisons test). CONCLUSIONS: These findings suggest that decontaminated surfaces were able to impair the counting of osteoblast-like cell adhesion and proliferation.

Evaluation of Regular Market Ethyl Cyanoacrylate Cytotoxicity for Human Gingival Fibroblasts and Osteoblasts.

Background: The aim of this study was to evaluate the cytotoxicity of cyanoacrylate adhesives in an indirect contact assay in human gingival fibroblast (FGH) and oral osteoblasts (GO) lineages. Methods: Cover glasses were glued with adhesives following the ISO 10993-2012 protocol. The groups were: C (control with cells and regular Dulbecco Modified Eagle Medium; LC (liquid ethyl-cyanoacrylate); GC (ethyl-cyanoacrylate gel); EGC (easy gel [ethyl-cyanoacrylate]); and D (Dermabond [octyl-cyanoacrylate]). Each cell linage was plated in the sixth passage using 104 cells. Cell viability was measured by the MTT test at 24, 48, 72, and 96 hours. Data were analyzed by two-way analysis of variance complemented by the Tukey test, with p < 0.05 being significant. Results: Dermabond stimulated osteoblast viability at 72 h (p < 0.05). All other groups were similar to the control cells (p > 0.05). For the fibroblasts, there was no difference in the groups, including the control except that EGC was cytotoxic for these cells (p < 0.05). Conclusions: Ethyl-cyanoacrylate gel and liquid forms available on the general chemical market were not cytotoxic for oral osteoblasts and fibroblasts in most cases. However, the easy gel form was cytotoxic for fibroblasts.

Experimental surgical and orthodontic extrusion of teeth in dogs.

The purpose of this study was to evaluate histologically the root surfaces of teeth submitted to orthodontic and surgical extrusion procedures in a dog model. Eighteen adult male dogs, divided into six groups of three dogs each, were used in the study. Each animal underwent two procedures: rapid orthodontic extrusion and surgical extrusion of the maxillary lateral incisors. The animals were sacrificed to produce samples at 7, 14, 45, 90, 120, and 180 days after surgery for assessment of cross sections of the coronal, medial, and apical thirds of the treated teeth. At early time points, some active surface and inflammatory resorption was observed exclusively in the surgical extrusion group; however, samples collected at later times demonstrated functional repair of the resorption gaps in both groups. Ankylosis was observed as a minor event and was apparently of a transient nature in samples of the surgical extrusion group. The results demonstrate the importance of maintaining the periodontal ligament and cementum surface; both are vital for the prevention of root resorption. It may be postulated that orthodontic extrusion is more conservative and physiologic than surgical extrusion; however, the results showed that function was restored in both groups.

A combined regenerative approach for the treatment of aggressive periodontitis: long-term follow-up of a familial case.

This article reports the longitudinal follow-up of a familial case of aggressive periodontitis treated by a combined regenerative approach that consisted of root conditioning, bone grafting, and membrane positioning. Treatment resulted in attachment level gain, reduction of probing depth, absence of bleeding on probing, and complete bone filling of the defect. The short-term results obtained after surgery were maintained after 6 years, suggesting that the combined regenerative approach is able to completely arrest the disease with long-term stability.

Blue photosensitizers for aPDT eliminate Aggregatibacter actinomycetemcomitans in the absence of light: An in vitro study.

The main treatment of periodontal disease is the mechanical removal of supra and subgingival biofilm. Adjuvant therapies as antimicrobial photodynamic therapy (aPDT) may offer improved clinical and microbiological results. The aim of this in vitro study was to evaluate the effect of toluidine and methylene blue dyes, associated with red laser and LED, on elimination of a suspension of Aggregatibacter actinomycetemcomitans (A.a). Experimental groups (n = 29) consisted of positive (broth) and negative (gentamicin) controls, three different dyes concentrations (0.05; 0.1; 10 mg/ml) alone or associated with laser (660 nm) at two power settings (70 and 100 mW) and LED (627 ±â€¯10 nm). Bacterial suspension received all treatments, and after serial dilutions they were cultured for 24 h in petri dishes for colony forming unit counts. Data were analyzed by ANOVA complemented by Tukey's test (p < 0.05). The results showed that both dyes, at a concentration of 10 mg/ml, alone or associated with laser and LED, caused 100% of death similar to the negative control (p > 0.05). It can be concluded that blue dyes for aPDT, at high concentration (10 mg/ml), are capable of eliminating A.a without adjuvant use of light sources.

Root coverage stability with acellular dermal matrix in multiple gingival recessions in esthetic zone: A clinical case report with 12-year follow-up.

The importance of surgical intervention for the maintenance of long-term results by root coverage of multiple gingival recessions in an esthetic area treated with a tissue substitute (acellular dermal matrix [ADM]-Alloderm®) is widely required. The present case report highlights the effectiveness of the ADM for the treatment of multiple recession defects in a female patient with Class I and II gingival recession in relation to maxillary anterior associated with esthetics and dentin hypersensitivity demands. The root coverage rate for the anterior area showed greater value with 70% of coverage; at 90 days and 2 and 12 years of follow-up, it showed 70.5%, 79%, and 77%, respectively. Conversely, for the posterior area, these rates were 68.5%, 63%, and 57% for the same follow-up periods. Results regarding gain of keratinized tissue demonstrated superior values for the anterior area, namely 3.92, 3.34 and 3.22 mm at 90 days and 2 and 12 years. These values for the posterior area were 0.54 mm, 2.41, and 1.87 mm, respectively. These findings suggest that the tissue substitute Alloderm® showed excellent long-term results for areas of multiple recessions, providing root coverage and stability of keratinized tissue gain. After this long period of evaluation, some local factors associated with recurrence of gingival recessions were detected, and a lack of proper periodontal maintenance care was observed. However, esthetic and functional outcomes achieved with surgical procedure were maintained.

Root surface demineralization by citric acid/tetracycline gel and aPDT associated to subepithelial connective tissue graft improves root coverage outcomes. A 12-month preliminary randomized clinical trial.

Root conditioners are used to promote root surface biomodification to increase the success rate of root coverage. Citric acid and tetracycline are commonly used. There is recent indication for using antimicrobial photodynamic therapy (aPDT) with this purpose. The aim of this study is to evaluate the effects of citric acid/tetracycline gel and aPDT in root coverage of gingival recessions using subepithelial connective tissue graft. This parallel, double-blinded clinical trial enrolled 17 patients (60 recession defects; 20/group). Experimental groups were: Control group (SRP) - scaling and root planing only; Citric acid/tetracycline gel (CAT) group - SRP plus citric acid/tetracycline gel; aPDT - SRP, toluidine blue O (100 µl/ml) and red laser. At baseline and after 3, 6 and 12 months, the clinical parameters were evaluated: recession depth (RD), percentage of root coverage (%RC), keratinized tissue width (KTW), soft tissue thickness (STT), probing depth (PD), clinical attachment level (CAL), dentin hypersensitivity (HYPER) and esthetic perception by patient (EST). CAT group presented reduction in PD, CAL, RD, increase in KTW and STT, higher %RC (81.6%) and better esthetic in relation to SRP group (p < .05). aPDT treatment promoted CAL reduction, gain of KTW and STT and higher %RC (82.1%) in relation to SRP (57.7%) (p < .05). There was a reduction in dentin sensitivity in all groups. Complete root coverage was higher for CAT group (65%) and aPDT group (70%) in relation to SRP (30%) (p < .05). Root conditioning, with citric acid/tetracycline gel and aPDT, promotes better long-term clinical outcomes and root coverage after SCTG procedures.

Clinical parameters, histological analysis, and laser Doppler flowmetry of different subepithelial connective tissue grafts.

Subepithelial connective tissue graft (SCTG) presents favorable outcomes. However, the harvesting technique can influence the anatomical and histological composition of the SCTG. Within the limitations of a case report, the behavior of SCTGs removed by two techniques was evaluated bilaterally in one patient using double blade scalpel (DBS) and de-epithelialized graft (DE). Clinical parameters, laser Doppler flowmetry (LDF) and histological analysis were assessed. Complete root coverage was observed bilaterally, as well as improvement in width and thickness of keratinized tissue 2 years postoperatively. The LDF analysis demonstrated better revascularization in the DBS recipient area compared to DE. The histological evaluation showed differences in tissue composition and organization of collagen fibers. Similar clinical outcomes were observed bilaterally, nevertheless greater morbidity and aesthetic was reported in the DE harvesting area.

Isolation and characterization of progenitor cells from surgically created early healing alveolar defects in humans: A preliminary study.

BACKGROUND: The granulation tissue present in surgically-created early healing sockets has been considered as a possible source of osteoprogenitor cells for periodontal regeneration, as demonstrated in animal studies. However, the in vitro osteogenic properties of tissue removed from human surgically-created early healing alveolar defects (SC-EHAD) remains to be established, being that the aim of this study. METHODS: Surgical defects were created in the edentulous ridge of two systemically healthy adults. The healing tissue present in these defects was removed 21 days later for the establishment of primary culture. The in vitro characteristics of the cultured cells were determined by Armelin method, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, immunohistochemistry, alkaline phosphatase (ALP) activity, mineralization assay, and flow cytometry for detection of stem cells/osteoprogenitor cell markers. RESULTS: Cells were able to adhere to the plastic and assumed spindle-shaped morphology at earlier passages, changing to a cuboidal one with increasing passages. Differences in the proliferation rate were observed with increasing passages, suggesting osteogenic differentiation. ALP and mineralization activities were detected in conventional and osteogenic medium. Fresh samples of SC-EHAD tissue exhibited CD34- and CD45- phenotypes. Cells at later passages (14th) exhibited CD34- , CD45- , CD105- , CD166- , and collagen type I+ phenotype. CONCLUSION: Tissue removed from SC-EHAD is a possible source of progenitor cells.

Cross-sectional evaluation of the presence of gingival recession in individuals with cleft lip and palate.

BACKGROUND: In dentistry, investigations in the field of periodontics in individuals with cleft lip and palate are scarce, with few studies on the prevalence, incidence, extension, and severity of periodontal alterations and their treatment in adults. METHODS: This study analyzed the prevalence, extension, and severity of recessions and the relationship with some etiologic factors in 200 individuals with cleft lip and palate aged 13 to 56 years. Examination comprised measurements of gingival recession, amount of keratinized mucosa, frenum insertions, and tooth positioning in the dental arch. RESULTS: Analysis of the results revealed that the prevalence of recession was high, because 75% of individuals in the sample had at least one tooth with gingival recession; 100% of individuals aged >43 years presented at least one recession, and most recessions were Miller Class I. The multiple linear regression demonstrated that recession is directly proportional to age (i.e., it increases with age); concerning the amount of keratinized mucosa, the proportion is inverse (i.e., the larger the recession, the smaller the amount of mucosa); and gingival inflammation also presented a positive correlation. CONCLUSIONS: Gingival recessions are caused by many etiologic factors, which usually act in combination. Thus, all factors causing recession should be analyzed so that planning and treatment of this clinical condition may be established, for achievement of optimal outcomes.

Stimulation of human gingival fibroblasts viability and growth by roots treated with high intensity lasers, photodynamic therapy and citric acid.

OBJECTIVE: The aim of this study was to compare the effect of root biomodification by lasers, citric acid and antimicrobial photodynamic therapy (aPDT) on viability and proliferation of human gingival fibroblasts (FGH). DESIGN: Groups were divided in control (CC - only cells), and root fragments treated by: scaling and root planing (positice control - SC), Er:YAG (ER-60mJ,10pps,10Hz,10s,2940nm), Nd:YAG (ND-0.5W,15Hz,10s,1640nm), antimicrobial photodynamic therapy (PDT-InGaAIP,30mW,45J/cm2,30s,660nm,toluidine blue O), citric acid plus tetracycline (CA). Fibroblasts (6th passage, 2×103) were cultivated in a 24-h conditioned medium by the treated root fragments. Cell viability was measured by MTT test at 24, 48, 72 and 96h. In a second experiment, FGH cells (104) were cultivated on root fragments which received the same treatments. After 24, 48, 72h the number of cells was counted in SEM pictures. In addition, chemical elements were analyzed by energy dispersive spectroscopy (EDS). Data was analyzed by two-way ANOVA (first experiment), repeated measures ANOVA (second experiment) and ANOVA (EDS experiment) tests complemented by Tukey's test (p<0.05). RESULTS: ND, PDT and CA promoted higher cell viability (p<0.05). ND and ER groups presented higher number of cells on root surfaces (p<0.05). ER group presented higher calcium and CA group a higher carbon percentages (p<0.05). CONCLUSIONS: All treatments but scaling and root planing stimulated fibroblast viability while Er:YAG and Nd:YAG treated root surfaces presented higher number of cells.