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.

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.

Cyanoacrylate Adhesive as an Alternative Tool for Membrane Fixation in Guided Tissue Regeneration.

AIM: To report a well succeeded use of cyanoacrylate adhesive for fixating a resorbable membrane during a guided tissue regeneration procedure (GTR). BACKGROUND: The immobilization of membranes in GTR is essential for establishing proper environment for cell differentiation and tissue regeneration. However, some membranes are very difficult to be kept in position by sutures and its fixation by mini screws or pins may be time consuming and expensive. CASE DESCRIPTION: A 47-year-old woman presenting a vertical bone defect at the palatal aspect of the left central incisor was treated by GTR using particulate autogenous bone graft associated to a collagen membrane. The membrane was glued to the bone surrounding the defect and to the tooth surface with cyanoacrylate adhesive. The postoperative period was uneventful and 4 years later, excellent results in terms of radiographic filling of the defect and reduction of the probing depth were seen. For illustrative purposes, histological findings obtained during a previous experiment in calvaria of guinea pigs is shown, characterizing a foreign body granuloma and proving that the cyanoacrylate adhesive is a safe tool in GTR. CONCLUSION: The use of a membrane glued with cyanoacrylate to immobilize membranes in GTR is viable and safe from both technical and biological standpoints and may be advantageous for clinical and research purposes. CLINICAL SIGNIFICANCE: The alternative method for membrane fixation shown in this case report can contribute to simplify the technique in GTR procedures.

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.

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.

Impact of Subepithelial Connective Tissue for Root Coverage on Brazilian Patients' Quality of Life: A Longitudinal Clinical Study.

Aims: Gingival recession has been associated with dentin hypersensitivity and aesthetic impairment. The impact of gingival recession and periodontal surgical procedures on adult patients' quality of life are scarce. The aim of this study was to evaluate the quality of life of patients submitted to root coverage procedures with subepithelial connective tissue grafts and coronally advanced flap. Materials and methods: Patients were asked to use a numerical rating scale to classify their dentin hypersensitivity, aesthetics, pain/discomfort, chewing, and brushing abilities in gingival recession sites treated with subepithelial connective tissue grafts plus coronally advanced flap. The patients answered a self-administered questionnaire about quality of life-related to oral health (OHIP-14) after 7, 14, 30, 90, and 180 days. Descriptive statistics were used to synthesize the data recorded. Results: Mean percentage of root coverage was positively related to OHIP-14 (dimension 2- physical pain) in 90 days postoperatively. The quality of life (OHIP-14 total score) significantly improved from baseline to 90 and 180 days postoperatively. The numerical rating score analysis revealed significant improvement in the chewing and brushing abilities when period of 7 days was compared to 90 and 180 days and from 14 to 180 days. Conclusions: Root coverage procedures with subepithelial connective tissue grafts plus coronally advanced flap result in a positive effect on adult patients' quality of life.

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.

The concentration of citric acid as dental root conditioner influences the behavior of fibroblasts from human periodontal ligament.

OBJECTIVE: This study investigated the behavior of fibroblasts from human periodontal ligament (hPLF) cultured on dental roots subjected to different protocols of citric acid conditioning. MATERIALS AND METHODS: 32 human teeth extracted due to advanced periodontal disease provided 63 radicular fragments, which were randomly divided in groups according to the treatment given to the surface: rinsing with saline solution for 90 s (C), 10 % citric acid (CA10), or 50 % citric acid (CA50). The treatments were applied during 90 s, 120 s and 180 s (n = 9). hPLF were cultured for 24, 48 and 72 h (n = 3) on the treated samples and analyzed by scanning electron microscopy (SEM) for surface area covered by cells and dentinal tubules widening. RESULTS: Excepting group C, all the other groups showed almost complete coverage of root surface by hPLF with time. At 24 h of cell culture, the largest area of coverage was seen in the samples treated with CA10-90 (98 ± 0.89 %) at 24 h of cell culture and this difference was significant (p < 0.05) in comparison to CA10-180 (84.04 ± 5.01 %), CA50-90 (63.28 ± 12.46 %), CA50-180 (56.59 ± 8.76 %) and C (0.06 ± 0.11 %). In all the other comparisons, there was no statistically significant differences between CA10 and CA50 (p > 0.05). Cells grown on surfaces treated with CA10 were more spread and flatten than in the CA50 specimens. CONCLUSIONS: Periodontally compromised roots surfaces conditioned with 10 % citric acid for 90 s resulted in better substrate for hPLF proliferation, in initial periods of culture than 50 % citric acid. The enlargement of the dentinal tubules did not seem to be influenced by the acid concentration.

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.

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.

Periapical Lesion on an Implant after Socket Shield Technique: A Case Report.

A 47-year-old Caucasian male presented with a radiolucent area around the apical region of an implant placed using the socket shield technique. A second surgical procedure was performed to curette the lesion and fill the defect with a xenogeneic bone graft. Twenty months after implant placement and 10 months after the second surgery, there was no sign of recurrence of the lesion and radiographic evaluation was consistent with new bone formation in the region. Thus, although numerous studies have demonstrated the effectiveness of the socket shield technique, this case report illustrates the need for further randomized clinical studies for a better understanding of the clinical complications and indications for the technique.

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.

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.

Bone demineralization promotes superior spread of preosteoblast in culture.

Previous studies have shown substances capable of similar effects of demineralization, accelerating the process of bone remodeling. This study investigated preosteoblasts behavior in cell culture after bone demineralization with citric acid and tetracycline. Seventy-four Wistar rats provided 144 calvarial bone samples, 126 of which were randomly divided in seven groups according to the treatment given to the surface: no demineralization (C), citric acid (CA), tetracycline (TCN) during 15, 30, and 60 s. Each group received preosteoblasts cultured for 24, 48, and 72 hr. Eighteen remaining samples were analyzed for the atomic percentage (A%) by energy dispersive spectroscopy (EDS) before and after demineralization. The average percentage of bone area covered by cells increased with time and it was significantly higher after 24 and 48 hr of culture in groups CA15s, CA30s, CA60s, TCN15s, and TCN30s than in groups TCN60 and C (p < 0.05). The cell morphology in all CA and TCN groups was shown to be compatible with more advanced stages of differentiation than in C group. The A% changed after demineralization. We conclude that demineralization with citric acid or tetracycline for 15-30 s increased the area of bone surface covered by preosteoblasts. The A% changes were not sufficient to impair the cells spreading and morphology. Bone demineralization may promote potential benefits in bone regenerative procedures. HIGHLIGHTS: Low pH effects did not interfere on cell growth. Bone demineralization favored the preosteoblasts growth. A possible alternative to improve graft consolidation.

Candida albicans-Cell Interactions Activate Innate Immune Defense in Human Palate Epithelial Primary Cells via Nitric Oxide (NO) and ß-Defensin 2 (hBD-2).

The presence of Candida albicans in the biofilm underlying the dental prosthesis is related to denture stomatitis (DS), an inflammatory reaction of the oral mucosa. The oral epithelium, a component of the innate immune response, has the ability to react to fungal invasion. In this study, we evaluated the in vitro effect of viable C. albicans on the apoptosis, nitric oxide (NO) production, and ß-defensin 2 (hBD-2) expression and production of human palate epithelial cells (HPECs). We further determined whether or not these effects were correlated with fungal invasion of epithelial cells. Interaction between HPEC primary culture and C. albicans was obtained through either direct or indirect cell-cell contact with a supernatant from a hyphal fungus. We found that the hyphae supernatants were sufficient to induce slight HPEC apoptosis, which occurred prior to the activation of the specific mechanisms of epithelial defense. The epithelial defense responses were found to occur via NO and antimicrobial peptide hBD-2 production only during direct contact between C. albicans and HPECs and coincided with the fungus's intraepithelial invasion. However, although the hBD-2 levels remained constant in the HPEC supernatants over time, the NO release and hBD-2 gene expression were reduced at a later time (10 h), indicating that the epithelial defense capacity against the fungal invasion was not maintained in later phases. This aspect of the immune response was associated with increased epithelial invasion and apoptosis maintenance.

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.

Effects of TGF-beta1, PDGF-BB, and IGF-1 on the rate of proliferation and adhesion of a periodontal ligament cell lineage in vitro.

BACKGROUND: Considering the role of growth factors in periodontal regeneration, the aim of this study was to evaluate the influence of platelet-derived growth factor (PDGF)-BB, insulin-like growth factor (IGF)-1, and transforming growth factor-beta 1 (TGF-beta1), alone or in combination, on the rate of proliferation and adhesion of periodontal ligament (PDL) cells in vitro. METHODS: After establishment and characterization of a primary culture of PDL cells, 72 culture dishes were plated with 10(3) cells distributed among four test groups and a control group. Test groups had PDGF-BB, TGF-beta1, IGF-1, or a combination of all three added to the culture medium, whereas the control group received no growth factor. The samples were counted in triplicate 1, 3, 5, and 7 days after seeding. For the adhesion assay, 14 patients provided 30 root fragments distributed among 10 groups: scaling and root planing (SRP), SRP + growth factors, SRP + citric acid plus tetracycline (CA+T), and SRP + (CA+T) + growth factors. The data were evaluated statistically by analysis of variance complemented by Tukey, Dunnett, and Student-Newman-Keuels methods. RESULTS: Maximum rates of proliferation were observed at day 3 for all groups. TGF-beta1 induced a 344.17% +/- 58.80% increased proliferation rate over control (P < 0.05), followed by the combination (277.5% +/- 29.38%), PDGF-BB (238.79% +/- 5.79%), and IGF-1 (233.16% +/- 19.19%). Groups treated by (CA+T) showed increased numbers of cells attached to root fragments, especially SRP + (CA+T) + combination (13.25 +/- 1.79), with significant differences (P < 0.05) from groups treated only by SRP. CONCLUSION: This combination of growth factors stimulated a mitogenic response and favored the adhesion of PDL cells in vitro, suggesting its possible role in periodontal regeneration.