Investigating the diagnostic potential of IL-1ß, IL-10, and IL-36γ in gingival crevicular fluid in patients with different periodontal conditions.

BACKGROUND: The study aimed to analyze cytokine levels, including interleukin (IL)-1ß, IL-10, and IL-36γ, to investigate the link between pro- and anti-inflammatory responses in periodontal conditions and assess their potential as diagnostic biomarkers for distinguishing between different types of periodontal conditions. METHODS: 80 systemically healthy non-smokers (25 periodontally healthy, 25 with gingivitis, 30 with periodontitis) were included. Clinical periodontal parameters were recorded, and gingival crevicular fluid (GCF) samples were obtained. Receiver operating characteristic (ROC) curve analysis was applied to determine the diagnostic value of cytokines. RESULTS: IL-36γ had the highest sensitivity for diagnosing periodontitis, although its specificity for identifying those without periodontitis was relatively low. The combination of IL-1ß and IL-36γ was the most effective in differentiating periodontitis from periodontal health. IL-10 was found to be an acceptable discriminator for distinguishing gingivitis from healthy conditions. However, its sensitivity and specificity for identifying gingivitis were lower. The combination of the three cytokines showed the highest ability to distinguish between periodontitis and gingivitis. CONCLUSION: The levels of IL-1ß, IL-10, and IL-36γ in GCF may provide insights into periodontal health and disease status. Further studies are needed to validate these results and explore the potential of these cytokines in periodontal disease management.

All three of these cytokines exhibit exceptional diagnostic accuracy, particularly in distinguishing between chronic periodontitis and periodontal health.Moreover, the combination of IL-1ß and IL-36γ stands out as the most accurate diagnostic indicator for periodontitis. This combination could serve as a robust biomarker panel for the early detection and monitoring of periodontal disease, potentially allowing for timely interventions to prevent disease progression.

Netrin-1 and its receptor Unc5b as markers of periodontal disease.

OBJECTIVES: The aim of this present study was to evaluate the levels of Netrin-1 and Unc5b in periodontal health and disease. BACKGROUND: Netrin-1, acting via its receptor UNC5b, regulates the inflammatory response and takes apart in bone destructive diseases. METHODS: Samples of gingival crevicular fluid (GCF), whole saliva, and serum were taken from systemically healthy, nonsmoking 20 periodontitis , 20 gingivitis patients, and 20 periodontally healthy subjects at baseline and 4 weeks after nonsurgical periodontal treatment (NSPT). Whole-mouth and site-specific clinical periodontal parameters were recorded. Netrin-1 and Unc5b levels were measured by enzyme-linked immunosorbent assay. Data were analyzed by nonparametric tests. RESULTS: Total amount of Netrin-1 in GCF was significantly higher in periodontitis than the others, and the levels were significantly reduced after NSPT. Salivary and serum concentrations of Netrin-1 were significantly different among the study groups (P = .000), and NSPT significantly increased the concentration levels of both salivary and serum Netrin-1 (P < .05). Healthy subjects had significantly lower GCF (P = .001) and conversely, higher salivary and serum levels of Unc5b than the other groups (P = .002). The GCF levels of Unc5b were significantly reduced (P < .01), and conversely, serum concentrations were significantly increased after NSPT (P < .05). GCF Netrin-1 and Unc5b total amounts were positively correlated with clinical parameters (P < .01 and P < .05) whereas salivary Netrin-1 and Unc5b concentrations were negatively correlated with clinical parameters (P < .01 and P < .05). CONCLUSIONS: The results of this study indicate that Netrin-1 and its receptor Unc5b may have essential roles in periodontal inflammation and those can be assumed as useful therapeutic agent to control inflammation and periodontal breakdown.

Clinical importance of neuregulin-4 and its receptor ErbB4 in periodontal disease pathogenesis.

OBJECTIVE: The aim of this clinical trial was to evaluate the levels of Neuregulin-4 (Nrg4), Erb-b2 receptor tyrosine kinase 4 (ErbB4), interleukin (IL)-6, IL-10, nitric oxide synthase (NOS)-2, and arginase (Arg)-1 in periodontal health and disease. MATERIALS AND METHODS: This study includes systemically healthy 20 periodontally healthy (H), 20 gingivitis (G), 20 stage II periodontitis (P1), and 20 stage III periodontitis (P2) subjects. Periodontal clinical measurements and samples of gingival crevicular fluid (GCF) and serum were obtained at baseline and 4 weeks after non-surgical periodontal treatment (NSPT). Enzyme-linked immunosorbent assay (ELISA) was used to determine ErbB4, Nrg4, IL-6, IL-10, NOS2, and Arg1 levels in all samples. RESULTS: GCF ErbB4 and Nrg4 total amounts and IL-6/IL-10 ratio were significantly higher in G, P1, and P2 groups than H group. Serum NOS2 levels were significantly lower, whereas serum Arg1 levels were higher in H group than the others. The GCF levels of ErbB4 and Nrg4 were significantly decreased after NSPT in G, P1, and P2 groups. Additionally, the GCF levels of ErbB4 and Nrg4 were positively correlated with all clinical parameters and IL-6/IL-10 ratio. CONCLUSIONS: Nrg4 and its receptor ErbB4 might have crucial roles in the pathogenesis of periodontal disease. These results should be verified with future prospective studies to further clarify the exact role of those biomarkers.

The effect of different cleaning methods on the surface and temperature of failed titanium implants: an in vitro study.

The aims of this in vitro study are to compare the efficacy of different cleaning methods in removing debris of failed implants and to detect thermal changes of the implants treated by various scaling instruments. Twenty-seven failed implants and two unused implants as control were included to this study-group 1: plastic curette (P), group 2: titanium curette (T), group 3: carbon curette (C), group 4: titanium brush (TB), group 5: Er:YAG laser (laser 1 (L1) 100 mJ/pulse at 10 Hz), group 6: Er:YAG laser (laser 2 (L2) 150 mJ/pulse at 10 Hz), group 7: Er:YAG laser (laser 3 (L3) 200 mJ/pulse at 10 Hz), group 8: ultrasonic scaler appropriate for titanium (US), group 9: air abrasive method (AA) + citric acid, and group 10: implantoplasty (I). The changes on the treated/untreated titanium surfaces and remnant debris were observed by scanning electron microscopy (SEM). Temperature of the implants before and after treatment was detected using a thermocouple. The use of air abrasive and citric acid combination and Er:YAG laser groups was found as the best methods for the decontamination of titanium surfaces of failed implant. When the hand instruments were compared, titanium curette was found better than both the plastic and the carbon curettes which leave plastics and carbon remnants on the titanium surface. The temperature was higher after hand instrumentation when compared to other experimental groups (p < 0.05). Within the limitations of the present in vitro model, it can be concluded that the best method for decontamination of the implant surface is the use of air abrasives and Er:YAG laser.

Biostimulation with diode laser positively regulates cementoblast functions, in vitro.

The aim of this study was to evaluate the effects of diode laser biostimulation on cementoblasts (OCCM.30). A total of 40 root plates were obtained from healthy third molar teeth and assigned to the following two groups: (1) control group and (2) laser-treated group. Root plates were placed into the cell culture inserts, and OCCM.30 cells were seeded onto root plates. Cells were irradiated with a low level of diode laser (power: 0.3 W in continuous wave, 60 s/cm2). Proliferation and mineralized tissue-associated gene's and BMP's messenger RNA (mRNA) expressions of cementoblasts were evaluated. Total RNAs were isolated on day 3 and integrin-binding sialoprotein (Ibsp), bone gamma-carboxyglutamate protein (Bglap), Type I collagen (Col1a1), osteoblastic transcription factor, runt-related transcription factor (Runx2), and Bone Morphogenetic Protein (BMP)-2, 3, 4, 6, and 7 mRNA expressions were determined using quantitative RT-PCR. von Kossa staining was performed to evaluate biomineralization of OCCM.30 cells. In the proliferation experiment, while there was no significant difference until 96 h, laser irradiation retarded the decrease in cell proliferation trend after 96 h compared to the untreated control group. Statistically significant increase in Ibsp, Bglap, and BMP-2,3,6,7 mRNA expressions were noted in the laser groups when compared to the untreated control group (p < 0.05). Laser irradiation induced mineralized nodule formation of cementoblasts. The results of this study reveal that the biostimulation setting of diode laser modulates the behavior of cementoblasts inducing mineralized tissue-associated gene's mRNA expressions and mineralization. Therefore, biostimulation can be used during regenerative periodontal therapies to trigger cells with periodontal attachment apparatus.

Nonsurgical periodontal therapy with/without diode laser modulates metabolic control of type 2 diabetics with periodontitis: a randomized clinical trial.

In order to evaluate whether nonsurgical periodontal treatment with/without diode laser (DL) decontamination improves clinical parameters, the levels of IL-1ß, IL-6, IL-8, intercellular adhesion molecule (ICAM), and vascular cell adhesion molecule (VCAM) in gingival crevicular fluid and metabolic control (HbA1c) in chronic periodontitis (CP) patients with diabetes mellitus type 2 (DM2). Sixty patients with DM2 and CP were randomly assigned into two groups to receive scaling and root planing (SRP, n = 30) or SRP followed by diode laser application (SRP + DL, n = 30). Clinical periodontal and gingival crevicular fluid (GCF) parameters were assessed at baseline, 1, and 3 months after periodontal treatment. HbA1c levels were evaluated at baseline and 3 months post-therapy. Total amounts of cytokines and molecules were analyzed by ELISA. Nonsurgical periodontal treatment with/without DL appeared to improve clinical, biochemical parameters, and glycemic control in DM2 patients (BMI < 25 kg/m(2)) with CP. The SRP + DL group provided better reductions in probing depth (PD) and clinical attachment level (CAL) parameters compared to the SRP group (P < 0.05). Significant reductions were found in the total amounts of GCF levels of IL-1, IL-6, IL-8, ICAM, and VCAM after treatment (P < 0.05). HbA1c levels decreased significantly at 3 months after treatment (P < 0.05). SRP + DL reduced HbA1c levels more significantly compared to SRP alone (0.41 vs. 0.22 %, P < 0.05). SRP, especially in combination with DL, shows improvement of glycemic control for DM2 patients with CP.

Clinical and biochemical effects of diode laser as an adjunct to nonsurgical treatment of chronic periodontitis: a randomized, controlled clinical trial.

The aim of this randomized, parallel, controlled clinical trial was to examine the clinical and biochemical efficacy of diode laser as an adjunct to scaling and root planing (SRP). Thirty chronic periodontitis patients were randomly assigned into two groups to receive SRP alone (control) or SRP followed by diode laser (test). Plaque index, gingival index, bleeding on probing, probing depth, and clinical attachment level were measured at baseline and at 1, 3, and 6 months after treatment. The gingival crevicular fluid levels of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-8 (IL-8), matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-8 (MMP-8) and tissue inhibitor matrix metalloproteinase-1 (TIMP-1) were analyzed by enzyme-linked immunosorbent assay. Test group showed significantly a better outcome compared to the control group in full-mouth clinical parameters. MMP-1, MMP-8, and TIMP-1 showed significant differences between groups after treatment compared to baseline (p < 0.05). The total amount of IL-1ß, IL-6, MMP-1, MMP-8, and TIMP-1 decreased (p < 0.05) and IL-8 increased after treatment in both test and control groups (p < 0.05). Diode laser provided significant improvements in clinical parameters and MMP-8 was significantly impacted by the adjunctive laser treatment at first month providing an insight to how lasers can enhance the outcomes of the nonsurgical periodontal therapy.

Characterization of Klebsiella pneumoniae strains isolated from urinary tract infections: detection of ESBL characteristics, antibiotic susceptibility and RAPD genotyping.

In this study, a hundred Klebsiella pneumoniae strains isolated from urinary tract infections were evaluated in terms of genotyping, susceptibility to certain antibiotics and detection of extended spectrum of beta lactamase (ESBL) production. The random amplified polymorphic DNA (RAPD-PCR) method was used to identify the genetic differentiation of K. pneumoniae isolates. A total of 26 different DNA bands ranging between 334 bp and 28033 bp were detected among the strains. It was found that 100 K. pneumoniae strains revealed 11 different RAPD profiles. Antibiotic susceptibility tests were conducted using a disc diffusion method against 16 antibiotics. Fifty-five different resistance profiles were determined among the strains. ESBL-productions of the strains were determined by the double disc synergy test (DDST) and ESBL E-test methods. ESBL production rates among the strains were found to be 55% by E-test method and 45% by DDST method. While ESBL-producing K. pneumoniae strains showed the greatest resistance to penicillin G (100%), followed by piperacillin (92.7%) and erythromycin (85.4%),the resistance rates of non ESBLproducing strains to those antibiotics were determined as 97.8%, 88.8% and 88.8%, respectively. Both groups of strains showed the highest sensitivity to meropenem. Based on the results obtained from the study, it was concluded that the detection of ESBL-producing strains by the E-test method was more sensitive than by the DDST method. Phenotypic and genotypic identification methods should be used together to detect ESBL presence. The RAPD-PCR method alone will not be adequate in the genotyping of the strains and alternative DNA-based methods should be used.

Nail Mineral Composition Changes Do Not Reflect Bone Mineral Changes Caused by Boron Supplementation.

Nails have been found to be a non-invasive and readily available tissue whose mineral content can change because of a change in dietary mineral intake. Thus, this study was undertaken to determine whether boron (B) supplementation would change the concentrations of some mineral elements in nails and whether these changes correlated with changes induced in bone. Female New Zealand White rabbits (aged 8 months, 2-2.5 kg weight) were fed a grain-based, high-energy diet containing 3.88 mg B/kg. The rabbits were divided into four treatment groups: controls receiving no supplemental B (N: 7; C) and three groups supplemented with 30 mg B/L in drinking water as borax decahydrate (Na2B4O7∙10H2O, N: 10; BD), borax anhydrous (Na2B4O7, N: 7; Bah), and boric acid (H3BO3, N: 7; BA). Boron, calcium (Ca), copper (Cu), iron (Fe), magnesium (Mg), phosphorus (P), potassium (K), sodium (Na), sulfur (S), and zinc (Zn) concentrations in nails were determined by inductively coupled plasma atomic emission spectroscopy. Parametric and non-parametric multiple group comparisons and post hoc tests were performed and whether a correlation between nail and tibia and femur mineral elements concentrations were determined. A p-value of < 0.05 was considered statistically significant. Boron was not detectable in control nails but was found in the nails of the three B supplemented groups. Boron supplementation markedly increased the Ca concentration in nails with the effect greatest in the BA and BD groups. The P and Mg concentrations also were increased by B supplementation with the effect most marked in the BA group. In contrast, B supplementation decreased the Na concentration with the effect most noticeable in the BD and Bah groups. The Zn concentration in nails was not affected by BA and BD supplementation but was decreased by Bah supplementation. Boron supplementation did not significantly affect the concentrations of Cu, Fe, Mo, K, and S in nails. No meaningful significant correlations were found between nail mineral elements and tibia and femur mineral elements found previously. Nails can be an indicator of the response to boron supplementation but are not useful to indicate changes in mineral elements in bone in response to B supplementation.

Borate and boric acid supplementation of drinking water alters teeth and bone mineral density and composition differently in rabbits fed a high protein and energy diet.

The reported beneficial effects of boron on mineralized tissues in animals and humans vary. Thus, a study was performed to assess whether the variability was the result of different forms of boron supplementation, method of supplementation, and increased adiposity of the rabbit experimental model. Thirty-one female New Zealand White rabbits, (aged 8 months, 2-2.5 kg weight) were fed a grain-based high energy diet containing 11.76 MJ/kg (2850 kcal/kg) and 3.88 mg boron/kg. The rabbits were randomly divided into four treatment groups: Control group was not supplemented with boron (n:7; C), and three groups supplemented with 30 mg boron/L in drinking water in the forms of borax decahydrate (Na2O4B7 10H2O, n:10; BD), borax anhydrous (Na2O4B7, n:7; Bah) or boric acid (H2BO3, n:7; BA). Cone beam micro computed tomographic (micro-CT), histological and elemental analysis was used to evaluate the bones/teeth. Results of the experiments demonstrated that boron supplementation had beneficial effects on mineralized tissue but varied with the type of treatment. Mineral density of the femur was increased by the Bah and BA treatments (p < 0.001), but only BA increased mineral density in the tibia (p = 0.015). In incisor teeth, mineral density of dentin was increased by all boron treatments (p < 0.001), and mineral density of enamel was increased by the BD and Bah treatments. Mineral analysis found that all boron treatments increased the boron concentration in tibia and femur. In the tibia, both the BD and Bah treatments decreased the iron concentration, and the BD treatment decreased the magnesium concentration. Sodium and zinc concentrations in the tibia were decreased by the Bah and BA treatments. The boron treatments did not significantly affect the calcium, copper, molybdenum, potassium phosphorus, and sulfur concentrations. The findings show that boron supplementation can have beneficial effects on mineralized tissues in an animal model with increased adiposity, which is a model of increased inflammatory stress. However, this effect varies with the form of boron supplemented, the method of supplementation, and the mineralized tissue examined.

Effects of root planing procedures with hand instrument or erbium, chromium:yttrium-scandium-gallium-garnet laser irradiation on the root surfaces: a comparative scanning electron microscopy study.

The purpose of this study was to investigate the efficiency of hand instrumentation and laser irradiation on calculus removal from the root surfaces, in vitro. Thirty-two human teeth, extracted for periodontal reasons, were used in this study. Root surfaces of single-rooted teeth were treated by different methods including (1) conventional hand instruments; (2) hand instruments and tetracycline-hydrochloride (Tet-HCl); (3) erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation, setting I (short pulse); (4) Er,Cr:YSGG laser irradiation, setting II (long pulse). Three premolar teeth, extracted for orthodontic reasons, served as control. The morphology of the root surfaces was evaluated by light and scanning electron microscopy. Energy dispersive X-ray (EDX) analysis was performed to compare the mineral content of root surfaces treated with hand instrumentation and lasing procedures. The results of this study demonstrated that all treatments were efficient in calculus removal from the root surfaces. Thermal changes, including melting and carbonization, were not observed in either lasing procedure. The surface was rougher in the laser groups than in the groups treated with hand instruments. Moreover, roughness was greater in the long-pulse laser setting than in the short-pulse setting. While increased calcium (Ca) and decreased phosphate (P) (weight concentration percent) were observed in all treatments when compared with the control, laser procedures resulted in a more similar mineral content than in the groups treated with hand instruments. Based on these findings, laser procedures, when used in appropriate settings, are capable of performing scaling and root planing in the treatment of periodontitis. It may be concluded that short pulse laser may be more suitable for the micro-morphology of the root surface. However, additional in vitro and clinical studies are necessary to clarify the success of laser in periodontal therapy.

Dietary boron does not affect tooth strength, micro-hardness, and density, but affects tooth mineral composition and alveolar bone mineral density in rabbits fed a high-energy diet.

The objective of this study was to determine whether dietary boron (B) affects the strength, density and mineral composition of teeth and mineral density of alveolar bone in rabbits with apparent obesity induced by a high-energy diet. Sixty female, 8-month-old, New Zealand rabbits were randomly assigned for 7 months into five groups as follows: (1) control 1, fed alfalfa hay only (5.91 MJ/kg and 57.5 mg B/kg); (2) control 2, high energy diet (11.76 MJ and 3.88 mg B/kg); (3) B10, high energy diet + 10 mg B gavage/kg body weight/96 h; (4) B30, high energy diet + 30 mg B gavage/kg body weight/96 h; (5) B50, high energy diet + 50 mg B gavage/kg body weight/96 h. Maxillary incisor teeth of the rabbits were evaluated for compression strength, mineral composition, and micro-hardness. Enamel, dentin, cementum and pulp tissue were examined histologically. Mineral densities of the incisor teeth and surrounding alveolar bone were determined by using micro-CT. When compared to controls, the different boron treatments did not significantly affect compression strength, and micro-hardness of the teeth, although the B content of teeth increased in a dose-dependent manner. Compared to control 1, B50 teeth had decreased phosphorus (P) concentrations. Histological examination revealed that teeth structure (shape and thickness of the enamel, dentin, cementum and pulp) was similar in the B-treated and control rabbits. Micro CT evaluation revealed greater alveolar bone mineral density in B10 and B30 groups than in controls. Alveolar bone density of the B50 group was not different than the controls. Although the B treatments did not affect teeth structure, strength, mineral density and micro-hardness, increasing B intake altered the mineral composition of teeth, and, in moderate amounts, had beneficial effects on surrounding alveolar bone.

MMP-1, MMP-9, and TIMP-1 levels in oral lichen planus patients with gingivitis or periodontitis.

OBJECTIVE: Oral lichen planus (OLP) is a chronic inflammatory disease of unknown aetiology. The aim of this study was to investigate matrix metalloproteinase (MMP)-1, MMP-9, and MMP inhibitor-1 (TIMP-1) levels in gingival crevicular fluid (GCF) by enzyme-linked immunosorbent assay and to investigate MMP-1, MMP-9, and TIMP-1 levels in gingival tissue by immunohistochemical staining of samples from patients with and without OLP. DESIGN: Twenty-seven patients with OLP (gingivitis, OLPG; chronic periodontitis, OLPP) and thirty healthy non-OLP patients (gingivitis, HG; chronic periodontitis, HP) were included in this study. The MMP-1, MMP-9, and TIMP-1 levels in GCF were determined by enzyme-linked immunosorbent assay. The MMP-1, MMP-9, and TIMP-1 levels in gingival tissue were determined by immunohistochemical staining. RESULTS: The mean levels of MMP-1 and MMP-9 in the GCF of OLPP patients were significantly higher and TIMP-1 was significantly lower than in HP patients; similarly, the mean levels of MMP-1 and MMP-9 were higher and TIMP-1 was significantly lower in OLPG patients than in HG patients. Our findings illustrate that tissue MMP-9 levels were statistically higher and TIMP-1 level were significantly lower in the OLPP group in comparison to the HP group, and the OLPG group in comparison to the HG group. The tissue MMP-1 level in the non-OLP group was found to be lower when compared with the OLP groups. But not statistically significant. CONCLUSIONS: Increased levels of MMP-1 and MMP-9 with decreased levels of TIMP-1 in GCF and increased MMP-1, MMP-9 levels and decreased TIMP-1 levels in the gingival tissue of OLP patients in combination with poor oral hygiene may cause increased tissue breakdown. The results of our study provide information about the effects of the periodontal status on the enzyme profiles in GCF and gingival tissue of OLP and non-OLP patients.

Boron enhances strength and alters mineral composition of bone in rabbits fed a high energy diet.

An experiment was performed to determine whether boron had a beneficial effect on bone strength and composition in rabbits with apparent adiposity induced by a high energy diet. Sixty female New Zealand rabbits, aged 8 months, were randomly divided into five groups with the following treatments for seven months: control 1, fed alfalfa hay only (5.91 MJ/kg); control 2, high energy diet (11.76 MJ and 3.88 mg boron/kg); B10, high energy diet+10 mg/kg body weight boron gavage/96 h; B30, high energy diet+30 mg/kg body weight boron gavage/96 h; B50, high energy diet+50mg/kg body weight boron gavage/96 h. Bone boron concentrations were lowest in rabbits fed the high energy diet without boron supplementation, which suggested an inferior boron status. Femur maximum breaking force was highest in the B50 rabbits. Tibia compression strength was highest in B30 and B50 rabbits. All boron treatments significantly increased calcium and magnesium concentrations, and the B30 and B50 treatments increased the phosphorus concentration in tibia of rabbits fed the high energy diet. The B30 treatment significantly increased calcium, phosphorus and magnesium concentrations in femur of rabbits fed the high energy diet. Principal component analysis of the tibia minerals showed that the three boron treatments formed a separate cluster from controls. Discriminant analysis suggested that the concentrations of the minerals in femur could predict boron treatment. The findings indicate boron has beneficial effects on bone strength and mineral composition in rabbits fed a high energy diet.

Comparison of Er,Cr:YSGG laser and hand instrumentation on the attachment of periodontal ligament fibroblasts to periodontally diseased root surfaces: an in vitro study.

BACKGROUND: This study investigates the effects of erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation and hand instrumentation on the attachment of periodontal ligament (PDL) fibroblasts to periodontally involved root surfaces. METHODS: Twenty-four single-rooted periodontally involved human teeth (test groups), and six healthy premolar teeth extracted for orthodontic reasons (control group) were included in this study. A total of 45 root slices were obtained from all selected teeth and assigned to the following five groups: 1) untreated healthy group (+control); 2) untreated periodontally diseased group (-control); 3) hand instrumentation group (scaled Gracey); 4) laser I, Er,Cr:YSGG laser irradiation setting-I (short pulse); and 5) laser II, Er,Cr:YSGG laser irradiation setting-II (long pulse). All of the root slices were autoclaved in phosphate buffered saline and slices were placed onto cell culture inserts. PDL fibroblasts were placed at the density of 80,000 cells on the root plate (5 x 6 mm) and incubated for 48 hours and transferred to 24-well plates. The attachment PDL fibroblasts on the root plates were observed using confocal microscopy (at 12 hours and on days 3 and 7) and scanning electron microscopy (at 12 hours and day 3). 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay was performed on day 5 for PDL fibroblast survival. RESULTS: 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay shows that whereas laser-treated specimens showed a significantly higher cell density, the Gracey-treated group showed a lower cell density compared to the positive control group (P <0.05). Based on confocal microscopy, apparent reduction was observed in the attachment of PDL cells to the periodontally diseased root surfaces. In the laser and Gracey groups, cells looked well-oriented to the root surfaces. Laser-treated groups provided suitable environment for cell adhesion and growth. Laser I treatment was more favorable for the attachment of PDL compared to scaled Gracey, laser II, and even healthy root surfaces. CONCLUSION: The results of the study indicate that short-pulse laser setup (laser I) looks more promising regarding the attachment, spreading, and orientation of PDL cells.