Clinical, immunological and microbiological evaluation of experimental peri-implant mucositis and gingivitis in subjects with Grade C, stage III/IV periodontitis background.

AIM: To compare individuals with a periodontitis background (Grade C, stage III/IV-formerly generalized aggressive periodontitis) (H-GAP) with periodontally healthy subjects (H-Health) in terms of molecular changes (immunological/microbiological) accompanying experimental peri-implant mucositis and gingivitis. MATERIALS AND METHODS: H-GAP and control (H-Health) subjects were recruited, and experimental mucositis/gingivitis was induced around a single screw-retained implant and one contralateral tooth. Participants refrained from oral hygiene for 21 days in the selected areas, followed by professional prophylaxis and hygiene instructions for 21 days. Clinical parameters, immunological markers (multiplex analysis) and microbial data (16S rRNA gene sequencing) were collected at baseline, during induction (7, 14 and 21 days) and following remission (42 days). RESULTS: Clinically, no significant differences were observed between the groups (n = 10/each group) (H-GAP vs. H-Health) (p > .05, Mann-Whitney test) and the type of site (tooth vs. implant) (p > .05, Wilcoxon test) at the time of onset and resolution, or severity of gingival/mucosal inflammation. H-GAP displayed lower concentrations of the cytokines interleukin (IL)-1B, IL-4, IL-17, tumor necrosis factor-α and interferon-γ around implants than H-Health at baseline and during induction of mucositis (p < .05, Mann-Whitney test). In both groups, implants showed significantly higher inflammatory background at baseline and all subsequent visits when compared with teeth (p < .05, Wilcoxon test). Alpha and ß-diversity metrics showed a significant shift in the microbiome composition and abundances of core species during induction and resolution of peri-implant mucositis and gingivitis (p < .05, restricted maximum likelihood method of Shannon and Bray-Curtis indices, respectively). Differences were not significant for these parameters between the H-Health and H-GAP groups when the periodontal and peri-implant microbiomes were compared separately; however, at each time point, the peri-implant microbiome differed significantly from the periodontal microbiome. CONCLUSIONS: Within the limitations of this pilot study (e.g. low power), it can be concluded that different microbial shifts contribute to the onset and progression of inflammatory responses around teeth and implants and that history of periodontal disease experience plays an additional role in modulating the immune response of peri-implant and periodontal tissues to biofilm accumulation.

Microbial colonization in the partially exposed nonabsorbable membrane during alveolar ridge preservation.

AIM: This study evaluated the impact of the partial exposition of the nonabsorbable membrane (dPTFE) on microbial colonization during bone healing. MATERIALS AND METHODS: Patients indicated for tooth extraction were randomized to dPTFE group (n = 22) - tooth extraction and alveolar ridge preservation (ARP) using an intentionally exposed dPTFE membrane and USH group (n = 22) - tooth extraction and unassisted socket healing. Biofilm samples were collected at the barrier in the dPTFE and on the natural healing site in the USH after 3 and 28 days. Samples from the inner surface of the dPTFE barrier were also collected (n = 13). The microbiome was evaluated using the Illumina MiSeq system. RESULTS: Beta diversity was different from 3 to 28 days in both groups, and at 28 days, different microbial communities were identified between therapies. The dPTFE was characterized by a higher prevalence and abundance of gram-negative and anaerobic species than USH. Furthermore, the inner surface of the dPTFE membrane was colonized by a different community than the one observed on the outer surface. CONCLUSION: Intentionally exposed dPTFE membrane modulates microbial colonization in the ARP site, creating a more homogeneous and anaerobic community on the inner and outer surfaces of the membrane. CLINICAL RELEVANCE: DPTFE promoted faster biofilm colonization and enrichment of gram-negative and anaerobes close to the regenerated site in the membrane's inner and outer surfaces. dPTFE membrane can be used exposed to the oral site, but approaches for biofilm control should still be considered. The study was retrospectively registered at Clinicaltrials.gov (NCT04329351).

Minimal invasiveness in nonsurgical periodontal therapy.

Periodontal treatment is quickly moving towards a philosophy consisting of a less invasive approach. In this context, minimally invasive nonsurgical therapy (MINST) is a promising option. This paper reviews the concepts behind minimal invasiveness in nonsurgical periodontology and reports the state-of the art evidence for this topic. Instruments used and protocols suggested for these applications are introduced and discussed. The original papers reviewed show probing pocket depth (PPD) reductions and clinical attachment level (CAL) gains ranging from 2 to 4 mm between baseline and 6 months to 5 years posttreatment for intrabony defects and from 1.5 to 3 mm between baseline and 2-6 months of follow-up for full-mouth results. These clinical outcomes are accompanied by statistically significant reductions in radiographic bone defect depth and increases in intrabony defect angles posttreatment. Wound healing mechanisms following MINST are presented, and clinical applications and directions for future research are suggested.

Parents with periodontitis drive the early acquisition of dysbiotic microbiomes in their offspring.

AIM: To evaluate the microbial colonization in different dentition phases on individuals from 0 to 18 years of age belonging to families with a history of periodontitis compared to descendants of periodontally healthy parents. MATERIALS AND METHODS: The offspring of subjects with periodontitis ('Perio' group) and the offspring of periodontally healthy subjects ('Healthy' group), matched for gender and age, were included in this cross-sectional study and divided according to the dentition phase: pre-dentate, primary, mixed and permanent. The patients were clinically assessed, and their saliva was collected. DNA was extracted, and V1-V3 and V4-V5 regions of the 16S rRNA gene were sequenced. RESULTS: Fifty children of parents with periodontitis and 50 from healthy parents were included in the study and divided according to the dentition phase: pre-dentate (n = 5/group), primary dentition (n = 15/group), mixed dentition (n = 15/group) and permanent dentition (n = 15/group) in each group. The microbiome composition was different between dentitions for both groups. Children of the Perio group presented a microbial diversity different from that of the Healthy group in mixed and permanent dentitions. The more intense shift in the community occurred between primary and mixed dentition in the Perio group, while the transition between mixed and permanent dentition was the period with greater changes in the microbiome for the Healthy group. Furthermore, a pathogen-rich environment-higher prevalence and abundance of periodontitis-associated species such as Prevotella spp., Selenomonas spp., Leptotrichia spp., Filifactor alocis, Prevotella intermedia, Treponema denticola and Tannerella forsythia- was observed in the Perio group. CONCLUSIONS: The parents' periodontal status significantly affects the microbiome composition of their offspring from an early age. The mixed dentition was the phase associated with establishing a dysbiotic and pathogen-rich microbiome in descendants of parents with periodontitis.

Resveratrol for preventing medication-related osteonecrosis of the jaws in rats.

This study evaluated the effect of resveratrol (RES) on the prevention of medication-related osteonecrosis of the jaws (MRONJ) in ovariectomized (OVX) rats treated with zoledronate (ZOL). Fifty rats were distributed in five groups: SHAM (n = 10): non-ovariectomy + placebo; OVX (n = 10):ovariectomy + placebo; OVX + RES (n = 10):ovariectomy + resveratrol; OVX + ZOL (n = 10):ovariectomy + placebo + zoledronate; and OVX + RES + ZOL (n = 10):ovariectomy + resveratrol + zoledronate. The mandibles left sides were analyzed with micro-CT, histomorphometry, and immunohistochemistry. On the right side, bone markers gene expression was analyzed by qPCR. ZOL increased the percentage of necrotic bone and reduced the neo-formed bone compared to groups not receiving ZOL (p < 0.05). RES impacted the tissue healing pattern in OVX + ZOL + RES, reduced inflammatory cell infiltrate, and improved bone formation in the extraction site. Osteoblasts, alkaline phosphatase (ALP)-, and osteocalcin (OCN)-immunoreactive cells were lower in OVX-ZOL than in SHAM, OVX, and OVX-RES. The OXV-ZOL-RES had fewer osteoblasts and ALP- and OCN-cells than the SHAM and OVX-RES. The tartrate-resistant acid phosphatase (TRAP)-positive cells were reduced in the presence of ZOL (p < 0.05), while the TRAP mRNA levels increased with ZOL treatment, with or without resveratrol, compared with the other groups (p < 0.05). RES alone increased superoxide dismutase levels compared to OVX + ZOL and OVX + ZOL + RES (p < 0.05). In conclusion, resveratrol reduced the tissue impairment severity induced by ZOL; however, it could not prevent the occurrence of MRONJ.

Comparison between flapless-guided and conventional surgery for implant placement: a 12-month randomized clinical trial.

OBJECTIVES: The study was aimed at comparing implants installed with guided and conventional surgery. MATERIAL AND METHODS: Twenty-nine total edentulous patients were selected, and maxillary contralateral quadrants were randomly assigned to static computer-aided implant surgery (S-CAIS): flapless computer-guided surgery, and conventional surgery (CS): flap surgery with conventional planning. Tomography scans were performed at baseline and 10 days after the surgery for deviation measurement, and radiography was done at baseline and after 6 and 12 months, for peri-implant bone level (PIBL) analysis. Peri-implant fluid and subgingival biofilm were collected to evaluate bone markers and periodontal pathogens. RESULTS: S-CAIS showed less linear deviation at the apical point and the midpoint and less angular deviation (p < 0.05), with greater depth discrepancy in the positioning of the platform (p < 0.05). Higher values of vertical PIBL were observed for the S-CAIS group at baseline (p < 0.05), while lower values of horizontal PIBL were observed for CS (p < 0.05). Bone markers and Tf presented higher levels in CS (p < 0.05). Flapless S-CAIS allowed smaller linear and angular deviations than the conventional technique. CONCLUSION: However, PIBL was higher in S-CAIS; the conventional technique led to a greater angiogenic and bone remodeling activity by elevating the angiogenic levels and bone markers. CLINICAL RELEVANCE: Evaluating the different implant insertion techniques can guide clinical and surgical regarding the accuracy, the release pattern of bone markers, and the peri-implant bone level. TRIAL REGISTRATION: ReBEC-RBR-8556fzp.

Smoking negatively impacts the clinical, microbiological, and immunological treatment response of young adults with Grade C periodontitis.

OBJECTIVE: This study aimed to investigate the influence of smoking on clinical, microbiological and immunological parameters in young adult with stage III-IV Grade C periodontitis after full-mouth ultrasonic debridement (FMUD) associated with Amoxicillin and Metronidazole (AMX + MTZ), comparing smokers (PerioC-Y-Smk) with non-smokers (PerioC-Y-NSmk). MATERIALS AND METHODS: Fifteen PerioC-Y-NSmk and 14 PerioC-Y-Smk patients underwent FMUD associated with AMX + MTZ for 10 days. All parameters were collected at baseline and 3 and 6 months after treatment. Plaque index (PI), bleeding on probing (BoP), probing depth (PD), clinical attachment level (CAL)- the primary variable-, and gingival recession (GR) were clinically assessed. The impact of PI on CAL change at 6-month was verified by a regression analysis. Samples of the subgingival biofilm was collected for detection of levels of Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), Porphyromonas gingivalis (P.gingivalis), Tannerella forsythia (T. forsythia), and Fusobacterium nucleatum ssp (F. nucleatum), and were analyzed by real-time qPCR; gingival crevicular fluid was collected for detection of levels of interleukin (IL)-1ß, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ, which were analyzed using an enzyme immunoassay. RESULTS: PerioC-Y-Smk had significantly higher PI, BOP, and GR at baseline compared to non-smokers (p < .05). PerioC-Y-Smk presented higher PD, CAL, and GR at 3 and 6 months (p < .05) compared with PerioC-Y-NSmk in the same periods; PI negatively affected CAL gain in PerioC-Y-NSmk at 6-month follow-up (p = .052) and did not impact on clinical response in PerioC-Y-Smk (p = .882). Lower levels of IFN-γ, IL1-ß, and IL-4 were observed at 3 months in the PerioC-Y-NSmk (p < .05) compared with PerioC-Y-Smk. Lower proportions of P. gingivalis were observed in PerioC-Y-NSmk at baseline and at 3 months (p < .05) and lower proportions of F. nucleatum were observed at 6 months, in the PerioC-Y-NSmk (p < .05). CONCLUSIONS: PerioC-Y-Smk presents an unfavorable clinical, microbiological, and immunological response after 3 and 6 months after FMUD associated with AMX + MTZ. CLINICAL RELEVANCE: Smoking worsens periodontal condition of young treated adults presenting stage III/IV Grade C periodontitis.

The familial trend of the local inflammatory response in periodontal disease.

OBJECTIVES: The imbalanced host response in front of a dysbiotic biofilm is one of the major aspects of severe periodontitis, which also presents a strong familial aggregation related to the susceptibility factors transmission within family members. This study hypothesized that aggressive periodontitis (GAgP) patients and their descendants could present a similar trend of a local inflammatory response that is different from healthy controls. METHODS: Fifteen GAgP subjects and their children and fifteen healthy subjects and their children were clinically assessed, and the concentration of interferon (IFN)-γ, interleukin (IL)-10, IL-17, IL-1ß, IL-4, IL-6, IL-8, and tumor necrosis factor (TNF)-α was evaluated in the gingival fluid using the multiplexed bead immunoassay. RESULTS: Children from the GAgP group presented lower IL-10 and IFN-γ subgingival concentration than Health children, despite no difference in the clinical parameters. GAgP parents showed a lower IFN-γ, IL-10, and IL-6 than healthy subjects. IL-10/IL-1ß and IFN-γ/IL-4 ratios were reduced in GAgP dyads, suggesting a familial trend in the subgingival cytokine's profile. The cytokines correlated to the clinical data and were predictors of probing depth increase. CONCLUSION: GAgP parents and their children presented a similar cytokine profile and an imbalance in the subgingival response characterized by decreased IFN-γ/IL-4 and IL10/IL-1ß ratios.

Salivary IL-4: A Bleeding Predictor on Probing in Descendants of Severe Periodontitis Patients.

OBJECTIVE: Periodontitis in younger patients can cause severe periodontal destruction, and cases are usually more numerous in members of the same family due to the sharing of susceptibility factors. Thus, the use of a familial study design could improve our understanding of initial alterations in periodontal tissue. This observational study aimed to evaluate the salivary inflammatory pattern in descendants of periodontitis patients and identify any correlation with the clinical periodontal condition. STUDY DESIGN: Fifteen children of Generalized Aggressive Periodontitis (GAgP) patients and 15 children with periodontally healthy parents were evaluated for their plaque index (PI), gingival index (GI), bleeding on probing (BoP), and probing depth (PD). The concentrations of interferon (IFN)-γ, interleukin (IL)-10, IL-17, IL-1ß, IL-4, and tumor necrosis factor (TNF)-α were measured in unstimulated saliva using the Luminex MAGPix platform. RESULTS: Children from the GAgP group presented higher probing depth (PD) and bleeding on probing (BoP) (p<0.05) and lower release of IL-4 in saliva (p<0.05) than the periodontally healthy group. The cytokines IL-10, IFN-γ, IL-17, and IL-4 were negatively correlated with the gingival index, while IL-4 was negatively correlated with BoP. A regression analysis revealed that salivary IL-4 and plaque were predictors of BoP. CONCLUSIONS: Children of GAgP parents presented lower salivary IL-4 and higher BoP and PD than children from periodontally healthy families. Additionally, salivary IL-4 was a predictor of bleeding on probing in the children, suggesting that the lower presence of this anti-inflammatory cytokine is related to higher clinical inflammation.

Subgingival endotoxin and lipoteichoic acid modulate cytokine production in diabetic subjects: A Case-control Study.

BACKGROUND: Periodontal disease and diabetes mellitus (DM) are highly prevalent and interrelated diseases, resulting in altered host response microbiota. Thus, this study aimed to evaluate the impact of DM on local levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) and their relationship with cytokines and matrix metalloproteinases' (MMPs) profile. METHODS: This case-control study included diabetic (n = 15) and non-diabetic (n = 15) subjects presenting Stage 3-4, Grade C, Periodontitis. Gingival crevicular fluid (GCF) was collected, and LPS and LTA levels were analyzed by enzyme-linked immunosorbent assay (ELISA), while IFN-γ, IL-10, IL-17, IL-1ß, IL-4, MMP-2, and MMP-9 were measured by LUMINEX/MAGpix. Mann-Whitney and Spearman's correlation tests were used to compared and to correlate variables (p < 0.05). RESULTS: Higher levels of LTA, LPS, IL-10, IL-1ß, and MMP-2 (p < 0.05) and lower levels of IL-17 were found in the DM group (p < 0.05). Non-diabetic subjects presented higher LPS, IFN-γ, IL-17, and MMP-2 levels and lower IL-10 concentration (p < 0.05). No significant correlation was seen between LPS and cytokine profile in non-diabetic. Local levels of LTA were positively correlated with IL-17 and MMP-2 and negatively with IL-10. CONCLUSION: LTA and LPS drove the inflammatory profile through the modulation of cytokines and MMPs in a different manner in DM and non-diabetic subjects.

Surgical and non-surgical debridement for the treatment of peri-implantitis: a two-center 12-month randomized trial.

OBJECTIVES: To compare surgical (ST) and non-surgical (NST) debridement for the treatment of peri-implantitis in a two-center randomized trial. MATERIALS AND METHODS: Forty-five individuals with 63 implants with probing depth (PPD) ≥5mm, bleeding on probing (BOP), and radiographic bone loss ≥2mm were included. In the NST (30 implants), submucosal debridement was performed. In the ST (33 implants), a mucoperiosteal flap was raised and surfaces were decontaminated only by debridement as performed in NST. Clinical parameters and radiographs were compared at baseline and after 12 months. Means and standard errors were reported. RESULTS: PPD considering all implant sites reduced significantly in NST from 4.14±0.25 to 3.25±0.18mm. In ST, PPD also significantly changed (3.74±0.22 to 3.00±0.29mm). No significant differences were observed between the two groups. For deep sites (≥7mm), PPD was 7.82±0.20mm at baseline and reduced to 5.10±0.30mm in NST, while in ST group, it was 7.11±0.11mm and changed to 5.22±0.91mm (between-groups p value=0.51). BOP significantly reduced from ~60 to 35% of all sites in both groups, without significant differences between them. When sites with radiographic bone level ≥3mm at baseline were analyzed, there was a significant difference between groups in bone gain after 12 months in favor of ST (ST=0.78±0.30mm compared to NST=0.25mm±0.13; p=0.03). CONCLUSIONS: Surgical and non-surgical debridement for the treatment of peri-implantitis present similar clinical outcomes. Bone levels were better improved in ST than NST for sites with higher initial bone loss. CLINICAL RELEVANCE: The treatment of peri-implantitis is still a challenge in clinical practice, since less than half of affected implants achieve health after surgical or non-surgical debridement. Considering the lack of clinically relevant differences between these two treatments, non-surgical debridement should be considered the first therapeutic choice for peri-implantitis, mainly mild to moderate cases.

Impact of resveratrol in the reduction of the harmful effect of diabetes on peri-implant bone repair: bone-related gene expression, counter-torque and micro-CT analysis in rats.

OBJECTIVE: Investigate the impact of resveratrol (RESV) on peri-implant repair and its effect on bone-related markers in rats with induced diabetes mellitus (DM). MATERIAL AND METHODS: Ninety rats were divided into: DM + RESV (n = 18); DM + placebo (PLAC) (n = 18); DM + insulin (INS) (n = 18); DM + RESV + INS (n = 18); Non-DM (n = 18). Diabetes was induced by streptozotocin. One screw-shaped titanium implant was inserted in each tibiae of animals. Treatments were administered during 30 days. After, one of the implants was removed for counter-torque and the peri-implant tissue was collected for mRNA quantification of BMP-2, OPN, Runx2, Lrp-5, Osx, ß-catenin, Dkk1, OPG, and RANKL by Real-time PCR. The other tibia was submitted to MicroCT analysis to measure: bone volume (BV/TV), trabecular thickness (Tb.Th) and bone-implant contact (BIC). RESULTS: Higher counter-torque values were observed for implant removal in DM + RESV, DM + RESV + INS and Non-DM groups when compared to DM + PLAC (p < .05). Augmented Tb.Th was observed in DM + RESV and Non-DM when compared to DM + PLAC group (p < .05), whereas higher BIC was detected in DM + RESV, DM + RESV + INS and Non-DM animals when compared to DM + PLAC (p < .05). Levels of RANKL were downregulated by the RESV and/or INS therapy, whereas only the association of RESV and INS upregulated the levels of Runx2 (p < .05). CONCLUSIONS: The therapy with RESV may favour peri-implant bone repair improving bone formation around implants.

Periodontitis is an inflammatory disease of oxidative stress: We should treat it that way.

Periodontitis is a highly prevalent disease. As it progresses, it causes serious morbidity in the form of periodontal abscesses and tooth loss and, in the latter stages, pain. It is also now known that periodontitis is strongly associated with several nonoral diseases. Thus, patients with periodontitis are at greater risk for the development and/or exacerbation of diabetes, chronic obstructive pulmonary disease, and cardiovascular diseases, among other conditions. Although it is without question that specific groups of oral bacteria which populate dental plaque play a causative role in the development of periodontitis, it is now thought that once this disease has been triggered, other factors play an equal, and possibly more important, role in its progression, particularly in severe cases or in cases that prove difficult to treat. In this regard, we allude to the host response, specifically the notion that the host, once infected with oral periodontal pathogenic bacteria, will mount a defense response mediated largely through the innate immune system. The most abundant cell type of the innate immune system - polymorphonuclear neutrophils - can, when protecting the host from microbial invasion, mount a response that includes upregulation of proinflammatory cytokines, matrix metalloproteinases, and reactive oxygen species, all of which then contribute to the tissue damage and loss of teeth commonly associated with periodontitis. Of the mechanisms referred to here, we suggest that upregulation of reactive oxygen species might play one of the most important roles in the establishment and progression of periodontitis (as well as in other diseases of inflammation) through the development of oxidative stress. In this overview, we discuss both innate and epigenetic factors (eg, diabetes, smoking) that lead to the development of oxidative stress. This oxidative stress then provides an environment conducive to the destructive processes observed in periodontitis. Therefore, we shall describe some of the fundamental characteristics of oxidative stress and its effects on the periodontium, discuss the diseases and other factors that cause oxidative stress, and, finally, review potentially novel therapeutic approaches for the management (and possibly even the reversal) of periodontitis, which rely on the use of therapies, such as resveratrol and other antioxidants, that provide increased antioxidant activity in the host.

Impact of natural curcumin on the progression of experimental periodontitis in diabetic rats.

OBJECTIVE: To evaluate the role of natural curcumin (CURC) on experimental periodontitis (EP) in animals with diabetes mellitus (DM). MATERIAL AND METHODS: One hundred rats were assigned to DM + placebo (PLA); DM + CURC; DM + insulin (INS); DM + CURC + INS; and Non-DM. Diabetes was induced by streptozotocin. After 3 days, they were initiated CURC and PLAC solutions and insulin administrations, daily for 30 days. This included a period of 19 days prior to EP induction (ligature at the first mandibular and the second maxillary molar) and then additional 11 days. Specimens from the mandible were processed for morphometric examination of bone level. Gingival tissues from mandibular molars were collected for quantification of IL-1ß, IL-4, IL-6, IL-17, IFN-γ, and TNF-α using a Luminex/MAGpix assay. Gingivae from maxillary molars were subjected to RT-PCR for assessment of Runx2, RANKL, OPG, SIRT, Dkk1, and Sost levels. RESULTS: Lower linear bone loss was detected in ligated molars of DM + CURC + INS vs DM + PLAC and DM + INS groups (P < 0.05). In ligated sites from DM rats treated with CURC + INS, IL-6, IL-1ß, INF-γ, and TNF-α levels were the lowest in comparison with PLAC and/or INS and CURC as monotherapies (P < 0.05). CURC, independently of INS, increased Runx2 and SIRT when compared to DM + PLAC (P < 0.05) in ligated sites, whereas only CURC + INS reduced the RANKL/OPG ratio when compared to DM + PLAC (P < 0.05). CONCLUSION: Natural CURC, when associated with INS, reduces the DM-induced loss of supporting alveolar bone and promotes favorable modulation on osteo-immune-inflammatory mediators.

Effect of resveratrol on the progression of experimental periodontitis in an ovariectomized rat model of osteoporosis: Morphometric, immune-enzymatic, and gene expression analysis.

OBJECTIVE: This study aimed to determine the role of resveratrol (RESV) on the progression of experimental periodontitis (EP) in ovariectomy rats (OVT). BACKGROUND: Estrogen deficiency is the main cause of osteoporosis and is related to higher periodontal attachment loss and reduction of alveolar bone. Zoledronate (ZLD) is an antiresorptive drug used to control osteoporosis but can lead to osteonecrosis of the jaw. RESV, a natural product, can reduce bone loss and control and prevent osteoporosis. Thus, this study aimed at investigating the effect of RESV on the progression of EP in estrogen-deficient rats. MATERIAL AND METHODS: The animals were subjected to the OVT or sham surgery to induce estrogen-deficiency and then were divided into the groups: OVT + RESV (n: 10); OVT + PLAC (n: 10): OVT + placebo; OVT + ZLD +PLA (n: 10); OVT + RESV +ZLD (n: 10): OVT + RESV and ZLD; SHAM (n: 10): non-ovariectomized animals + placebo. To induce estrogen deficiency, the rats were subjected to ovariectomy. Experimental periodontitis was induced by the placement of a ligature at the second maxillary molars. Daily administration of the placebo solution, resveratrol (10 mg/kg), and ZLD (0.1 mg/kg) was carried out for a period 42 days prior to initiation of EP, and then for another 28 days following ligature placement. After euthanasia, the specimens were processed for micro-CT and morphometric analysis of bone loss (linear measurement), and the gingival tissue surrounding the maxillary second molar was collected for the quantification of inflammatory markers using Luminex/MAGPix, of oxidative stress markers using ELISA assay, and gene expression analysis of bone markers, by real-time PCR. RESULTS: Morphometric and micro-CT analysis showed higher bone loss and lower bone density, respectively, in OVT + PLAC when compared to the other groups (P < .05). ZLD treated groups had lower alveolar bone loss, as well as, higher density and percentage of bone volume, when compared to OVT + RESV and SHAM + PLAC groups (P < .05). IL-4 levels were significantly lower in the OVT + PLAC group versus OVT + ZLD +RESV and SHAM + PLAC (P < .05). NADPH oxidase (nicotinamide adenine dinucleotide phosphate oxidase) levels were significantly lower OVT + RESV group when compared to OVT + PLAC (P < .05). OPG mRNA levels were lower in OVT + PLAC compared with the SHAM + PLAC group (P < .05). CONCLUSION: It can be concluded that resveratrol modulated alveolar bone loss during experimental periodontitis progression in estrogen-deficient rats by downregulating NADPH oxidase levels.

Novel 20% doxycycline-loaded PLGA nanospheres as adjunctive therapy in chronic periodontitis in type-2 diabetics: randomized clinical, immune and microbiological trial.

OBJECTIVE: This study evaluated the clinical, microbiological, and immunological results of poly lactic-co-glycolic acid (PLGA) nanospheres containing 20% doxycycline (DOXY) in the treatment of type-2 diabetic patients (DM-2) with chronic periodontitis (CP). MATERIAL AND METHODS: A parallel, double-blind, randomized, placebo-controlled clinical trial was conducted in DM-2 presenting severe and generalized CP. All patients received one-stage full-mouth ultrasonic debridement (FMUD) and they were randomly divided into two groups: PLAC (n = 20)-local application of placebo PLGA nanospheres, and DOXY (n = 20)-local application of doxycycline-loaded nanospheres; both in six non-contiguous sites. Clinical, metabolic (fasting plasma glucose level-FPG and glycated hemoglobin-HbA1c), cytokine pattern (multiplexed bead immunoassay) and microbiological assessments were performed at baseline, and 1, 3, and 6 months after treatment. RESULTS: Both groups showed clinical improvement in all parameters after treatment (p < 0.05). Deep pockets showed improvements in bleeding on probing-BoP (3 and 6 months), PD (at 3 months), and CAL gain (at 1 and 3 months) favoring DOXY (p < 0.05). The percentage of sites presenting PD reduction and CAL gain ≥ 2 mm was higher in DOXY at 3 months (p < 0.05). DOXY group exhibited a significant increase in the levels of anti-inflammatory interleukin (IL)-10 and a reduction in IL-8, IFN-y, IL-6, and IL-17 (p < 0.05), significant reduction in periodontal pathogens (p < 0.05), and a lower mean percentage of HbA1C at 3 months (p < 0.05). CONCLUSION: DOXY nanospheres may be considered a potential adjunct to mechanical debridement in the therapy of periodontitis in DM-2, offering additional benefits in deep pockets, improving the cytokine profile, and reducing periodontal pathogen levels. CLINICAL RELEVANCE: The use of locally applied doxycycline nanospheres may represent an adjunctive therapeutic approach in the treatment of periodontal disease in type-2 diabetic patients, achieving additional benefits in the local modulation of cytokines, microbial reduction, and clinical parameters, especially in deep pockets.

Triclosan toothpaste as an adjunct therapy to plaque control in children from periodontitis families: a crossover clinical trial.

OBJECTIVES: Studies have demonstrated that children from aggressive periodontitis (AgP) parents presented precocious alterations in their periodontal condition, and the use of chemical agents in association to plaque control could be useful to control these alterations. This study aimed to evaluate the effect of Triclosan toothpaste to modulate the clinical and subgingival condition in children from AgP parents. METHODS: Fifteen children from AgP parents and 15 from periodontally healthy parents were included in this crossover placebo study. Children were randomly allocated into triclosan or placebo therapy, using selected toothpaste for 45 days. After 15 days of wash-out, groups were crossed, changing the used toothpaste. Clinical examination and saliva, crevicular gingival fluid (GCF), and subgingival biofilm collection were performed at baseline and 45 days of each phase. GCF cytokines' levels were analyzed by Luminex/MAGpix platform and subgingival and salivary periodontal pathogens' levels by qPCR. RESULTS: At baseline, AgP group presented higher plaque index (PI), gingival index (GI), and bleeding on probing (BoP), higher Aggregatibacter actinomycetemcomitans (Aa) abundance in saliva and subgingival biofilm, and lower levels of INF-É£, IL-4, and IL-17 in GCF. Placebo therapy only reduced PI in both groups. Triclosan toothpaste reduced PI and GI in both groups. Triclosan promoted reduction of BoP and probing depth (PD), Aa salivary, and IL-1ß levels in AgP group. In health group, triclosan reduced INF-É£ and IL-4 concentration. CONCLUSION: Triclosan toothpaste demonstrated to be more effective than placebo toothpaste to control the periodontal condition in children from AgP parents, by reducing the BoP, PD, salivary Aa, and IL-1ß. CLINICAL RELEVANCE: Triclosan toothpaste can improve oral conditions in higher-risk population for AgP. TRIAL REGISTRATION: This study was registered at ClinicalTrials.gov with the identifier NCT03642353.

Local IL-10 level as a predictive factor in generalized aggressive periodontitis treatment response.

Generalized aggressive periodontitis (GAgP) presents a reduced response to non-surgical therapy. However, it is not clear if the initial clinical, microbiological or immunological characteristics are impacting the worse response to treatment. This study aimed to identify the predictive value of clinical, microbiological and immunological patterns on the clinical response to therapy in GAgP patients. Twenty-four GAgP patients were selected, and gingival crevicular fluid (GCF) and subgingival biofilm were collected. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia levels were evaluated by qPCR, and IL-1ß and IL-10 concentration by ELISA. Twelve patients were treated with SRP (scaling and root planning), and twelve with SRP plus 375 mg amoxicillin and 250 mg metronidazole (8/8 hours, 7 days) (SRP + AM). The clinical changes (Probing Pocket Depth [PPD] reduction and Clinical Attachment Level [CAL] gain) 6 months post-treatment were correlated to the initial clinical, inflammatory and microbiological variables using stepwise logistic regression (α = 5%). CAL gain at 6 months was 1.16 ± 0.77 for SRP and 1.74 ± 0.57 mm for SRP + AM (P > .05). PPD reduction was 1.96 ± 0.82 for SRP and 2.45 ± 0.77 mm for SRP + AM (P < .05). In the SRP group, IL-10 showed a predictive value for clinical response. The higher the IL-10 concentration at baseline, the higher the reduction in PPD at 6 months (P = .01, r = .68). However, when antimicrobials were administered, no significant influence was detected (P > .05). It can be concluded that the IL-10 levels in GFC act as a predictor of clinical response to GAgP. Moreover, the intake of antimicrobials appears to overlap the influence of the inflammatory response on clinical response to treatment. Clinical trial registration number: NCT03933501.

Resveratrol attenuates oxidative stress during experimental periodontitis in rats exposed to cigarette smoke inhalation.

OBJECTIVES: This study aimed at investigating the effect of the systemic administration of resveratrol (RESV) on oxidative stress during experimental periodontitis in rats subjected to cigarette smoke inhalation. MATERIAL AND METHODS: Experimental periodontitis (EP) was induced in 26 male Wistar rats by the insertion of a ligature around one of the first mandibular and maxillary molars. The animals were assigned randomly to the following groups: cigarette smoke inhalation (CSI; 3 times/d, 8 minutes/d) + resveratrol (10 mg/Kg), that is, SMK + RESV (n = 13) and cigarette smoke inhalation + placebo, that is, SMK + PLAC (n = 13). The substances were administered daily for 30 days (19 days prior and 11 days following EP induction), and then, the animals were euthanized. The maxillary specimens were processed for morphometric analysis of bone loss, and the tissue surrounding the first maxillary molars was collected for mRNA quantification of Sirtuin 1 (SIRT1) by real-time PCR. The gingival tissues surrounding the mandibular first molars were collected for quantification of superoxide dismutase 1 (SOD1) and nicotinamide adenine dinucleotide phosphatase oxidase (NADPH) using an ELISA assay. RESULTS: Reduced bone loss was demonstrated in animals in the SMK + RESV group as compared to those in the SMK + PLAC (P < 0.05) group on the basis of morphometric analysis. Resveratrol promoted higher levels of SIRT and SOD (P < 0.05) as well as reduced levels of NADPH oxidase (P < 0.05) were found in tissues derived from animals in the SMK + RESV group when compared to those in the SMK + PLAC group. CONCLUSION: Resveratrol is an efficient therapeutic agent that reduces exacerbation of bone loss found in animals with EP that were also exposed to smoke. The results suggest that its effects could be mediated, at least in part, by its antioxidant and anti-inflammatory properties which attenuate the effects of oxidative stress on EP in the presence of cigarette smoke.

Resveratrol reverses the negative effect of smoking on peri-implant repair in the tibia of rats.

OBJECTIVES: Innovative approaches capable to improve peri-implant bone repair are relevant in the presence of smoking, a risk factor for healing around implants. This study investigated the effect of resveratrol (RESV) on peri-implant repair, and its influence on bone-related markers in rats submitted to cigarette smoking inhalation (CSI). MATERIALS AND METHODS: One titanium implant was inserted in each tibiae of rats assigned to CSI+RESV (n:18); CSI+placebo (n:18); and non-CSI (n:18). One implant was removed for counter torque, and the peri-implant tissue was collected for mRNA quantification of BMP-2, OPN, Runx2, Lrp-5, Osx, ß-catenin, Dkk1, OPG, and RANKL. The other tibia was submitted to microCT to measure: bone volume, bone porosity, trabecular spacing, trabecular thickness, and bone-implant contact (BIC). RESULTS: No differences were detected between counter torque in CSI+RESV and non-CSI group (p > 0.05), whereas CSI+placebo group presented lower values when compared to the others (p < 0.05). RESV improved the BIC in CSI rats without differences when compared to non-CSI group (p > 0.05), whereas CSI+placebo showed reduced BIC when compared to the other groups (p < 0.05). RESV reduced RANKL/OPG and Lrp-5 levels and increased ß-catenin in CSI rats when compared to CSI+placebo (p < 0.05). CONCLUSION: Although further investigations should be considered using oral models of dental implants, within the limits of the present study, it was concluded that RESV reverses the negative effects of smoking in the peri-implant repair, benefiting the modulation of bone-related markers.