Efficacy of a novel three-step decontamination protocol for titanium-based dental implants: An in vitro and in vivo study.

AIM: The aim of the study was to evaluate several mechanical and chemical decontamination methods associated with a newly introduced biofilm matrix disruption strategy for biofilm cleaning and preservation of implant surface features. MATERIALS AND METHODS: Titanium (Ti) discs were obtained by additive manufacturing. Polymicrobial biofilm-covered Ti disc surfaces were decontaminated with mechanical [Ti curette, Teflon curette, Ti brush, water-air jet device, and Er:YAG laser] or chemical [iodopovidone (PVPI) 0.2% to disrupt the extracellular matrix, along with amoxicillin; minocycline; tetracycline; H2 O2 3%; chlorhexidine 0.2%; NaOCl 0.95%; hydrocarbon-oxo-borate-based antiseptic] protocols. The optimal in vitro mechanical/chemical protocol was then tested in combination using an in vivo biofilm model with intra-oral devices. RESULTS: Er:YAG laser treatment displayed optimum surface cleaning by biofilm removal with minimal deleterious damage to the surface, smaller Ti release, good corrosion stability, and improved fibroblast readhesion. NaOCl 0.95% was the most promising agent to reduce in vitro and in vivo biofilms and was even more effective when associated with PVPI 0.2% as a pre-treatment to disrupt the biofilm matrix. The combination of Er:YAG laser followed by PVPI 0.2% plus NaOCl 0.95% promoted efficient decontamination of rough Ti surfaces by disrupting the biofilm matrix and killing remnants of in vivo biofilms formed in the mouth (the only protocol to lead to ~99% biofilm eradication). CONCLUSION: Er:YAG laser + PVPI 0.2% + NaOCl 0.95% can be a reliable decontamination protocol for Ti surfaces, eliminating microbial biofilms without damaging the implant surface.

BMP-2 and asporin expression regulate 5-aza-dC-mediated osteoblast/cementoblast differentiation of periodontal dental ligament mesenchymal progenitor cells.

Periodontal dental ligament (PDL) is composed of heterogeneous population of mesenchymal progenitor cells. The mechanisms that regulate the differentiation of these cells towards osteoblast/cementoblast phenotype are not fully understood. Some studies have demonstrated that is possible to change the pattern of cell differentiation via epigenetic mechanisms. The proposal of this study was to investigate whether 5-aza-2'-deoxycytidine (5-aza-dC) treatment would stimulate the osteoblast/cementoblast differentiation of periodontal ligament mesenchymal progenitor cells (PDL-CD105+ enriched cells), characterized as low osteoblast potential, through bone morphogenetic protein-2 (BMP-2) modulation. PDL-CD105+ cells from a single donor were cloned and characterized in two populations as high osteoblast/cementoblast potential (HOP) and low osteoblast/cementoblast potential (LOP) by mineralization in vitro and expression of osteogenic gene markers, such as runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), bone morphogenetic protein 2 (BMP-2) and asporin (ASPN). Next, two LOP clones (L1 and L2) were pretreated with 5-aza-dC (10 µM) for 48 h, cultured under osteogenic condition and evaluated for mineralized matrix in vitro, transcription modulation of osteogenic gene markers, methylated and hydroxymethylated DNA levels of BMP-2 and ASPN and intracellular/extracellular expression of BMP-2 protein. LOP clones showed high expression of ASPN transcripts associated with low mRNA levels of BMP-2, RUNX2, ALP, and OCN. 5-aza-dC treatment raised hydroxymethylated DNA levels of BMP-2 and increased the expression of BMP-2 transcripts in both LOP clones. However, BMP-2 protein (intracellular and secreted forms) was detected only in L1 cell clones, in which it was observed an increased expression of osteoblast/cementoblast markers (RUNX2, ALP, OCN) associated with higher mineralization in vitro. In L2 cell clones, 5-aza-dC increased gene expression of ASPN, with no great change in for osteoblast/cementoblast differentiation potential. These data show that 5-aza-dC improves osteoblast/cementoblast differentiation of PDL-CD105+ cells via BMP-2 secretion, and this effect depends on low levels of ASPN expression.

Do smokers have a different gingival crevicular fluid cytokine/chemokine profile than nonsmokers in clinically healthy periodontal sites? A systematic review and meta-analysis.

OBJECTIVES: Assessing the evidence and comparing the levels of cytokines in gingival crevicular fluid (GCF) of periodontal healthy sites of smokers and nonsmokers. MATERIALS AND METHODS: Seven databases were surveyed for observational studies up to April 8, 2021. Studies comparing cytokine levels on GCF in periodontally healthy sites of smokers vs. nonsmokers were included in the study. The risk of bias was evaluated using NIH (2014) tool. For meta-analyses, levels in GCF were analyzed, followed by evidence certainty assessment using the GRADE approach. RESULTS: Eighteen studies were included for qualitative evaluation, and eight were included in meta-analysis. Qualitatively, despite high heterogeneity and risk of bias observed among the studies, most of them presented no significant difference in the gingival crevicular cytokine fluid levels between groups. Regarding meta-analyses, interleukin-8 (IL-8) and superoxide dismutase (SOD) levels in GCF were analyzed. The significant difference was observed only in SOD levels, where heavy smokers had lower levels compared to nonsmokers (MD - 30.06 [- 40.17, - 19.96], p = 0.07, 95%CI), as well as light smokers had lower levels compared to nonsmokers (MD - 15.22 [- 16.05, - 14.39], p < 0.00001, 95%CI). CONCLUSION: No distinct GCF cytokine profiles were detected for smokers and non-smokers. However, despite the limitations observed in the included studies, lower levels of SOD were identified in smokers. CLINICAL RELEVANCE: Indicating a distinct GCF profile of cytokines in periodontal healthy smokers may help to understand the mechanism whereby smoking may affect the host response.

Targeting Biomechanical Endurance of Dental-Implant Abutments Using a Diamond-Like Carbon Coating.

Abutment components (i.e., fixtures associated with oral implants) are essentially made of titanium (Ti), which is continuously exposed to the hash oral environment, resulting in scratching. Thus, such components need to be protected, and surface treatments are viable methods for overcoming long-term damage. Diamond-like carbon (DLC), an excellent protective material, is an alternative surface-treatment material for Ti abutments. Here, we demonstrate that a silicon interlayer for DLC film growth and the pulsed-direct current plasma-enhanced chemical vapor deposition (DC-PECVD) method enables the deposition of an enhanced protective DLC film. As a result, the DLC film demonstrated a smooth topography with a compact surface. Furthermore, the DLC film enhanced the mechanical (load-displacement, hardness, and elastic modulus) and tribological properties of Ti as well as increased its corrosion resistance (16-fold), which surpassed that of a bare Ti substrate. The biofilm formed (Streptococcus sanguinis) after 24 h exhibited an equal bacterial load (∼7 Log colony-forming units) for both the groups (Ti and DLC). In addition, the DLC film exhibited good cytocompatibility, owing to its noncytotoxicity toward human gingival fibroblast cells. Therefore, DLC deposition via DC-PECVD can be considered to be a promising protective and cytocompatible alternative for developing implant abutments with enhanced mechanical, tribological, and electrochemical properties.

Sorption, solubility and cytotoxicity of novel antibacterial nanofilled dental adhesive resins.

Dental adhesives hydrolyze in the mouth. This study investigated the water sorption (SOR), solubility (SOL) and cytotoxicity (CYTO) of experimental adhesives containing nitrogen-doped titanium dioxide nanoparticles (N_TiO2). Specimens (n = 15/group [SOR, SOL]; n = 10/group [CYTO]) of unaltered Clearfil SE Protect (CSP), OptiBond Solo Plus (OSP), Adper Scotchbond (ASB) and experimental adhesives (OSP + 25% or 30% of N_TiO2) were fabricated, desiccated (37 °C) and tested for SOR and SOL according to ISO Specification 4049 (2009). CYTO specimens were UV-sterilized (8 J/cm2) and monomer extracted in growth medium (1, 3 or 7 days). Human pulp cells were isolated and seeded (0.5 × 104) for MTT assay. SOR and SOL data was analyzed using GLM and SNK (α = 0.05) and CYTO data was analyzed with Kruskal-Wallis and SNK tests (α = 0.05). SOR and SOL values ranged from 25.80 µg/mm3 (30% N_TiO2) to 28.01 µg/mm3 (OSP) and 23.88 µg/mm3 (30% N_TiO2) to 25.39 µg/mm3 (25% N_TiO2). CYTO results indicated that pulp cells exposed to experimental materials displayed comparable viabilities (p > 0.05) to those of OSP. Experimental materials displayed comparable SOR, SOL and CYTO values (p > 0.05) when compared to unaltered materials. N_TiO2 incorporation have not adversely impacted SOR, SOL and CYTO properties of unaltered adhesives.

Enhancing adhesion and alignment of human gingival fibroblasts on dental implants.

BACKGROUND: Promoting the directional attachment of gingiva to the dental implant leads to the formation of tight connective tissue which acts as a seal against the penetration of oral bacteria. Such a directional growth is mostly governed by the surface texture. MATERIAL AND METHODS: In this study, three different methods, mechanical structuring, chemical etching and laser treatment, have been explored for their applicability in promoting cellular attachment and alignment of human primary gingival fibroblasts (HGFIBs). RESULTS: The effectiveness of mechanical structuring was shown as a simple and a cost-effective method to create patterns to align HGIFIBs. CONCLUSION: Combining mechanical structuring with chemical etching enhanced both cellular attachment and the cellular alignment.