Chronic wounds have harmful effects on both patients and healthcare systems. Wound chronicity is attributed to an impaired healing process due to several host and local factors that affect healing pathways. The resulting ulcers contain a wide variety of microorganisms that are mostly resistant to antimicrobials and possess the ability to form mono/poly-microbial biofilms. The search for new, effective and safe compounds to handle chronic wounds has come a long way throughout the history of medicine, which has included several studies and trials of conventional treatments. Treatments focus on fighting the microbial colonization that develops in the wound by multidrug resistant pathogens. The development of molecular medicine, especially in antibacterial agents, needs an in vitro model similar to the in vivo chronic wound environment to evaluate the efficacy of antimicrobial agents. The Lubbock chronic wound biofilm (LCWB) model is an in vitro model developed to mimic the pathogen colonization and the biofilm formation of a real chronic wound, and it is suitable to screen the antibacterial activity of innovative compounds. In this review, we focused on the characteristics of chronic wound biofilms and the contribution of the LCWB model both to the study of wound poly-microbial biofilms and as a model for novel treatment strategies.

Chronic wounds represent an increasing problem worldwide. Graphene oxide (GO) has been reported to exhibit strong antibacterial activity toward both Gram-positive and Gram-negative bacteria. The aim of this work was to investigate the in vitro antimicrobial and antibiofilm efficacy of GO against wound pathogens. Staphylococcus aureus PECHA 10, Pseudomonas aeruginosa PECHA 4, and Candida albicans X3 clinical isolates were incubated with 50 mg/liter of GO for 2 and 24 h to evaluate the antimicrobial effect. Optical and atomic force microscopy images were performed to visualize the effect of GO on microbial cells. Moreover, the antibiofilm effect of GO was tested on biofilms, both in formation and mature. Compared to the respective time controls, GO significantly reduced the S. aureus growth both at 2 and 24 h in a time-dependent way, and it displayed a bacteriostatic effect in respect to the GO t = 0; an immediate (after 2 h) slowdown of bacterial growth was detected for P. aeruginosa , whereas a tardive effect (after 24 h) was recorded for C. albicans . Atomic force microscopy images showed the complete wrapping of S. aureus and C. albicans with GO sheets, which explains its antimicrobial activity. Moreover, significant inhibition of biofilm formation and a reduction of mature biofilm were recorded for each detected microorganism. The antibacterial and antibiofilm properties of GO against chronic wound microorganisms make it an interesting candidate to incorporate into wound bandages to treat and/or prevent microbial infections.

Background: Two‐piece implants present gaps and cavities between the implant and the abutment, and these hollow spaces can act as a trap for bacteria. The aim of the present in vitro study was to evaluate the migration of two different microbial species from the inside to the outside of the implant–abutment assembly in three different connection types.Methods: A total of 30 implants (10 implants per group) were used. The implants presented a screwed trilobed connection (group 1), a cemented connection (group 2), and an internal conical connection (group 3). The inner parts of five implants, per group, were inoculated with Pseudomonas aeruginosa suspension and the remaining five implants, per group, with Aggregatibacter actinomycetemcomitans. The penetration of bacteria into the surrounding solution was determined by the observation of turbidity of the broth.Results: In group 1, bacterial contamination was found in six of 10 implants. In group 2, no contaminated samples were found. In group 3, bacterial contamination was found in one implant of 10. Statistically significant differences were detected between group 1 versus group 3 (P <0.05) and between group 1 versus group 2 (P 0.05).Conclusion: The present study confirms previous results about the hermeticity of the cement‐retained implant–abutment assembly, the very low permeability to bacteria of the conical implant–abutment connection, and the high prevalence of bacterial penetration of screw‐retained implant–abutment assemblies.

Background: The main therapeutic approach for periodontal diseases is mechanical treatment of root surfaces via scaling and root planing (SRP). Multicenter clinical trials have demonstrated that the adjunctive use of a chlorhexidine (CHX) chip is effective in improving clinical results compared to SRP alone. However, some recent studies failed to confirm these clinical results, and conflicting results were reported regarding the effects of the CHX chip on subgingival microflora. The aim of this study was to provide further data on the clinical and microbiologic effects of CHX chips when used as an adjunct to SRP.Methods: A total of 116 systemically healthy individuals with moderate to advanced periodontitis, aged 33 to 65 years, were recruited from the Departments of Periodontology of four Italian universities. For each subject, two experimental sites were chosen that had probing depths (PD) ≥5 mm and bleeding on probing (BOP) and were located in two symmetric quadrants. These two sites were randomized at the split‐mouth level, with one receiving SRP treatment alone and the other receiving treatment with SRP plus one CHX chip (SRP + CHX). PD, relative attachment level (RAL), and BOP were evaluated at baseline, prior to any treatment, and after 3 and 6 months. Supragingival plaque and the modified gingival index were evaluated at baseline and after 15 days and 1, 3, and 6 months. Subgingival microbiologic samples were harvested at baseline and after 15 days and 1, 3, and 6 months, cultured for total bacterial counts (TBCs), and investigated by polymerase chain reaction analysis for the identification of eight putative periodontopathogens.Results: When all of the pockets were considered, the PD and RAL were significantly less at 3 and 6 months compared to the baseline scores (P <0.01) for both treatments. Moreover, the PD was reduced in the SRP + CHX treatment group compared to the SRP treatment group at 3 and 6 months, whereas the RAL was similar for both treatments at 3 months and was reduced in the SRP + CHX treatment group at 6 months. The differences in PD reductions between the treatments were 0.30 and 0.55 mm at 3 and 6 months, respectively (P <0.01); for the RAL gain, the differences were 0.28 and 0.64 mm, respectively (P <0.001). The TBCs decreased significantly with both treatments. A similar, although less evident, pattern was noted when only the pockets with an initial PD ≥7 mm were considered. The percentage of sites positive for BOP was similar between the treatments at each time point. At 15 days and 1 month, the TBC for the SRP + CHX treatment group was significantly lower than for the SRP treatment group (P <0.01 and P <0.05, respectively). Over time, both treatments generally reduced the percentages of sites positive for the eight putative periodontopathic bacteria, although greater reductions were seen often for the SRP + CHX treatment group.Conclusions: The adjunctive use of the CHX chip resulted in a significant PD reduction and a clinical attachment gain compared to SRP alone. These results were concomitant with a significant benefit of SRP + CHX treatment on the subgingival microbiota.

Background: The use of locally delivered antibacterials containing chlorhexidine (CHX) was proposed to improve the effectiveness of non‐surgical periodontal treatment. The present multicenter randomized study investigated the effects of a xanthan‐based chlorhexidine (Xan‐CHX) gel used as an adjunct to scaling and root planing (SRP) in the treatment of chronic periodontitis.Methods: Ninety‐eight systemically healthy subjects with moderate to advanced periodontitis were recruited in four centers (59 females and 39 males; aged 24 to 58 years). For each subject, two experimental sites located in two symmetric quadrants were chosen with probing depths (PD) ≥5 mm and positive for bleeding on probing (BOP). These two sites were randomized at the split‐mouth level with one receiving a single SRP treatment and the other receiving a single SRP + Xan‐CHX gel treatment. Supragingival plaque, modified gingival index, PD, clinical attachment level (CAL), and BOP were evaluated at baseline (prior to any treatment) and after 3 and 6 months. At the same times, subgingival microbiologic samples and gingival crevicular fluid (GCF) were collected for the analysis of total bacterial counts (TBCs), including the identification of eight putative periodontopathogens, and alkaline phosphatase (ALP) activity, respectively.Results: The Xan‐CHX treatment group showed greater improvements compared to the SRP group for PD and CAL at 3 and 6 months (P <0.001). The differences in PD reduction between the treatments were 0.87 and 0.83 mm at 3 and 6 months, respectively (P <0.001); for CAL, these were 0.94 and 0.90 mm, respectively (P <0.001). Similar behavior was seen when the subgroup of pockets ≥7 mm was considered. The percentage of sites positive for BOP was similar between the treatments at each time point. For the comparisons between the treatment groups, no differences were seen in the TBCs and GCF ALP activity at baseline and 6 months; in contrast, slightly, but significantly, lower scores were recorded for the Xan‐CHX treatment group at 3 months (P = 0.018 and P = 0.045, respectively). Moreover, greater reductions in the percentages of sites positive for the eight putative periodontopathic bacteria were generally seen for the Xan‐CHX treatment group compared to SRP alone.Conclusions: The adjunctive use of Xan‐CHX gel promoted greater PD reductions and CAL gains compared to SRP alone. These results were concomitant with better microbiologic and biochemical outcomes when Xan‐CHX gel use was added to SRP, particularly up to 3 months after treatment.

Background: Microbial penetration inside an implant's internal cavity results in a bacterial reservoir that has been associated with an area of inflamed connective tissue facing the fixture–abutment junction. The aim of the present clinical trial was to evaluate the effectiveness of a 1% chlorhexidine gel on the internal bacterial contamination of implants with screw‐retained abutments.Methods: Thirty subjects (age range: 27.3 to 54.2 years) underwent single implant restoration. Three months after prosthodontic restoration, the modified sulcus bleeding index, modified plaque index, full‐mouth plaque score, and full‐mouth bleeding score were recorded. Microbiologic samples were also collected from the internal part of each fixture. Subjects were then divided into two equal groups: control and test groups (CG and TG, respectively). The CG had the abutment screwed and the crown cemented without any further intervention. Conversely, the TG had the internal part of the fixture filled with a 1% chlorhexidine gel before the abutment placement and screw tightening. Six months later, microbiologic and clinical procedures were repeated in both groups. Total bacterial count and multiplex polymerase chain analysis were performed to detect specific pathogens.Results: Clinical parameters remained stable throughout the study. From baseline to the 6‐month examination, the total bacterial counts underwent a significant reduction in the TG (P <0.05). Detection of the single pathogen species did not show any significant differences. However, periopathogens were detected more frequently in the CG.Conclusion: The application of a 1% chlorhexidine gel seemed to be an effective method to reduce bacterial colonization of the implant cavity over a 6‐month period.