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Rafał Pokrowiecki,1,2 Urszula Szałaj,3,4 Damian Fudala,3 Tomasz Zaręba,5 Jacek Wojnarowicz,3 Witold Łojkowski,3 Stefan Tyski,5 Krzysztof Dowgierd,6 Agnieszka Mielczarek7 1Department of Cranio- Maxillofacial Surgery, Oral Surgery and Implantology, Medical University of Warsaw, Warsaw, Poland; 2Head and Neck Surgery Department—Maxillofacial Surgery Department, Craniofacial Center, Regional Specialized Children’s Hospital, Olsztyn, 10-561, Poland; 3Institute of High Pressure Physics, Polish Academy of Sciences, Warsaw, Poland; 4Faculty of Materials Engineering, Warsaw University of Technology, Warsaw, Poland; 5Department of Antibiotics and Microbiology, National Medicines Institute, Warsaw, Poland; 6Head and Neck Surgery Clinic for Children and Young Adults, Department of Clinical Pediatrics, University of Warmia and Mazury, Olsztyn, 10-719, Poland; 7Department of Conservative Dentistry, Medical University of Warsaw, Warsaw, PolandCorrespondence: Agnieszka Mielczarek, Department of Conservative Dentistry, Medical University of Warsaw, Warsaw, Poland, Tel +48 691226414 ; +48 22 116 64 46, Email agnieszka.mielczarek.wum@gmail.comBackground: Periimplantitis is continuously one of major threats for the uneventful functioning of dental implants. Current approaches of drug delivery systems are being more commonly implemented into oral- and maxillofacial biomaterials in order to decrease the risk of implant failure due to bacterial infection. Silver nanoparticles and their compounds have been proven in eradicating oral bacteria responsible for peri-implant infections. Nevertheless, their evaluation as coating for implant abutments has not been extensively evaluated so far. This article describes a novel coating consisting of zinc oxide (ZnO) and silver (Ag) nanoparticles (NPs). This coating was used to modify healing abutments that could be used as drug delivery systems in oral implantology.Materials and Method: Nanoparticles with a ZnO + 0.1% Ag composition were produced by microwave solvothermal synthesis and then incorporated into the surface of titanium healing abutments by high-power ultrasonic deposition. Surface morphology, roughness, wettability were evaluated. Ability of biofilm formation inhibition was tested against S. mutans, S. oralis, S. aureus and E. coli.Results: ZnO+0.1%Ag NPs were sufficiently deposed on the surface of the abutments creating nanostructured coating which increased surface roughness and decreased wettability. Modified abutments significantly decreased bacterial biofilm formation. Bacteria present in SEM studies were unlikely to settle and replicate on the experimental abutments as their cells were rounded, insufficiently spread on the surface and covered with released NPs.Conclusion: Experimental nanostructured abutments were easily manufactured by high-power ultrasonic deposition and provided significant antibacterial properties. Such biomaterials could be used as temporary drug delivery abutments for prevention and treatment of intra- and extraoral peri-implant infections in the area of the head and neck.Keywords: dental implants, periimplantitis, nanotechnology, nanoparticles, NPs, drug delivery, zinc oxide, ZnO, silver, Ag

Rafal Pokrowiecki,1–3 Jacek Wojnarowicz,4 Tomasz Zareba,2 Iwona Koltsov,4 Witold Lojkowski,4 Stefan Tyski,2,5 Agnieszka Mielczarek,6 Pawel Zawadzki1 1Department of Cranio-Maxillofacial Surgery, Oral Surgery and Implantology, Medical University of Warsaw, Warsaw, Poland; 2Department of Antibiotics and Microbiology, National Medicines Institute, Warsaw, Poland; 3Private Practice, Warsaw, Poland; 4Institute of High Pressure Physics, Polish Academy of Sciences, Warsaw, Poland; 5Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland; 6Department of Conservative Dentistry, Medical University of Warsaw, Warsaw, PolandCorrespondence: Agnieszka MielczarekDepartment of Conservative Dentistry, Medical University of Warsaw, Binieckiego 6, Warsaw 02-097, PolandTel +48 22 116 6446Fax +48 572 0154Email sekretariat.zachowawcza@wum.edu.plAim: The aims of this study were to investigate new nano-formulations based on ZnO and Ag nanoparticle (NP) compounds when used against clinical strains of oral gram-positive and gram-negative bacteria, and to examine the stability and behaviour of nano-formulation mixtures in saliva based on different compositions of Ag NPs, ZnO NPs and ZnO+x·Ag NPs. Methods: ZnO NPs with and without nanosilver were obtained by microwave solvothermal synthesis. Then, antibacterial activity was evaluated against bacteria isolated from human saliva. Behavior and nanoparticle solutions were evaluated in human saliva and control (artificial saliva and deionized water). Results were statistically compared. Results: The NP mixtures had an average size of 30±3 nm, while the commercial Ag NPs had an average size of 55±5 nm. The suspensions displayed differing antibacterial activities and kinetics of destabilisation processes, depending on NPs composition and fluid types.Conclusion: The present study showed that all NPs suspensions displayed significant destabilisation and high destabilisation over the 24 h of the analyses. The agglomeration processes of NPs in human saliva can be reversible.Keywords: nanomaterials, controlled drug release, nanoparticles, metal nanoparticles, bacteria

Rafal Pokrowiecki,1 Agnieszka Mielczarek,2 Tomasz Zareba,3 Stefan Tyski3,4 1Department of Head and Neck Surgery-Maxillofacial Surgery, Otolaryngology and Ophthalmology, Prof Stanislaw Popowski Voivoid Children Hospital, Olsztyn, 2Department of Conservative Dentistry, Medical University of Warsaw, 3Department of Antibiotics and Microbiology, National Medicines Institute, 4Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland Abstract: Peri-implant infective diseases (PIIDs) in oral implantology are commonly known as peri-implant mucositis (PIM) and periimplantitis (PI). While PIM is restricted to the peri-implant mucosa and is reversible, PI also affects implant-supporting bone and, therefore, is very difficult to eradicate. PIIDs in clinical outcome may resemble gingivitis and periodontitis, as they share similar risk factors. However, recent study in the field of proteomics and other molecular studies indicate that PIIDs exhibit significant differences when compared to periodontal diseases. This review aims to elucidate the current knowledge of PIIDs, their etiopathology and diversified microbiology as well as the role of molecular studies, which may be a key to personalized diagnostic and treatment protocols of peri-implant infections in the near future. Keywords: dental plaque, infection, titanium, microbiome, periimplantitis

Rafał Pokrowiecki,1,2 Tomasz Zaręba,3 Barbara Szaraniec,4 Krzysztof Pałka,5 Agnieszka Mielczarek,6 Elżbieta Menaszek,7 Stefan Tyski3,8 1Center for Cranio-Maxillo-Facial Surgery, Voivodeship Children’s Hospital, Olsztyn, 2Department of Oral Surgery, Jagiellonian Medical University, Kraków, 3Department of Antibiotics and Microbiology, National Medicines Institute, Warsaw, 4Faculty of Material Science and Ceramics, AGH University of Science and Technology, Kraków, 5Department of Materials Engineering, Lublin University of Technology, Lublin, 6Department of Conservative Dentistry, Medical University of Warsaw, Warsaw, 7Department of Cytobiology, Collegium Medicum, Jagiellonian University, Kraków, 8Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland Abstract: The addition of an antibacterial agent to dental implants may provide the opportunity to decrease the percentage of implant failures due to peri-implantitis. For this purpose, in this study, the potential efficacy of nanosilver-doped titanium biomaterials was determined. Titanium disks were incorporated with silver nanoparticles over different time periods by Tollens reaction, which is considered to be an eco-friendly, cheap, and easy-to-perform method. The surface roughness, wettability, and silver release profile of each disc were measured. In addition, the antibacterial activity was also evaluated by using disk diffusion tests for bacteria frequently isolated from the peri-implant biofilm: Streptococcus mutans, Streptococcus mitis, Streptococcus oralis, Streptococcus sanguis, Porphyromonas gingivalis, Staphylococcus aureus, and Escherichia coli. Cytotoxicity was evaluated in vitro in a natural human osteoblasts cell culture. The addition of nanosilver significantly increased the surface roughness and decreased the wettability in a dose-dependent manner. These surfaces were significantly toxic to all the tested bacteria following a 48-hour exposure, regardless of silver doping duration. A concentration of 0.05 ppm was sufficient to inhibit Gram-positive and Gram-negative species, with the latter being significantly more susceptible to silver ions. However, after the exposure of human osteoblasts to 0.1 ppm of silver ions, a significant decrease in cell viability was observed by using ToxiLight™ BioAssay Kit after 72 hours. Data from the present study indicated that the incorporation of nanosilver may influence the surface properties that are important in the implant healing process. The presence of nanosilver on the titanium provides an antibacterial activity related to the bacteria involved in peri-implantitis. Finally, the potential toxicological considerations of nanosilver should further be investigated, as both the antibacterial and cytotoxic properties may be observed at similar concentration ranges. Keywords: biomaterials, dental plaque, peri-implantitis, peri implant mucositis, silver, nanotechnology, nanomedicine