Cold Atmospheric Plasma Improves the Colonization of Titanium with Primary Human Osteoblasts: An In Vitro Study.

Several studies have shown that cold atmospheric plasma (CAP) treatment can favourably modify titanium surfaces to promote osteoblast colonization. The aim of this study was to investigate the initial attachment of primary human osteoblasts to plasma-treated titanium. Micro-structured titanium discs were treated with cold atmospheric plasma followed by the application of primary human osteoblasts. The microwave plasma source used in this study uses helium as a carrier gas and was developed at the Leibniz Institute for Surface Modification in Leipzig, Germany. Primary human osteoblasts were analyzed by fluorescence and cell biological tests (alkaline phosphatase activity and cell proliferation using WST-1 assay). The tests were performed after 4, 12, and 24 h and showed statistically significant increased levels of cell activity after plasma treatment. The results of this study indicate that plasma treatment improves the initial attachment of primary human osteoblasts to titanium. For the first time, the positive effect of cold atmospheric plasma treatment of micro-structured titanium on the initial colonization with primary human osteoblasts has been demonstrated. Overall, this study demonstrates the excellent biocompatibility of micro-structured titanium. The results of this study support efforts to use cold atmospheric plasmas in implantology, both for preimplantation conditioning and for regeneration of lost attachment due to peri-implantitis.

Effects of Cold Atmospheric Plasma Pre-Treatment of Titanium on the Biological Activity of Primary Human Gingival Fibroblasts.

Cold atmospheric plasma treatment (CAP) enables the contactless modification of titanium. This study aimed to investigate the attachment of primary human gingival fibroblasts on titanium. Machined and microstructured titanium discs were exposed to cold atmospheric plasma, followed by the application of primary human gingival fibroblasts onto the disc. The fibroblast cultures were analyzed by fluorescence, scanning electron microscopy and cell-biological tests. The treated titanium displayed a more homogeneous and denser fibroblast coverage, while its biological behavior was not altered. This study demonstrated for the first time the beneficial effect of CAP treatment on the initial attachment of primary human gingival fibroblasts on titanium. The results support the application of CAP in the context of pre-implantation conditioning, as well as of peri-implant disease treatment.

[The G8 questionnaire as a geriatric screening tool in urooncology]. / Der G8-Fragebogen als geriatrisches Screening-Tool in der Uroonkologie.

The G8 questionnaire is a geriatric screening tool recommended by the SIOG and the EAU. To save time and resources, it can precede a comprehensive geriatric assessment (CGA) in a two-step evaluation. Based on our experience, this 8-item questionnaire is easy and fast to perform (4 - 5 minutes), even by medical staff untrained in geriatrics. The G8 questionnaire has become an established screening tool also in geriatric oncology. It has been shown in several studies that it provides an independent prognostic indicator for the overall survival of cancer patients. However, some critical aspects remain: its low specificity (60 %), its focus on nutritional aspects, and possible interobserver differences. These aspects should be known and taken into account by clinicians.

Side effects by oral application of atmospheric pressure plasma on the mucosa in mice.

Cold atmospheric pressure plasma (CAP) has been investigated with promising results for peri-implant diseases treatment. However, prior to in-vivo applications of CAP sources in humans, short-term harmful mucosal damage or other unwanted side effects have to be reviewed. 180 male mice (B6C3F1) were divided into twelve treatment groups (n = 15). The right buccal cheek mucosa was treated with CAP. The first and second group each received continuous 10 sec irradiation with 2 different plasma sources (kINPen09, PS-MWM). The third group was treated with the kINPen09 for one minute. Control groups were treated with a corresponding dose of ultraviolet light for 8 seconds or 48 seconds and the other one was left untreated. The animals were weighed before and after treatment. The animals were sacrificed one day or one week after exposure. Stained tissue samples were histologically examined for tissue damage independently by two experienced pathologists. One day after CAP treatment histological analysis showed focal mucosal erosion with superficial ulceration and necrosis accompanied by a mild inflammatory reaction. One week after CAP treatment, the mucosal defects were completely re-epithelialized, associated with remnants of granulation tissue in the stroma irrespective of treatment duration. Furthermore, no cytological atypia was found and no severe weight loss occurred. The control groups did not show any alterations at all. CAP treatment led to a superficial mucosal damage that healed within few days. Nonetheless, further long-term experiments are necessary to exclude undesirable side effects after longer observation time. Particularly, potential carcinogenic effects must be ruled out prior to the application of CAP treatment in daily dental practice.

Destruction of oral biofilms formed in situ on machined titanium (Ti) surfaces by cold atmospheric plasma.

The decontamination of implant surfaces represents the basic procedure in the management of peri-implant diseases, but it is still a challenge. The study aimed to evaluate the degradation of oral biofilms grown in situ on machined titanium (Ti) discs by cold atmospheric plasma (CAP). ~200 Ti discs were exposed to the oral cavities of five healthy human volunteers for 72 h. The resulting biofilms were divided randomly between the following treatments: CAP (which varied in mean power, treatment duration, and/or the gas mixture), and untreated and treated controls (diode laser, air-abrasion, chlorhexidine). The viability, quantity, and morphology of the biofilms were determined by live/dead staining, inoculation onto blood agar, quantification of the total protein content, and scanning electron microscopy. Exposure to CAP significantly reduced the viability and quantity of biofilms compared with the positive control treatments. The efficacy of treatment with CAP correlated with the treatment duration and plasma power. No single method achieved complete biofilm removal; however, CAP may provide an effective support to established decontamination techniques for treatment of peri-implant diseases.

Removing biofilms from microstructured titanium ex vivo: a novel approach using atmospheric plasma technology.

The removal of biofilms from microstructured titanium used for dental implants is a still unresolved challenge. This experimental study investigated disinfection and removal of in situ formed biofilms from microstructured titanium using cold atmospheric plasma in combination with air/water spray. Titanium discs (roughness (Ra): 1.96 µm) were exposed to human oral cavities for 24 and 72 hours (n = 149 each) to produce biofilms. Biofilm thickness was determined using confocal laser scanning microscopy (n = 5 each). Plasma treatment of biofilms was carried out ex vivo using a microwave-driven pulsed plasma source working at temperatures from 39 to 43°C. Following plasma treatment, one group was air/water spray treated before re-treatment by second plasma pulses. Vital microorganisms on the titanium surfaces were identified by contact culture (Rodac agar plates). Biofilm presence and bacterial viability were quantified by fluorescence microscopy. Morphology of titanium surfaces and attached biofilms was visualized by scanning electron microscopy (SEM). Total protein amounts of biofilms were colorimetrically quantified. Untreated and air/water treated biofilms served as controls. Cold plasma treatment of native biofilms with a mean thickness of 19 µm (24 h) to 91 µm (72 h) covering the microstructure of the titanium surface caused inactivation of biofilm bacteria and significant reduction of protein amounts. Total removal of biofilms, however, required additional application of air/water spray, and a second series of plasma treatment. Importantly, the microstructure of the titanium discs was not altered by plasma treatment. The combination of atmospheric plasma and non-abrasive air/water spray is applicable for complete elimination of oral biofilms from microstructured titanium used for dental implants and may enable new routes for the therapy of periimplant disease.

Killing of adherent oral microbes by a non-thermal atmospheric plasma jet.

Atmospheric plasma jets are being intensively studied with respect to potential applications in medicine. The aim of this in vitro study was to test a microwave-powered non-thermal atmospheric plasma jet for its antimicrobial efficacy against adherent oral micro-organisms. Agar plates and dentin slices were inoculated with 6 log(10) c.f.u. cm(-2) of Lactobacillus casei, Streptococcus mutans and Candida albicans, with Escherichia coli as a control. Areas of 1 cm(2) on the agar plates or the complete dentin slices were irradiated with a helium plasma jet for 0.3, 0.6 or 0.9 s mm(-2), respectively. The agar plates were incubated at 37 degrees C, and dentin slices were vortexed in liquid media and suspensions were placed on agar plates. The killing efficacy of the plasma jet was assessed by counting the number of c.f.u. on the irradiated areas of the agar plates, as well as by determination of the number of c.f.u. recovered from dentin slices. A microbe-killing effect was found on the irradiated parts of the agar plates for L. casei, S. mutans, C. albicans and E. coli. The plasma-jet treatment reduced the c.f.u. by 3-4 log(10) intervals on the dentin slices in comparison to recovery rates from untreated controls. The microbe-killing effect was correlated with increasing irradiation times. Thus, non-thermal atmospheric plasma jets could be used for the disinfection of dental surfaces.

Combined surface-focused acoustic microscopy in transmission and scanning ultrasonic holography.

Employment of ultrasound techniques in nondestructive testing may require identification of the acoustic modes contributing to imaging. Such identification can be achieved, with some restrictions, by time-of-flight analysis. Another approach is acoustic holography that reveals the propagation properties of any selected mode. In anisotropic media, the propagation features are distinct and allow for a reliable classification of the selected mode. Both techniques were applied for classification of bonded, disbonded, and weakly bonded areas in directly bonded semiconductor wafers.

Application of spatially and temporally apodized non-confocal acoustic transmission microscopy to imaging of directly bonded wafers.

Application of a line-shaped point spread function (PSF) to imaging of void defects in directly bonded wafers is considered. Two non-confocally adjusted spherical transducers are employed to implement an acoustic microscope operating in transmission with a time dependent point spread function, whose shape is optimized by both temporal apodization of the received signal and spatial apodization of the transducer aperture. Strong imaging artifacts resulting from the generation and detection of edge waves are eliminated in this way. It is shown by several examples that only a broadband system can be utilized in order to obtain a line-shaped PSF suitable for imaging.