Tissue Adhesives in Reconstructive and Aesthetic Surgery-Application of Silk Fibroin-Based Biomaterials.

Tissue adhesives have been successfully used in various kind of surgeries such as oral and maxillofacial surgery for some time. They serve as a substitute for suturing of tissues and shorten treatment time. Besides synthetic-based adhesives, a number of biological-based formulations are finding their way into research and clinical application. In natural adhesives, proteins play a crucial role, mediating adhesion and cohesion at the same time. Silk fibroin, as a natural biomaterial, represents an interesting alternative to conventional medical adhesives. Here, the most commonly used bioadhesives as well as the potential of silk fibroin as natural adhesives will be discussed.

Success and patient satisfaction of immediately loaded zirconia implants with fixed restorations one year after loading.

BACKGROUND: There is limited evidence for the use of zirconium dioxide implants in immediate implant placement as well as for related immediate loading protocols. The aim of this retrospective study was to investigate the survival rate, success and patient satisfaction of immediately placed zirconia implants compared to delayed placed implants. METHODS: The study included 58 partially edentulous patients who were treated between 2013 and 2015 with immediate and delayed transgingival healing zirconium dioxide implants (SDS, Kreuzlingen/ Switzerland). In addition to survival and success rate, marginal bone loss was assessed using radiographs and soft tissue was evaluated using Pink Esthetic Score. Oral health-related quality of life was investigated prospectively using a modified OHIP questionnaire. RESULTS: The cumulative survival rate of all implants included was 92% with 88% classified as full success. No significant difference was found between the bone levels of immediately and delayed placed and immediately and delayed loaded implants. The mean Pink Esthetic Score after final prosthetic rehabilitation was 12.2/14 points indicating excellent esthetic clinical results. Analysis of the OHIP questionnaire showed a mean value of 0.54/100 points reflecting a high patient satisfaction. CONCLUSIONS: Immediate and delayed placed as well as loaded zirconium dioxide implants showed excellent results regarding implant success and survival in this study. Zirconium dioxide implants may ensure excellent esthetic results and high patient satisfaction.

Clinical performance of zirconia implant abutments luted to a titanium base - a retrospective cross-sectional study.

AIM: To evaluate the survival of implant-retained restorations fabricated on CAD/CAM-derived zirconia abutments luted to a titanium base. MATERIALS AND METHODS: 153 patients who received a total of 310 dental implants (Camlog Promote plus or Xive S) and all-ceramic restorations on yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP) abutments luted to a titanium base during the last 10 years were included. Patients were examined for technical complications during routine visits. Crestal bone level changes were randomly analyzed based on periapical radiographs of 75 implants. RESULTS: Among the included 153 patients, 17 ceramic chippings (5.5%), 6 abutment loosenings (1.9%), and 2 abutment fractures (0.6%) were identified. The mean follow-up time was 4.7 years (standard deviation [SD]: 1.94), with a follow-up period of up to 10 years (maximum). Kaplan-Meier estimation resulted in a survival rate without complications of 91.6% for the restoration and 97.4% for the abutment. There was no statistically significant difference between the two implant systems, either between implant location or regarding the complication rate of the type of restoration. For the 75 implants included in the radiographic analysis, the mean bone level change was 0.384 mm (SD: 0.242, 95% CI: 0.315 to 0.452) for the Camlog implant system and 0.585 mm (SD: 0.366, 95% CI: 0.434 to 0.736) for the Xive system (P = 0.007). CONCLUSION: The results of the present retrospective study demonstrate acceptable clinical outcomes for zirconia abutments luted to a titanium base in combination with all-ceramic restorations. The assessed abutment design does not appear to have a negative impact on peri-implant hard tissue.

Temporomandibular Joint Osteoarthritis: Regenerative Treatment by a Stem Cell Containing Advanced Therapy Medicinal Product (ATMP)-An In Vivo Animal Trial.

Temporomandibular joint osteoarthritis (TMJ-OA) is a chronic degenerative disease that is often characterized by progressive impairment of the temporomandibular functional unit. The aim of this randomized controlled animal trial was a comparative analysis regarding the chondroregenerative potency of intra-articular stem/stromal cell therapy. Four weeks after combined mechanical and biochemical osteoarthritis induction in 28 rabbits, therapy was initiated by a single intra-articular injection, randomized into the following groups: Group 1: AB Serum (ABS); Group 2: Hyaluronic acid (HA); Group 3: Mesenchymal stromal cells (STx.); Group 4: Mesenchymal stromal cells in hyaluronic acid (HA + STx.). After another 4 weeks, the animals were euthanized, followed by histological examination of the removed joints. The histological analysis showed a significant increase in cartilage thickness in the stromal cell treated groups (HA + STx. vs. ABS, p = 0.028; HA + ST.x vs. HA, p = 0.042; STx. vs. ABS, p = 0.036). Scanning electron microscopy detected a similar heterogeneity of mineralization and tissue porosity in the subchondral zone in all groups. The single intra-articular injection of a stem cell containing, GMP-compliant advanced therapy medicinal product for the treatment of iatrogen induced osteoarthritis of the temporomandibular joint shows a chondroregenerative effect.

The Impact of Lidocaine on Adipose-Derived Stem Cells in Human Adipose Tissue Harvested by Liposuction and Used for Lipotransfer.

The local anesthetic lidocaine, which has been used extensively during liposuction, has been reported to have cytotoxic effects and therefore would be unsuitable for use in autologous lipotransfer. We evaluated the effect of lidocaine on the distribution, number, and viability of adipose-derived stem cells (ASCs), preadipocytes, mature adipocytes, and leukocytes in the fatty and fluid portion of the lipoaspirate using antibody staining and flow cytometry analyses. Adipose tissue was harvested from 11 female patients who underwent liposuction. Abdominal subcutaneous fat tissue was infiltrated with tumescent local anesthesia, containing lidocaine on the left and lacking lidocaine on the right side of the abdomen, and harvested subsequently. Lidocaine had no influence on the relative distribution, cell number, or viability of ASCs, preadipocytes, mature adipocytes, or leukocytes in the stromal-vascular fraction. Assessing the fatty and fluid portions of the lipoaspirate, the fatty portions contained significantly more ASCs (p < 0.05), stem cells expressing the preadipocyte marker Pref-1 (p < 0.01 w/lidocaine, p < 0.05 w/o lidocaine), and mature adipocytes (p < 0.05 w/lidocaine, p < 0.01 w/o lidocaine) than the fluid portions. Only the fatty portion should be used for transplantation. This study found no evidence that would contraindicate the use of lidocaine in lipotransfer. Limitations of the study include the small sample size and the inclusion of only female patients.

Influence of the Casting Concentration on the Mechanical and Optical Properties of FA/CaCl2-Derived Silk Fibroin Membranes.

In this study, we describe the manufacturing and characterization of silk fibroin membranes derived from the silkworm Bombyx mori. To date, the dissolution process used in this study has only been researched to a limited extent, although it entails various potential advantages, such as reduced expenses and the absence of toxic chemicals in comparison to other conventional techniques. Therefore, the aim of this study was to determine the influence of different fibroin concentrations on the process output and resulting membrane properties. Casted membranes were thus characterized with regard to their mechanical, structural and optical assets via tensile testing, SEM, light microscopy and spectrophotometry. Cytotoxicity was evaluated using BrdU, XTT, and LDH assays, followed by live-dead staining. The formic acid (FA) dissolution method was proven to be suitable for the manufacturing of transparent and mechanically stable membranes. The fibroin concentration affects both thickness and transparency of the membranes. The membranes did not exhibit any signs of cytotoxicity. When compared to other current scientific and technical benchmarks, the manufactured membranes displayed promising potential for various biomedical applications. Further research is nevertheless necessary to improve reproducible manufacturing, including a more uniform thickness, less impurity and physiological pH within the membranes.

The Influence of Local Pamidronate Application on Alveolar Dimensional Preservation after Tooth Extraction-An Animal Experimental Study.

The aim of this randomized, controlled animal exploratory trial was to investigate the influence of local application of aminobisphosphonate pamidronate during the socket preservation procedure. Mandibular premolars were extracted in five Göttingen minipigs. Two animals underwent socket preservation using BEGO OSS (n = 8 sockets) and three animals using BEGO OSS + Pamifos (15 mg) (n = 12 sockets). After jaw impression, cast models (baseline, eight weeks postoperative) were digitized using an inLab X5 scanner (Dentsply Sirona) and the generated STL data were superimposed and analyzed with GOM Inspect 2018 (GOM, Braunschweig). After 16 weeks, the lower jaws were prepared and examined using standard histological methods. In the test group (BEGO OSS + pamidronate), buccooral dimensional loss was significantly lower, both vestibulary (0.80 ± 0.57 mm vs. 1.92 ± 0.63 mm; p = 0.00298) and lingually (1.36 ± 0.58 mm vs. 2.56 ± 0.65 mm; p = 0.00104) compared with the control group (BEGO OSS). The test group showed a significant difference between vestibular and lingual dimensional loss (p = 0.04036). Histology showed cortical and cancellous bone in the alveolar sockets without signs of local inflammation. Adjuvant application of pamidronate during socket preservation reduces alveolar dimensional loss significantly. Further investigations with regard to dose-response relationships, volume effects, side effects, and a verification of the suitability in combination with other bone substitute materials (BSMs) are necessary.

Triterpenes for Well-Balanced Scar Formation in Superficial Wounds.

Triterpenes are demonstrably effective for accelerating re-epithelialisation of wounds and known to improve scar formation for superficial lesions. Among the variety of triterpenes, betuline is of particular medical interest. Topical betuline gel (TBG) received drug approval in 2016 from the European Commission as the first topical therapeutic agent with the proven clinical benefit of accelerating wound healing. Two self-conducted randomized intra-individual comparison clinical studies with a total of 220 patients involved in TBG treatment of skin graft surgical wounds have been screened for data concerning the aesthetic aspect of wound healing. Three months after surgery wound treatment with TBG resulted in about 30% of cases with more discreet scars, and standard of care in about 10%. Patients themselves appreciate the results of TBG after 3 months even more (about 50%) compared to standard of care (about 10%). One year after surgery, the superiority of TBG counts for about 25% in comparison with about 10%, and from the patients' point of view, for 25% compared to 4% under standard of care. In the majority of wound treatment cases, there is no difference visible between TBG treatment and standard of care after 1 year of scar formation. However, in comparison, TBG still offers a better chance for discreet scars and therefore happens to be superior in good care of wounds.

Multiple central giant cell granuloma of the jaws: diagnostic signposts of Noonan syndrome and RASopathy.

Noonan syndrome (NS) is a phenotypically variable inherited multi-system disorder. Maxillofacial findings can be diagnostic, especially in the evaluation of discrete facial dysmorphia. Diagnostic landmark findings of therapeutic relevance for the jaws such as central giant cell granuloma (CGCG) are rare in NS. However, recent molecular genetic studies indicate that these rare, benign lesions are neoplasms and more common in specific syndromes grouped under the umbrella term RASopathies. A specialist surgical diagnosis can be helpful in identifying the underlying disease. This report outlines diagnosis and treatment of a case of CGCG for which jaw diagnosis became the key to identifying a syndromic disease.

Risk factors and injury patterns of e-scooter associated injuries in Germany.

Since the introduction of widely available e-scooter rentals in Hamburg, Germany in June of 2019, our emergency department has seen a sharp increase in the amount of e-scooter related injuries. Despite a rising number of studies certain aspects of e-scooter mobility remain unclear. This study examines the various aspects of e-scooter associated injuries with one of the largest cohorts to date. Electronic patient records of emergency department admissions were screened for e-scooter associated injuries between June 2019 and December 2021. Patient demographic data, mechanism of injury, alcohol consumption, helmet usage, sustained injuries and utilized medical resources were recorded. Overall, 268 patients (57% male) with a median age of 30.3 years (IQR 23.3; 40.0) were included. 252 (94%) were e-scooter riders themselves, while 16 (6%) were involved in crashes associated with an e-scooter. Patients in non-rider e-scooter crashes were either cyclists who collided with e-scooter riders or older pedestrians (median age 61.2 years) who tripped over parked e-scooters. While e-scooter riders involved in a crash sustained an impact to the head or face in 58% of cases, those under the influence of alcohol fell on their head or face in 84% of cases. This resulted in a large amount of maxillofacial soft tissue lacerations and fractures. Extremity fractures and dislocations were more often recorded for the upper extremities. This study comprises one of the largest cohorts of e-scooter associated injuries to date. Older pedestrians are at risk to stumble over parked e-scooters. E-scooter crashes with riders who consumed alcohol were associated with more severe injuries, especially to the head and face. Restricted e-scooter parking, enforcement of drunk driving laws for e-scooters, and helmet usage should be recommended.

Osseointegration of Zirconia Implants after UV-Light or Cold Atmospheric Plasma Surface Treatment In Vivo.

The influence of UV light and non-thermal plasma on the osseointegration of yttria-stabilized zirconia implants (Y-TZP) comparing the two methods is unclear. The aim of this study was to show the influence of these methods on the osseointegration of dental zirconia implants in an animal model. A total of 54 implants were either untreated, treated with UV light (UV), or non-thermal oxygen plasma for 12 min and inserted into the parietal bones of six domestic pigs. The animals were sacrificed after a healing interval of two, four, and nine weeks. The degree of osseointegration was determined using histomorphometric determination of bone-to-implant contact values (BIC) and the bone-to-implant contact values within the retentive parts of the implants (BAFO). BIC values decreased in all groups after four weeks of healing and re-increased after nine weeks in all groups. BAFO increased significantly over time in all groups. However, there were no statistically significant differences in BIC and BAFO values between the control group and the test groups and over time. Clinical studies may follow to confirm the influence of cold plasma and UV light on the healing and survival of zirconia implants.

Potential Risk Factors Influencing the Formation of Postoperative Seroma After Breast Surgery - A Prospective Study.

BACKGROUND/AIM: This trial intended to identify patient- and therapy-specific risk factors influencing the genesis of seroma and the extent of its formation. PATIENTS AND METHODS: Within a prospective randomized controlled trial, 70 patients (n=35 TissuGlu®; n=35 drain) underwent a mastectomy with or without sentinel lymphonodectomy. Specific seroma-associated risk factors were recorded. Regular outpatient aftercare was performed during a 90-day postoperative follow-up. RESULTS: A statistically significant increase in the postoperative seroma rate was demonstrated for those with pre-adiposity compared to normal body weight (p=0.016), as well as for the state of health evaluated by the score of American Society of Anesthesiologists (ASA) (ASA III compared to I; p=0.046), the presence of diabetes mellitus (p<0.001) and the reduction of the length of the surgical procedure (p=0.044). CONCLUSION: A high body mass index, a poor state of health (ASA score), and diabetes mellitus, as well as a shorter duration of surgery, favor the incidence of postoperative seroma.

Long-term Risk Assessment for Medical Application of Cold Atmospheric Pressure Plasma.

Despite increasing knowledge gained based on multidisciplinary research, plasma medicine still raises various questions regarding specific effects as well as potential risks. With regard to significant statements about in vivo applicability that cannot be prognosticated exclusively based on in vitro data, there is still a deficit of clinical data. This study included a clinical follow-up of five probands who had participated five years previously in a study on the influence of cold atmospheric pressure plasma (CAP) on the wound healing of CO2 laser-induced skin lesions. The follow-up included a complex imaging diagnostic involving dermatoscopy, confocal laser scanning microscopy (CLSM) and hyperspectral imaging (HSI). Hyperspectral analysis showed no relevant microcirculatory differences between plasma-treated and non-plasma-treated areas. In summary of all the findings, no malignant changes, inflammatory reactions or pathological changes in cell architecture could be detected in the plasma-treated areas. These unique in vivo long-term data contribute to a further increase in knowledge about important safety aspects in regenerative plasma medicine. However, to confirm these findings and secure indication-specific dose recommendations, further clinical studies are required.

Impact of the Lysine-urethane Adhesive TissuGlu® on Postoperative Complications and Interventions After Drain-free Mastectomy.

BACKGROUND/AIM: The influence of a polyurethane-based tissue adhesive (TissuGlu®) on common complications following breast surgery was investigated. PATIENTS AND METHODS: Within a Randomized-Controlled-Trial 70 women (n=35 TissuGlu®, n=35 drain) underwent a mastectomy with or without sentinel lymph node excision (SLNE), followed by a 90-day postoperative follow-up. RESULTS: Postoperative interventions: Non-inferiority of the application of TissuGlu® was seen. Pain-Level/ Hospitalization: A statistically significant pain reduction from day four onwards (p<0.001) and a shorter hospitalization period (p<0.001) was observed. In contrast, the TissuGlu® group showed increased mean puncture incidence (p=0.013), and increased puncture volume (p=0.021). CONCLUSION: Application of the polyurethane-based tissue adhesive TissuGlu® after mastectomy, with or without SLNE, showed potential for improvement of the clinical outcome. In contrast, high intervention rates and increased puncture volume, caused by recurring seromas following application of the surgical adhesive TissuGlu®, have a negative impact on the patient-specific convalescence.

Effect of process parameters on additive-free electrospinning of regenerated silk fibroin nonwovens.

Silk fibroin is a biomaterial with multiple beneficial properties for use in regenerative medicine and tissue engineering. When dissolving and processing the reconstituted silk fibroin solution by electrospinning, the arrangement and size of fibers can be manifold varied and according fiber diameters reduced to the nanometer range. Such nonwovens show high porosity as well as potential biocompatibility. Usually, electrospinning of most biomaterials demands for the application of additives, which enable stable electrospinning by adjusting viscosity, and are intended to evaporate during processing or to be washed out afterwards. However, the use of such additives increases costs and has to be taken into account in terms of biological risks when used for biomedical applications. In this study, we explored the possibilities of additive-free electrospinning of pure fibroin nonwovens and tried to optimize process parameters to enable stable processing. We used natural silk derived from the mulberry silkworm Bombyx mori. After degumming, the silk fibroin was dissolved and the viscosity of the spinning solution was controlled by partial evaporation of the initial solving agent. This way, we were able to completely avoid the use of additives and manufacture nonwovens, which potentially offer higher biocompatibility and reduced immunogenicity. Temperature and relative humidity during electrospinning were systematically varied (25-35 °C, 25-30% RH). In a second step, the nonwovens optionally underwent methanol treatment to initiate beta-sheet formation in order to increase structural integrity and strength. Comprehensive surface analysis on the different nonwovens was performed using scanning electron microscopy and supplemented by additional mechanical testing. Cytotoxicity was evaluated using BrdU-assay, XTT-assay, LDH-assay and live-dead staining. Our findings were, that an increase of temperature and relative humidity led to unequal fiber diameters and defective nonwovens. Resistance to penetration decreased accordingly. The most uniform fiber diameters of 998 ± 63 nm were obtained at 30 °C and 25% relative humidity, also showing the highest value for resistance to penetration (0.20 N). The according pure fibroin nonwoven also showed no signs of cytotoxicity. However, while the biological response showed statistical evidence, the material characteristics showed no statistically significant correlation to changes of the ambient conditions within the investigated ranges. We suggest that further experiments should explore additional ranges for temperature and humidity and further focus on the repeatability of material properties in dependency of suitable process windows.

Production and Characterization of Porous Fibroin Scaffolds for Regenerative Medical Application.

BACKGROUND/AIM: Silk is a natural biomaterial with several superior features for applications in regenerative medicine. In the present study an optimized process for manufacturing porous scaffolds out of the silk protein fibroin was developed. MATERIALS AND METHODS: The silk protein fibroin was dissolved in Ajisawa's reagent and the resulting fibroin solution was used to produce scaffolds by means of freeze-thawing cycling. Porosity, pressure and stab resistance as well as degradation behavior were assessed in order to characterize the physical properties of the resulting scaffolds. RESULTS: The resulting sponge-like fibroin scaffolds were highly porous while the porosity correlated inversely with the concentration of the starting fibroin solution. Increased initial fibroin concentrations of the scaffolds resulted in increased compressive and cannulation resistance. The majority of the fibroin scaffolds were digested by 1 mg/ml protease XIV in 3 weeks, indicating their biodegradability. CONCLUSION: The production of scaffolds made of varying fibroin concentrations by means of freeze-thawing, following dissolution using Ajisawa's reagent, provides a simple and straightforward strategy for adjusting the physical and chemical properties of fibroin scaffolds for various medical applications.

The Use of TissuGlu® Surgical Adhesive for Mastectomy With or Without Lymphonodectomy.

BACKGROUND/AIM: Using drains after breast surgery is a preventive, but invasive measure to reduce seroma formation. A polyurethane-based tissue adhesive (TissuGlu®) might facilitate drainage-free wound healing after mastectomy in a non-invasive manner. PATIENTS AND METHODS: Retrospectively, data from 84 patients (42 receiving TissuGlu®, 42 receiving a drainage) who underwent mastectomy, were collected (90 days postoperative follow-up). Study endpoints were defined as the number of fluid-related postoperative clinical interventions, cumulative volume of postoperative wound fluid, duration of hospitalization and postoperative complications. RESULTS: In the entirety of postoperative interventions, no significant difference could be demonstrated (p=0.298). The drainage arm showed significantly less seroma aspirations (p=0.024) and complications (p=0.012). A significantly reduced length of hospitalization (p<0.001) and less cumulative wound secretion volume (p<0.001) appeared in the TissuGlu® group. CONCLUSION: The polyurethane-based tissue adhesive is a less invasive alternative to drain use in mastectomy.

Hyperspectral imaging as a possible tool for visualization of changes in hemoglobin oxygenation in patients with deficient hemodynamics - proof of concept.

There is a lack of imaging tools for the evaluation of spatial alterations in microcirculation including blood oxygen saturation and hemoglobin distribution but recent innovative developments in hyperspectral technology may offer a solution. We examined different hemodynamic disorders in patients suffering from scleroderma, Dupuytren surgery, chronic foot ulcera and skin infections. Superficial and deeper blood oxygen saturation, hemoglobin distribution and water content were determined using hyperspectral imaging (HSI). In the patient with scleroderma, distinct cutaneous low perfused regions correlated with macroscopic skin aspects and seem to be potential therapy control marker. With HSI accurate clinical evaluation of a macroscopic conspicuous wound after Dupuytren surgery was possible and influenced further surveillance decisions. HSI clearly revealed the spatial geometry and also the clinically related perfusion parameters of abscess formation and chronic ulcer wounds. The hemodynamically relevant parameters like blood oxygen saturation (1 mm to approx. 6 mm subcutaneous), total hemoglobin distribution and tissue water content can be easily determined and visualized with HSI in near real time. Hence, this technique seems to be suitable for routine diagnostics of acute and chronic wounds as well as for the examination of systemic hemodynamic disturbances. Special indications may be transplant surveillance and monitoring of therapeutical interventions.

Hyperspectral imaging: innovative diagnostics to visualize hemodynamic effects of cold plasma in wound therapy.

An important clinical potential of cold atmospheric plasma (CAP) lies in tumor and wound treatment, whereby the last-mentioned is well-referenced already. However, the underlying mechanisms of improved wound healing have not been sufficiently clarified yet, in particular the influence of CAP on microcirculation. Hyperspectral imaging (HSI) enables the visualization of microcirculation of large tissue areas, thus this technique seems to be a candidate to examine CAP effects on perfusion and oxygen saturation in wounds. During clinical wound management, one chronic wound caused by peripheral arterial occlusive disease and one acute wound after surgical removal of cervical lymph nodes were examined using HSI before and after CAP treatment. HSI was able to demonstrate CAP effects on microcirculation showing a relevant increase of superficial and deeper cutaneous oxygen saturation together with elevated hemoglobin concentration in treated and also surrounding wound area. For the first time, it was shown that CAP improves the superficial and deeper oxygenation and hemoglobin perfusion in and around the treated area of acute and chronic wounds. This effect may contribute to healing support by CAP in wounds. HSI seems suitable for evaluating and monitoring CAP effects in clinical settings.

Visible tumor surface response to physical plasma and apoptotic cell kill in head and neck cancer.

The aim of the study was to learn, whether clinical application of cold atmospheric pressure plasma (CAP) is able to cause (i) visible tumor surface effects and (ii) apoptotic cell kill in squamous cell carcinoma and (iii) whether CAP-induced visible tumor surface response occurs as often as CAP-induced apoptotic cell kill. Twelve patients with advanced head and neck cancer and infected ulcerations received locally CAP followed by palliative treatment. Four of them revealed tumor surface response appearing 2 weeks after intervention. The tumor surface response expressed as a flat area with vascular stimulation (type 1) or a contraction of tumor ulceration rims forming recesses covered with scabs, in each case surrounded by tumor tissue in visible progress (type 2). In parallel, 9 patients with the same kind of cancer received CAP before radical tumor resection. Tissue specimens were analyzed for apoptotic cells. Apoptotic cells were detectable and occurred more frequently in tissue areas previously treated with CAP than in untreated areas. Bringing together both findings and placing side by side the frequency of clinical tumor surface response and the frequency of analytically proven apoptotic cell kill, detection of apoptotic cells is as common as clinical tumor surface response. There was no patient showing signs of an enhanced or stimulated tumor growth under influence of CAP. CAP was made applicable by a plasma jet, kINPen(®) MED (neoplas tools GmbH, Greifswald, Germany).