Cold Atmospheric Plasma Treatment of Chondrosarcoma Cells Affects Proliferation and Cell Membrane Permeability

Chondrosarcoma is the second most common malign bone tumor in adults. Surgical resection of the tumor is recommended because of its resistance to clinical treatment such as chemotherapy and radiation therapy. Thus, the prognosis for patients mainly depends on sufficient surgical resection. Due to this, research on alternative therapies is needed. Cold atmospheric plasma (CAP) is an ionized gas that contains various reactive species. Previous studies have shown an anti-oncogenic potential of CAP on different cancer cell types. The current study examined the effects of treatment with CAP on two chondrosarcoma cell lines (CAL-78, SW1353). Through proliferation assay, the cell growth after CAP-treatment was determined. A strong antiproliferative effect for both cell lines was detected. By fluorescein diacetate (FDA) assay and ATP release assay, alterations in the cell membrane and associated translocation of low molecular weight particles through the cytoplasmic membrane were observed. In supernatant, the non-membrane-permeable FDA and endogenously synthesized ATP detected suggest an increased membrane permeability after CAP treatment. Similar results were shown by the dextran-uptake assay. Furthermore, fluorescence microscopic G-/F-actin assay was performed. G-and F-actin were selectively dyed, and the ratio was measured. The presented results indicate CAP-induced changes in cell membrane function and possible alterations in actin-cytoskeleton, which may contribute to the antiproliferative effects of CAP. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

New Approach against Chondrosoma Cells—Cold Plasma Treatment Inhibits Cell Motility and Metabolism, and Leads to Apoptosis

(1) Background: Chondrosarcoma (CS) is a malignant primary bone tumor with a carti-laginous origin. Its slow cell division and severely restricted vascularization are responsible for its poor responsiveness to chemotherapy and radiotherapy. The decisive factor for the prognosis of CS patients is the only adequate therapy—surgical resection. Cold atmospheric pressure plasma (CAP) is emerging as a new option in anti-cancer therapy. Its effect on chondrosarcomas has been poorly investigated. (2) Methods: Two CS cell lines—SW 1353 and CAL 78—were used. Various assays, such as cell growth kinetics, glucose uptake, and metabolic activity assay, along with two different apoptosis assays were performed after CAP treatment. A radius cell migration assay was used to examine cell motility. (3) Results: Both cell lines showed different growth behavior, which was taken into account when using the assays. After CAP treatment, a reduction in metabolic activity was observed in both cell lines. The immediate effect of CAP showed a reduction in cell numbers and in influence on this cell line’s growth rate. The measurement of the glucose concentration in the cell culture medium showed an increase after CAP treatment. Live-dead cell imaging shows an increase in the proportion of dead cells over the incubation time for both cell lines. There was a significant increase in apoptotic signals after 48 h and 72 h for both cell lines in both assays. The migration assay showed that CAP treatment inhibited the motility of chondrosarcoma cells. The effects in all experiments were related to the duration of CAP exposure. (4) Conclusions: The CAP treatment of CS cells inhibits their growth, motility, and metabolism by initiating apoptotic processes

Inhibition of Angiogenesis by Treatment with Cold Atmospheric Plasma as a Promising Therapeutic Approach in Oncology

Background: Cold atmospheric plasma (CAP) is increasingly used in the field of oncology. Many of the mechanisms of action of CAP, such as inhibiting proliferation, DNA breakage, or the destruction of cell membrane integrity, have been investigated in many different types of tumors. In this regard, data are available from both in vivo and in vitro studies. Not only the direct treatment of a tumor but also the influence on its blood supply play a decisive role in the success of the therapy and the patient’s further prognosis. Whether the CAP influences this process is unknown, and the first indications in this regard are addressed in this study. Methods: Two different devices, kINPen and MiniJet, were used as CAP sources. Human endothelial cell line HDMEC were treated directly and indirectly with CAP, and growth kinetics were performed. To indicate apoptotic processes, caspase-3/7 assay and TUNEL assay were used. The influence of CAP on cellular metabolism was examined using the MTT and glucose assay. After CAP exposure, tube formation assay was performed to examine the capillary tube formation abilities of HDMEC and their migration was messured in separate assays. To investigate in a possible mutagenic effect of CAP treatment, a hypoxanthine-guanine-phosphoribosyl-transferase assay with non malignant cell (CCL-93) line was performed. Results: The direct CAP treatment of the HDMEC showed a robust growth-inhibiting effect, but the indirect one did not. The MMT assay showed an apparent reduction in cell metabolism in the first 24 h after CAP treatment, which appeared to normalize 48 h and 72 h after CAP application. These results were also confirmed by the glucose assay. The caspase 3/7 assay and TUNEL assay showed a significant increase in apoptotic processes in the HDMEC after CAP treatment. These results were independent of the CAP device. Both the migration and tube formation of HDMEC were significant inhibited after CAP-treatment. No malignant effects could be demonstrated by the CAP treatment on a non-malignant cell line. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

An Innovative Therapeutic Option for the Treatment of Skeletal Sarcomas: Elimination of Osteo- and Ewing’s Sarcoma Cells Using Physical Gas Plasma

Osteosarcoma and Ewing’s sarcoma are the most common malignant bone tumors. Conventional therapies such as polychemotherapy, local surgery, and radiotherapy improve the clinical outcome for patients. However, they are accompanied by acute and chronic side effects that affect the quality of life of patients, motivating novel research lines on therapeutic options for the treatment of sarcomas. Previous experimental work with physical plasma operated at body temperature (cold atmospheric plasma, CAP) demonstrated anti-oncogenic effects on different cancer cell types. This study investigated the anti-cancer effect of CAP on two bone sarcoma entities, osteosarcoma and Ewing’s sarcoma, which were represented by four cell lines (U2-OS, MNNG/HOS, A673, and RD-ES). A time-dependent anti-proliferative effect of CAP on all cell lines was observed. CAP-induced alterations in cell membrane functionality were detected by performing a fluorescein diacetate (FDA) release assay and an ATP release assay. Additionally, modifications of the cell membrane and modifications in the actin cytoskeleton composition were examined using fluorescence microscopy monitoring dextran-uptake assay and G-/F-actin distribution. Furthermore, the CAP-induced induction of apoptosis was determined by TUNEL and active caspases assays. The observations suggest that a single CAP treatment of bone sarcoma cells may have significant anti-oncogenic effects and thus may be a promising extension to existing applications. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Development of coronary atherosclerosis in older donor hearts after heart transplantation

Einleitung: Die Herztransplantation (HTX) ist heute eine der erfolgreichsten Therapieformen der terminalen Herzinsuffizienz. Allerdings ist die Zahl der Patienten auf den HTX-Wartelisten stets deutlich größer als die Zahl der verfügbaren Spenderherzen. Dies führt zur zunehmenden Verwendung älterer Spenderherzen die altersbedingte Veränderungen haben können. So beträgt die Prävalenz der koronaren Arteriosklerose in einer gesunden 20-25-jährigen Bevölkerung bereits 20%. Der europäische Spenderpool ist im Durchschnitt 10-15 Jahre älter und das Risiko für ein versehentliches Übertragen koronarer Arteriosklerose liegt bei 7%, mit allen ihren Folgen, wie einer erhöhten Empfängermortalität. Weiterhin ist wenig darüber bekannt, wie sich eine übertragene Koronarosklerose nach HTX entwickelt. Patienten und Methoden: Die Studie wurde an älteren, 40-65-jährigen Spenderherzen zur Untersuchung der Entwicklung von übertragener koronarer Arteriosklerose durchgeführt. Zwischen Januar 2001 und Dezember 2009 wurden in der Deutsche Stiftung Organtransplantation Nord-Ost insgesamt 549 Herzspender registriert. Davon waren 328 40-65 Jahre alt (Mittelwert 49,5 J.) und erfüllten die HTX- Kriterien. Herzkatheterisierung vor Explantation und nach HTX erfolgte in 42 Fällen (21,9%, Durchschnittsalter 49 J.). Es wurde die Progression oder Reduktion der koronaren Gefäßveränderungen untersucht. Ergebnisse: Vor HTX waren 26 Herzen (62%) frei von Arteriosklerose und 16 (38%) nicht. Die Arteriosklerosehäufigkeit war altersabhängig (p=0,05). Nach HTX entwickelten 14 (53,8%) von den 26 gesunden Herzen Gefäßveränderungen. In der Gruppe der 16 Herzen mit Arteriosklerose verschlechterten sich zwei (12,5%), fünf (31,2%) verbesserten sich und neun Herzen (56,2%) zeigten keine Veränderung. Fazit: Es ist bekannt, dass die spendervermittelte Arteriosklerose das HTX Ergebnis verschlechtert, aber es ist wenig über ihre weitere Entwicklung in den Empfänger bekannt. Auch in einer kleinen Untersuchungsgruppe wurden nach HTX sowohl Verschlechterungen als auch Verbesserungen des koronaren Zustandes beobachtet. Die jüngeren Spenderherzen (40-49 J.) zeigten sowohl vor als auch nach HTX weniger Arteriosklerose als die älteren (50-65 J.). Nach HTX verschlechterten sich die älteren Spenderherzen stärker als die jüngeren. Vor allem in älteren weiblichen Spenderherzen verschlechterten sich die Befunde. Somit bargen ältere weibliche Spenderherzen ein höheres Risiko für die Empfänger als die Herzen der Vergleichsgruppen. Zusätzlich konnte aus Empfängersicht festgestellt werden, dass männliche Empfänger das schlechteste Milieu für ältere weibliche Spenderherzen waren.Introduction: Today, heart transplantation (HTX) is a routine therapy for end- stage heart failure. However, the number of patients on the HTX waiting list always exceeds the number of available donor hearts, leading to increasing use of older donor hearts. Coronary atherosclerosis (CAS) prevalence already amounts to 20% in a healthy, 20 to 25-year-old population. The European donor pool is 10-15 years older, bearing considerable risk for inadvertently transmitted CAS in about 7% of transplanted hearts. Little is known about how inadvertently transmitted CAS develops after HTX. Patients and Methods: This study was carryed out in older, 40 to 65-year-old donor hearts to study CAS development after HTX. Between January 2001 and December 2009 549 heart donors were registered in the German Foundation for Organ Transplantation North East; 328 were 40-65 years old (mean 49.5 years) and fulfilled HTX criteria. Coronary catheterization was performed in 42 (21.9%, mean age 49 years) before and after HTX. CAS progression or reduction was studied. Results: Before HTX, 26 (62%) hearts were CAS free and 16 (38%) were not. CAS frequency was age dependent (p=0.05). After HTX, 14 (53.8%) out of 26 healthy hearts developed CAS. In the group of 16 hearts with CAS, two (12.5%) showed CAS aggravation, five (31.25%) improvement and nine (56.25%) no change. Conclusion: Inadvertently transmitted donor heart CAS is known to worsen HTX outcome. Little is known about CAS development in the recipient. We showed that CAS aggravation or improvement is observable even in a small population. Younger donor hearts (40-49 years) had less CAS before and after HTX than the older hearts (50-65 years). Older donor hearts worsened after HTX faster than the younger. The fastest CAS deterioration was observed in older female donor hearts. Thus, female older donor hearts involved the highest risk for the recipient. From the recipients’ point of view male recipients were the least favorable environment for older female donor hearts

Why Kinematic Alignment Makes Little Sense in Valgus Osteoarthritis of the Knee: A Narrative Review

There is a debate about the best alignment strategies in total knee arthroplasty (TKA). Mechanical alignment (MA) targets in combination with necessary soft tissue releases are the gold standard for TKA in end-stage valgus osteoarthritis. Some authors propagate kinematic alignment (KA) with the aim of restoring the patient’s native alignment and minimizing the need for soft tissue releases. Our previous studies showed that MA with standardized soft tissue release produces reproducible results, and that the preoperative phenotype does not influence the results of patients with valgus osteoarthritis. These data suggest that there is no functional advantage to preserving valgus alignment in patients with valgus osteoarthritis. Many patients with valgus osteoarthritis present with a compromised medial collateral ligament and leaving the knee in valgus could increase the risk of secondary instability. The current literature supports MA TKA with soft tissue release as the gold standard. While using more sophisticated enabling technologies like robotic surgery might allow for aiming for very slight (1–2°) valgus alignment on the femoral side, any valgus alignment outside this range should be avoided. This review paper summarizes our current knowledge on the surgical techniques of TKA in patients with valgus osteoarthritis

Medial injuries of the clavicle: more prevalent than expected? A big data analysis of incidence, age, and gender distribution based on nationwide routine data

Purpose!#!Although shoulder-girdle injuries occur frequently, injuries of the medial part remain widely unexplored. This study overviews these rare injuries with a focus on incidence, age, and sex distribution in Germany.!##!Methods!#!The data are based on diagnoses according to ICD-10 in all German hospitals provided by the German Federal Statistical Office. ICD-10 codes S42.01 (medial clavicle fracture, MCF) and S43.2 (sternoclavicular joint dislocation, SCJD) were evaluated in detail between 2012 and 2014.!##!Results!#!We identified 14,264 cases with medial clavicle injuries (MCIs). MCFs occurred more often (11.6% of all clavicle-related shoulder-girdle injuries vs. 0.6% for SCJD). Mean ages of MCI were significantly different between males (43.7 years) and females (57.1 years) (p < 0.01). Age demonstrated a bimodal distribution with peaks at 20 and 50 years, which were predominantly associated with males. Females showed more injuries at age beyond 70 years. This applies to both SCJD and MCF. The incidence rate of these shoulder-girdle injuries was 47.0 per 100,000 person-years, for MCIs overall 5.9 (4.1 for men, 1.8 for women). This indicates disparity with a significant predominance of male patients over females as for all shoulder-girdle injuries (p < 0.01). Among children (< 16 years old), the incidence rate showed no significant difference in gender ratio.!##!Conclusion!#!MCIs appear more frequently than estimated so far and are distinguished from other clavicle fractures in that they occur more at higher age and peaking around 50 years. Further work on possible prevention strategies should focus on the most frequently affected groups of men around 20 and 50 years old

Shoulder girdle injuries involving the medial clavicle differ from lateral clavicle injuries with a focus on concomitant injuries and management strategies

Introduction Although shoulder girdle injuries are frequent, those of the medial part are widely unexplored. Our aim is to improve the knowledge of this rare injury and its management in Germany by big data analysis. Methods The data are based on ICD-10 codes of all German hospitals as provided by the German Federal Statistical Office. Based on the ICD-10 codes S42.01 (medial clavicle fracture, MCF) and S43.2 (sternoclavicular joint dislocation, SCJD), anonymized patient data from 2012 to 2014 were evaluated retrospectively for epidemiologic issues. We analyzed especially the concomitant injuries and therapy strategies. Results A total of 114,003 cases with a clavicle involving shoulder girdle injury were identified with 12.5% of medial clavicle injuries (MCI). These were accompanied by concomitant injuries, most of which were thoracic and craniocerebral injuries as well as injuries at the shoulder/ upper arm. A significant difference between MCF and SCJD concerning concomitant injuries only appears for head injuries (p = 0.003). If MCI is the main diagnosis, soft tissue injuries typically occur as secondary diagnoses. The MCI are significantly more often associated with concomitant injuries (p < 0.001) for almost each anatomic region compared with lateral clavicle injuries (LCI). The main differences were found for thoracic and upper extremity injuries. Different treatment strategies were used, most frequently plate osteosynthesis in more than 50% of MCF cases. Surgery on SCJD was performed with K-wires, tension flange or absorbable materials, fewer by plate osteosynthesis. Conclusions We proved that MCI are rare injuries, which might be why they are treated by inhomogeneous treatment strategies. No standard procedure has yet been established. MCI can occur in cases of severely injured patients, often associated with severe thoracic or other concomitant injuries. Therefore, MCI appear to be more complex than LCI. Further studies are required regarding the development of standard treatment strategy and representative clinical studies

Cold Atmospheric Plasma Treatment of Chondrosarcoma Cells Affects Proliferation and Cell Membrane Permeability

Chondrosarcoma is the second most common malign bone tumor in adults. Surgical resection of the tumor is recommended because of its resistance to clinical treatment such as chemotherapy and radiation therapy. Thus, the prognosis for patients mainly depends on sufficient surgical resection. Due to this, research on alternative therapies is needed. Cold atmospheric plasma (CAP) is an ionized gas that contains various reactive species. Previous studies have shown an anti-oncogenic potential of CAP on different cancer cell types. The current study examined the effects of treatment with CAP on two chondrosarcoma cell lines (CAL-78, SW1353). Through proliferation assay, the cell growth after CAP-treatment was determined. A strong antiproliferative effect for both cell lines was detected. By fluorescein diacetate (FDA) assay and ATP release assay, alterations in the cell membrane and associated translocation of low molecular weight particles through the cytoplasmic membrane were observed. In supernatant, the non-membrane-permeable FDA and endogenously synthesized ATP detected suggest an increased membrane permeability after CAP treatment. Similar results were shown by the dextran-uptake assay. Furthermore, fluorescence microscopic G-/F-actin assay was performed. G- and F-actin were selectively dyed, and the ratio was measured. The presented results indicate CAP-induced changes in cell membrane function and possible alterations in actin-cytoskeleton, which may contribute to the antiproliferative effects of CAP

Orthopaedic applications of cold physical plasma

Cold physical plasma (CPP) technology is of high promise for various medical applications. The interplay of specific components of physical plasma with living cells, tissues and organs on a structural and functional level is of paramount interest with the aim to induce therapeutic effects in a controlled and replicable fashion. In contrast to other medical disciplines such as dermatology and oromaxillofacial surgery, research reports on CPP application in orthopaedics are scarce. The present implementation of CPP in orthopaedics involves surface modifications of orthopaedic materials and biomaterials to optimize osseointegration. In addition, the influence of CPP on musculoskeletal cells and tissues is a focus of research, including possible adverse reactions and side effects. Its bactericidal aspects make CPP an attractive supplement to current treatment regimens in case of microbial inflammations such as periprosthetic joint infections. Attributed anticancerogenic and pro-apoptotic effects underline the clinical relevance of CPP as an additive in treating malignant bone lesions. The present review outlines ongoing research in orthopaedics involving CPP; it distinguishes considerations for safe application and the need for more evidence-based research to facilitate robust clinical implementation