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- Klinik und Poliklinik für Chirurgie Abt. für Unfall- und Wiederherstellungschirurgie (7) (remove)
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on-healing wounds continue to be a clinical challenge for patients and medical staff.
These wounds have a heterogeneous etiology, including diabetes and surgical trauma wounds. It is
therefore important to decipher molecular signatures that reflect the macroscopic process of wound
healing. To this end, we collected wound sponge dressings routinely used in vacuum assisted therapy
after surgical trauma to generate wound-derived protein profiles via global mass spectrometry.
We confidently identified 311 proteins in exudates. Among them were expected targets belonging to
the immunoglobulin superfamily, complement, and skin-derived proteins, such as keratins. Next to
several S100 proteins, chaperones, heat shock proteins, and immune modulators, the exudates
presented a number of redox proteins as well as a discrete neutrophil proteomic signature, including
for example cathepsin G, elastase, myeloperoxidase, CD66c, and lipocalin 2. We mapped over 200
post-translational modifications (PTMs; cysteine/methionine oxidation, tyrosine nitration, cysteine
trioxidation) to the proteomic profile, for example, in peroxiredoxin 1. Investigating manually
collected exudates, we confirmed presence of neutrophils and their products, such as microparticles
and fragments containing myeloperoxidase and DNA. These data confirmed known and identified
less known wound proteins and their PTMs, which may serve as resource for future studies on
human wound healing
Osteosarcoma and Ewing’s sarcoma are the most common malignant bone tumors.Conventional therapies such as polychemotherapy, local surgery, and radiotherapy improve theclinical outcome for patients. However, they are accompanied by acute and chronic side effectsthat affect the quality of life of patients, motivating novel research lines on therapeutic optionsfor the treatment of sarcomas. Previous experimental work with physical plasma operated atbody temperature (cold atmospheric plasma, CAP) demonstrated anti-oncogenic effects on differentcancer cell types. This study investigated the anti-cancer effect of CAP on two bone sarcomaentities, 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 lineswas observed. CAP-induced alterations in cell membrane functionality were detected by performinga fluorescein diacetate (FDA) release assay and an ATP release assay. Additionally, modifications ofthe cell membrane and modifications in the actin cytoskeleton composition were examined usingfluorescence 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 significantanti-oncogenic effects and thus may be a promising extension to existing applications.
: Human osteosarcoma (OS) is the most common primary malignant bone tumor occurring
most commonly in adolescents and young adults. Major improvements in disease-free survival have
been achieved by implementing a combination therapy consisting of radical surgical resection of the
tumor and systemic multi-agent chemotherapy. However, long-term survival remains poor, so novel
targeted therapies to improve outcomes for patients with osteosarcoma remains an area of active
research. This includes immunotherapy, photodynamic therapy, or treatment with nanoparticles.
Cold atmospheric plasma (CAP), a highly reactive (partially) ionized physical state, has been shown
to inherit a significant anticancer capacity, leading to a new field in medicine called “plasma oncology.”
The current article summarizes the potential of CAP in the treatment of human OS and reviews the
underlying molecular mode of action.
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.
AbstractCold physical plasma is a partially ionized gas that generates various components identified as potential anticancer compounds. Due to its topical application, cold plasmas are suitable, especially in dermatological applications. We, therefore, tested the cold plasma effects in skin cancer cells in vitro. An atmospheric pressure argon plasma jet was used as the plasma source. The plasma exposure alone reduced the metabolic activity and induced lethal effects in a treatment time-dependent fashion in both cell lines investigated. This was accompanied by executioner caspases 3 and 7, cleavage indicative of apoptosis and reduced cell migration and proliferation. Recent research also indicated roles of novel indirubin derivatives with potent anticancer effects. Three candidates were tested, and reduced metabolic activity and viability in a dose-dependent manner were found. Strikingly, one compound exerted notable synergistic toxicity when combined with plasma in skin cancer cells, which may be promising for future in vivo experiments.
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.
(1) Background: Chondrosarcoma (CS) is a malignant primary bone tumor with a cartilaginous 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.