Publikationen am Institut für Chemie und Biotechnik

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Radivoievych, Aleksandar; Kolp, Benjamin; Grebinyk, Sergii; Prylutska, Svitlana; Ritter, Uwe; Zolk, Oliver; Glökler, Jörn Felix; Frohme, Marcus; Grebinyk, Anna
Silent death by sound: C60 fullerene sonodynamic treatment of cancer cells. - In: International journal of molecular sciences, ISSN 1422-0067, Bd. 24 (2023), 2, 1020, S. 1-17

The acoustic pressure waves of ultrasound (US) not only penetrate biological tissues deeper than light, but they also generate light emission, termed sonoluminescence. This promoted the idea of its use as an alternative energy source for photosensitizer excitation. Pristine C60 fullerene (C60), an excellent photosensitizer, was explored in the frame of cancer sonodynamic therapy (SDT). For that purpose, we analyzed C60 effects on human cervix carcinoma HeLa cells in combination with a low-intensity US treatment. The time-dependent accumulation of C60 in HeLa cells reached its maximum at 24 h (800 ± 66 ng/106 cells). Half of extranuclear C60 is localized within mitochondria. The efficiency of the C60 nanostructure’s sonoexcitation with 1 MHz US was tested with cell-based assays. A significant proapoptotic sonotoxic effect of C60 was found for HeLa cells. C60′s ability to induce apoptosis of carcinoma cells after sonoexcitation with US provides a promising novel approach for cancer treatment.
Schuett, Timo; Anufriev, Ilya; Endres, Patrick; Stumpf, Steffi; Nischang, Ivo; Hoeppener, Stephanie; Bode, Stefan; Schubert, Ulrich Sigmar; Geitner, Robert
A user-guide for polymer purification using dialysis. - In: Polymer chemistry, ISSN 1759-9962, Bd. 14 (2023), 1, S. 92-101

Dialysis diffusion kinetics are investigated via in situ NMR spectroscopy for numerous different raw polymeric solutions to result in a general guideline for polymer purification using dialysis. In several approaches, a polymer was on purpose contaminated with its respective monomer, regenerated conducting conventional dialysis and monitored online utilizing in situ NMR spectroscopy. Consequently, polymer type and molar mass, monomer type, molar mass cut-off of the dialysis tubing and type of solvent were varied resulting in 29 different purification approaches and over 40 000 NMR-spectra. As a result, several major parameters were identified affecting the purification process significantly such as the chosen solvent, viscosity and alpha value. On the contrary, parameters such as dialysis tubing molar mass cut-off and molar mass of the polymer did not affect the purification in a significant manner. Furthermore, physical properties such as density, viscosity, alpha value, and dipole moment of the ingredients were combined in a principal component analysis in order to identify the most important parameters.
Karagianni, Alexandra; Tsierkezos, Nikos; Prato, Maurizio; Terrones, Mauricio; Kordatos, Konstantinos V.
Application of carbon-based quantum dots in photodynamic therapy. - In: Carbon, ISSN 0008-6223, Bd. 203 (2023), S. 273-310

Photodynamic Therapy (PDT) is a non-invasive therapeutic modality that can treat a wide variety of cancer types by means of photosensitizer drug, light, and oxygen. Due to enhanced specificity and fewer side effects, PDT can be an alternative approach for cancer treatments. However, conventional photosensitizers (PSs) exhibit low selectivity, hydrophobicity, and limited photophysical properties. Nanotechnology emerges as a potential solution to these issues and improves PDT efficiency. Nanomaterials such as Carbon Quantum Dots (CQDs) and Graphene Quantum Dots (GrQDs) have been widely applied on PDT research recently, regarding their excellent photoluminescence properties, biocompatibility, as well as their hydrophilicity. The present review article summarizes the main features of PDT and carbon-based quantum dots with an emphasis on used PSs and methods for synthesis of carbon dots. Additionally, the most recent applications of CQDs and GrQDs in PDT have been extensively discussed. The main conclusion that arises is that carbon-based quantum dots seem to be a powerful tool in cancer diagnosis and treatment.
Xie, Ting; Köhler, Michael; Heyder, Stefan; Günther, Mike; Cao-Riehmer, Jialan
Microfluidically-assisted isolation and characterization of Achromobacter spanius from soils for microbial degradation of synthetic polymers and organic solvents. - In: Environments, ISSN 2076-3298, Bd. 9 (2022), 12, 147, S. 1-17

A micro segmented-flow approach was utilized for the isolation soil bacteria that can degrade synthetic polymers as polyethylene glycols (PEG) and polyacrylamide (PAM). We had been able to obtain many strains; among them, five Achromobacter spanius strains from soil samples of specific sampling sites that were connected with ancient human impacts. In addition to the characterization of community responses and isolating single strains, this microfluidic approach allowed for investigation of the susceptibility of Achromobacter spanius strains against three synthetic polymers, including PEG, PAM, and Polyvinylpyrrolidone (PVP) and two organic solvents known as 1,4-dioxane and diglyme. The small stepwise variation of effector concentrations in 500 nL droplets provides a detailed reflection of the concentration-dependent response of bacterial growth and endogenous autofluorescence activity. As a result, all five strains can use PEG600 as carbon source. Furthermore, all strains showed similar dose-response characteristics in 1,4-dioxane and diglyme. However, significantly different PAM- and PVP-tolerances were found for these strains. Samples from the surface soil of prehistorical rampart areas supplied a strain capable of degradation of PEG, PVP, and PAM. This study demonstrates on the one hand, the potential of microsegment flow for miniaturized dose-response screening studies and its ability to detect novel strains, and on the other hand, two of five isolated Achromobacter spanius strains may be useful in providing optimal growth conditions in bioremediation and biodegradation processes.
Geitner, Robert;
Physikalische Chemie : Trendbericht. - In: Nachrichten aus der Chemie, ISSN 1868-0054, Bd. 70 (2022), 5, S. 64-67

Die Aufklärung von Reaktionsmechanismen ist in der Katalyse wichtig, um die geschwindigkeitsbegrenzende Schritte zu verstehen und zu beschleunigen. Mit maschinellem Lernen lassen dann sich auf Basis der Mechanismen neue Katalysatoren entwickeln. Photochemische Umsetzungen in weichen Membranen folgen einer anderen Kinetik als Reaktionen in Lösung. Mikroschwimmer, Mikromotoren oder Phototaxis zählen zu aktiver Materie. Sie wandeln kontinuierlich Energie aus ihrer Umgebung um und bewegen sich autonom.
Prylutskyy, Yuriy; Nozdrenko, Dmytro; Gonchar, Olga; Prylutska, Svitlana; Bogutska, Kateryna; Franskevych, Daria; Hromovyk, Bohdan; Scharff, Peter; Ritter, Uwe
C60 fullerene attenuates muscle force reduction in a rat during fatigue development. - In: Heliyon, ISSN 2405-8440, Bd. 8 (2022), 12, e12449, S. 1-9

C60 fullerene (C60) as a nanocarbon particle, compatible with biological structures, capable of penetrating through cell membranes and effectively scavenging free radicals, is widely used in biomedicine. A protective effect of C60 on the biomechanics of fast (m. gastrocnemius) and slow (m. soleus) muscle contraction in rats and the pro- and antioxidant balance of muscle tissue during the development of muscle fatigue was studied compared to the same effect of the known antioxidant N-acetylcysteine (NAC). C60 and NAC were administered intraperitoneally at doses of 1 and 150 mg kg−1, respectively, daily for 5 days and 1 h before the start of the experiment. The following quantitative markers of muscle fatigue were used: the force of muscle contraction, the level of accumulation of secondary products of lipid peroxidation (TBARS) and the oxygen metabolite H2O2, the activity of first-line antioxidant defense enzymes (superoxide dismutase (SOD) and catalase (CAT)), and the condition of the glutathione system (reduced glutathione (GSH) content and the activity of the glutathione peroxidase (GPx) enzyme). The analysis of the muscle contraction force dynamics in rats against the background of induced muscle fatigue showed, that the effect of C60, 1 h after drug administration, was (15-17)% more effective on fast muscles than on slow muscles. A further slight increase in the effect of C60 was revealed after 2 h of drug injection, (7-9)% in the case of m. gastrocnemius and (5-6)% in the case of m. soleus. An increase in the effect of using C60 occurred within 4 days (the difference between 4 and 5 days did not exceed (3-5)%) and exceeded the effect of NAC by (32-34)%. The analysis of biochemical parameters in rat muscle tissues showed that long-term application of C60 contributed to their decrease by (10-30)% and (5-20)% in fast and slow muscles, respectively, on the 5th day of the experiment. At the same time, the protective effect of C60 was higher compared to NAC by (28-44)%. The obtained results indicate the prospect of using C60 as a potential protective nano agent to improve the efficiency of skeletal muscle function by modifying the reactive oxygen species-dependent mechanisms that play an important role in the processes of muscle fatigue development.
Khan, Nida Zaman; Martin, Daniel; Pliquett, Uwe; Zaikou, Yahor; Thomas, Nacke; Heinrich, Doris; Köhler, Michael; Nguyen, Thi-Huong
High-frequency contactless sensor for the detection of Heparin-induced thrombocytopenia antibodies via platelet aggregation. - In: International journal of molecular sciences, ISSN 1422-0067, Bd. 23 (2022), 22, 14395, insges. 13 S.

Heparin-induced thrombocytopenia (HIT), a severe autoimmune disorder, occurs in patients undergoing heparin therapy. The presence of platelet-activating antibodies against platelet factor 4/Heparin in the blood confirms patients suffering from HIT. The most widely used methods for HIT diagnosis are immunoassays but the results only suit to rule out HIT as the assays provide only around 50% specificity. To confirm HIT, samples with positive results in immunoassays are retested in functional assays (>98% specificity) that track platelet-activating antibodies via platelet aggregation. However, the protocols in functional assays are either time-consuming (due to the requirement of the detection of serotonin release) or require highly trained staff for the visualization of platelets. Here, we applied a cheap and easy-to-use contactless sensor, which employs high-frequency microwaves to detect the changes in the resonant frequency caused by platelet aggregation/activation. Analysis of change in conductivity and permittivity allowed us to distinguish between HIT-like (KKO) and non-HIT-like (RTO) antibodies. KKO caused a stronger reduction of conductivity of platelet samples than RTO. Our results imply that the high-frequency contactless sensor can be a promising approach for the development of a better and easier method for the detection of HIT.
Grebinyk, Anna; Prylutska, Svitlana; Grebinyk, Sergii; Ponomarenko, Stanislav; Virych, Pavlo; Chumachenko, Vasyl; Kutsevol, Nataliya; Prylutskyy, Yuriy; Ritter, Uwe; Frohme, Marcus
Drug delivery with a pH-sensitive star-like dextran-graft polyacrylamide copolymer. - In: Nanoscale advances, ISSN 2516-0230, Bd. 4 (2022), 23, S. 5077-5088

The development of precision cancer medicine relies on novel formulation strategies for targeted drug delivery to increase the therapeutic outcome. Biocompatible polymer nanoparticles, namely dextran-graft-polyacrylamide (D-g-PAA) copolymers, represent one of the innovative non-invasive approaches for drug delivery applications in cancer therapy. In this study, the star-like D-g-PAA copolymer in anionic form (D-g-PAAan) was developed for pH-triggered targeted drug delivery of the common chemotherapeutic drugs - doxorubicin (Dox) and cisplatin (Cis). The initial D-g-PAA copolymer was synthesized by the radical graft polymerization method, and then alkaline-hydrolyzed to get this polymer in anionic form for further use for drug encapsulation. The acidification of the buffer promoted the release of loaded drugs. D-g-PAAan nanoparticles increased the toxic potential of the drugs against human and mouse lung carcinoma cells (A549 and LLC), but not against normal human lung cells (HEL299). The drug-loaded D-g-PAAan-nanoparticles promoted further oxidative stress and apoptosis induction in LLC cells. D-g-PAAan-nanoparticles improved Dox accumulation and drugs’ toxicity in a 3D LLC multi-cellular spheroid model. The data obtained indicate that the strategy of chemotherapeutic drug encapsulation within the branched D-g-PAAan nanoparticle allows not only to realize pH-triggered drug release but also to potentiate its cytotoxic, prooxidant and proapoptotic effects against lung carcinoma cells.
Richter, Felix; Chen, Minqian; Schaub, Patrick; Wüst, Florian; Zhang, Di; Schneider, Steffen; Groß, Gregor Alexander; Mäder, Patrick; Dovzhenko, Oleksandr; Palme, Klaus; Köhler, Michael; Cao-Riehmer, Jialan
Induction of embryogenic development in haploid microspore stem cells in droplet-based microfluidics. - In: Lab on a chip, ISSN 1473-0189, Bd. 22 (2022), 22, S. 4292-4305

This work presents the application of droplet-based microfluidics for the cultivation of microspores from Brassica napus using the doubled haploid technology. Under stress conditions (e.g. heat shock) or by chemical induction a certain fraction of the microspores can be reprogrammed and androgenesis can be induced. This process is an important approach for plant breeding because desired plant properties can be anchored in the germline on a genetic level. However, the reprogramming rate of the microspores is generally very low, increasing it by specific stimulation is, therefore, both a necessary and challenging task. In order to accelerate the optimisation and development process, the application of droplet-based microfluidics can be a promising tool. Here, we used a tube-based microfluidic system for the generation and cultivation of microspores inside nL-droplets. Different factors like cell density, tube material and heat shock conditions were investigated to improve the yield of vital plant organoids. Evaluation and analysis of the stimuli response were done on an image base aided by an artificial intelligence cell detection algorithm. Droplet-based microfluidics allowed us to apply large concentration programs in small test volumes and to screen the best conditions for reprogramming cells by the histone deacetylase inhibitor trichostatin A and for enhancing the yield of vital microspores in droplets. An enhanced reprogramming rate was found under the heat shock conditions at 32 &ring;C for about 3 to 6 days. In addition, the comparative experiment with MTP showed that droplet cultivation with lower cell density (<10 cells per droplet) or adding media after 3 or 6 days significantly positively affects the microspore growth and embryo rate inside 120 nL droplets. Finally, the developed embryos could be removed from the droplets and further grown into mature plants. Overall, we demonstrated that the droplet-based tube system is suitable for implementation in an automated, miniaturized system to achieve the induction of embryogenic development in haploid microspore stem cells of Brassica napus.
Marx-Blümel, Lisa; Marx, Christian; Schober, Andreas; Beck, James F.
In vitro-Amplifikation humaner hämatopoetischer Stammzellen im 3D-System. - In: Biospektrum, ISSN 1868-6249, Bd. 28 (2022), 5, S. 489-492

A promising strategy to increase the numbers of hematopoietic stem cells (HSCs) for clinical applications, like stem cell transplantation, is offered by advanced in vitro culture systems. We developed artificial 3D bone marrow-like scaffolds made of polydimethylsiloxane (PDMS) mimicking the natural HSC niche in vitro. These 3D PDMS scaffolds in combination with an optimized culture medium allow the amplification of high numbers of undifferentiated HSCs by activating specific molecular signaling pathways.