An atrial septal defect (ASD) is one of the most common congenital heart defects in children and the most frequent congenital defect found in adults. Currently, several types of kits are available for percutaneous closure of ASD. The design of these implants is based on the Nitinol alloy. Despite the good biocompatibility of Nitinol alloys, the use of these materials for long-term implantation is questionable due to the high nickel content and the risk of releasing nickel ions as a result of corrosion in the body’s environment. A way to improve the hemocompatibility of Nitinol alloys is to modify their surface. As part of this work, the conditions for the production of SiO2 surface layers using the atomic layer deposition (ALD) method and laser surface modification with three different laser cutting speeds were developed to improve biocompatibility. This allowed us for the comparison of different surface modifications (ALD and laser modification) in terms of their impact on cell-material interactions. The general analysis concerning biocompatibility confirmed the biological usability of the designed ALD deposited coatings. Surface nanostructuring had a positive effect on the natural biological layer formation. The analysis performed indicated the appropriate behaviour of the natural biological layer, known in the literature as pseudointima, in contact with blood. It was evident that platelet activation on the surface was reduced.
Bacterial infections pose a serious threat to human health. For many years, there has been a search for materials that would inhibit their development. It was decided to take a closer look at various elastomeric materials with the addition of chitosan. Mixtures based on silicone, silicone with a platinum catalyst, acrylonitrile-butadiene rubber, natural rubber, and ethylene-propylene-diene rubber were developed and tested for antibacterial and physico-mechanical properties. The dispersion of chitosan in the elastomer was also investigated using a scanning electron microscope. Of the tested mixtures, three were selected, characterised by the best antibacterial and physico-mechanical properties and a very good dispersion of chitosan in the matrix. The mixtures were based on silicone, silicone with a platinum catalyst and natural rubber. Tests were performed to measure the release of compounds into water for these mixtures. Furthermore, cytotoxicity with L929 cells and cytocompatibility in direct contact with MG63 cells were investigated for silicone samples. The results showed that these materials were not toxic to mammalian cells and supported their growth. The best bactericidal properties against E. coli and S. aureus strains compared to the other tested materials (>99.0–99.9% of killed bacteria) were shown by samples made of silicone and silicone with a platinum catalyst and added chitosan. At the same time, the best physico-mechanical properties were found for the samples with chitosan based on silicone with added platinum and natural rubber. Developed materials appeared to be good candidates for manufacturing medical equipment on which the adhesion and growth of bacteria should be prevented.
Burn wounds are a unique type of injury that can affect the entire body and cause irreversible damage. They are characterized by significant morbidity and mortality due to the pathophysiology of the healing process manifested by unremitting inflammation, leading to a critical need to search for new treatments. This study focuses on the development of drug delivery systems in the form of lipid microparticles loaded with quercetin, as an agent to combat acute inflammation in burn wounds. We aimed to explore the effect of quercetin in modulating macrophage polarization from proinflammatory (M1) to anti-inflammatory (M2) phenotype. The absence of a cytotoxic effect of the produced particles on macrophages, as well as the lack of negative effects on their morphology was proven. The study confirmed the ability of quercetin and quercetin-loaded lipid microparticles to modulate macrophage polarization in an anti-inflammatory direction, based on the analysis of their surface markers expression performed with the use of flow cytometry. With the use of quercetin, the expression of M2 specific marker increased. Furthermore, better results were obtained for encapsulated quercetin, confirming the necessity of encapsulation to increase the therapeutic potential.
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The plant mediated biogenic synthesis of nanoparticles is of magnificent concern due to its eco-benign and single pot nature. Here, Cinnamomum tamala (C. tamala) aqueous leaf extract was utilised for the silver nanoparticles’ (Ag NPs) synthesis. The phytoconstituents in the leaf extract were analysed by standard methods. These metabolites, especially carbohydrate polymers reduce Ag ions to Ag NPs accompanied by a reddish-brown coloration of the reaction mixture. The visual observation of intense brown colour is the first indication of the formation of Ag NPs. Various spectro-analytical techniques further characterise the Ag NPs. The green synthesised spherical Ag NPs were crystalline with an average size of 38 nm. The Ag NPs were scrutinised for antioxidant, antimicrobial and cytotoxic activity and obtained good results. The free radical scavenging was studied by 2, 2-Diphenyl-l-picrylhydrazyl (DPPH) assay. The antibacterial activity of Ag NPs was assessed against human pathogens, and it shown to have good antibacterial potency against a wide spectrum of bacteria. The cytotoxic activity against HEK-293T (human embryonic kidney) cell line was evaluated by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay. These potent biological activities enable C. tamala capped Ag NPs to be suitable candidates for the future applications in various fields, predominantly clinical and biomedical.
In this study, the bio state of the alloy produced in the modified metal injection system was monitored after sintering. A new system operating with high gas pressure, far from the traditional injection model, has been established for material production. In this system, 316L stainless steel powders were molded using a PEG/PMMA/SA polymer recipe. During molding, approximately 60% 316L and 40% binder by volume were used. The samples obtained were sintered at different temperatures (1100-1300°C) after de-binding. Density measurement (Archimedes) and hardness tests (HV1) of the samples were measured as 6.74 g/cm3 and ~285 HV1, respectively. A potentiodynamic corrosion test was applied to monitor the effect of the amount of oxide in the structure of the 316L stainless steel produced. Corrosion tests were carried out in artificial body solutions. The corrosion rate was measured at the level of 17.08×10-3 mm/y. In terms of biocompatibility, a cytotoxicity test was applied to the samples and the life course of the bacteria was monitored. For the 316L alloys produced, the % vitality reached approximately 103%.
This study investigated the influence of three types of metallic microfillers, spherical silver and spherical, and dendritic copper, on the ability of polyamide 12 (PA12) to inhibit microorganism growth on the surfaces of samples produced using laser-based powder bed fusion of polymers (PBF-LB/P). The aim of this study was to initially characterize these materials regarding their potential applicability for parts dedicated to use in the hospitals, which surfaces are periodically disinfected using chemical and/or physical measures.
The research was focused on determining basic mechanical properties, surface, and phase structure taking into consideration basic cytotoxicity analysis towards human cells. Methods: Biological tests were performed on human C-12302 fibroblasts cells using 3D-printed Ti6Al4V alloy (Ti64), produced by laser-based powder bed fusion (LB-PBF) and Alumina Toughened Zirconia 20 (ATZ20), produced by lithography-based ceramic manufacturing (LCM). Surface modifications included electropolishing and hydroxyapatite or hydroxyapatite/zinc coating. Structure analysis was carried out using a variety of techniques such as X-Ray diffraction, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM), followed by mechanical properties evaluation using nanoindentation testing. Results: Samples subjected to surface modifications showed diversity among surface and phase structure and mechanical properties. However, the cytotoxicity towards tested cells was not significantly higher than the control. Though, a trend was noted among the materials analysed, indicating that HAp/Zn coating on Ti64 and ATZ20 resulted in the best biological performance increasing cell survivability by more than 10%. Conclusions: Hydroxyapatite coating on Ti64 and ATZ20 resulted in the best biological properties. Tested materials are suitable for in vivo toxicity testing.
The aim of the presented work was to characterize the new obtained bioglasses and assess their biological performance in vitro. Bioglasses were produced using the sol-gel method in the SiO2-P2O5-CaO system, for the purpose as composite ingredients. Their chemical composition was enriched with ZnO to introduce antibacterial properties and SrO with osteoinductive effect. The properties of bioglasses were compared and the effect of chemical composition and particle size on their biological properties was assessed. Methods: The bioglasses were evaluated via TG-DTA, FTIR, SEM-EDS analyses before and after incubation in SBF solution. LDH and WST-1 tests were used to determine the level of cytotoxicity of the tested bioglasses on hFOB1.19 osteoblasts. Results: The results show that the developed bioglasses release Ca2+, are bioactive in SBF solution, not cytotoxic and show antibacterial activity in contact with Pseudomonas aeruginosa and Staphylococcus aureus strains. Bioglasses enriched with ZnO show the highest bactericidal activity. All tested bioglasses enhanced hFOB 1.19 cells proliferation. Particle size has a lower effect on biological performance of the bioglasses than their chemical composition. Conclusions: The conducted research showed that bioglass modification with SrO and ZnO can be considered particularly for the development of biomaterials supporting bone regeneration and the treatment of infected bone defects.
W artykule przedstawiono wyniki badań in vitro dotyczące cytotoksyczności i działania bakteriobójczego materiału do stosowania w endodoncji – cementu typu MTA (ang. Mineral Trioxide Aggregate) o nowym składzie chemicznym. Cement wytworzono na bazie krzemianu trójwapnia wzbogaconego ZnO i płynu będącego 15% roztworem CaCl2 , a jako czynnik wprowadzający kontrast RTG zastosowano ZrO2 . Wytworzony cement MTA oceniono poprzez scharakteryzowanie właściwości fizykochemicznych obejmujących: czas wiązania, wytrzymałość na ściskanie, a także kontrast RTG i bioaktywność w roztworze symulującym osocze (SBF). Do zbadania wpływu ziaren ZrO2 na ilość i wielkość porów w cemencie wykorzystano technikę mikrotomografii komputerowej (µCT). Działanie cytotoksyczne cementów oceniano przez zastosowanie referencyjnej linii komórkowej L-929. Warunki hodowli komórkowej w kontakcie z badanymi materiałami lub ekstraktami z cementów oceniano za pomocą analizy obrazu lub testu kolorymetrycznego MTT. Do badania aktywności przeciwbakteryjnej wykorzystano dwa szczepy paciorkowców: Streptococcus mutans i Streptococcus sanguinis. Opracowany cement typu MTA charakteryzuje się odpowiednimi właściwościami użytkowymi i spełnia wymagania zawarte w normie PN-EN ISO 10993-5:2009 „Biologiczna ocena wyrobów medycznych, część 5: Badania cytotoksyczności in vitro”. Jednak, aby mógł być rozważany jako bezpieczny wyrób medyczny do wypełniania wstecznego kanałów korzeniowych, wymaga dalszych badań.
EN
The article presents the results of in vitro cytotoxicity and bactericidal activity of MTA-type cement (Mineral Trioxide Aggregate) for endodontics with a new chemical composition. The cement was made on the basis of ZnO-enriched tricalcium silicate and a liquid being a 15% CaCl2 solution. ZrO2 was used as an X-ray radiopacity agent in the cement. The produced MTA-type cement was assessed by characterizing the physicochemical properties including: setting time, compressive strength, as well as X-ray radiopacity and bioactivity in a simulated body fluid (SBF). Microtomography technique (µCT) was used to investigate the effect of ZrO2 grains on the number and size of pores in the cement. The cytotoxic activity of the cements was assessed by using the reference L-929 cell line. Cell culture conditions in contact with test materials or cement extracts were assessed by image analysis or MTT colorimetric test. Two strains of streptococci were used to test the antibacterial activity: Streptococcus mutans and Streptococcus sanguinis. The developed MTA cement has appropriate functional properties and meets the requirements of the PNEN ISO 10993-5:2009 standard "Biological evaluation of medical devices, Part 5: In vitro cytotoxicity tests". However, in order to be considered a safe medical device for retrograde root canal filling, it requires further research.
Opracowanie nowej formulacji produktu leczniczego, wyrobu medycznego czy suplementu diety to niezwykle skomplikowany i złożony proces, który obejmuje szereg niezbędnych analiz potwierdzających jakość i przydatność tych produktów. Rozważania dotyczące bezpieczeństwa, a w przypadku produktów leczniczych również skuteczności działania (w rozumieniu mocy/aktywności) są nieodzownym elementem w trakcie opracowywania formulacji. Proces kontrolny powinien dotyczyć wszystkich etapów rozwoju produktu, począwszy od wytypowania substancji czynnej o charakterze leczniczym lub wspomagającej działanie lecznicze, aż po ostatni etap badań przedklinicznych przed testami na ludziach. Badania powinny być również kontynuowane po etapie komercjalizacji, dla produktów z rynku, w celu potwierdzenia odpowiedniej jakości, bezpieczeństwa, czystości i mocy produktu. Metody in vitro są kluczowymi testami stosowanymi przez środowisko naukowe oraz firmy branży farmaceutycznej do generowania wysokiej jakości informacji na temat bezpieczeństwa i skuteczności cząsteczek o potencjale terapeutycznym i nie tylko. Jaki jest cel badań in vitro? Jakich informacji dostarczają w procesie opracowywania nowych produktów? Jaki mają wpływ na branżę farmaceutyczną i w jaki sposób jej pomagają?
EN
The development of a new formulation of a drug, medical device or dietary supplement is an extremely complicated and complex process that includes a number of necessary analyses confirming the quality and potential of these products. Safety considerations, and in case of drugs also the potency (activity) considerations, are necessary steps in the formulation development. The control process should cover all stages of product development. From selecting on active ingredient with therapeutic purposes or supporting the therapeutic effect, up to the last stage of preclinical studies before clinical trials. Control activities should continue after the commercialization stage, for products from the market, in order to confirm the appropriate quality, purity and potency of the product in the production process. In vitro methods are key tests used by the scientific community and pharmaceutical companies to generate high-quality information on the safety and efficacy of drug candidates and more. What is the purpose of in vitro research? What information do they provide in the drug development process? What is their impact on the pharmaceutical industry and how are they helping it?
Preformed stainless steel crowns are used in pediatric dentistry to obtain full crown restoration of primary molar teeth. They are consider the best restoration in terms of durability and effectiveness. The purpose of this study is to evaluate microbial, cytological and physio-chemical properties to determine whereas stainless steel crown are biocompatible, safe for surrounding tissue and helpful in avoiding micro-organisms influence on the tooth tissue. Based on the results, it was determined that stainless steel crowns used in pediatric dentistry represent no cytotoxic risk to the surrounding tissues, have a low probability of developing hypersensitivity to the coronal material and also that their biological properties make them suitable to use in pediatric dentistry for the reconstruction of damaged primary molar tissue.
Synthesis of ethyl 7-hydroxy-1-azacoumarin-3-carboxylate (3) was developed using ethyl-7-hydroxy coumarin-3-carboxylate and ammonium solution as the key synthons. Condensation of ethyl 7-hydroxy-1-azacoumarin-3-carboxylate with ammonium acetate and aniline to give N-substituted-7-hydroxy-1-azacoumarin-3-carboxamides (7-Hydroxy -1-azacoumarin-3-carboxamide (4) and N-phenyl 7-Hydroxy-1-azacoumarin-3-carboxamide (5)). Bromo derivative (N-phenyl 6, 8-dibromo-7-hydroxy-1-azacoumarin-3-carboxamide (6)) was obtained from halogenation of compound N-phenyl 7-Hydroxy-1-azacoumarin-3-carboxamide (5) with bromine in glacial acetic acid. N-phenyl-2,5-diacetoxy-6, 8-disubstituted-Quinoline-3-carboxamides (N-phenyl 2,7-diacetoxy-Quinoline-3-carboxamide (7) and N-phenyl 2,7-diacetoxy-6,8-dibromo-Quinoline-3-carboxamide (8)) were prepared via the acetylation of compounds 5 and 6 with acetic anhydride. Five compounds 4–8 were evaluated in vitro against more than one human tumor cell lines. Among the selected compounds, 6 showed the best in vitro cytotoxicity against the human cancer cell line; MCF-7 (with IC50 = 10.12 μM). In addition, cell cycle analysis of compound 6 demonstrated cell cycle arrest at G2/M phase and Pre-G1 apoptosis.
The absence of even a single finger results in a major impairment in the hand function (precise grasping, grip power), therefore significantly affecting the social and professional life of victims who are frequently young people. Finger amputation is a surgical treatment for ~69.000 patients in the EU after traumatic injury, in which replantation microsurgery fails due to the severity of tissue damage. The surgical reconstruction is currently possible only via autograft transplantation, e.g. a toe-to-hand transfer, thus leading to foot impairment. Some motion functional restoration is also possible using a bone-anchored silicone prosthesis but without the sense revalidation. Our current research focuses on alternatives for surgical reconstruction by means of novel patient- -specific, durable, biomimetic, bioactive and antibacterial implants for reconstructing lost bone and joints. The implant design – and the improved micro(neuro) surgery (beyond the project) – will consist in the fast successful rehabilitation, including the soft-tissue related mobility, the implantation of state-of-the-art nerve conduits as well as the aesthetic appearance. Key issues for the long-term functionality of the biomaterial-based reconstruction of hard tissue are based on surgical demands, such as: (1) perfect integration of a bone-substituting metal with the surrounding bone tissue (a) with no signs of loosening due to stress shielding at the interface and (b) enhanced with protective activity against bacterial inflammation (antimicrobial properties and formation of vascularized bone tissue (ossification)) even months to years after the injury; (2) biomimetic finger joints based on non-wearing materials without ossification meant to prevent the loss of the motion function.
The problem of treating chronic wounds is widespread throughout the world and carries a huge cost. Biomaterials engineering tries to solve this problem by creating innovative bioactive dressings dedicated to specific types of wounds. Both synthetic and natural polymers are the main base to produce wound dressings. Biopolymers have the advantage over synthetic polymers by being more biocompatible, non-toxic, biodegradable, and eco-friendly. The aim of this work was to produce a bioactive biomaterial based on natural polymers with potential applications to manage chronic highly exuding and infected wounds. A newly developed method for chemical synthesis of the curdlan/agarose biomaterial at high temperature combined with freeze-drying process resulted in a superabsorbent dressing material with antibiotic immobilized. The obtained biomaterial was subjected to basic microbiological in vitro tests and a cytotoxicity assay according to ISO 10993-5. Moreover, the experimental treatment of the infected wound in a veterinary patient was performed using the developed material. Based on the conducted research, it was proved that the produced dressing is not toxic to normal human skin fibroblasts. An additional advantage of the biomaterial is its ability to inhibit the growth of harmful microorganisms, such as Staphylococcus aureus and Pseudomonas aeruginosa. Furthermore, the experimental treatment confirmed the validity of using the produced biomaterial as a dressing dedicated to the treatment of difficult-to-heal infected wounds. To summarize, the produced biomaterial possesses great potential to be used as a wound dressing for infected wounds.
This work aims to analyze the effects of niobium on the bioactivity of a titanium, nickel, aluminum, and niobium alloy obtained by the Plasma Skull Push Pull process (PSPP). Titanium alloys, such as NiTinol (NiTi), are metallic biomaterials that have wide application in health and surgical prostheses. In this work the microstructural and bioactivity characteristics of the alloys are evaluated. The addition of aluminum improves alloy ductility and reduces its cost. The addition of niobium favors the hydroxyapatite nucleation. Therefore, the addition of the combination of the two elements contributes to lower cost and better alloy bioactivity.
In this work, we examined the in vitro cytotoxicity of new biodegradable surgical wires. The wires made of zinc with the addition of a small amount of magnesium (pure zinc, ZnMg 0.0026, ZnMg 0.0068, and ZnMg 0.08) have been investigated. The wires were produced using a technology based on extrusion and subsequent drawing. The resulting wires with a diameter of 0.8–1.0 mm are designed to be used in surgical operations related to bone joints. For cytotoxicity studies, we have selected human dental pulp stem cells (hDPSC) as the cell population representing normal osteoprogenitor cells. Considering that, after bone surgeries, the chance of osteosarcoma increases, we have compared the results obtained in hDPSC to those obtained with Saos-2 human osteosarcoma cell line. Cultured cells were exposed to the extracts obtained from the materials incubated in culture medium for 24 h with and without preincubation. Extracts of different ratios were examined. The results showed that the extracts obtained from wires made of ZnMg 0.0026 alloy exhibit high toxicity to Saos-2 osteosarcoma cells and low toxicity to hDPSC cells. This was in contrast to all reference materials, i.e., commercial surgical sutures made of steel and polymers, that did not display cytotoxicity toward osteosarcoma cells. Thus, the detected phenomenon for the ZnMg 0.0026 alloy can become the basis for creating biodegradable Zn-Mg surgical wires with antitumor activity.
The impact of the addition of the nanofiller – halloysite – on the mechanical, physicochemical and biological properties of a nanocomposite, in which thermoplastic polyurethane fulfilled the role of the matrix was investigated. The nanocomposite was obtained by extrusion in three variants with 1, 2 and 3 wt % halloysite. The nanostructure of the obtained materials was confirmed using Atomic Force Microscopy (AFM). Based on the mechanical tests carried out, it was proven that the obtained nanocomposites were characterized by a tensile modulus greater than the polyurethane constituting the matrix. The density and hardness of the nanocomposites had changed within error limits compared to unmodified polyurethane. Biological tests showed no cytotoxicity of all the tested materials to normal human dermal fibroblasts (NHDF). Degradation tests were carried out in artificial plasma and showed that samples with 2 wt % halloysite addition had the best ratio of tensile strength and elongation at break to elasticity modulus.
PL
Zbadano wpływ naturalnego nanonapełniacza – haloizytu – na właściwości mechaniczne, fizykochemiczne oraz biologiczne kompozytu wytworzonego na bazie termoplastycznego poliuretanu. Nanokompozyt o zawartości 1, 2 i 3% mas. haloizytu otrzymywano na drodze wytłaczania. Nanostrukturę wytworzonych materiałów potwierdzono za pomocą mikroskopii sił atomowych (AFM). Na podstawie przeprowadzonych badań mechanicznych wykazano, że uzyskane nanokompozyty charakteryzują się modułem sprężystości większym niż poliuretan stanowiący osnowę. Zmiany gęstości i twardości niemodyfikowanego poliuretanu po dodaniu haloizytu mieściły się w granicach błędu pomiaru. Testy biologiczne nie wykazały cytotoksyczności wszystkich badanych materiałów wobec prawidłowych fibroblastów ludzkiej skóry (NHDF). Badania degradacji przeprowadzone w środowisku syntetycznego osocza wykazały, że próbki z dodatkiem 2% mas. haloizytu mają najlepszy stosunek wytrzymałości na rozciąganie i wydłużenia przy zerwaniu do modułu sprężystości.
Purpose: The present research aimed to determine whether and how the aluminium chloride – based materials affect the cell line of the bacterial line and fungi. Methods: Cytotoxicity of haemostatic astringents: Alustat (liquid), Alustat (gel), Alustat (foam), Alustin, Hemostat, Racestyptine and Traxodent containing AlCl3 was conducted on L929 cell line with the use of MTT and SRB assays. The antimicrobial activity (CFU and MIC) against C. albicans, S. mutans, L. rhamnosus was determined. Results: In the MTT results, cell viability for all agents were very low. In SRB, the lowest cytotoxicity was demonstrated for Hemostat and Alustat (foam), Traxodent and Racestyptine. Total reduction of the CFU of S. mutans was observed. Alustat (gel) and Alustat (liquid) completely inhibited the growth of C. albicans, S. mutans and L. rhamnosus. Conclusions: The viability of L929 cells obtained in the SRB assay is more reliable than that obtained in the MTT assay, in the case of gingival haemostatic agents.
The present investigation was aimed to study the cytotoxic response induced by molybdenum disulfide in nano- (MoS2-NPs) and micro- (MoS2-MPs) size of particle in human bronchial (BEAS-2B) and alveolar (A549) cells. The cells were exposed with different particle size of MoS2 in concentrations range 1-200 μg/mL for 24, 48, and 72 h, and then the cytotoxicity assays (MTT and NRU) was performed. Afterwards, long-term toxicity was assessed by colony forming efficiency assay (CFEA) during 10 days exposure of the cells. Both MoS2-NPs and MoS2-MPs showed similar, weak cytotoxic effects on BEAS-2B and A549 cells assessed by MTT assay, that is reduction of cell viability to approx. 60-70% at concentrations of 2.5 and 5 μg/mL. The percentage viability remained relatively constant at this level across all concentrations above 5 μg/mL. In long-term exposure, both MoS2 inhibited colony formation in a wider range of concentrations to 200 μg/mL. MoS2-NPs were slightly more cytotoxic than MoS2-MPs. The data suggest the low potential hazardous nature of both MoS2 tested with an indication of higher toxicity of MoS2-NPs.
PL
Celem badań była ocena cytotoksycznego działania nano- (MoS2-NPs) i mikrometrycznego (MoS2-MPs) disiarczku molibdenu na ludzkie komórki nabłonka oskrzelików (BEAS-2B) i pęcherzyków (A549) płuc. Komórki narażano na różnej wielkości cząstki MoS2 w zakresie stężeń 1-200 μg/ml przez 24, 48 i 72 h. Badano wpływ obu rodzaju cząstek MoS2 na integralność błon komórkowych (test NRU) oraz aktywność metaboliczną komórek (test MTT). Oceniano również zdolność komórek do proliferacji po długotrwałym (10 dni) narażeniu na podstawie testu efektywności formowania kolonii (CFEA). Oba MoS2 powodowały podobne, słabe działanie cytotoksyczne na komórki oceniane testem MTT, tj. obniżenie przeżywalności komórek do ok. 60-70% w stężeniach of 2,5 and 5 μg/mL po 24 h, które w niewielkim stopniu nasilało się z czasem narażenia, natomiast nie obserwowano wzrostu cytotoksyczności ze wzrostem stężenia. Po długotrwałej ekspozycji (10 dni) oba MoS2 hamowały zdolność tworzenia kolonii w szerszym zakresie stężeń tj. do 200 μg/mL, przy czym MoS2-NPs wykazywały silniejsze działanie ograniczające zdolność komórek do proliferacji niż MoS2-MPs. Dane wskazują na niski potencjał toksyczny obu badanych MoS2 ze wskazaniem wyższej toksyczności MoS2-NP.
The aim of the study was to compare the cytotoxic effects of (methyl and propyl) parabens likely to be environmental contaminants on early and late passage fibroblasts isolated from human skin. The study was carried out on senescent diploid cell lines, namely normal (senescent) dermal fibroblasts CCD-1136Sk (ATCC® CRL-2697TM). In order to assess the cytotoxic effect, the MTT test which determines the cells' metabolic activity and the neutral red uptake assay which assesses the cell membrane integrity (NRU test) were applied. Propyl paraben (PrPB) appeared to be more cytotoxic to the analysed dermal fibroblasts (since it reached lower IC50 values); however, none of the tests found a consistent trend for a change in the cells’ sensitivity to the analysed parabens as they age, that would be reflected in the determined IC50 values. On the other hand, having analysed the course of the curves of relationships between the cell viability and the preservative concentration, it was found that at the exposure of CCD-1136Sk fibroblasts to propyl paraben, none of the two tests observed different sensitivity of late and early cell passages; however, the exposure of dermal fibroblasts to methyl paraben, determined using the NRU test, indicated a stronger effect of the preservative at a concentration of 50 and 200 µg/ml on the earliest cells passage (no. 8), compared to the subsequent passages of these cells. At the same time, the compound with a concentration of 50 and 800 µg/ml was the least toxic to the latest cells passage (no. 20). The presented results indicate that the analysed parabens used in the cosmetic and pharmaceutical industries may be toxic to skin cells; moreover, it is not excluded that with consecutive passaging, differences in the susceptibility to cytotoxic effects may occur. It therefore appears necessary to take into account the possibility of different later and earlier passages cells' reactivity when interpreting the results of studies into the effects of preservatives on the living body. Furthermore, it should be borne in mind that humans may be exposed to preservatives due to environmental contamination.
PL
Celem badań było porównanie cytotoksycznego działania parabenów (metylu i propylu), mogących stanowić zanieczyszczenie środowiska naturalnego, na wczesne oraz późne pasaże fibroblastów wyprowadzonych ze skóry człowieka. Badania wykonano na starzejących się diploidalnych liniach komórkowych: fibroblastach skórnych CCD-1136Sk (ATCC® CRL-2697TM). Do oceny cytotoksycznego działania zastosowano test MTT, który określa aktywność metaboliczną komórek oraz test pochłaniania czerwieni obojętnej oceniający integralność błon komórkowych (test NRU). Bardziej cytotoksyczny (osiągający niższe wartości IC50) dla badanych fibroblastów skórnych okazał się propyloparaben (PrPB), jednak w żadnym z testów nie stwierdzono spójnej tendencji zmiany wrażliwości komórek na badane parabeny w miarę ich starzenia się, która znalazłaby odzwierciedlenie w wyznaczonych wartościach IC50. Analizując natomiast przebieg krzywych zależności żywotności komórek od stężenia konserwantu stwierdzono, że przy narażeniu fibroblastów CCD-1136Sk na paraben propylowy nie zaobserwowano zróżnicowanej wrażliwości komórek późnych i wczesnych pasaży w żadnym z dwóch testów, natomiast narażenie fibroblastów skórnych na paraben metylowy oceniane testem NRU wskazało na silniejsze działanie konserwantu o stężeniu 50 i 200 μg/ml na komórki najwcześniejszego pasażu (nr 8) w porównaniu z kolejnymi pasażami tych komórek. Jednocześnie, związek o stężeniu 50 i 800 μg/ml był najmniej toksyczny dla komórek najpóźniejszego pasażu (nr 20). Przedstawione wyniki wskazują, iż badane parabeny stosowane w przemyśle kosmetycznym i farmaceutycznym mogą działać toksycznie na komórki skóry, nie wyklucza się także, że wraz z ich starzeniem się mogą być występować różnice w podatności na działanie cytotoksyczne. Konieczne zatem wydaje się uwzględnianie przy interpretacji wyników badań nad odziaływaniem substancji konserwujących na żywy organizm również możliwości odmiennej reaktywności komórek późniejszych i wcześniejszych pasaży. Nie należy ponadto zapominać o możliwości narażenia człowieka na substancje konserwujące wynikające z zanieczyszczenia środowiska naturalnego.
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