Publications by authors named "Mustafa Türk"

31 Publications

Phosphorus-nitrogen compounds: part 53-synthesis, characterization, cytotoxic and antimicrobial activity, DNA interaction and molecular docking studies of new mono- and dispirocyclotriphosphazenes with pendant arm(s).

Mol Divers 2021 May 14. Epub 2021 May 14.

Department of Physics, Hacettepe University, 06800, Ankara, Turkey.

Mono-/dispirocyclotriphosphazenes with pendant arm(s) are robust, but they are less investigated inorganic ring systems. In this study, a series of mono (3 and 4)- and dispirocyclotriphosphazenes with 4-chloro-benzyl pendant arm(s) (13-16) was obtained from the Cl exchange reactions of hexachlorocyclotriphosphazene with sodium (N-benzyl)aminopropanoxides (1 and 2). When compound (3) reacted with excess pyrrolidine, morpholine, tetra-1,4-dioxa-8-azaspiro[4,5]decane (DASD) and piperidine, the fully substituted monospirocyclotriphosphazenes (7, 9, 10 and 12) occurred. But, the reactions of 4 with excess piperidine and morpholine produced the gem-piperidino (5)- and morpholino (6)-substituted monospirocyclotriphosphazenes, whereas the reactions of 4 with excess pyrrolidine and DASD gave the fully substituted monospirocyclotriphosphazenes (8) and (11). However, it should be indicated that these derivatives were obtained to be used for the investigation of their spectral, stereogenic and biological properties. The structures of 5, 7 and 14 were determined crystallographically. X-ray data of 5 and 14 displayed that both of compounds were chiral in solid state, and their absolute configurations were assigned as R and RR. Additionally, the antimicrobial activities of phosphazenes were investigated. Minimum inhibitory concentrations, minimal bacterial concentrations and minimum fungicidal concentrations of phosphazenes were determined. The interactions of phosphazenes with plasmid DNA were evaluated by agarose gel electrophoresis. The cytotoxic activities of compounds were studied against L929 fibroblast and DLD-1 colon cancer cells. In addition, density functional theory calculations of 5, 7 and 14 were reported, and their molecular docking studies with DNA, E. coli DNA gyrase and topoisomerase IV were presented.
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http://dx.doi.org/10.1007/s11030-021-10231-5DOI Listing
May 2021

omparison of nanofibrillar and macroporous-spongious composite tissue scaffolds for periodontal tissue engineering.

Connect Tissue Res 2021 Apr 26:1-15. Epub 2021 Apr 26.

Department of Periodontology, Hacettepe University, Ankara, Turkey.

: The ultimate goal of periodontal treatment is to regenerate the lost periodontal tissues. The interest in nanomaterials in dentistry is growing rapidly and has focused on improvements in various biomedical applications, such as periodontal regeneration and periodontal tissue engineering. To enhance periodontal tissue regeneration, hydroxyapatite (HA) was used in conjunction with other scaffold materials, such as Poly lactic-co-glycolic-acid (PLGA) and collagen (C). The main target of this study was to compare the effects of nano and macrostructures of the tissue scaffolds on cell behavior for periodontal tissue engineering.: Nanofibrillar and macroporous-spongious composite tissue scaffolds were produced using PLGA/C/HA. Subgroups with BMP-2 signal molecule and without HA were also created. The scaffolds were characterized by FTIR, SEM/EDX techniques, and mechanical tests. The scaffolds were compared in the periodontal ligament (PDL) and MCT3-E1 cell cultures. The cell behaviors; adhesions by SEM, proliferation by WST-1, differentiation by ALP and mineralization with Alizarin Red Tests were determined.: Cell adhesion and mineralization were higher in the nanofibrillar scaffolds compared to the macroporous-spongious scaffolds. Macroporous-spongious scaffolds seemed better for the proliferation of PDL cells and differentiation of MC3T3-E1-preosteoblastic cells, while nanofibrillar scaffolds were more convenient for the differentiation of PDL cells and proliferation of MC3T3-E1-preosteoblastic cells.: In general, nanofibrillar scaffolds showed more favorable results in cell behaviors, compared to the macroporous-spongious scaffolds, and mostly, BMP-2 and HA promoted the activities of the cells.
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http://dx.doi.org/10.1080/03008207.2021.1912029DOI Listing
April 2021

The Pimpled Gold Nanosphere: A Superior Candidate for Plasmonic Photothermal Therapy.

Int J Nanomedicine 2020 24;15:2903-2920. Epub 2020 Apr 24.

Department of Chemical Engineering, Northeastern University, Boston, MA 02115, USA.

Background: The development of highly efficient nanoparticles to convert light to heat for anti-cancer applications is quite a challenging field of research.

Methods: In this study, we synthesized unique pimpled gold nanospheres (PGNSs) for plasmonic photothermal therapy (PPTT). The light-to-heat conversion capability of PGNSs and PPTT damage at the cellular level were investigated using a tissue phantom model. The ability of PGNSs to induce robust cellular damage was studied during cytotoxicity tests on colorectal adenocarcinoma (DLD-1) and fibroblast cell lines. Further, a numerical model of plasmonic (COMSOL Multiphysics) properties was used with the PPTT experimental assays.

Results: A low cytotoxic effect of thiolated polyethylene glycol (SH-PEG-SH-) was observed which improved the biocompatibility of PGNSs to maintain 89.4% cell viability during cytometry assays (in terms of fibroblast cells for 24 hrs at a concentration of 300 µg/mL). The heat generated from the nanoparticle-mediated phantom models resulted in ΔT=30°C, ΔT=23.1°C and ΔT=21°C for the PGNSs, AuNRs, and AuNPs, respectively (at a 300 µg/mL concentration and for 325 sec). For the in vitro assays of PPTT on cancer cells, the PGNS group induced a 68.78% lethality (apoptosis) on DLD-1 cells. Fluorescence microscopy results showed the destruction of cell membranes and nuclei for the PPTT group. Experiments further revealed a penetration depth of sufficient PPTT damage in a physical tumor model after hematoxylin and eosin (H&E) staining through pathological studies (at depths of 2, 3 and 4 cm). Severe structural damages were observed in the tissue model through an 808-nm laser exposed to the PGNSs.

Conclusion: Collectively, such results show much promise for the use of the present PGNSs and photothermal therapy for numerous anti-cancer applications.
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http://dx.doi.org/10.2147/IJN.S248327DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7188077PMC
July 2020

Synthesis and biological evaluation of novel urea, thiourea and squaramide diastereomers possessing sugar backbone.

Carbohydr Res 2020 Jun 29;492:107991. Epub 2020 Mar 29.

Department of Bioengineering, Kırıkkale University, 71450, Yahşihan, Kırıkkale, Turkey.

A series of novel chiral 14 urea, thiourea and squaramide stereoisomers possessing carbohydrate backbones as well as amide functional groups was synthesized and characterized by their, H NMR, C NMR, FT-IR, HRMS, optical rotation, and melting points. Their antiproliferative activities were investigated against HeLa and PC3 cell lines. The compounds 9, 11 and 12 showed better activities at 25 μM against PC3 cell line with respect to the standard 5-fluorouracil (5-FU). Especially, the compounds 9 and 11 showed higher activities than the standard 5-FU even at low concentration (5 μM) against HeLa cell line. IC results also confirm these activities. The compounds 9, 10 and 11 have the IC values of 1.10 μM, 1.51 μM and 1.02 μM, respectively while 5-FU has 2.51 μM. Moreover, their cytotoxicity tests have proven that their viabilities were in between 50% and 100%.
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http://dx.doi.org/10.1016/j.carres.2020.107991DOI Listing
June 2020

Synthesis and characterization of antibacterial drug loaded β-tricalcium phosphate powders for bone engineering applications.

J Mater Sci Mater Med 2020 Jan 21;31(2):16. Epub 2020 Jan 21.

Center for Nanotechnology and Biomaterials Application and Research (NBUAM), Marmara University, 34722, Istanbul, Turkey.

Powders of β-tricalcium phosphate [β-TCP, β-Ca(PO)] and composite powders of β-TCP and polyvinyl alcohol (PVA) were synthesized by using wet precipitation methods. First, the conditions for the preparation of single phase β-TCP have been delineated. In the co-precipitation procedure, calcium nitrate and diammonium hydrogen phosphate were used as calcium and phosphorous precursors, respectively. The pH of the system was varied in the range 7-11 by adding designed amounts of ammonia solution. The filtered cakes were desiccated at 80 °C and subsequently calcined at different temperatures in the range between 700-1100 °C. Later on, rifampicin form II was used to produce drug-loaded β-TCP and PVA/β-TCP powders. All the synthesized materials have been characterized from morphological (by scanning electron microscopy) and structural-chemical (by X-ray diffraction and Fourier transform infrared spectroscopy) point of view. The drug loading capacity of the selected pure β-TCP powder has been assessed. The biological performance (cytocompatibility in fibroblast cell culture and antibacterial efficacy against Escherichia coli and Staphylococcus aureus) has been tested with promising results. Application perspectives of the designed drug-bioceramic-polymer blends are advanced and discussed.
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http://dx.doi.org/10.1007/s10856-019-6356-1DOI Listing
January 2020

Autologous stem cell-derived chondrocyte implantation with bio-targeted microspheres for the treatment of osteochondral defects.

J Orthop Surg Res 2019 Nov 28;14(1):394. Epub 2019 Nov 28.

Department of Orthopedics and Traumatology, Ankara Yildirim Beyazit University, 06800, Ankara, Turkey.

Background: Chondral injury is a common problem around the world. Currently, there are several treatment strategies for these types of injuries. The possible complications and problems associated with conventional techniques lead us to investigate a minimally invasive and biotechnological alternative treatment. Combining tissue-engineering and microencapsulation technologies provide new direction for the development of biotechnological solutions. The aim of this study is to develop a minimal invasive tissue-engineering approach, using bio-targeted microspheres including autologous cells, for the treatment of the cartilage lesions.

Method: In this study, a total of 28 sheeps of Akkaraman breed were randomly assigned to one of the following groups: control (group 1), microfracture (group 2), scaffold (group 3), and microsphere (group 4). Microspheres and scaffold group animals underwent adipose tissue collection prior to the treatment surgery. Mesenchymal cells collected from adipose tissue were differentiated into chondrocytes and encapsulated with scaffolds and microspheres. Osteochondral damage was conducted in the right knee joint of the sheep to create an animal model and all animals treated according to study groups.

Results: Both macroscopic and radiologic examination showed that groups 3 and 4 have resulted better compared to the control and microfracture groups. Moreover, histologic assessments indicate hyaline-like cartilage formations in groups 3 and 4.

Conclusion: In conclusion, we believe that the bio-targeted microspheres can be a more effective, easier, and safer approach for cartilage tissue engineering compared to previous alternatives.
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http://dx.doi.org/10.1186/s13018-019-1434-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6883666PMC
November 2019

imaging/detection of MRSA bacterial infections in mice using fluorescence labelled polymeric nanoparticles carrying vancomycin as the targeting agent.

J Biomater Sci Polym Ed 2020 02 25;31(3):293-309. Epub 2019 Nov 25.

Bioengineering Division, Institute of Graduate Studies, Hacettepe University, Beytepe, Ankara, Turkey.

This study aims to develop fluorescence labelled polymeric nanoparticle (NP) carrying vancomycin as the targeting agent for imaging of Methicillin-resistant bacterial infections in animal models. Maleimide functionalized 1,2-distearoyl-sn-glycero-3-phosphoethanolamine--[maleimide (polyethylene glycol)-2000] as the main was carrier matrix to prepare the NPs. A fluorescence probe, namely, poly[9,9'-bis (6″-,,-trimethylammonium) hexyl) fluorene-co-alt-4,7-(2,1,3-benzothiadiazole) dibromide] was encapsulated within these NPs by ultrasonication successfully. UV-Vis spectro- photometry of the NPs showed the characteristic shifting on the peak of conjugated polymers indicating successful packaging of this compound with lipid bilayers in nanoscales. Zeta-sizer and TEM analysis showed that the prepared NPs have a diameter of 80-100 nm in a narrow size distribution. Thiolated vancomycin was synthesized and attached to the NPs as the targeting agent. FTIR and MALDI-TOF spectroscopy analysis confirmed the immobilization. The specific targeting properties of the vancomycin conjugated NPs to the target bacteria were first confirmed in bacterial cultures in which was the non-target bacteria - using confocal microscopy and TEM. Imaging of bacterial infections was investigated in mice model using a non-invasive live animal fluorescence imaging technique. The results confirmed that bacterial infections can be detected using these novel polymeric NPs carrying fluorescence probes for imaging and vancomycin as the targeting agent - successfully.
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http://dx.doi.org/10.1080/09205063.2019.1692631DOI Listing
February 2020

Effects of B2O3 (boron trioxide) on colon cancer cells: our first-step experience and in vitro results.

Turk J Biol 2019 13;43(3):209-223. Epub 2019 Jun 13.

Department of Bioengineering, Faculty of Engineering, Kırıkkale University, Kırıkkale, Turkey.

Boron oxide (B2O3) is derived from dehydration of boric acid and is a colorless, semitransparent, crystalline compound that is moderately soluble in water. On the other hand, boron oxide is chemically hygroscopic. This gives the molecule the ability to soak up water and adhere to tissues. Boron oxide can be used locally after tumor debulking in inoperable tumors and especially when the tumor-free margin distance cannot be provided. For all these reasons we aimed to evaluate the in vitro test results of B2O3 in terms of cytotoxicity, genotoxicity, apoptosis, and necrotic effects on L929 fibroblast cells and DLD-1 colorectal adenocarcinoma cells. Our studies demonstrated that boron oxide compounds appear to be highly cytotoxic for both cell lines according to WST cell viability assay (44.22% and 18.36% on DLD-1 and L929, respectively). Although no genotoxic effects were observed, boron oxide compounds showed antiproliferative effects for both cell lines. The prepared boron oxide compounds may hold the potential to be applied locally to the remaining tissue after surgery and further research and evaluation will be needed to determine its effectiveness.
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http://dx.doi.org/10.3906/biy-1901-34DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6620035PMC
June 2019

Transscleral Delivery of Bevacizumab-Loaded Chitosan Nanoparticles.

J Biomed Nanotechnol 2019 Apr;15(4):830-838

: The aim of this study was to synthesize bevacizumab-loaded nanoparticles and evaluate their effects on the treatment of posterior segment diseases via subtenon injections. : Bevacizumab-loaded chitosan nanoparticles (BLCNs) were synthesized by the ionic gelation method, and their physicochemical characteristics and release profile were studied. The BLCNs were characterized using atomic force microscopy (AFM), FTIR spectroscopy, dynamic light scattering, and scanning electron microscopy. The BLCNs were delivered into rabbits' eyes via posterior subtenon injections. An immunohistochemical evaluation of the ocular tissues was performed, and the vitreous humor and serum bevacizumab levels were measured by ELISA. : Bevacizumab-loaded chitosan nanoparticles with a diameter of 80 to 380 nm were prepared and characterized. studies showed that after the first 5 days of the experiment, a significant increase in the drug release maintained the desired drug dosage for 3 weeks. Immunohistochemical studies revealed that there were BLCNs penetrating through the sclera. Furthermore, the intravitreal bevacizumab concentration reached a maximum concentration of 18 g/ml, and it decreased to 6 g/ml after only a week. : The results revealed that subtenon injection of BLCNs is a promising alternative to intravitreal injections. In addition to the ELISA studies, immunohistochemical experiments confirmed that BLCNs enable transscleral bevacizumab penetration, and BLCN usage may provide the required bevacizumab levels for the treatment of posterior segment diseases.
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http://dx.doi.org/10.1166/jbn.2019.2716DOI Listing
April 2019

Pectin-conjugated magnetic graphene oxide nanohybrid as a novel drug carrier for paclitaxel delivery.

Artif Cells Nanomed Biotechnol 2018 3;46(sup1):264-273. Epub 2018 Jan 3.

b Department of Bioengineering , Kırıkkale University , Kırıkkale , Turkey.

Recent studies have shown that graphene oxide (GO) drug carrier functionalized with biocompatible natural polymers lead to higher loading efficacy and better stability with diminished cellular toxicity. Pectin (PEC) is one of the polysaccharide natural polymers, which has the potential to be used for drug delivery. In this work, we have successfully developed a novel PEC-conjugated magnetic GO nanocarrier for effective delivery of paclitaxel. The structure, surface morphology and thermal stability of the nanohybrid were investigated using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM) and zeta-sizer. Moreover, drug loading and release performance were studied by UV-vis absorption spectra. The cytotoxicity test was also performed by MTT test using L-929 fibroblast normal cell and MCF-7 cancer lines. The prepared nanocarrier showed an improved stability with enhanced drug loading capacity. Additionally, pH-responsive release analysis of the nanohybrid illustrated higher drug release at endosomal pH of cancer cell than that of normal physiological environment. Besides, cytotoxicity test demonstrated the synthesized nanohybrid is biocompatible, having very high relative cell viability. Bearing in mind these findings, the designed multifunctional nanohybrid drug carrier will be a good candidate for cancer drug delivery.
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http://dx.doi.org/10.1080/21691401.2017.1421211DOI Listing
May 2019

Modified gold-based siRNA nanotherapeutics for targeted therapy of triple-negative breast cancer.

Nanomedicine (Lond) 2017 Aug 26;12(16):1961-1973. Epub 2017 Jul 26.

Department of Nanotechnology & Nanomedicine, Hacettepe University, Ankara, Turkey.

Aim: In this study, we aimed to therapeutically target eukaryotic elongation factor 2 kinase (eEF-2K) in an in vivo triple-negative breast cancer (TNBC) tumor model.

Materials & Methods: We synthesized a highly monodisperse nanoformulation using polyethylenimine-modified gold nanoparticles (AuNP-PEI) as siRNA delivery vehicle and evaluated gene downregulation.

Results: We found that AuNP-PEI/eEF-2K nanoformulation was highly effective for in vitro and in vivo gene downregulation and showed remarkable antitumor efficacy that was associated with eEF-2K knockdown, inhibition of Src and MAPK-ERK signaling pathways in a TNBC orthotopic tumor model.

Conclusion: Our study suggests that eEF-2K plays an important role in TNBC tumorigenesis and its inhibition by AuNP-PEI/eEF-2K siRNA-based nanotherapeutics may be a potential therapeutic strategy for TNBC.
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http://dx.doi.org/10.2217/nnm-2017-0081DOI Listing
August 2017

Multiwalled Carbon Nanotube-Chitosan Scaffold: Cytotoxic, Apoptoti c, and Necrotic Effects on Chondrocyte Cell Lines.

Curr Pharm Biotechnol 2017 ;18(4):327-335

Faculty of Pharmacy, Department of Analytical Chemistry, Gazi University, Ankara. Turkey.

Background: Carbon nanotubes (CNTs) have been considered highly successful and proficient in terms of their mechanical, thermal and electrical functionalization and biocompatibility. In regards to their significant extent in bone regeneration, it has been determined that CNTs hold the capability to endure clinical applications through bone tissue engineering and orthopedic procedures. In the present study, we report on a composite preparation, involving the use of CNT-chitosan as scaffold for bone repair and regeneration. Through the use of water-soluble tetrazolium salt (WST-1) and double staining methods, the cytotoxic, necrotic, and apoptotic effects of chitosan-multiwalled carbon nanotube nanocomposites on the chondrocyte ATTC cell line have been exhibited.

Methods: The chitosan-multiwalled carbon nanotube scaffolds were prepared. Chondrocytes differentiation tool (ATCC) cell line was prepared. WST-1 assay for cytotoxicity studies were performed by using chondrocytes cells in 12.5-200 μL concentration range. The samples of membranes (chitosan- multiwalled carbon nanotube scaffold) were measured at 2 mg/mL and further prepared amongst chitosan- multiwalled carbon nanotube scaffold's which were placed into separate wells. While in the process of incubation, in the four-hour time range, the plates were immediately read in an Elisa microplate Reader. To predict the number of apoptotic and necrotic cells in culture, the technique of double staining with Hoechst dye was performed with PI on the basis of scoring cell nuclei. The mechanical properties such as tensile strength and elongation at break values of the chitosan only and chitosan/CNT scaffolds were evaluated on Texture Analyzer.

Results: Based on the results of the WST-1 assay procedure, the amount of cell viability was not significantly affected by nanocomposite concentrations and the lowest mortality rate of cells was obtained at a concentration of 12.5 μg/mL, whereas the highest mortality rate was obtained at a rate of 200 μg/mL. In addition, the effects of chitosan-CNT nanocomposites were not found to cytotoxic on chondrocyte cells. The double staining method has been able to determine the apoptotic and necrotic effects of chitosan MWCNT nanocomposites. The apoptotic and necrotic effects of the combined compounds had varied within the concentrations. In a similar manner to the outcome of the control groups, apoptosis was obtained at a percentage of 2.67%. Under a fluorescent inverted microscope, the apoptotic cell nuclei were stained with a stronger blue fluorescence in comparison to non-apoptotic cells, which may have had an effect. We also compared the strain-stress curve measurements results. The results indicated that the mechanical properties of scaffold were not different. Elongation at break values increased by addition of CNT.

Conclusion: CNTs as a biomaterial hold the potential to be used for applications in future regenerative medicine. By using the components of chondrocytes (ATTC) cell lines, the cytotoxicity evaluations were made for the chitosan-multiwalled carbon nanotube scaffold. The chitosan-MWCNT nanocomposites do not seem to induce drastic cytotoxicity to the chondrocyte cells.
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http://dx.doi.org/10.2174/1389201018666170127105555DOI Listing
December 2017

Bi-layered constructs of poly(glycerol-sebacate)-β-tricalcium phosphate for bone-soft tissue interface applications.

Mater Sci Eng C Mater Biol Appl 2017 Mar 27;72:316-324. Epub 2016 Nov 27.

Environmental Engineering Department, Bioengineering Division, Centre for Bioengineering, Hacettepe University, Ankara, Turkey. Electronic address:

This study aims to establish a facile protocol for the preparation of a bi-layered poly(glycerol-sebacate) (PGS)/β-tricalcium phosphate (β-TCP) construct and to investigate its potential for bone-soft tissue engineering applications. The layered structure was prepared by distributing the ceramic particles within a prepolymer synthesized in a microwave reactor followed by a cross-linking of the final construct in vacuum (<10mbar). The vacuum stage led to the separation of cross-linked elastomer (top) and ceramic (bottom) phases. Results showed that addition of β-TCP particles to the elastomer matrix after the polymerization led to an increase in compression strength (up to 14±2.3MPa). Tensile strength (σ), Young's modulus (E), and elongation at break (%) values were calculated as 0.29±0.03MPa and 0.21±0.03; 0.38±0.02 and 1.95±0.4; and 240±50% and 24±2% for PGS and PGS/β-TCP bi-layered constructs, respectively. Morphology was characterized by using Scanning Electron Microscopy (SEM) and micro-computed tomography (μ-CT). Tomography data revealed an open porosity of 35% for the construct, mostly contributed from the ceramic phase since the elastomer side has no pore. Homogeneous β-TCP distribution within the elastomeric structure was observed. Cell culture studies confirmed biocompatibility with poor elastomer-side and good bone-side cell attachment. In a further study to investigate the osteogenic properties, the construct were loaded with BMP-2 and/or TGF-β1. The PGS/β-TCP bi-layered constructs with improved mechanical and biological properties have the potential to be used in bone-soft tissue interface applications where soft tissue penetration is a problem.
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http://dx.doi.org/10.1016/j.msec.2016.11.082DOI Listing
March 2017

Synthesis, characterization and modification of Gum Arabic microgels for hemocompatibility and antimicrobial studies.

Carbohydr Polym 2017 Jan 16;156:380-389. Epub 2016 Sep 16.

Faculty of Science & Arts, Chemistry Department, Canakkale Onsekiz Mart University, Terzioglu Campus, 17100 Canakkale, Turkey; Nanoscience and Technology Research and Application Center (NANORAC), Canakkale Onsekiz Mart University, Terzioglu Campus, 17100 Canakkale, Turkey. Electronic address:

Gum Arabic (GA) microgels were successfully prepared via reverse micellization method with high yield (78.5±5.0%) in 5-100μm size range using divinyl sulfone (DVS) as a crosslinker. The GA microgels were degraded hydrolytically 22.8±3.5% at pH 1 in 20days, whereas no degradation was observed at pH 7.4 and pH 9 at 37°C. By using diethylenetriamine (DETA), and taurine (TA) as chemical modifying agents, GA microgels were chemically modified as GA-DETA and GA-TA, and the zeta potential values of 5.2±4.1 and -24.8±1.3mV were measured, respectively in comparison to -27.3±4.2mV for GA. Moreover, blood compatibility of GA, GA-TA, and GA-DETA microgels was tested via in vitro protein adsorption, % hemolysis ratio, and blood clotting index. All the microgels were hemocompatible with% hemolysis ratio between 0.23 to 2.05, and the GA microgels were found to be highly compatible with a blood clotting index of 81±40. The biocompatibility of GA, GA-DETA and GA-Taurine microgels against L929 fibroblast cells also revealed 84.4, 89.1, and 67.0% cell viability, respectively, at 25.0μg/mL concentration, suggesting great potential in vivo biomedical applications up to this concentration. In addition, 5 and 10mg/mL minimum inhibition concentrations of protonated GA-DETA microgels (GA-DETA-HCl) were determined against E. coli and S. aureus, respectively.
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http://dx.doi.org/10.1016/j.carbpol.2016.09.052DOI Listing
January 2017

The effect of calcium chloride concentration on alginate/Fmoc-diphenylalanine hydrogel networks.

Mater Sci Eng C Mater Biol Appl 2016 Sep 25;66:221-229. Epub 2016 Apr 25.

Bioengineering Department, Hacettepe University, Ankara 06800, Turkey; Department of Chemistry, Faculty of Science, Hacettepe University, Ankara 06800, Turkey. Electronic address:

Peptide based hydrogels gained a vast interest in the tissue engineering studies thanks to great superiorities such as biocompatibility, supramolecular organization without any need of additional crosslinker, injectability and tunable nature. Fmoc-diphenylalanine (FmocFF) is one of the earliest and widely used example of these small molecule gelators that have been utilized in biomedical studies. However, Fmoc-peptides are not feasible for long term use due to low stability and weak mechanical properties at neutral pH. In this study, Fmoc-FF dipeptides were mechanically enhanced by incorporation of alginate, a biocompatible and absorbable polysaccharide. The binary hydrogel is obtained via molecular self-assembly of FmocFF dipeptide in alginate solution followed by ionic crosslinking of alginate moieties with varying concentrations of calcium chloride. Hydrogel characterization was evaluated in terms of morphology, viscoelastic moduli and diffusional phenomena and the structures were tested as 3D scaffolds for bovine chondrocytes. In vitro evaluation of scaffolds lasted up to 14days and cell viability, sulphated glycosaminoglycan (sGAG) levels, collagen type II synthesis were determined. Our results showed that alginate incorporation into FmocFF hydrogels leads to better mechanical properties and higher stability with good biocompatibility.
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http://dx.doi.org/10.1016/j.msec.2016.04.084DOI Listing
September 2016

Fabrication and characterization of PVA/ODA-MMT-poly(MA-alt-1-octadecene)-g-graphene oxide e-spun nanofiber electrolytes and their response to bone cancer cells.

Mater Sci Eng C Mater Biol Appl 2016 Apr 23;61:257-68. Epub 2015 Dec 23.

Department of Biology, Faculty of Science, Division of Bioengineering, Kirikkale University, 71450 Kirikkale, Turkey.

This work presents a new approach to fabrication and characterization of novel polymer nanofiber electrolytes from intercalated PVA/ODA-MMT nanocomposite as a matrix polymer and encapsulated graphene oxide (GO) nanosheets with amphiphilic reactive copolymer as partner polymers using electrospinning method. The chemical and physical structures, surface morphology, thermal behaviors and electric conductivity of nanocomposites and nanofibers were investigated using analyses methods including FTIR, XRD, SEM, DSC-TGA and conductivity analysis. Significant improvements in nanofiber morphology and size distribution were observed when GO and reactive organoclay were incorporated as reinforcement fillers into various matrix/partner solution blends. The structural factors of matrix-partner polymer nanocomposite particles with higher zeta-potential play important roles in both chemical and physical interfacial interactions and phase separation processing and also lead to the formation of nanofibers with unique surface morphologies and good conductivities. The cytotoxic, necrotic and apoptotic effects of chosen nanofibers on osteocarcinoma cells were also investigated. These multifunctional, self-assembled, nanofibrous surfaces can serve as semi-conductive and bioactive platforms in various electrochemical and bio-engineering processes, as well as reactive matrices used for the immobilization of various biopolymer precursors.
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http://dx.doi.org/10.1016/j.msec.2015.12.045DOI Listing
April 2016

Antisense oligonucleotide delivery to cancer cell lines for the treatment of different cancer types.

Artif Cells Nanomed Biotechnol 2016 Dec 27;44(8):1938-1948. Epub 2015 Nov 27.

e Department of Chemistry, Division of Biochemistry , Hacettepe University , Beytepe , Ankara , Turkey.

Amphiphilic poly(3-hydroxylalkanoate) (PHA) copolymers find interesting applications in drug delivery. The aim of this study was to prepare nucleic acid adsorbed on (PHB-b-PEG-NH) nanoparticle platform for gene delivery. For this purpose, PHB-b-PEG-NH block copolymers were synthesized via transesterification reactions. The copolymers obtained were characterized by Proton Nuclear Magnetic Resonance (H-NMR), Fourier Transform Infrared Spectrometer (FTIR), Thermogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC) techniques. The cytotoxic, apoptotic and necrotic effects of these nanoparticles in the MDA 231 human breast cancer cell, the A549 human lung cancer cell and the L929 fibroblast cell lines were also investigated.
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http://dx.doi.org/10.3109/21691401.2015.1115409DOI Listing
December 2016

Biocompatible and biodegradable poly(Tannic Acid) hydrogel with antimicrobial and antioxidant properties.

Int J Biol Macromol 2016 Jan 23;82:150-9. Epub 2015 Oct 23.

Bioengineering Department, Engineering Faculty, Kirikkale University, Yahsihan, Kırıkkale 71450, Turkey.

A novel resourceful bulk poly(Tannic Acid) (p(TA)) hydrogel was prepared by crosslinking TA molecules with an epoxy crosslinker, trimethylolpropane triglycidyl ether (TMPGDE), in an autoclave at 90°C for 2h. The obtained p(TA) hydrogels were in disk form and have highly porous morphology. The swelling characteristics of p(TA) hydrogels were investigated in wound healing pH conditions of pH 5.4, 7.4, and 9 at 37.5°C, and the hydrogels showed good swelling and moisture content behavior. Especially, p(TA) hydrogels were found to be sensitive to pH 9 with 1669% maximum swelling. P(TA) hydrogels were completely degraded at pH 9 hydrolytically in 9 days. Total phenol contents and the effects of scavenging ABTS(+) radicals of degraded p(TA) hydrogels at pH 5.4, 7.4, and 9 were evaluated and calculated in terms of gallic acid equivalent and trolox equivalent antioxidant capacity, respectively, and found to be very effective. Moreover, degraded p(TA) hydrogels display strong antimicrobial behavior against gram positive Staphylococcus aureus, Bacillus subtilis, gram negative Pseudomonas aeruginosa bacteria strains and Candida albicans fungus strain. The WST-1 results indicated that bulk p(TA) hydrogels have no cyctotoxicity to the L929 fibroblast cell line in vitro.
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http://dx.doi.org/10.1016/j.ijbiomac.2015.10.057DOI Listing
January 2016

Novel multifunctional colloidal carbohydrate nanofiber electrolytes with excellent conductivity and responses to bone cancer cells.

Carbohydr Polym 2015 Nov 14;133:624-36. Epub 2015 Jul 14.

Department of Biology, Division of Bioengineering, Kirikkale University, 71450 Kirikkale, Turkey.

This work presents a new approach to fabricating novel polymer nanofiber composites (NFCs) from water solution blends of PVA (hydrolyzed 89%)/ODA-MMT and Na-CMC/ODA-MMT nanocomposites as well as their folic acid (FA) incorporated modifications (NC-3-FA and NC-4-FA) through green electrospinning nanotechnology. The chemical and physical structures and surface morphology of the nanofiber composites were confirmed. Significant improvements in nanofiber morphology and size distribution of the NFC-3-FA and NFC-4-FA nanofibers with lower average means 110 and 113nm compared with those of NFC-1/NFC-2 nanofibers (270 and 323nm) were observed. The structural elements of polymer NFCs, particularly loaded partner NC-2, plays an important role in chemical and physical interfacial interactions, phase separation processing and enables the formation of nanofibers with unique morphology and excellent conductivity (NFC-3-FA 3.25×10(-9)S/cm and NFC-4-FA 8.33×10(-4)S/cm). This is attributed to the higher surface contact areas and multifunctional self-assembled supramacromolecular nanostructures of amorphous colloidal electrolytes. The anticancer activity of FA-containing nanofibers against osteocarcinoma cells were evaluated by cytotoxicity, apoptotic and necrotic analysis methods.
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http://dx.doi.org/10.1016/j.carbpol.2015.06.106DOI Listing
November 2015

Downregulation of ABCE1 via siRNA affects the sensitivity of A549 cells against chemotherapeutic agents.

Med Oncol 2015 Apr 6;32(4):103. Epub 2015 Mar 6.

Chemistry Department, Biochemistry Division, Hacettepe University, Beytepe, 06800, Ankara, Turkey.

ATP-binding cassette E1 (ABCE1) is involved in several biological functions in cancer cells such as tumor proliferation, antiapoptotic pathway and chemoresistance mechanism. This work aimed to investigate the alterations in chemosensitivity of A549 lung cancer cells for 5-Fluorouracil (5-FU) and irinotecan by silencing ABCE1 using specific small interfering RNAs (siRNA). The cells were treated with low doses of drugs, alone and also their combinations with ABCE1 siRNA. Cytotoxicity, cell proliferation and apoptosis/necrosis evaluations were performed in order to examine the effects of the combined treatment. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to confirm the downregulation of ABCE1. We also investigated the levels of B cell lymphoma 2 (Bcl-2) and mammalian target of rapamycin (mTOR) after the treatments by RT-PCR. Downregulation of ABCE1 improved the anticancer effects of 5-FU in inducing cell viability/proliferation inhibition and apoptosis/necrosis, whereas interestingly, almost did not change or slightly reduced the anticancer effects of irinotecan. ABCE1 expression significantly decreased by transfecting the cells with ABCE1 siRNA. Moreover, Bcl-2 and mTOR levels changed after the single or combined therapy in parallel with the apoptotic and antiproliferation effect. In conclusion, the simultaneous treatment of lung cancer cells with ABCE1 siRNA and 5-FU exhibited synergistic or additive effects; however, ABCE1 siRNA and irinotecan had unexpected antagonistic effects. Our findings demonstrate that the strategy of downregulation of ABCE1 may be included in conventional 5-FU chemotherapy for lung cancer, minimizing the usage of 5-FU at high dosages.
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http://dx.doi.org/10.1007/s12032-015-0557-3DOI Listing
April 2015

Poly(sucrose) micro particles preparation and their use as biomaterials.

Int J Biol Macromol 2014 May 26;66:236-44. Epub 2014 Feb 26.

Bioengineering Department, Engineering Faculty, Kirikkale University, 06450 Kirikkale, Turkey.

Crosslinked p(sucrose) micro particles were synthesized for the first time from sucrose in water-in-oil microemulsion. Using divinyl sulfone (DVS) as crosslinker via reverse micelles of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) p(sucrose) micro particles formed in a single step with very high yield (>90%). The particles have wide size distributions, and negative zeta potential, -27.30 mV, and can be made magnetic field responsive. P(sucrose) particles were shown to be degradable at pHs of 2.5 and 11. Dopamine and gallic acid were used as model drugs for absorption/release studies from p(sucrose) particles. Interestingly, it was shown that p(sucrose) microparticles can be a nutrient for Escherichia coli, and maybe used as a growth medium for other cells, bacteria and organisms. Additionally, the cytotoxic effect of p(sucrose) particles were determined as 26 and 32.5% dead cells against MDA MB-231 cancerous cells and L929 fibroblast cells at 100 ug/ml concentration, respectively. P(sucrose) particles can be safely used for in vivo applications.
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http://dx.doi.org/10.1016/j.ijbiomac.2014.02.012DOI Listing
May 2014

Nanoemulsions and nonwoven fabrics carrying AgNPs: antibacterial but may be cytotoxic.

Artif Cells Nanomed Biotechnol 2014 Dec 15;42(6):392-9. Epub 2013 Oct 15.

Division of Nanotechnology and Nanomedicine, Hacettepe University , Ankara , Turkey.

Abstract The aim of this study is to prepare nonwoven fabrics carrying silver nanoparticles (AgNPs), and to investigate their antibacterial activities and cytotoxicities in parallel. AgNPs were impregnated from their nanoemulsions onto two commercially available nonwoven fabrics: pure-cotton fabrics (PCF) and polyester/viscous fabrics (PVF), by a simple adsorption (dipping) and were then heat stabilized. PCF exhibited stronger antibacterial effects on both Staphylococcus aureus and Escherichia coli. In-vitro cell culture studies demonstrated that AgNPs nanoemulsions and also fabrics carrying them were cytotoxic on L929-fibroblasts in all concentrations used here (6.25-400 ppm) in different extends. Only the fabrics loaded with AgNPs using nanoemulsion with the lowest concentration of 6.25 ppm exhibited low cytotoxicity but were still antibacterial.
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http://dx.doi.org/10.3109/21691401.2013.834908DOI Listing
December 2014

Fabrication and characterization of gold-nanoparticles/chitosan film: a scaffold for L929-fibroblasts.

Artif Cells Nanomed Biotechnol 2013 Dec 18;41(6):395-401. Epub 2013 Jan 18.

Faculty of Life Sciences , Department of Biology, Kırıkkale University , Kırıkkale Turkey.

The objective of the present study was to fabricate a gold nanoparticle crosslinked chitosan (Ch/AuNPs) composite film simple and to evaluate its use as a carrier matrix for L929- fibroblasts. L929- fibroblasts were seeded either onto Ch or Ch/AuNPs scaffolds. The Ch/AuNPs scaffold exhibited a higher cell proliferation and growth rate. The cytotoxicity test determined trypan blue staining indicated that Ch scaffolds devoid of AuNPs expressed almost no toxicity while the Ch/AuNPs composite scaffolds expressed a very limited toxicity only at higher doses. The Ch/AuNPs scaffold promotes cell attachment, growth and proliferation with almost no cytotoxicity.
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http://dx.doi.org/10.3109/21691401.2012.761228DOI Listing
December 2013

Characterization of chondrocytes cultured on catechin-loaded alginate-chitosan scaffolds.

Artif Cells Nanomed Biotechnol 2013 Aug 19;41(4):240-8. Epub 2012 Sep 19.

Department of Biology, Faculty of Art-science, Kirikkale University, Kirikkale, Turkey.

Bovine chondrocytes were seeded into scaffolds of a high molecular weight chitosan and alginate with a pore size of 50-350 μm with or without catechin. In polymerase chain reaction (PCR), unlike type II, collagen type I was no longer expressed at day 14. The DNA content increased until day 8 and began declining, indicating cell detachment. The GAG content increased during the first 12 days. The percentage of round and collagen type II immunoreactive cells increased over the time. Catechin has some protective properties on chondrocytes seeded on the alginate-chitosan scaffolds during the first 12 days by means of DNA and chondrocyte morphology (p < 0.05).
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http://dx.doi.org/10.3109/10731199.2012.718283DOI Listing
August 2013

Bioengineering functional copolymers. XXI. Synthesis of a novel end carboxyl-trithiocarbonate functionalized poly(maleic anhydride) and its interaction with cancer cells.

Bioorg Med Chem 2012 Aug 5;20(16):5053-61. Epub 2012 Jun 5.

Institute of Science & Engineering, Division of Nanotechnology and Nanomedicine, Hacettepe University, Beytepe 06800, Ankara, Turkey.

A novel carboxyl-trithiocarbonate functionalized polymer with a highly selective antitumor activity was synthesized by a reversible addition-fragmentation chain transfer (RAFT) polymerization of maleic anhydride (MA) with benzoyl peroxide as an initiator and S-1-dodecyl-S-(α,α'-dimethyl-α"-acetic acid)trithiocarbonate as a RAFT agent with the aim to design and synthesize an effective anticancer agent with minimum side effects. The structure, molecular weights and composition of synthesized polymers were investigated by (1)H ((13)C) NMR, MALDI-TOF-MS and GPC analyzes. It was demonstrated that RAFT polymerization of MA was accompanied by a partially controlled decarboxylation of anhydride units and the formation of conjugated double bond fragments in backbone macromolecular chains. The mechanism of interaction of pristine RAFT agent and PMA-RAFT polymer with cancer (HeLa human cervix carcinoma) and normal (L929 Fibroblast) cells was investigated by using a combination of chemical, biochemical, statistical, spectroscopic (SEM and fluorescence inverted microscope) and real-time analysis (RTCA) methods. PMA-RAFT exhibited higher and selective cytotoxicity, apoptotic and necrotic effects toward HeLa cells at relatively low concentrations (around 7.5-75 μg mL(-1), IC(50) = 11.183 μg mL(-1)) and toward Fibroblast cells at high concentrations (IC(50) > 100 μg mL(-1)). The observed highly selective antitumor activity render PMA-RAFT polymers as promising candidates for the utilization in cancer chemotherapy.
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http://dx.doi.org/10.1016/j.bmc.2012.05.058DOI Listing
August 2012

Preparation and characterization of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHX) based nanoparticles for targeted cancer therapy.

Eur J Pharm Sci 2011 Oct 23;44(3):310-20. Epub 2011 Aug 23.

Karaelmas University, Chemistry Department, Physical Chemistry Division, Zonguldak, Turkey.

Targeted drug delivery systems are one of the most promising alternatives for the cancer therapy. Rapid developments on nanomedicine facilitated the creation of novel nanotherapeutics by using different nanomaterials. Especially polymer based nanoparticles are convenient for this purpose. In this study; a natural polymer (poly(3-hydroxybutyrate-co-3-hydroxyhexanoate), PHBHHX) was used as a base matrix for the production of a novel nanotherapeutic including antineoplastic agent, Etoposide and attached folic acid as a ligand on the nanoparticles. Modified solvent evaporation technique was used for the production of PHBHHX nanoparticles and the average size of the obtained PHBHHX nanoparticles were observed in the range of 180 nm and 1.5 μm by the change in experimental conditions (i.e., homogenization rate, surfactant concentration and polymer/solvent ratio). By the increase in homogenization rate and surfactant concentration, size of the nanoparticles was decreased, while the size was increased by the increase in polymer/solvent ratio. Drug loading ratio was also found to be highly affected by polymer/drug ratio. Surface charge of the prepared nanoparticles was also investigated by zeta potential measurements. In the cytotoxicity tests; Etoposide loaded and folic acid attached PHBHHX nanoparticles were observed as more effective on HeLa cells than Etoposide loaded PHBHHX nanoparticles without attached folic acid. The cytotoxicity of folic acid conjugated PHBHHX nanoparticles to cancer cells was found to be much higher than that of normal fibroblast cells, demonstrating that the folate conjugated nanoparticles has the ability to selectively target to cancer cells. In addition, apoptotic/necrotic activities were evaluated for all formulations of the PHBHHX nanoparticles and parallel results with cytotoxicity tests were obtained. These studies demonstrate that the folic acid attached and Etoposide loaded PHBHHX nanoparticles seem as promising for the targeted cancer therapy.
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http://dx.doi.org/10.1016/j.ejps.2011.08.013DOI Listing
October 2011

Cytotoxicity and apoptotic effects of nickel oxide nanoparticles in cultured HeLa cells.

Folia Histochem Cytobiol 2010 Dec;48(4):524-9

Department of Chemistry, Kirikkale University Faculty of Art and Sciences, Kirikkale, Turkey.

The aim of this study was to observe the cytotoxicity and apoptotic effects of nickel oxide nanoparticles on human cervix epithelioid carcinoma cell line (HeLa). Nickel oxide precursors were synthesized by an nickel sulphate-excess urea reaction in boiling aqueous solution. The synthesized NiO nanoparticles (<200 nm) were investigated by X-ray diffraction analysis and transmission electron microscopy techniques. For cytotoxicity experiments, HeLa cells were incubated in 50-500 μg/mL NiO for 2, 6, 12 and 16 hours. The viable cells were counted with a haemacytometer using light microscopy. The cytotoxicity was observed low in 50-200 μg/mL concentration for 16 h, but high in 400-500 μg/mL concentration for 2-6 h. HeLa cells' cytoplasm membrane was lysed and detached from the well surface in 400 μg/mL concentration NiO nanoparticles. Double staining and M30 immunostaining were performed to quantify the number of apoptotic cells in culture on the basis of apoptotic cell nuclei scores. The apoptotic effect was observed 20% for 16 h incubation.
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http://dx.doi.org/10.2478/v10042-010-0045-8DOI Listing
December 2010

Bioengineering functional copolymers. XIV. Synthesis and interaction of poly(N-isopropylacrylamide-co-3,4-dihydro-2H-pyran-alt-maleic anhydride)s with SCLC cancer cells.

Bioorg Med Chem 2010 Nov 22;18(22):7975-84. Epub 2010 Sep 22.

Department of Biology, Faculty of Science and Art, Kirikkale University, Turkey.

Novel antitumor active functional polymers with supramacromolecular structures were synthesized by a complex-radical terpolymerization of N-isopropylacrylamide (NIPAm), 3,4-dihydro-2H-pyran (DHP), and maleic anhydride (MA) with 2,2'-azoisobisbutyronitrile as a radical initiator in 1,4-dioxane at 65°C under nitrogen atmosphere. The structure and composition of terpolymers were investigated by (1)H ((13)C) NMR spectroscopy. Interaction of terpolymers with human lung small cell carcinoma (SCLC) were investigated by using different methods such as cytotoxicity, statistical, apoptotic and necrotic cell indexes, double staining and caspase-3 immunostaining, light and fluorescence inverted microscopy analyses. Investigations into structure, composition, and antitumor activity relationships revealed that terpolymers containing a combination of ionisable amide-pyran linkages and H-bonded carboxylic groups exhibited higher cytotoxicity. It was observed that terpolymer with nearly alternating structure provides a maximum concentration of ionisable and H-bonded antitumor sites, and therefore, exhibits higher in vitro cytotoxicity, apoptotic and necrotic effects towards SCLC cancer cells.
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http://dx.doi.org/10.1016/j.bmc.2010.09.031DOI Listing
November 2010

Potential c-myc antisense oligonucleotide carriers: PCl/PEG/PEI and PLL/PEG/PEI.

Artif Cells Blood Substit Immobil Biotechnol 2011 Jun 2;39(3):143-54. Epub 2010 Aug 2.

Yıldız Technical University, Bioengineering Department, Davutpasa, Istanbul, Turkey.

In this work, positively charged, micelle-forming polymers were synthesized and used as a model vector to deliver antisense oligodeoxynucleotide (ASODN) into melanoma cells. Polymers and polymer/ASODN complexes were characterized by DLS according to size, charge, and critical micelle concentration. Nanosize and spherical complexes were observed by AFM. Complexes did not reveal significant toxicity to melanoma cells. Antiproliferative effect of the complexes was observed by immunocytochemical staining and estimated as 56.8% with N/P:9. High amount of apoptosis and very small amount of necrosis were estimated. According to the results, these positively charged polymers forming micelle-like structures seem promising as ASODN carriers.
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http://dx.doi.org/10.3109/10731199.2010.506852DOI Listing
June 2011

Alkaline phosphatase, cytokeratin 7, cytokeratin 8 in the diagnosis of primary lung adenocarcinoma from 148 pleura fluids specimens.

Folia Histochem Cytobiol 2009 ;47(1):87-92

Department of Biology, Kirlkkale University, 71450 Yahşihan-Kirlkkale, Turkey.

Adenocarcinomas are the most common cause of malignancy in pleura fluids. Usual primary sites include the lung, breast, gastrointestinal tract, and genitourinary tracts. Predicting the site of origin of an adenocarcinoma can be difficult due to overlapping morphologic characteristics. We investigated the use of alkaline phosphatase (AP), Cytokeratin7 (CK7) Cytokeratin8 (CK8) to distinguish adenocarcinomas of lung in 148 body cavity fluid samples. Overall results for primary lung adenocarcinomas, demonstrated CK8 reactivity in 106 (72%) of 148 cases. 95 primary lung carcinoma samples (65%) were positive for CK7. AP was expressed in 81% of primary lung adenocarcinomas. Positive immunoreactivity for AP was characterized by a red, diffusely apical cytoplasmic staining in tumor cells that ocurred singly or in groups. There was a significant difference between AP, CK 7 and CK 8 expressions in primary lung adenocarcinomas (P=0.02; Chi-squared test). The sensitivity of AP, CK8, CK7 as a marker for primary lung adenocarcinomas were 82%, 72%, 64%, respectively. Thus the AP positive staining largely confirmed the cytologic diagnosis of lung adenocarcinoma.
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http://dx.doi.org/10.2478/v10042-009-0001-7DOI Listing
July 2009