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Öğe Enhancing gene delivery efficiency with amphiphilic chitosan modified by myristic acid and tertiary amino groups(Elsevier, 2024) Fidan, Emine Busra Eker; Bal, Kevser; Senturk, Sema; Kaplan, Ozlem; Demir, Kamber; Gok, Mehmet KorayThe aim of this study is to synthesize new amphiphilic chitosan containing myristic acid as the hydrophobic tail and tertiary amine groups as the hydrophilic head and to evaluate the gene delivery efficiency. In this context, the primary amine groups of chitosan were first modified with myristic acid (Chi-M), followed by the modification of the methylol groups with 3-dimethylamino-1-propyl chloride hydrochloride. The chemical characterization of this chitosan formulation (Chi-MA) was determined using nuclear magnetic resonance (NMR), Fouriertransform infrared spectroscopy (FTIR) analysis and gel permeation chromatography-size exclusion chromatography. Chi-MA nanoparticles were prepared via ionic gelation, and particle size, polydispersity and zeta potential were determined. The nanoparticles were evaluated for their proton buffering capacity and gene complexing capacity. Additionally, the cytotoxicity of Chi-MA on HEK293T cells was determined via MTT assay, and the transfection efficiency of Chi-MA was analyzed by a flow cytometer. The results indicate a significant increase in gene complexing capacity (8-fold) and nanoparticle formation ability of Chi-MA compared to other chitosan formulations. Chi-MA nanoparticles showed no toxicity against HEK293T cells and exhibited the highest transfection efficiency with significantly lower nanoparticle: gene ratios compared to previous studies. These findings demonstrate the effective use of amphiphilic Chi-MA as a gene carrier.Öğe Enhancing transfection efficiency of primary cell lines using different terminated PBAE structures without endcapping reaction(Springer, 2025) Demir, Irmak; Celik, Sibel Kucukertugrul; Bal, Kevser; Kaplan, Ozlem; Senturk, Sema; Demir, Kamber; Gok, Mehmet KorayGene therapy holds promise for a wide range of diseases, including Alzheimer's, diabetes, and cancer, and requires the efficient transfer of nucleic acids into cells. However, transfection in primary cells is still problematic and requires the development of new transfection agents. Poly (beta-amino ester) (PBAE) has attracted great attention in transfection research due to their low toxicity, high gene loading capacity, endosomal escape ability, and biodegradability properties. In this study, two new PBAEs with different molecular weights are synthesized that could provide high viability and transfection efficiency in primary cells. They are characterized using FTIR and 1H NMR analysis. GPC-SEC system is also used to calculate the average molecular weight (Mw) and polydispersity index. PBAE nanoparticle preparation is carried out using the nanoprecipitation technique. The gene loading capacity, protective ability against nuclease degradation, and proton buffering capacity of nanoparticles are determined. Additionally, the morphology of PBAEA:pEGFN1 complexes was investigated by STEM analysis. Finally, their cytotoxicity and transfection efficiency in primary ovine fibroblast (POF) cells are also investigated. The results reveal that the new PBAE with higher Mw achieves quite high transfection efficiency of about 87% and did not show any cytotoxic effects on these cells. These findings suggest that PBAE is a promising option to achieve high transfection efficiency in primary cells.Öğe Improving physiological solubility and gene transfer efficiency of chitosan via 3-nitrobenzaldehyde and amino acid conjugation(Elsevier, 2025) Bal, Kevser; Kaplan, Ozlem; Senturk, Sema; Celik, Sibel Kucukertugrul; Demir, Kamber; Gok, Mehmet KorayIn this study, chitosan was chemically modified with 3-nitrobenzaldehyde (3NBA) and three amino acids (arginine, cysteine, and histidine) to enhance its gene delivery performance. 3-NBA was selected for its known DNA binding properties, while the amino acids were chosen based on their functional groups, which can improve solubility, facilitate cellular uptake, and contribute to endosomal escape. The modified chitosan polymers were characterized using Fourier Transform Infrared Spectroscopy (FTIR) and Nuclear Magnetic Resonance Spectroscopy (NMR). Nanoparticles were prepared using the ionotropic gelation method, and their particle size, polydispersity index (PDI), zeta potential were analyzed by dynamic light scattering (DLS). The particle sizes ranged from 105.07 f 3.45 to 206.15 f 10.39 nm, with PDI values between 0.29 f 0.01 and 0.39 f 0.02. Zeta potentials were measured between 32.05 f 0.49 mV and 51.95 f 0.35 mV. The cysteine-modified chitosan (Chi3NBACys) exhibited approximately 8.4-fold higher solubility than unmodified chitosan. In vitro studies demonstrated that the modified chitosan nanoparticles exhibited low cytotoxicity in HEK293T cells. Among the tested formulations, Chi-3NBACys showed the highest transfection efficiency, comparable to commercial agent LipofectamineTM 2000. These findings suggest that chitosan nanoparticles modified with 3-NBA and amino acids can be safe and efficient non-viral gene delivery vectors.Öğe Mukoadhezyonun Temel Prensipleri Üzerine Bir Derleme: Mukoadhezif Biyopolimer Olarak Kitosan'ın Önemi(Alanya Alaaddin Keykubat Üniversitesi, 2024) Fidan, Emine Büşra Eker; Bal, Kevser; Pabuccuoğlu, Saadet KevserMukoadhezif polimerler, vücuttaki göz, burun, vajinal ve bukkal boşluklar ile gastrointestinal sistem gibi mukusla kaplı bölgelerde mukus/epitel yüzeyine yapışma özelliğine sahiptir. Bu nedenle, dozaj formunun mukozadaki kalış süresini arttırma ve ilaç uygulamasını önemli ölçüde iyileştirme özellikleri gösterirler. Kitosan ve modifiye edilmiş türevlerini içeren mukoadhezif ilaç taşıma sistemleri, hem lokal hem de sistemik ilaç taşıma için birçok avantaja sahiptir. Bu derlemenin amacı, fonksiyonel bir mukoadhezif ilaç taşıma sistemi olarak kitosan'ın önemini ortaya koymaktır.Öğe Optimization Studies of Mg/Al-NO3 Layered Double Hydroxide Nanoparticles by Hydrothermal Treatment(Alanya Alaaddin Keykubat University, 2023) Bal, Kevser; Şentürk, Sema; Kaplan, Özlem; Gök, Mehmet Koray; Pabuccuoğlu, Saadet KevserLayered Double Hydroxides based nanoparticles offer significant advantages in biological applications with high biocompatibility and low cytotoxicity. In this study, nanoparticles (nMg/Al-NO3-LDH) were synthesized by the co-precipitation method and synthesis optimization of the nanoparticles was carried out by hydrothermal treatment. The effect of hydrothermal treatment on Z-average and surface charge was evaluated. Experiments were performed at 80°C and 100°C during in the range of 2-48 h by using different stirring rates (250, 1000, and 1500 rpm) and without stirring. Dynamic Light Scattering (DLS) was used to assess the particle size (nm), polydispersity index (PDI), and zeta potential (mV) of the nanoparticles. The chemical structure of nanoparticles was characterized by Fourier Transform Infrared spectrometry (FTIR). As a result, nanoparticles with an optimum particle size of 86.87 nm, a PDI of 0.132 and a zeta potential (mV) of 44.4±8.74 were obtained at 80°C, 48h and 250 rpm. The data showed that Mg/Al-NO3-LDH nanoparticles have suitable physical properties for biological applications.Öğe Recent progress in chitosan-based nanoparticles for drug delivery: a review on modifications and therapeutic potential(Taylor & Francis Ltd, 2025) Bal, Kevser; Celik, Sibel Kucukertugrul; Senturk, Sema; Kaplan, Ozlem; Eker, Emine Busra; Gok, Mehmet KorayChitosan, obtained from chitin by deacetylation, is a versatile biopolymer known for its biocompatibility, biodegradability and environmental friendliness. Combined with its chemical and physical modifiability, these properties have made chitosan an important material in biomedical and pharmaceutical fields, especially in drug delivery systems. Chitosan-based nanomaterials exhibit enhanced functions through various chemical modifications such as thiolation, acetylation, carboxylation and phosphorylation, as well as through physical and enzymatic approaches. These modifications address inherent limitations such as poor solubility, limited acid resistance and insufficient mechanical strength, expanding the applications of chitosan in tissue engineering, gene therapy, vaccine delivery, wound healing and bioimaging. This review provides an in-depth analysis of the chemical structure, physicochemical properties and modification strategies of chitosan. It also explores current methodologies for preparing chitosan nanoparticles, along with drug loading and release techniques. Various targeting strategies employed in chitosan-based delivery systems are examined in detail. To illustrate the clinical relevance of these approaches, representative examples from recent therapeutic studies are included. Moreover, it highlights future research directions and the innovation potential of chitosan-based materials.Öğe Redox-responsive lipoic acid-modified poly ((3-amino ester) nanoparticles for enhanced gene delivery(Elsevier, 2025) Kaplan, Ozlem; Bal, Kevser; Senturk, Sema; Demir, Kamber; Gok, Mehmet KorayEfficient and safe delivery of genetic material into cells is a critical step in gene therapy applications. Poly ((3-amino ester) (P(3AE) polymers have garnered significant attention among non-viral gene delivery systems due to their biocompatibility, low toxicity, and structural versatility for modifications. However, the hydrolysisdependent nucleic acid release mechanism of P(3AE often results in uncontrolled release, thereby limiting transfection efficiency. In this study, two P(3AE polymers were modified with lipoic acid to develop redoxresponsive drug delivery systems, and their transfection efficiency was evaluated in cervical cancer cells (HeLa) and human embryonic kidney (HEK293T) cells. The modifications were confirmed via Fourier-transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) analyses. The proton buffering capacities of P(3AE and lipoic acid-modified P(3AE within the pH range of 5.0-7.4 exhibited similar profiles. Nanoparticles were prepared from these polymers using the nanoprecipitation technique, yielding particle sizes ranging from 102.0 +/- 2.5 nm to 212.2 +/- 8.8 nm, with polydispersity index (PDI) values between 0.166 +/- 0.043 and 0.280 +/- 0.019. The zeta potential of all nanoparticles ranged from +27.3 +/- 1.1 mV to +47.4 +/- 1.7 mV. While the particle sizes remained stable over six weeks, an increasing trend in PDI values was observed. A decrease in zeta potential was recorded, attributed to the hydrolysis of P(3AE. Redox sensitivity analyses using dithiothreitol (DTT) confirmed the redox responsiveness of the nanoparticles and validated their rapid degradation under reductive conditions. In vitro studies revealed that lipoic acid modifications enhanced transfection efficiency in HeLa cells. These findings suggest that lipoic acid-modified P(3AE polymers hold significant potential for developing more effective gene delivery systems targeting cancer cells.Öğe Silane and amino acid functionalized PBAEs for enhanced gene Delivery: Synthesis, characterization, and transfection efficiency(Elsevier, 2025) Kaplan, Ozlem; Bal, Kevser; Senturk, Sema; Demir, Kamber; Gok, Mehmet KorayThis study aimed to synthesize, characterize, and evaluate the transfection efficiency of end group-modified branched poly (R-amino ester) (PBAE) based polymers containing silane groups to be used as gene carriers. Branched PBAE polymers were synthesized using bisphenol A ethoxylate diacrylate, diethylenetriamine, and N-[3-(trimethoxysilyl)propyl] ethylenediamine, and various amino acids (histidine, isoleucine, serine, methionine, phenylalanine, arginine) were added for end group modification. The structures of the polymers were confirmed by nuclear magnetic resonance (NMR) and Fourier-transform infrared spectroscopy (FTIR) analysis. Nano-particles were prepared using the nanoprecipitation technique, and their plasmid DNA complexing and protection capacity and endosomal escape abilities were determined. Finally, the cytotoxicity of the nanoparticles on HeLa and HEK293T cell lines was tested by the MTT test, and their transfection efficiency was tested by flow cytometry and fluorescence microscopy. All the tested nanoparticles showed lower toxicity in HeLa cells than in HEK293T cells. Transfection efficiency followed an increasing order, starting from the linear PBAE, progressing to silane modification, and subsequently with modifications by histidine, isoleucine, serine, methionine, phenylalanine, and arginine. This study highlights the potential of silane and amino acid-functionalized PBAE polymers as gene carriers and the impact of these modifications on transfection efficiency.Öğe Thiolated layered double hydroxide-based nanoparticles: A study on mucoadhesiveness and cytotoxicity(Elsevier, 2025) Senturk, Sema; Kaplan, Ozlem; Bal, Kevser; Topcu, Kubra Sen Bas; Gok, Mehmet KorayThe aim of this study was the synthesis and characterization of thiolated Mg/Al Layered double hydroxide nanoparticles (nLDH) that can adhere to mucosal surfaces. For this purpose, initially, nLDH were synthesized and then their surface was modified with L-cysteine (Cys). Then the Cys-modified nanoparticle structure (nLDHCys) Cys ) was characterized by FTIR, XRD and FE-SEM/EDS, and the amount of thiol groups was determined using the iodometric method. The cytotoxicity properties of nLDH and LDHCys Cys were examined on cervical (HeLa), fibroblast (L929) and colon (HT29) cell lines. In addition, the adhesion of nanoparticles to mucosal surfaces has been evaluated through mucin interaction experiments and ex vivo tests on ewe vaginal tissue. The results from characterization studies prove that Cys molecule has been successfully modified on the surface of nLDH. Cytotoxicity studies demonstrated that Cys modification did not negatively affect the cell viability, and these results were extremely close to those of nLDH. Ex vivo tests on ewe vaginal tissue and mucin interaction demonstrated that nLDHCys Cys enhanced retention on the mucosal surface. Considering their retention ability to mucosal surface and their biocompatible nature, nLDHCys Cys is expected to play an important role as a promising option for a safe mucoadhesive nanoparticle system for biological applications in the future.
