Nanoparticles of deoxycholic acid,polyethylene glycol and folic acid-modified chitosan for targeted delivery of doxorubicin

Chitosan (CS) was first modified hydrophobically with deoxycholic acid (DCA) and then with polyethylene glycol (PEG) to obtain a novel amphiphilic polymer (CS–DCA–PEG). This was covalently bound to folic acid (FA) to develop nanoparticles (CS–DCA–PEG–FA) with tumor cell targeting property. The structure of the conjugates was characterised using Fourier transform infrared and ¹H nuclear magnetic resonance spectroscopy and X-ray diffraction. Based on self-aggregation, the conjugates formed nanoparticles with a low critical aggregation concentration of 0.035 mg/ml. The anti-cancer drug doxorubicin (DOX) was encapsulated into the nanoparticles with a drug-loading capacity of 30.2 wt%. The mean diameter of the DOX-loaded nanoparticles was about 200 nm, with a narrow size distribution. Transmission electron microscopy images showed that the DOX-loaded nanoparticles were spherical. The drug release was studied under different conditions. Furthermore, the cytotoxic activities of DOX in CS–DCA–PEG–FA nanoparticles against folate receptor (FR)-positive HeLa cells and FR-negative fibroblast 3T3 cells were evaluated. These results suggested that the CS–DCA–PEG–FA nanoparticles may be a promising vehicle for the targeting anticancer drug to tumor cells.     

Silica/potassium ferrite nanocomposite: Structural,morphological, magnetic,thermal and in vitro cytotoxicity analysis

The coating of silica on potassium ferrite (KFeO₂) nanoparticles has been reported in the present study. The X-ray diffraction pattern revealed the formation of orthorhombic structure of bare potassium ferrite nanoparticles, which was also retained after the silica coating, along with a broad band near 2θ ∼ 20–25° pertaining to the presence of amorphous silica. The size of bare and coated potassium ferrite nanoparticles was found to be 4–8 nm and 10–22 nm, respectively, as observed from transmission electron microscope. The presence of silica was also revealed by the Fourier transform infrared spectrum and high resolution transmission electron microscope. In vibrating sample magnetometer analysis, both bare as well as coated potassium ferrite nanoparticles exhibited superparamagnetic behaviour with magnetic saturation values, 49.01 and 21.17 emu/g, respectively. Dose-dependent cellular toxicity was observed in the in vitro MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole) – assay study on Jurkat cells, where both bare as well as silica coated nanoparticles exhibited non-toxicity below 250 μg/ml. An augmentation of cell viability was observed in case of silica coated potassium ferrite nanoparticles. The nanosize, superparamagnetic behaviour and enhanced cell viability make silica coated potassium ferrite nanoparticles a potential claimant for biomedical applications.     

On-chip evaluation of shear stress effect on cytotoxicity of mesoporous silica nanoparticles

In this work, nanotoxicity in the bloodstream was modeled, and the cytotoxicity of sub-50 nm mesoporous silica nanoparticles to human endothelial cells was investigated under microfluidic flow conditions. Compared to traditional in vitro cytotoxicity assays performed under static conditions, unmodified mesoporous silica nanoparticles show higher and shear stress-dependent toxicity to endothelial cells under flow conditions. Interestingly, even under flow conditions, highly organo-modified mesoporous silica nanoparticles show no significant toxicity to endothelial cells. This paper clearly demonstrates that shear stress is an important factor to be considered in in vitro nanotoxicology assessments and provides a simple device for pursuing this consideration.     

Fabrication of Coated/Uncoated Magnetic Nanoparticles to Determine Their Surface Properties

Fabrication of coated and uncoated magnetic nanoparticles (MNPs) was achieved in the present study. The preparation and characterization of MNPs were confirmed by Fourier transform infrared spectroscopy (FTIR) spectroscopy, streaming potential (SP), and magnetic force microscopy (MFM) techniques. Coated and uncoated nanoparticles were analyzed by dynamic light scattering method to obtain the mean size of nanoparticles. The SP was used to record the electrical surface charge of nanoparticles. The results obtained revealed that the bare nanoparticles were negative charged at higher pH (pH > 6.0) while coated nanoparticles were positive charged at lower pH (pH < 6.0). The porosity of surface of bare and coated nanoparticles was shown by MFM.     

Multifunctional Upconversion Mesoporous Silica Nanostructures for Dual Modal Imaging and In Vivo Drug Delivery

Incorporating the agents for magnetic resonance imaging (MRI), optical imaging, and therapy in one nanostructured matrix to construct multifunctional nanomedical platform has attracted great attention for simultaneous diagnostic and therapeutic applications. In this work, a facile methodology is developed to construct a multifunctional anticancer drug nanocarrier by combining the special advantages of upconversion nanoparticles and mesoporous silica. β‐NaYF₄:Yb³⁺, Er³⁺@β‐NaGdF₄:Yb³⁺ is chosen as it can provide the dual modality of upconversion luminescence and MRI. Then mesoporous silica is directly coated onto the upconversion nanoparticles to form discrete, monodisperse, highly uniform, and core–shell structured nanospheres (labeled as UCNPs@mSiO₂), which are subsequently functionalized with hydrophilic polymer poly(ethylene glycol) (PEG) to improve the colloidal stability and biocompatibility. The obtained multifunctional nanocomposites can be used as an anticancer drug delivery carrier and applied for imaging. The anticancer drug doxorubicin (DOX) is absorbed into UCNPs@mSiO₂‐PEG nanospheres and released in a pH‐sensitive pattern. In vitro cell cytotoxicity tests on cancer cells verify that the DOX‐loaded UCNPs@mSiO₂‐PEG has comparable cytotoxicity with free DOX at the same concentration of DOX. In addition, the T₁‐weighted MRI that measures in aqueous solutions reveals that the contrast brightening increases with the concentration of Gd³⁺ component. Upconversion luminescence images of UCNPs@mSiO₂‐PEG uptaken by cells show green emission under 980 nm infrared laser excitation. Finally, the nanocomposites show low systematic toxicity and high in vivo antitumor therapy efficacy. These findings highlight the fascinating features of upconversion‐mesoporous nanocomposites as multimodality imaging contrast agents and nanocarrier for drug molecules.     

Fabrication of Janus particles via a “photografting-from” method and gold photoreduction

Faster and simpler methods for the fabrication of Janus particles are of tremendous importance for a real implementation of these particles. By combining thiol-modified silica particles (SMPs) with the use of UV light, it is possible to rapidly fabricate Janus particles coated with polymer brushes and gold nanoparticles via photochemical emulsion-assisted route. From the silica particle surface, polymeric brushes of polyethylene(glycol), PEG, were grafted via a photografting-from method on one hemisphere by using the thiol groups as photoinitiator of the polymerization. The other hemisphere was coated with gold nanoparticles (AuNPs) generated in situ via photoreduction of chloroauric acid promoted by Norrish type I photoinitiator. PEG/AuNPs@SMPs coated with Au nanoparticles with average diameter of 12.7 or 22.5 nm were obtained by playing on the mass ratio between thiol-modified silica particles and gold precursor. The Janus PEG/AuNPs@SMPs were fully characterized by scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy and UV–Vis spectroscopy. This strategy merges the advantages of emulsion-based selective masking and UV-induced reactions, and it is proved to be a feasible and fast route (reactions are completed in dozens of minutes) for Janus particles fabrication.     

Cytotoxicity and Cell Cycle Effects of Bare and Poly(vinyl alcohol)‐Coated Iron Oxide Nanoparticles in Mouse Fibroblasts

Super‐paramagnetic iron oxide nanoparticles (SPIONs) are recognized as powerful biocompatible materials for use in various biomedical applications, such as drug delivery, magnetic‐resonance imaging, cell/protein separation, hyperthermia and transfection. This study investigates the impact of high concentrations of SPIONs on cytotoxicity and cell‐cycle effects. The interactions of surface‐saturated (via interactions with cell medium) bare SPIONs and those coated with poly(vinyl alcohol) (PVA) with adhesive mouse fibroblast cells (L929) are investigated using an MTT assay. The two SPION formulations are synthesized using a co‐precipitation method. The bare and coated magnetic nanoparticles with passivated surfaces both result in changes in cell morphology, possibly due to clustering through their magnetostatic effect. At concentrations ranging up to 80 × 10^?3 M , cells exposed to the PVA‐coated nanoparticles demonstrate high cell viability without necrosis and apoptosis. In contrast, significant apoptosis is observed in cells exposed to bare SPIONs at a concentration of 80 × 10^?3 M . Nanoparticle exposure (20–80 × 10^?3 M ) leads to variations in both apoptosis and cell cycle, possibly due to irreversible DNA damage and repair of oxidative DNA lesions, respectively. Additionally, the formation of vacuoles within the cells and granular cells indicates autophagy cell death rather than either apoptosis or necrosis.     

Synthesis of stable silica-dye hybrid nanomaterial as DNA carrier

A new method is proposed for the fabrication of fluorescence-labeled and amine-modified silica nanoparticles for application as nonviral vectors in gene delivery. Highly monodisperse, stable fluorescent silica nanoparticles were prepared using 2,5-bis(5-tert-butyl-2-benzoxazolyl)thiophene and the water-in-oil microemulsion method. The green-fluorescent-protein gene can be easily combined onto the positively charged surfaces of nanoparticles to form a nanoparticle-DNA complex. The nanoparticle-DNA complex successfully passed through various barriers into the HeLa and HEK 293 K cells. The cytotoxicity of the PEI-coated and BBOT-encapsulated silica nanoparticles on both the HeLa and HEK 293T cell lines was found to be at an acceptable level for use as gene carriers when the particle concentration was below 125 microg/ml. The fluorescence intracellular images confirm the successful delivery of the nanoparticle-DNA complex and gene expression. The present work suggests the potential use of dye-incorporated silica nanoparticles in nonviral gene delivery.     

Real‐Time Label‐Free Monitoring of Nanoparticle Cell Uptake

The surface plasmon resonance technique in combination with whole cell sensing is used for the first time for real‐time label‐free monitoring of nanoparticle cell uptake. The uptake kinetics of several types of nanoparticles relevant to drug delivery applications into HeLa cells is determined. The cell uptake of the nanoparticles is confirmed by confocal microscopy. The cell uptake of silica nanoparticles and polyethylenimine–plasmid DNA polyplexes is studied as a function of temperature, and the uptake energies are determined by Arrhenius plots. The phase transition temperature of the HeLa cell membrane is detected when monitoring cell uptake of silica nanoparticles at different temperatures. The HeLa cell uptake of the mesoporous silica nanoparticles is energy‐independent at temperatures slightly higher than the phase transition temperature of the HeLa cell membrane, while the uptake of polyethylenimine–DNA polyplexes is energy‐dependent and linear as a function of temperature with an activation energy of Ea = 62 ± 7 kJ mol^?1 = 15 ± 2 kcal mol^?1. The HeLa cell uptake of red blood cell derived extracellular vesicles is also studied as a function of the extracellular vesicle concentration. The results show a concentration dependent behavior reaching a saturation level of the extracellular vesicle uptake by HeLa cells.     

pH‐Responsive PEG‐Shedding and Targeting Peptide‐Modified Nanoparticles for Dual‐Delivery of Irinotecan and microRNA to Enhance Tumor‐Specific Therapy

Irinotecan is one of the main chemotherapeutic agents for colorectal cancer (CRC). MicroRNA‐200 (miR‐200) has been reported to inhibit metastasis in cancer cells. Herein, pH‐sensitive and peptide‐modified liposomes and solid lipid nanoparticles (SLN) are designed for encapsulation of irinotecan and miR‐200, respectively. These peptides include one cell‐penetrating peptide, one ligand targeted to tumor neovasculature undergoing angiogenesis, and one mitochondria‐targeting peptide. The peptide‐modified nanoparticles are further coated with a pH‐sensitive PEG‐lipid derivative with an imine bond. These specially‐designed nanoparticles exhibit pH‐responsive release, internalization, and intracellular distribution in acidic pH of colon cancer HCT116 cells. These nanoparticles display low toxicity to blood and noncancerous intestinal cells. Delivery of miR‐200 by SLN further increases the cytotoxicity of irinotecan‐loaded liposomes against CRC cells by triggering apoptosis and suppressing RAS/β‐catenin/ZEB/multiple drug resistance (MDR) pathways. Using CRC‐bearing mice, the in vivo results further indicate that irinotecan and miR‐200 in pH‐responsive targeting nanoparticles exhibit positive therapeutic outcomes by inhibiting colorectal tumor growth and reducing systemic toxicity. Overall, successful delivery of miR and chemotherapy by multifunctional nanoparticles may modulate β‐catenin/MDR/apoptosis/metastasis signaling pathways and induce programmed cancer cell death. Thus, these pH‐responsive targeting nanoparticles may provide a potential regimen for effective treatment of colorectal cancer.     

The pH-controlled nanoparticles size of polydopamine for anti-cancer drug delivery

A facile method was used to prepare polydopamine (PDA) nanoparticles. The effect of the initial pH of the dopamine solution on the formation kinetics, chemical structure, and biocompatibility of PDA nanoparticles was evaluated. Additionally, camptothecin (CPT) was chosen as a model anti-cancer drug with which to evaluate the efficiency of drug loading and release behavior of PDA nanoparticles. The results indicated that the size and yield of PDA nanoparticles, consisting of quinoid and indoline species, were closely related to the pH value of the precursor solution. At a reaction time of 6 h, the uniform particle sizes of PDA nanoparticles were ~400, 250, 150, and 75 nm in solutions with initial pH values of 7.5, 8, 8.5, and 9, respectively, and with corresponding yields of 3, 7, 20, and 34 %. The amounts of CPT loaded in 1 mg of PDA nanoparticles synthesized at pH values of 7.5, 8, 8.5, and 9 for 6 h were 10.85, 11.81, 10.17, and 6.19 μg, respectively. After the first day, 19, 20, 25, and 36 % of the CPT was released from PDA nanoparticles synthesized at pH values of 7.5, 8, 8.5, and 9, respectively, depending on the particle size. The PDA nanoparticles had excellent haemocompatibility: there was no apparent hemolysis, and they did not cause acute toxicity in A549 and HeLa cells. The loading of CPT into PDA nanoparticles significantly reduced the viability of A549 and HeLa cells, comparable to free CPT. It can be concluded that the PDA nanoparticles prepared by our facile method are potential carriers of anticancer drugs for cancer therapy.     

Linear and Hyperbranched Polyglycerol Derivatives as Excellent Bioinert Glass Coating Materials

The non‐specific adsorption of proteins to surfaces in contact with biofluids constitutes a major problem in the biomedical and biotechnological field, due to the initiation of biofilm formation and the resulting improper function of devices. Therefore, non‐fouling surfaces modified with poly(ethylene glycol) (PEG) are usually applied. In this study, we report the synthesis of triethoxysilane modified glycerol based polymers of linear and branched architecture for the preparation of covalently attached monolayers on glass. Evaluation of the biocompatibility of these surfaces was performed in comparison to bare non‐coated glass, hydrophobic hexadecane modified glass, and mPEG modified glass as the controls. Protein adsorption of BSA and fibrinogen (1 mg · mL^?1 in PBS) after 4 and 24 h immersion was reduced by more than 96 and 90%, respectively, compared to the adsorption on bare glass substrates. In addition, mouse NIH‐3T3 fibroblast cells showed only marginal adhesion on the polyglycerol and mPEG coated slides after 3 and 7 d incubation in cell suspension, which demonstrates the long‐term stability of the applied glass coatings. The non‐adhesive properties of these coatings were further reflected in bacterial adhesion tests of Escherichia coli K12 and three clinically relevant Gram‐positive and negative strains (Staphylococcus aureus, Pseudomonas aeruginosa, and Aeromonas hydrophila), since linear polyglycerol (LPG(OH)), linear poly(methyl glycerol) (LPG(OMe)), and hyperbranched polyglycerol (HPG) reduced the adhesion for all tested strains by more than 99% compared to bare glass. Therefore, polyglycerol derivatives present an excellent non‐fouling surface coating as an alternative to PEG with feasibility for surface modification of various substrates.     

Fumed silica/polymer hybrid nanoparticles prepared by redox-initiated graft polymerization in emulsions

Hybrid particles comprising aggregated fumed silica nanoparticles as the core and hydrophobic polymers existing around the nanoparticles were prepared by ‘grafting from’ polymerization in emulsions. The emulsion polymerization employed cetyltrimethylammonium bromide (CTAB) as a cationic surfactant and sodium dodecyl sulfate (SDS) as an anionic surfactant, respectively, to stabilize the emulsion polymerization. The polymerization was initiated by the redox reaction between ceric ion Ce(IV) and the amine groups on the surfaces of aminated fumed silica nanoparticles that were modified by 3-aminopropyltriethoxysilane. Infrared spectroscopy and thermogravimetric analysis demonstrated that both poly(methyl methacrylate) (PMMA) and polystyrene (PS) were successfully grafted onto the fumed silica surface. The type of surfactant greatly affected the grafting ratio, monomer-to-polymer conversion, and morphology of the product. When CTAB was used as the surfactant, both the grafting ratio and monomer-to-polymer conversion were lower than when SDS was used, but transmission electron microscopy and light scattering analysis indicated that most of the resultant particles were sub-100 nm hybrid nanoparticles with a non-spherical shape and particles sizes of 75–90 and 57–85 nm for PMMA and PS-grafted fumed silica, respectively. Whereas, when SDS was used as the surfactant, the particles agglomerated to form large irregular clusters or even networks, possibly due to the electrostatic attractions between SDS and Ce(IV) and/or the aminated fumed silica nanoparticles in aqueous solution.     

In vitro evaluation of folic acid modified carboxymethyl chitosan nanoparticles loaded with doxorubicin for targeted delivery

The development of smart targeted nanoparticle that can deliver drugs to direct cancer cells, introduces better efficacy and lower toxicity for treatment. We report the development and characterizations of pH-sensitive carboxymethyl chitosan modified folic acid nanoparticles and manifest their feasibility as an effective targeted drug delivery vehicle. The nanoparticles have been synthesized from carboxymethyl chitosan with covalently bonded bifunctional 2,2′-(ethylenedioxy)-bis-(ethylamine) (EDBE) through the conjugation with folic acid. The conjugation has been analyzed by Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. The resultant nanoparticles with an average size less then 200 nm measured by dynamic light scattering and transmission electron microscopy. Confocal microscopy and flow cytometric analysis have revealed that folate-mediated targeting significantly enhances the cellular uptake of the nanoparticle and thus facilitates apoptosis of cancer cells (HeLa, B16F1). For the application of the nanoparticles as a drug carrier, Doxorubicin a potent anticancer drug has been loaded into the nanoparticles, with the drug loading amount and the drug release pattern observed.     

Preparation of magnetic mesoporous silica nanoparticles as a multifunctional platform for potential drug delivery and hyperthermia

We report the preparation of magnetic mesoporous silica (MMS) nanoparticles with the potential multifunctionality of drug delivery and magnetic hyperthermia. Carbon-encapsulated magnetic colloidal nanoparticles (MCN@C) were used to coat mesoporous silica shells for the formation of the core-shell structured MMS nanoparticles (MCN@C/mSiO₂), and the rattle-type structured MMS nanoparticles (MCN/mSiO₂) were obtained after the removal of the carbon layers from MCN@C/mSiO₂ nanoparticles. The morphology, structure, magnetic hyperthermia ability, drug release behavior, in vitro cytotoxicity and cellular uptake of MMS nanoparticles were investigated. The results revealed that the MCN@C/mSiO₂ and MCN/mSiO₂ nanoparticles had spherical morphology and average particle sizes of 390 and 320 nm, respectively. The MCN@C/mSiO₂ nanoparticles exhibited higher magnetic hyperthermia ability compared to the MCN/mSiO₂ nanoparticles, but the MCN/mSiO₂ nanoparticles had higher drug loading capacity. Both MCN@C/mSiO₂ and MCN/mSiO₂ nanoparticles had similar drug release behavior with pH-controlled release and temperature-accelerated release. Furthermore, the MCN@C/mSiO₂ and MCN/mSiO₂ nanoparticles showed low cytotoxicity and could be internalized into HeLa cells. Therefore, the MCN@C/mSiO₂ and MCN/mSiO₂ nanoparticles would be promising for the combination of drug delivery and magnetic hyperthermia treatment in cancer therapy.     

Uptake Kinetics and Nanotoxicity of Silica Nanoparticles Are Cell Type Dependent

In this study, it is shown that the cytotoxic response of cells as well as the uptake kinetics of nanoparticles (NPs) is cell type dependent. We use silica NPs with a diameter of 310 nm labeled with perylene dye and 304 nm unlabeled particles to evaluate cell type‐dependent uptake and cytotoxicity on human vascular endothelial cells (HUVEC) and cancer cells derived from the cervix carcinoma (HeLa). Besides their size, the particles are characterized concerning homogeneity of the labeling and their zeta potential. The cellular uptake of the labeled NPs is quantified by imaging the cells via confocal microscopy in a time‐dependent manner, with subsequent image analysis via a custom‐made and freely available digital method, Particle_in_Cell‐3D. We find that within the first 4 h of interaction, the uptake of silica NPs into the cytoplasm is up to 10 times more efficient in HUVEC than in HeLa cells. Interestingly, after 10 or 24 h of interaction, the number of intracellular particles for HeLa cells by far surpasses the one for HUVEC. Inhibitor studies show that these endothelial cells internalize 310 nm SiO₂ NPs via the clathrin‐dependent pathway. Remarkably, the differences in the amount of taken up NPs are not directly reflected by the metabolic activity and membrane integrity of the individual cell types. Interaction with NPs leads to a concentration‐dependent decrease in mitochondrial activity and an increase in membrane leakage for HUVEC, whereas HeLa cells show only a reduced mitochondrial activity and no membrane leakage. In addition, silica NPs lead to HUVEC cell death while HeLa cells survive. These findings indicate that HUVEC are more sensitive than HeLa cells upon silica NP exposure.     

Biocompatible nanoclusters of <Emphasis Type="Italic">O</Emphasis>-carboxymethyl chitosan-coated Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanoparticles: synthesis,characterization and magnetic heating efficiency

In this work, we developed a polymer encapsulation of Fe₃O₄ nanoparticles as a core–shell nanocluster with different sizes to investigate the cluster structure effect on their magnetic properties and magnetic heating behavior. Well-dispersed nanoclusters of O-carboxymethyl chitosan-coated Fe₃O₄ nanoparticles were synthesized by microwave-assisted co-precipitation. The cluster sizes were tunable by varying the concentration of polymers used during synthesis. Nanoclusters present superparamagnetic behavior at room temperature with a reduction in saturation magnetization as a consequence of coating layer. The shift of blocking temperature to the higher value with increasing clusters size shows the stronger magnetic interaction in larger magnetic clusters. In a low alternating magnetic field with frequency of 178 Hz and amplitude of 103 Oe, nanoclusters offer a high heating efficiency. A maximum specific absorption rate of 204 W/g is observed in the sample with hydrodynamic size of 53 nm. In vitro cytotoxicity analysis performed on HeLa cells verified that nanoclusters show a good biocompatibility and can be an excellent candidate for applications in hyperthermia cancer treatment.     

Resonance energy transfer-amplifying fluorescence quenching at the surface of silica nanoparticles toward ultrasensitive detection of TNT

This paper reports a resonance energy transfer-amplifying fluorescence quenching at the surface of silica nanoparticles for the ultrasensitive detection of 2,4,6-trinitrotoluene (TNT) in solution and vapor environments. Fluorescence dye and organic amine were covalently modified onto the surface of silica nanoparticles to form a hybrid monolayer of dye fluorophores and amine ligands. The fluorescent silica particles can specifically bind TNT species by the charge-transfer complexing interaction between electron-rich amine ligands and electron-deficient aromatic rings. The resultant TNT-amine complexes bound at the silica surface can strongly suppress the fluorescence emission of the chosen dye by the fluorescence resonance energy transfer (FRET) from dye donor to the irradiative TNT-amine acceptor through intermolecular polar-polar interactions at spatial proximity. The quenching efficiency of the hybrid nanoparticles with TNT is greatly amplified by at least 10-fold that of the corresponding pure dye. The nanoparticle-assembled arrays on silicon wafer can sensitively detect down to approximately 1 nM TNT with the use of only 10 microL of solution (approximately 2 pg TNT) and several ppb of TNT vapor in air. The simple FRET-based nanoparticle sensors reported here exhibit a high and stable fluorescence brightness, strong analyte affinity, and good assembly flexibility and can thus find many applications in the detection of ultratrace analytes.     

Determining early adhesion of cells on polysaccharides/PCL surfaces by a quartz crystal microbalance

The early adhesions of cells to various biopolymers are important to their growths and proliferations. Here, the adhesion of cells (e.g., fibroblasts) on the electrode of a quartz crystal microbalance (QCM) that was coated by PCL or PEG/PCL and further adsorbed by chitosan (CS) or CS/hyaluronic acid (HA) layers, was examined by cell-counting technique, QCM method and MTS assay under a serum-free condition for 3 h. The surfaces on electrodes of the QCM were confirmed to have been modified by measuring their contact angles, FT-IR spectra and the weights of biopolymers affected the frequency shifts of the QCM. Among tested surfaces on electrodes, the adhesion of fibroblasts on a HA/CS/PCL surface was the most (e.g., 3.08 × 10⁵ cells/cm²) while that on a PEG/PCL surface was the least (e.g., 0.7 × 10⁵ cells/cm²), as determined by cell-counting technique. The frequency shift and the mass of adhering fibroblasts on HA/CS/PCL electrodes were ?3,537 ± 770 Hz and 3.78 ± 0.22 μg (n = 3), respectively, that were significantly exceeded those on other electrodes (?393 ± 58 Hz and 0.32 ± 0.06 μg, n = 3, respectively, for PEG/PCL electrodes). These results were consistent with cell-counting technique. Although MTS assay yielded similar results, it was less sensitive than the two aforementioned methods. In conclusion, modified electrodes of a QCM provide a convenient and sensitive method for examining the early adhesion of cells (e.g., 3 h) to biopolymer surfaces.