In this study, we report pH-responsive polysaccharidic nanogels for cytosolic peptide delivery. We conjugated starch to water-soluble glycol chitosan and pH-responsive 3-diethylaminopropylamine (starch–(glycol chitosan–3-diethylaminopropylamine)). Starch–(glycol chitosan–3-diethylaminopropylamine) self-organizes in aqueous solution, with the glycol chitosan blocks on the hydrophilic outer shell and starch and 3-diethylaminopropylamine blocks in the hydrogel inner core. The experimental results demonstrated that the protonation of 3-diethylaminopropylamine at pH 6.0 (endosomal pH) allowed for accelerated release of the encapsulated D-(KLAKLAK)2 proapoptotic peptide from the nanogels as a result of electrostatic repulsion between D-(KLAKLAK)2 and 3-diethylaminopropylamine. A hemolysis test using red blood cell membranes (as an endosomal membrane model) revealed the excellent endosomolytic activity of these nanogels, which likely stems from the proton-sponge effect of 3-diethylaminopropylamine at pH 6.0. As a result, these nanogels resulted in increased KB tumor cell ablation.
In this study, nanoliposome-loaded poly(hexamethylene biguanide) is introduced as a novel biocompatible antibacterial product with higher activity than microliposomes. Soy lecithin as a clean product was used to prepare various nanoliposomes through sonication, high-pressure homogenizer, and normal homogenizer and also microliposomes through two methods of lipid film hydration and incubation methods. The nanoliposomes were formed under sonication with the size of 50 nm. The prepared liposomes were then loaded with poly(hexamethylene biguanide chloride) and the inclusion percentage was measured. The release profile of liposomes in buffer showed a release of 92% for poly(hexamethylene biguanide) during 24 h. The loaded liposomes were characterized with particle size analyzer, nuclear magnetic resonance, X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy. The antibacterial properties of different micro and nanoliposomes were investigated against a Gram-negative (Escherichia coli) and a Gram-positive (Staphylococcus aureus) bacteria. The poly(hexamethylene biguanide)–loaded nanoliposomes indicated higher antibacterial activities than microliposomes. Nanoliposomes have the potential to entrap lower poly(hexamethylene biguanide) dosages while retaining optimum therapeutic efficacy in the target site having lower cytotoxicity with lower side effects. The cytotoxicity of poly(hexamethylene biguanide) entrapped in liposomes was studied in human dermal fibroblasts and compared with free poly(hexamethylene biguanide) and blank liposomes. The maximum cytotoxicity was observed for free poly(hexamethylene biguanide) that is substantially decreased through loading within liposomes structure. Overall, the encapsulation of poly(hexamethylene biguanide) in liposomes improved the biocompatibility and safety of the product introducing a useful biocompatible antibacterial polymer for treatments of infectious diseases.
Silk fibroin protein, gastrodia elata, and tea tree oil are naturally derived and have been used throughout human history. This work develops an all-natural and highly porous foam-containing silk fibroin protein and above herbal extract, as a dressing for wound management. Scanning electron microscopic analyses and measurements of porosity by Archimedes method revealed a highly porous structure with porosity ranging from 40%–80%, depending on the preparation condition. In vitro, cytotoxicity test of a series of gastrodia elata–containing silk fibroin protein and tea tree oil–containing silk fibroin protein foam dressings on 3T3 fibroblast cells showed 90%–100% cell viability, which indicated that the produced all-natural dressings have no significant cytotoxicity toward skin cells. In another anti-inflammatory assay using the lipopolysaccharide-induced inflammatory Raw 264.7 macrophages model, the produced two dressings exhibited up to 70% and 90.1% of reduction in the formation of nitrite, in comparison with the untreated group. In vivo studies showed that all herbal extract–containing foam dressings accelerated wound recovery and achieved full closure of the wound within 21 days, and the histological analysis of regenerative skin tissues indicated that the produced foam dressings enhance the generation of thicker, denser, and more abundant collagen fibers in the dermis layer in comparison with the positive and negative control groups.
Poly(lactic acid)-based polymers are highly suitable for temporary biomedical applications, such as tissue support or drug delivery systems. Copolymers of different molecular weight based on poly(lactic acid) and poly(ethylene glycol) were prepared by polycondensation, catalysed by hydrochloric acid. A chain-extension reaction with
In this article, properties of thermosensitive chitosan hydrogels prepared with the use of chitosan chloride with β-glycerophosphate disodium salt pentahydrate enriched with calcium glycerophosphate are presented and compared with chitosan hydrogels with β-glycerophosphate disodium salt pentahydrate. The study is focused on the determination of hydrogel structure and biological testing of hydrogels with human osteoblasts line Saos-2. The structure of gels was visualized by scanning electron microscopy and was investigated by infrared spectroscopy. The crystallinity of gel structure was determined by X-ray diffraction analysis and thermal effects were determined using differential scanning calorimetry thermograms.
This study deals with the development of bioactive poly(ethylene terephthalate) surgical suture by adopting the immobilization route with bioactive nanogels and chlorhexidine. Carbon dioxide plasma was used for the generation of carboxyl functionality on poly(ethylene terephthalate) surface for the immobilization of the bioactive components. The nanosilver nanogel was prepared using polyethylene glycol which helps in the reduction of silver ions into nanosilver as well as the stabilization of nanoparticles. The particle size of the nanogels, as evaluated by high-resolution transmission electron microscopy, was observed to be in the range of 10–50 nm. Surface functionalization of poly(ethylene terephthalate) filament was observed by attenuated total reflectance measurements and mechanical studies were investigated by Instron. Elemental analysis and surface topography were carried out by energy dispersive X-ray and atomic force microscopy. The cumulative release of silver from the dressing was found to be 68% of the total loading after 72 h. Coated sutures have excellent antimicrobial activity against both Escherichia coli and Staphylococcus aureus. In vivo wound healing and histopathology studies were carried out over a period of 72 h for skin wounds created on Swiss albino mice. Fast healing was observed in nanogel-treated wounds without any inflammatory effects on the newly generated skin. These sutures offer improved healing along with excellent antimicrobial properties and appear to be promising material against surgical infection.
Tissue engineering aims at fabricating biological substitutes to improve, repair, and regenerate failing human tissues or organs. Designing a nanocomposite scaffolds with tailored properties that enhance the development of functional tissue can be an appropriate approach to achieve this purpose. In this study, the uniform and bead-free nanofibers of poly(-caprolactone) composited with different graphene oxide nanosheet contents (ranging from 0.5 to 2 wt%) were successfully fabricated through electrospinning process. A decrease in the average diameter of poly(-caprolactone) nanofibers was observed with the addition of graphene oxide nanosheets. Moreover, the nanocomposite scaffolds containing 2 wt% of graphene oxide nanosheets exhibited superior mechanical properties compared to that of pure poly(-caprolactone). Compared with pure poly(-caprolactone) scaffold, the degradation rate of poly(-caprolactone)-graphene oxide nanosheet nanofibers was enhanced, while the integrity of fibers was preserved. The presence of graphene oxide nanosheets in poly(-caprolactone) fibers promoted in vitro biomineralization, indicating bioactive features of the nanocomposite scaffolds. Compared to the pure one, nanocomposite fibers also showed better ability in protein adsorption. The in vitro cell culture studies showed that the addition of graphene oxide nanosheets did not diminish the biocompatibility of the electrospun poly(-caprolactone) nanofiber. Furthermore, the adhesion and proliferation of MG63 cells were increased. Altogether, the results demonstrated that electrospun poly(-caprolactone)-graphene oxide nanosheet nanofiber may be a suitable candidate for tissue engineering scaffold applications.
The success of gene therapy mainly depends on the carriers for effective gene delivery. A non-viral vector using a cationic block co-polyelectrolyte, PEI-PLA-PEG polyethyleneimine-poly(lactic acid)-poly(ethylene glycol)) was developed as a potential gene carrier. The cationic PEI-PLA-PEG showed less toxicity compared to PEI and formed a gene nanocomplex (termed polyplex) by interaction with plasmid DNA or small interference RNA. The polyplex showed smaller particle size and greater positive zeta potential by increasing the high polymer nitrogen/DNA phosphate ratio. The polyplex with a nitrogen/DNA phosphate ratio of 16 or 32 demonstrated higher gene transfection by fluorescence imaging, flow cytometry measurement, and β-galactosidase activity. In particular, the polyplex with therapeutic histone deacetylase small interference RNA at nitrogen/DNA phosphate ratio 16 showed the most favorable properties with definite tumor growth inhibition. The synthetic PEI-PLA-PEG also showed less toxicity and would, therefore, be a great potential gene carrier, particularly given that small interference RNA delivery does not increase the charge density of small interference RNA due to the formation of a stable complex through conjugation with PLA-PEG.
Due to the presence of tripeptide arginine–glycine–aspartic acid, gelatin is considered a very promising additive material to improve the cytocompatibility of alginate-based hydrogels. Two different strategies, physical blending and chemical crosslinking with gelatin, are used in this study to modify alginate hydrogel. As the intermolecular interactions between the polysaccharide and protein in the resulting physically blended and chemically crosslinked hydrogels are different, significant differences in the properties of these hydrogel types, regarding especially their surface topography, degradation kinetics, mechanical properties, and protein release behavior, are observed. Cellular behavior on both types of alginate–gelatin hydrogels is investigated using primary human dermal fibroblasts to elucidate the effects of the different structural, mechanical, and degradation properties of the produced hydrogels on fibroblast attachment and growth. The hydrogel that is chemically crosslinked with gelatin exhibits the highest degree of cytocompatibility regarding adhesion, proliferation, metabolic activity, and morphology of growing fibroblasts.
Thymol, a naturally occurring antioxidant and antimicrobial, is commonly researched for active packaging applications to deter food spoilage and bacterial growth. However, the high temperature necessary for processing often volatilizes the thymol, reducing its utility. To overcome this processing limitation, sugar-based poly(anhydride-esters) comprising thymol and compounds generally regarded as safe (succinic and tartaric acid) were successful prepared via mild solution polymerization methods. In vitro release studies demonstrated a sustained thymol release over 3 weeks at therapeutically relevant concentrations. Furthermore, the released thymol displayed antioxidant and antimicrobial activities as indicated by a 2,2-diphenyl-1-picrylhydrazyl radical scavenging and Kirby–Bauer disk diffusion assays, respectively. High-temperature melt blending with low-density polyethylene revealed that the chemical incorporation of thymol into a polymer backbone overcame volatility issues and maintained relevant bioactivity.
This study investigated some physicochemical properties of keratin extracted from Merino wool using five chemical extraction methods: alkali hydrolysis, sulfitolysis, reduction, oxidation, and extraction using ionic liquid. The ionic liquid method produced the highest protein yield (95%), followed by sulfitolysis method (89%), while the highest extraction yield was obtained with the reduction method (54%). The lowest yield was obtained with the oxidation method (6%). The oxidation extract contained higher molecular weight (>40 kDa) protein components, whereas the alkali hydrolysis extract contained protein material of <10 kDa. The sulfitolysis, reduction, and ionic liquid extracts contained various protein components between 3.5 and 60 kDa. Keratin obtained from various extraction methods had different yield, morphology, and physicochemical properties. None of the samples were toxic to L929 fibroblast cells up to a concentration of 2.5 mg/mL. Apart from the alkali hydrolysis extract, all other keratin extracts (reduction, sulfitolysis, ionic liquid, and oxidation) showed Fourier transform infrared adsorption peaks attributed to the sulfitolysis–oxidation stretching vibrations of cysteine-S-sulfonated residues, with the oxidation extract showing the highest content of cysteine-S-sulfonated residues. This study indicates that the properties of the keratin extract obtained vary depending on the extraction method used, which has implications for use in structural biomaterial applications.
The usage of hollow nerve conduits shows inferior recovery effect on the repair of peripheral nerve defects. In this study, a biocompatible and biodegradable pH-induced injectable chitosan–hyaluronic acid hydrogel for nerve growth factor encapsulation and sustained release was developed as the fillers in the lumen of hollow nerve conduit to reform its microenvironment for peripheral nerve regeneration. The physicochemical properties of hydrogel were characterized by gelation time, Fourier transform infrared spectroscopy, scanning electron microscopy, compressive modulus, porosity, swelling ratio, and in vitro degradation. The in vitro nerve growth factor release profiles and cell evaluation were also investigated. The results show that the structure of chitosan–hyaluronic acid hydrogel is composed of interconnected channels with a controllable pore diameter ranging from 20 to 100 µm. The hydrogel can be degraded more than 70% within 8 weeks in vitro and is available for nerve growth factor sustained release. The chitosan–hyaluronic acid/nerve growth factor hydrogel is non-toxic and suitable for adhesion and proliferation of nerve cells and capable of maintaining nerve growth factor activity. Therefore, it could be a promising intraluminal filler of nerve conduits for peripheral nerve regeneration in neural tissue engineering.
Three-dimensional cell culturing provides an appealing biomimetic platform to probe the biological effects of a designed extracellular matrix on the behavior of seeded neural stem or neural progenitor cells. This culturing model serves as an important tool to investigate functional regulators involved in proliferation and differentiation of neural progenitor cells. This study aims to reconstruct a polypeptide hydrogel matrix functionally integrated with cyclo-RGD motif [c(RGDfK)] for initial exploration of neural progenitor cell behavior in three-dimensional culture. Three types of hydrogel scaffolds including Type I collagen, RADA16 self-assembly peptide, and RADA16-c(RGDfK) self-assembly peptide hydrogel were employed to serve as the culturing extracellular matrix of neonatal rat spinal neural progenitor cells. The neural adhesion of functionalized self-assembly peptide hydrogel was acquired prior to its RADA16 counterpart with neural progenitor cell seeding tests. The biophysiological properties of self-assembly peptide hydrogel scaffolds were then detected by scanning electron microscopy and rheology measurements. The biological behavior of embedded neural progenitor cells including cell proliferation and differentiation in three-dimensional niche were analyzed by MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)] tests and immunocytochemistry fluorescence staining. The 1% (w/v) RADA16-c(RGDfK) hydrogel scaffold [R16-c(RGDfK)HS] demonstrated an elastic modulus(312 ± 5.7 Pa) compatible with central neural cells, which significantly facilitated the proliferation of embedded neural progenitor cells. Compared to collagen hydrogel, both RADA16 and RADA16-c(RGDfK) hydrogel scaffold improved the cellular proliferation and neuronal differentiation of neural progenitor cells in a three-dimensional culture model. In order to model neuronal regeneration, introduction of neurotrophin-3 in the differentiation environment significantly increased the neuronal differentiation in which the ratio of Tuj-1-positive cell number increased to 72.5% ± 4.7% in the c(RGDfK)-functionalized three-dimensional matrix environment at 7 days in culture. Collectively, the present R16-c(RGDfK)HS displays excellent central neural biocompatibility and emerges as a promising bioengineered extracellular matrix niche of neural stem or progenitor cells, building a solid foundation for the subsequent in vitro and in vivo studies including neural repair, regeneration, and development.
Effective local delivery methods for sustained and stable release of protein drugs are urgently needed. Biodegradable elastomers based on star-shaped polycyclic esters have received attention for their drug-loading and drug-release kinetics. However, the long degradation periods resulting from their strong lipophilicity greatly hinder their application. In this study, we synthesized new cross-linked elastomers based on methyl-acrylic-star-poly(-caprolactone-co-
Polypyrrole chitosan core shell nanoparticles were synthesized by in situ oxidation polymerization of pyrrole using FeCl3 in chitosan aqueous solution. 5-Phenyl-4H-1,2,4-triazole-3-thiol (I) was prepared and loaded into polypyrrole chitosan core shell nanoparticles at two different temperatures (25°C and 80°C). These core shell nanoparticle systems are insoluble in acidic medium and have good adsorption capacity. The release of loaded triazole was studied in different pH media (2, 7.4). The mechanism of triazole release was determined by applying zero-order release, first-order release, Higuchi model, Hixson–Crowell, and Korsmeyer–Peppas kinetics equations. The antibacterial activities against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis were evaluated. The potential cytotoxicity against Ehrlich ascites carcinoma cells and liver (HEPG2) cell line in vitro was tested.
The development of different chitosan derivatives for medical applications has increased recently. Among these chitosan derivatives, quaternized chitosan was designed to improve the solubility of chitosan in biological fluids for oral drug delivery while retaining the cationic character for mucoadhesion. However, the biocompatibility of quaternized chitosan on the human intestine is unknown. In this study, we aimed to examine the potential biological effects of quaternized chitosan on the intestinal barrier, in terms of cell proliferation and cell differentiation, using the Caco-2 cell line as an in vitro model. The lower the degree of substitution of quaternized chitosan, the lower the cytotoxic and anti-proliferative effect on Caco-2 cells. In addition, the anti-proliferative effect of quaternized chitosan might induce a cell cycle disturbance and differentiation delay. Long-term continuous exposure (9 days) to quaternized chitosan caused a delay in differentiation of the Caco-2 cells even at non-cytotoxic quaternized chitosan doses (0.005% (w/v)), as shown by the low level of alkaline phosphatase in the quaternized chitosan–treated group compared to the control cells. In contrast, short-term discontinuous exposure to quaternized chitosan (0.005% (w/v) for 4 h/day over 9 days) that more realistically mimics the daily intestinal exposure did not inhibit the intestinal differentiation of Caco-2 cells. Thus, the use of a low degree of substitution and a low concentration of quaternized chitosan resulted in a good biocompatibility to the intestinal barrier supporting the potential usefulness of quaternized chitosan in the application of an oral drug delivery system.
In this study, poly(methacrylic acid-glycine)-grafted agar-based hydrogels with optimized process parameters were synthesized via a two-step green-radiation induced grafting process using microwave heating. Poly(methacrylic acid) chains were graft copolymerized onto an agar backbone using ammonium persulfate as a free radical initiator and N,N'-methylene-bis-acrylamide as a cross-linking means using microwave heating. The influence of different reaction parameters was investigated on the percentage swelling behavior of the cross-linked hydrogel networks. The prepared hydrogel networks with optimum percentage swelling were characterized by Fourier transform infrared spectroscopy, time-of-flight secondary ion mass spectrometry, scanning electron microscopy, X-ray diffraction, and thermogravimetric analysis, using agar as a reference. The anti-bacterial activities of the prepared hydrogels against Gram-positive Staphylococcus aureus bacteria and Gram-negative Escherichia coli bacteria were investigated. Staphylococcus aureus was found to be more susceptible to the compounds compared to Escherichia coli. These results indicate that the prepared hydrogels have the potential to be applied as anti-bacterial agents.
Gamma irradiation was used to fabricate crosslinked poly(vinyl alcohol)/sericin hydrogels with different sericin concentrations, and the physicochemical and biological properties of the gamma-irradiated poly(vinyl alcohol)/sericin hydrogels were characterized. Following gamma irradiation, the hydrogels had a high gel fraction (80%–95%), implying a high degree of crosslinking. Fourier transform infrared spectra confirmed the crosslinking bonds within the hydrogels, as seen by the characteristic shift in the peak. Furthermore, a low tensile modulus together with a high elongation percentage indicated that the hydrogels were easy to handle. We also showed that all hydrogels released sericin simultaneously. The poly(vinyl alcohol)/sericin hydrogels with high sericin content released more sericin, possibly due to less crosslinking of the hydrogels. When L929 cells were cultured with the hydrogel extracts, the cells were viable and could proliferate, particularly for the cells cultured with the hydrogels containing a high sericin content, which released more sericin. Migration assays also demonstrated that the cells migrated toward the medium extract of hydrogels containing high sericin. We suggest that sterile gamma-irradiated poly(vinyl alcohol)/sericin hydrogels could be used as a wound dressing for the treatment of dry and low-exudate wounds.
Superabsorbent polymer hydrogels with antibacterial activity were prepared by an ion exchange reaction as a feasible approach to induce high saline absorption without gel blockage. Hydroethanolic solutions of cetyltrimethylammonium bromide were used to modify surface particles of cross-linked sodium acrylate-co-acrylic acid copolymers which already synthesized under defined conditions. Fourier transform infrared spectroscopy was employed to study the structural characteristic of the finished products. The influence of cetyltrimethylammonium bromide on free (in water) and loaded (in saline) swelling capacity as well as antibacterial activity of superabsorbent polymer hydrogels against Staphylococcus aureus was investigated. Modified samples displayed an improved free and loaded swelling in water and saline, as well as no gel-blocking. These improvements were found to be affected by the reaction time, cetyltrimethylammonium bromide concentration, and water percentage in the solvent mixture. The results from energy dispersive X-ray analysis showed that cetyltrimethylammonium bromide was distributed uniformly in the superabsorbent polymer hydrogel particle surface. Moreover, the modified superabsorbent polymer hydrogels showed high antibacterial activity against S. aureus. Both bacteriostatic and bactericide effects were observed depending on the reaction conditions. Overall, several improvements were concurrently achieved via a single cost-effective post-treatment on the superabsorbent polymer hydrogel particles. Therefore, the results can effectively be used in designing larger scale production of antibacterial superabsorbent polymer hydrogels with desirable swelling properties in hygiene applications.
Fabrication of a pre-vascularized tissue in vitro is an extensive research activity. The idea behind this approach is that a network of newly formed micro-vessels may be engineered in vitro by the seeding of a scaffold with endothelial cells. To this aim, understanding the effect of physicochemical properties of the scaffolding material and the method of cell seeding, in regulating endothelial cells’ behavior in the in vitro constructs, is an emerging requirement. In this study, the effect of interfacial self-assembly and contact guidance for the endothelial cell behavior and angiogenic network formation have been studied. This has been done by the fabrication of in vitro three-dimensional tissue constructs, using multilayer surface-modified polymer fibers, and two different methods of cell seeding by human umbilical vein endothelial cells. In the first method, human umbilical vein endothelial cells, fibers, and fibrin gel matrix were combined simultaneously. In the second method, the human umbilical vein endothelial cells and fibers, having various surface coatings, were sandwiched between two layers of fibrin gel matrix with or without fibroblast cell monolayer over the fibrin gel. In the optimal conditions, the effect of fibers in conjunction with the interfacial self-assembly enhanced a tube-like and interconnected network structure formation. This design could therefore have a major impact in the generation of the pre-vascularized tissue-engineered constructs.
The scaffold component is a major barrier to the development of a clinically useful small-diameter tissue-engineered vascular graft. Scaffold requirements include matching the mechanical and structural properties with those of native vessels and optimizing the microenvironment for cell integration, adhesion, and growth. Trilayered sulfated silk fibroin graft was developed to mimic native tissue structure and function. Physical properties and cell studies were assessed to evaluate the viability of their usage in small-diameter tissue-engineered vascular grafts. Compared with previously fabricated silk fibroin vascular grafts, these trilayered grafts provided comparable water permeability, tensile strength, burst pressure, as well as suture retention strength, to saphenous veins for vascular grafts. In addition, the in vitro results showed good cytocompatibility of the trilayered grafts. These physical and cellular outcomes indicate potential utility of these trilayered sulfated silk fibroin grafts for small-diameter vascular grafts.
In this work, pH-sensitive amphiphilic macromolecules are designed to possess good biocompatibility and drug loading while employing an acid-sensitive linkage to trigger drug release at tumor tissues. Specifically, two pH-sensitive amphiphilic macromolecules were synthesized with a hydrazone linkage between the hydrophobic and hydrophilic segments. The chemical structure, molecular weight, critical micelle concentration, micelle size, and pH-triggered cleavage of the amphiphilic macromolecules were characterized via matrix-assisted laser desorption/ionization time-of-flight, nuclear magnetic resonance, and dynamic light scattering techniques. Drug loading and release as well as cytotoxicity studies were performed using doxorubicin. Hydrodynamic diameters of the micelles formed with pH-sensitive amphiphilic macromolecules were within an optimal range for cellular uptake. The critical micelle concentration values were 10–8–10–6 M, indicating micellar stability upon dilution. The degradation products of the amphiphilic macromolecules after acidic incubation were identified using mass spectrometry, nuclear magnetic resonance, and dynamic light scattering methods. A pH-dependent release profile of the doxorubicin-encapsulated amphiphilic macromolecules was observed. Cytotoxicity studies against two cancer cell lines, MDA-MB-231 human breast cancer cells and A549 lung cancer cells, showed that doxorubicin encapsulated in pH-sensitive amphiphilic macromolecules decreased cell viability more efficiently than free doxorubicin, possibly due to the toxicity of the amphiphilic macromolecule degradation products. Resulting from enhanced release at acidic pH due to hydrolysis of the hydrazone linkage, pH-sensitive amphiphilic macromolecules also had improved efficacy toward cancer cells compared to other carriers (e.g. Pluronics®). These findings indicate that pH-sensitive amphiphilic macromolecules can potentially be applied as anticancer drug delivery vehicles to achieve controlled release and improved therapeutic effects.
Thermosensitive poly(N-isopropylacrylamide) is widely used in various biomedical applications including drug delivery systems, gene delivery systems, switching devices, sensors, and diagnostic assays. To promote these clinical applications, it is essential to have a comprehensive understanding of the biosafety of poly(N-isopropylacrylamide) and the interaction of poly(N-isopropylacrylamide) with different cell lines, which has little research until now. In this work, we evaluated the biocompatibility of poly(N-isopropylacrylamide) including cell viability, nitric oxide production, and apoptosis of macrophages RAW264.7, human bronchial epithelial cells, A549, and human umbilical vein endothelial cells in the presence of poly(N-isopropylacrylamide). We have also examined the cellular uptake mechanisms of poly(N-isopropylacrylamide) using endocytic inhibitors and insighted into the intracellular co-localization of poly(N-isopropylacrylamide) using confocal laser scanning microscope. The results showed that poly(N-isopropylacrylamide) had good biocompatibility and could be internalized by these cells. It is macropinocytosis that poly(N-isopropylacrylamide) could be internalized in RAW264.7 cells and caveolae-mediated endocytosis in human bronchial epithelial cells, A549, and human umbilical vein endothelial cells. In addition, we also evidenced that intracellular poly(N-isopropylacrylamide) was co-localized with lysosome. The study provided important information for the development and clinical applications of poly(N-isopropylacrylamide) in the biomedical field.
In this study, sodium montmorillonite (Na-MMT) was successfully modified by using n-hexadecyl trimethyl ammonium bromide (CTAB) via cationic exchange to obtain an organophilic-montmorillonite (CTAB-MMT). The Na-MMT, CTAB-MMT, and a commercial montmorillonite, that is, Cloisite15A were incorporated into gellan gum (GG) hydrogel and their mechanical, physical, thermal properties, biocompatibility, and antibacterial activities were investigated. The mechanical performance results show that the GG hydrogels containing Cloisite15A required smallest volume to achieve optimum compression stress, modulus, and compression strain at 5% (w/w) compared to both Na-MMT and CTAB-MMT at 10% (w/w). Swelling ratio of GG hydrogels increased upon addition of MMT, and water vapor transmission rate (WVTR) values of all hydrogels were in the range of 1106–1890 g m–2 d–1, which were comparable to WVTR values of commercial wound dressings. Thermal behavior shows that the inclusion of Cloisite15A in GG hydrogel improved the thermal stability than its counterparts. Cell studies exhibit that the GG incorporated with Na-MMT is non-cytotoxic to human skin fibroblast cells (CRL2522), and in contrast, the GG hydrogels incorporated CTAB-MMT and Cloisite15A revealed that the cells were dying and the cell growth depleted after being cultured for 72 h. Qualitative antibacterial study revealed that GG hydrogel containing CTAB-MMT only in the sample exhibits inhibition against the Gram-positive bacteria, that is, Staphylococcus aureus and Bacillus cereus, while there was no inhibition exhibited against Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae).
In order to replace damaged or lost bone in the human body, it is necessary to produce ‘spare body parts’ which are dependent on the use of biomaterial and stem cells and are referred to as ‘tissue engineering’. Surface modification and stem cell interaction of orthopaedic implants offer a promising approach and are investigated here specifically to improve osseointegration of the biomaterial. Osseointegration of titanium implants used in orthopaedic surgery requires that osseo-progenitor cells attach and adhere to the surface, proliferate, then differentiate into osteoblasts and, finally, produce a mineralised matrix. The surface modification of titanium with anionic polymer combined with coating of platelet-rich plasma is provided to create a favourable environment to promote early and strong fixation of implants. The ability of progenitor cells to attach to the surface during early stages is important in the development of new tissue structures; therefore, we developed in our laboratory a strategy involving the grafting of titanium implants with a polymer of sodium styrene sulphonate (poly(sodium styrene sulphonate)) and a biofilm coating of platelet-rich plasma which enables human mesenchymal stem cell interactions. The resulting biomaterial, titanium-poly(sodium styrene sulphonate) and coating of platelet-rich plasma, Ti-poly(sodium styrene sulphonate)–platelet-rich plasma was developed in order to further improve the biomaterial. In this work, we studied and characterised the ‘in vitro’ response of human mesenchymal stem cells to titanium biomaterial grafted with poly(sodium styrene sulphonate) bioactive polymer and coated with platelet-rich plasma proteins (Ti-poly(sodium styrene sulphonate)–platelet-rich plasma). This study shows an increased cell proliferation with Ti-poly(sodium styrene sulphonate)–platelet-rich plasma compared to foetal calf serum and an enhancement of the Ti-poly(sodium styrene sulphonate)–platelet-rich plasma effects on osteoblast differentiation. The results suggest that Ti-poly(sodium styrene sulphonate)–platelet-rich plasma would be a suitable scaffold for bone tissue engineering.
Developing tissue-engineered constructs for clinical use must satisfy the fundamental biologic parameters of biocompatibility, cell adhesiveness, and biodegradability. Physical entrapment of bioactive agents into synthetic polymers, as three-dimensional scaffolds, holds great promise for cell culture applications. Here, in an attempt to elucidate the effects of physical interlocking of natural and synthetic gel networks on cell responses within three-dimensional microenvironments, gelatin (of different concentrations) was physically incorporated into macroporous polyethylene glycol (PEG) hydrogels to fabricate PEG-GEL1 (10:1, PEG:gelatin) and PEG-GEL5 (10:5, PEG:gelatin). The effect of the physically entrapped gelatin on primary chondrocytes was investigated in relation to cell distribution, morphology and viability, proliferation, gene expression, and extracellular matrix accumulation in vitro. Our findings have shown successful incorporation of two different concentrations of gelatin into polyethylene glycol macroporous hydrogels through physical mixing. These physical blends not only enhanced chondrocyte adhesion and proliferation but also boosted gene expression of collagen II and aggrecan after 14 days in culture. Although results demonstrated that gelatin levels dropped sharply in PEG-GEL1 and PEG-GEL5 in the first 7 days, however evidently, after days 14 and 21 gelatin levels in both groups remained substantially unchanged and in turn enhanced glycosaminoglycan formation in vitro. Thus, the modification of polyethylene-glycol-based scaffolds with physically entrapped gelatin may be sufficient for dictating three-dimensional microenvironments for chondrocyte cultures.
An injectable hydrogel was obtained from the high methyl-esterified plum Prunus domestica L. (PD) pectin and calcium ions (Ca2+). PD hydrogel showed a weak gel-like behavior and could be squeezed out of the syringe with an injection force of ca. 9 N. PD hydrogel was not suitable for the NIH/3T3 fibroblast cell adhesion in vitro. The live/dead fluorescence and MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide) assays indicated that the PD hydrogel had a low cytotoxicity in relation to both the adhered and gel surrounding fibroblasts. PD hydrogel was found to inhibit adhesion formation in the sidewall defect-cecum abrasion rat model. In the control group, the occurrence of adhesion of the cecum to the peritoneal wall was found in seven of the total seven rats operated. Only four of the seven animals that were treated with the PD hydrogel were noted to have any adhesions. These adhesions were of a minimum grade of 1, 2, and 3 and were represented by a thin film that could be easily broken. The protective effect of PD hydrogel was found to be comparable with that of hyaluronic acid hydrogel used as a positive control. PD hydrogel appeared to possess enhanced in vivo residence stability on the injury sites compared to hyaluronic acid hydrogel as measured by staining of healing tissue with periodic acid-Schiff reagent. The data obtained offered the prospect for the development of the pectin-based gels as new barrier materials for surgery.
Efficient immobilization of the antibody to the substrate is of crucial importance in the development of anti-CD34-based endothelial progenitor cells capturing cardiovascular devices. This should go along with precise control of the antibody orientation by appropriate immobilization technology for retaining antibody activity, like in immunosensors. Recently, great attention was paid to immobilization of anti-CD34 antibody onto substrates by covalent binding, but at random orientation. Here, to investigate the biological effect of antibody orientation, we have prepared two kinds of anti-CD34 antibody coated surfaces, with random immobilization and oriented immobilization. The immunological binding activity (IBA) of the antibody at oriented immobilization was 3.48 times higher than at random immobilization, indicating that the two different surfaces were successfully prepared. The endothelial progenitor cell-capturing capability of oriented antibody-immobilized surface was 1.35 and 1.64 times higher than for the random immobilized surface after seeding for 2 and 12 h under flow condition, respectively. The endothelial progenitor cell-capturing efficiency per antibody by oriented immobilization was 5.16 and 6.26 times higher than for the random after seeding for 2 and 12 h under flow condition, respectively. In addition, the oriented antibody-immobilized surface possessed better blood-compatibility. These results clearly revealed the significance of antibody orientation which could retain its biological effect and may revolutionize the antibody-immobilization protocols used in cardiovascular and other blood-contacting biomedical devices.
The objective of this work was the preparation and evaluation of a bioresorbable bilayered system for application in the treatment of dermal lesions. The system was based on a polyesterurethane as the external layer and a gelatin membrane as the internal layer. The polyesterurethane was synthesized from poly(-caprolactone), polyethylene glycol of 1 or 10 kDa as a hydrophilic component or Pluronic F127 as an amphiphilic component and
Polyhydroxybutyrate-co-hydroxyvalerate has been identified as a useful polymer for biomedical application due to its biocompatibility and processability. Polyhydroxybutyrate-co-hydroxyvalerate nanoparticles loaded with quercetin, an antimicrobial, anti-inflammatory, and antiviral polyphenol with limited solubility, were obtained using a high-speed double-emulsion technique. The nanoparticle size and the dissolution of quercetin were controlled simultaneously through high-speed stirring (15,000 r/min) in the emulsification process. The size range of quercetin-loaded polyhydroxybutyrate-co-hydroxyvalerate nanoparticles was between 250 and 650 nm. Spherical shape with no aggregation of nanoparticles was confirmed by electron microscopy. Loaded nanoparticles showed less thermal degradation than unloaded nanoparticles. An encapsulation efficiency of 51% was found. Most of the quercetin was released from the nanoparticles within the first 5 h of water immersion. A biocompatibility analysis of the nanoparticles showed no cytotoxicity and no significant difference between loaded and unloaded nanoparticles.
The aim of this study was to test alginate beads and silk fibroin non-woven mats as stromal vascular fraction delivery systems to support cell implantation for tissue repair and regeneration, through trophic and immunomodulant paracrine signaling. Furthermore, in vivo scaffold biocompatibility was histologically analyzed in a murine model at different time endpoints, with particular focus on construct-induced vascularization and neoangiogenesis. The fibroin mat induced a typical foreign body reaction, recruiting macrophages and giant cells and concurrently promoted neovascularization of the implanted construct. Conversely, alginate beads triggered a more circumscribed, chronic inflammatory reaction, which decreased over time. The combined in vivo implantation of alginate beads and fibroin mat with stromal vascular fraction promoted vascularization and integration of scaffolds into the surrounding subcutaneous environment. The new blood vessel ingrowth should, hopefully, support engineered cell viability and functionality, as well as the transport of soluble bioactive molecules. Due to their neovascularization properties, stromal vascular fraction administration, using alginate or fibroin scaffolds, is a new, promising, cost-effective tissue engineering approach.
Currently, the main focus on tissue engineering strategies is to mimic the extracellular matrix of the related tissues. Many studies accomplished to build tissue scaffolds to act as the natural surroundings of the specific interest, which can be established to behave like either healthy or unhealthy tissues. The latter one of these conditions is a quite new approach and crucial for the design of three-dimensional in vitro disease models. This study investigates the potential of a composite scaffold consisting hydroxyapatite-integrated fluorenyl-9-methoxycarbonyl diphenylalanine hydrogels by focusing on the optimization of this hybrid scaffold for the development of an in vitro model of degenerative cartilage. Cell growth, chondrocyte proliferation, extracellular matrix production, hypertrophy marker monitoring, scaffold mechanical properties, and morphological analysis were evaluated. Fluorenyl-9-methoxycarbonyl diphenylalanine dipeptides were dissolved in null cell culture media and pH decreased sequentially to compel peptides to self-organize into fibrous hydrogel scaffolds. Nano-hydroxyapatite crystals were incorporated into fluorenyl-9-methoxycarbonyl diphenylalanine hydrogels during the gelation to investigate the effect on chondrocytes. It is observed that hydroxyapatite incorporation into peptide hydrogels significantly increased the alkaline phosphatase activity and assymetrical cell divisions, which is appraised as an outcome of chondrocyte hypertrophy. It is concluded that chondrocytes develop a hypertrophic potential when they are cultured in a media with nano-hydroxyapatites in a three-dimensional cell culture matrix mimicking the extracellular matrix conditions of degenerative cartilage.
The application of additive manufacturing principles to hydrogel processing represents a powerful route to develop porous three-dimensional constructs with a variety of potential biomedical applications, such as scaffolds for tissue engineering and three-dimensional in vitro tissue models. The aim of this study was to develop novel porous hydrogels based on a microstructured polyelectrolyte complex between chitosan and poly(-glutamic acid) by applying a computer-aided wet-spinning technique. The developed fabrication process was used to build up three-dimensional porous hydrogels by collecting microstructured layers made of chitosan/poly(-glutamic acid) on top of the other. Microstructured polyelectrolyte complex hydrogels were characterized and compared to chitosan/poly(-glutamic acid) porous hydrogels with similar composition prepared by conventional freeze-drying technique. Fourier transform infrared analysis confirmed the formation of an electrostatic interaction between the two processed polymers in all the developed chitosan/poly(-glutamic acid) hydrogels. The composition of the porous constructs as well as the employed processing techniques had a significant influence on the resulting swelling, thermal, and mechanical properties. In particular, the combination of the ionic interaction between chitosan/poly(-glutamic acid) and the defined internal microarchitecture of microstructured polyelectrolyte complex hydrogels provided a significant improvement of the compressive mechanical properties. Preliminary in vitro biological investigations revealed that microstructured polyelectrolyte complex hydrogels were suitable for the adhesion and proliferation of Balb/3T3 clone A31 mouse embryo fibroblasts. The encouraging results in terms of cytocompatibility and stability of the microstructure in aqueous solutions due to the ionic crosslinking suggest the investigation of the developed microstructured polyelectrolyte complex hydrogels as suitable scaffolds for three-dimensional cells’ culture.
Polymeric nanogels have been sophisticatedly designed promising a new generation of vaccine delivery/adjuvant systems capable of boosting immune response, a strategic priority in vaccine design. Here, nanogels made of mannan or dextrin were evaluated for their potential as carriers/adjuvants in vaccine formulations. Since lymph nodes are preferential target organs for vaccine delivery systems, nanogels were biotin-labeled, injected in the footpad of rats, and their presence in draining lymph nodes was assessed by immunofluorescence. Nanogels were detected in the popliteal and inguinal lymph nodes by 24 h upon subcutaneous administration, indicating entrapment in lymphatic organs. Moreover, the model antigen ovalbumin was physically encapsulated within nanogels and physicochemically characterized concerning size, zeta potential, ovalbumin loading, and entrapment efficiency. The immunogenicity of these formulations was assessed in mice intradermally immunized with ovalbumin–mannan or ovalbumin–dextrin by determining ovalbumin-specific antibody serum titers. Intradermal vaccination using ovalbumin–mannan elicited a humoral immune response in which ovalbumin-specific IgG1 levels were significantly higher than those obtained with ovalbumin alone, indicating a TH2-type response. In contrast, dextrin nanogel did not show adjuvant potential. Altogether, these results indicate that mannan nanogel is a material that should be explored as a future antigen delivery system.
Collagen–carboxymethyl cellulose–tricalcium phosphate cryogels were prepared for diverse biomedical applications. Further chemical and structural characterizations were performed by Fourier transform infrared spectra, thermogravimetric analysis, X-ray crystallography, and scanning electron microscopy. The mechanical properties were tested by unconfined compression test. Moreover, hemocompatibility of the cryogels was also evaluated by basic biochemical blood testing. Chemical and structural analysis results demonstrate the achievement of the cross-linking without any major alteration in collagen and carboxymethyl cellulose with a thermally and structurally stable blend formation. Scanning electron micrographs demonstrate the multi-lamellar formation with macro- and micro-pore compositions which can correlate with water uptake results of the cryogels. Hemocompatibility evaluations exhibited that the cryogels are non-toxic and blood-compatible. The overall results including mechanical testing of these tricalcium phosphate–consisting collagen/carboxymethyl cellulose cryogels may have potential use as a material for hard tissue regeneration.
Here, we have investigated in vitro antimicrobial efficacy of a quaternized cationic polymer, poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylamino)propyl]urea] (polyquaternium-2), against gram-positive as well as gram-negative bacteria along with several multi-drug-resistant bacterial strains. The antimicrobial efficacy of this polymer was first tested against some clinical pathogens followed by microorganisms isolated from acne lesions. Interestingly, polyquaternium-2 exhibited significant antimicrobial activity against methicillin-resistant Staphylococcus aureus, for which very limited drugs are available. Most importantly, the polymer displayed low haemolytic activity and non-toxic behaviour against mammalian cells. The results showed the promising potential of the projected polymer to be utilized as an antibacterial agent for various biomedical applications.
Formation of scar tissue may be reduced or prevented if wounds are locally treated with a combination of molecules tuned to the different healing phases, guiding tissue regeneration along a scar free path. To this end, drug delivery devices made of cellulose acetate phthalate and Pluronic F-127 were loaded with either quercetin or pirfenidone and plasticized with either triethyl citrate or tributyl citrate. Quercetin inhibits oxidative stress, and pirfenidone has been shown to reduce production of pro-inflammatory and fibrogenic molecules. The combined effects of drug and plasticizer on erosion, release, and mechanical properties of the drug delivery films were investigated. Triethyl citrate-plasticized films containing quercetin released drug at a slower rate than did tributyl citrate films. Pirfenidone-loaded films released drug at a faster rate than erosion occurred for both types of plasticizers. Higher plasticizer contents of both triethyl citrate and tributyl citrate increased the elongation and decreased the elastic modulus. In contrast, increased pirfenidone loading in both triethyl citrate and tributyl citrate films resulted in a significantly higher modulus, an antiplasticizer effect. Adding pirfenidone significantly decreased elongation for all film types, but quercetin-loaded samples had significantly greater elongation with increasing drug content. Films containing quercetin elongated more than did pirfenidone-loaded films. Quercetin is over 1.5 times larger than pirfenidone, has water solubility over 12 times lower, and has 6 times more bonding sites than pirfenidone. These differences affected how the two drugs interacted with cellulose acetate phthalate and Pluronic F-127 and thereby determined polymer properties. Drug release, erosion, and mechanical properties of association polymer films can be tailored by the characteristics of the drugs and plasticizers included in the system.
Biological interaction between cells and scaffolds is mediated through events at surfaces. Proteins present in the culture medium adsorb on substrates, generating a protein adlayer that triggers further downstream events governing cell adhesion. Polymer blends often combine the properties of the individual components, for example, can provide mechanical as well as surface properties in one fibre. Therefore, mixtures of synthetic polycaprolactone and gelatin as a denatured form of collagen were electrospun at selected conditions and polymer weight ratios. Fibre morphologies and chemical properties of the surfaces were analysed. These scaffolds were seeded with human mesenchymal stromal cells and their viability was studied. Gelatin addition to polycaprolactone leads to a reduction in fibre diameter. A linear increase in gelatin at the fibre surface was observed in function of the weighed polymers, except for polycaprolactone/gelatin fibres incorporating equal weight ratios. Thereby, a depletion of gelatin at the fibre surface is stated for equally mixed polymers. The depletion of gelatin at the fibre surface is most probably due to hydrophobic interactions between hydrophobic segments of polycaprolactone and gelatin, affecting the spinning mechanism and thus fibre structure. Furthermore, polycaprolactone/gelatin blends show enhanced wettability properties compared to pure gelatin, at least partly due to molecular segregation. Results of in vitro studies reveal an increase in cellular viability and proliferation for cells cultivated on nanofibres containing gelatin, caused by the cell-attractive surface composition as well as the hydrophilic nature of the scaffolds. Contact guidance of cells seeded on parallelised fibres is observed, and DNA tests show evidently enhanced cell numbers on nanofibres containing 20 wt% of gelatin.
A simple, rapid, and economical method to fabricate micropatterned thermoresponsive chitosan membranes was developed. Porous polystyrene films were prepared by liquid-induced phase separation. The size of pores on polystyrene films could be regulated by adjusting the composition of coagulation bath and changing the solvent evaporation rate. Subsequently, chitosan-based thermoresponsive membranes with island protrusions were fabricated using porous polystyrene films as templates. The effects of the micropatterns on the behaviors of mouse fibroblast L929 were investigated. The presence of micropatterns altered the cell cycle distribution and enhanced the gene expression of cyclin D1 and integrin β1. The micro-convex surface could promote the adhesion and proliferation of L929 cells. These results provided valuable guidance to design appropriate topographic surfaces for tissue engineering applications.
In this study, we developed a new approach in the preparation of chitosan-based polymeric nitric oxides. Chitosan film (unreacted chitosan) reacted with glutaraldehyde to introduce aldehyde groups onto the material surface (glutaraldehyde-treated chitosan). Glutaraldehyde-treated chitosan reacted with a small-molecule nitric oxide donor, 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene, to covalently immobilize nitric oxide–releasing moieties onto the polymer (chitosan-based polymeric nitric oxide). Chitosan-based polymeric nitric oxide showed sustained release of nitric oxide. The activation energies and rate constants of nitric oxide release were determined. The released nitric oxide provided potent antimicrobial effects against Gram-positive and Gram-negative bacteria living in biofilms, and the chitosan-based polymeric nitric oxide film showed added/synergistic effects with common antibiotics. At 4°C, the chitosan-based polymeric nitric oxide could be stored for more than 1 month, without significantly losing nitric oxide–releasing capabilities. Furthermore, chitosan-based polymeric nitric oxide showed excellent biocompatibility with mammalian cells, pointing to great potentials of the new materials for a wide range of biomedical applications.
In the field of drug delivery, many different carrier systems have been described to date, including nanoparticles, micelles, liposomes, and water-soluble polymer conjugates, where the active compound could be either incorporated non-covalently or linked to its carrier by a degradable chemical bond. In this study, we synthesized, characterized, and investigated the in vivo fate of N-(2-hydroxypropyl)methacrylamide-based polymer conjugates with a drug model bound via a disulfide bond, which is frequently cited in the literature as being completely stable in the bloodstream but readily cleaved after cell internalization. The concept was based on a "dual" labeling of N-(2-hydroxypropyl)methacrylamide copolymers with two different fluorescent dyes, where the first dye was linked via a disulfide bond, thus representing a model drug, while the second dye was attached as an amide and served as a label for the polymer carrier. Two conjugates, differing in their molecular weights (30 and 104 kDa), were examined using a multispectral optical imaging technique in athymic nude mice inoculated with HT-29 and DLD-1 human colon carcinoma xenografts. Additionally, necropsied organs and tumors were examined ex vivo to obtain more detailed information about polymer and model drug biodistribution. In vivo results confirmed preferential tumor accumulation for both conjugates. Moreover, different fluorescence patterns for the polymer and drug model were observed in both mice and necropsied tumors, indicating tumor-specific "drug" release.
Circulating tumor cells have received attention for their role in cancer diagnosis and the decision on which chemotherapeutic course to take. For these purposes, the isolation of circulating tumor cells has been important. Previously, we reported that non-blood cells can adhere on blood-compatible polymer substrates, such as poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate). In this study, we examined whether blood-compatible poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) allow the adhesion and growth of A549 lung cancer cells for isolating circulating tumor cells by adhesion-mediated manner to diagnose metastatic cancer and to decide on the chemotherapeutic course. A549 cells can adhere on poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates via an integrin-dependent mechanism after 1 h of incubation, suggesting that blood-compatible poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates possess the ability to capture circulating tumor cells selectively from peripheral blood. After 1 day of culture, A549 cells started to spread on poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates. A549 can also grow on poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates. Additionally, the chemoresistance of A549 cells against 5-fluorouracil on poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates was similar to that on the conventional cell culture substrate, tissue culture polystyrene. These results indicate that circulating tumor cells can be cultured on poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates after they are isolated from peripheral blood, and poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates can be used as circulating tumor cell culture substrates for screening anti-cancer drugs. Therefore, poly(2-methoxyethyl acrylate) and poly(tetrahydrofurfuryl acrylate) substrates might be able to be applied to the development of a new device for a circulating tumor cell–based diagnosis of metastatic cancer and a personalized medicine approach regarding the decision of which chemotherapeutic course should be taken.
The objective was to assess the immunoequivalence and protective efficacy of the novel, relatively safe dendrigraft poly-
Novel temperature-sensitive micelles, possessing a core-shell structure, were successfully fabricated and evaluated as possible systems for targeting anticancer drugs to solid tumors. The amphiphilic block copolymer poly(N-isopropylacrylamide-co-acrylamide)-b-poly(n-butyl methacrylate) was used to achieve a stimuli-responsive on/off release and spatial specificity. The anticancer drug methotrexate, which is poorly water soluble, was used as the model. Fourier transform–infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, gel-permeation chromatography, and critical micelle concentration were used to evaluate the successful synthesis of block copolymers with a lower critical solution temperature ~40°C. Based on transmission electron microscope images, the micelles are spherical particles with narrow size distribution. The thermally triggered release of methotrexate was observed in vitro. Quartz crystal microbalance with dissipation was used to investigate the interactions of the polymeric micelles with bovine serum albumin, to illustrate protein adsorption and cell attachment. Cytotoxicity studies were conducted on Lewis lung carcinoma cells, and the anticancer activity of methotrexate-loaded micelles was significantly enhanced in combination with hyperthermia. The thermo-sensitive characteristics of the micelles make them applicable as smart drug delivery systems, when combined with localized hyperthermia.
Nanoscale structures with large surface area-to-volume ratios are used as biomaterial scaffolds for vascular grafts, wound dressings, and air purifying filters. Using electrospinning, nanofibers containing an antibacterial agent, cetyltrimethylammonium bromide, were prepared for wound healing application. Polyvinylpyrrolidone, known as a biocompatible additive in food and drug industries, has been used as fiber processing agent with the organic active ingredient, cetyltrimethylammonium bromide. A series of samples with different polyvinylpyrrolidone/ cetyltrimethylammonium bromide ratios were successfully prepared by this method. The morphology and electroactive characteristics of nanofibers were investigated using scanning electron microscopy, atomic force microscopy, and electrochemical impedance spectroscopy. Fiber diameters and charge transfer resistances were found to decrease with salt content, while the double-layer capacitance increased with no apparent effect on the specific capacitance providing favorable conditions for the fabrication of biomaterials. In addition, the quaternary ammonium compound (cetyltrimethylammonium bromide) with a minimum ratio of 2.5 wt% showed reduction in bacterial activity of Klebsiella pneumonia, Staphylococcus aureus, and Escherichia coli.
A new class of pH-responsive multivalent host–guest interactions to manipulate polypeptide-based nano-vehicles was developed. Poly(
Biomaterials are extensively used in bone defect recovery caused by bone diseases. Multi-walled carbon nanotubes have been reported to reinforce synthetic polymeric materials. The aim of the study is to test poly(3-hydroxybutyrate-co-3-hydroxyvalerate) loaded with different amounts of multi-walled carbon nanotubes to fabricate nanocomposites. Mechanical, mineralization, and degradation properties were studied in vitro. The proliferation and differentiation of rat bone marrow stem cells were studied to determine biocompatibility in vivo. The incorporation of multi-walled carbon nanotubes greatly increased the mechanical properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and the strongest composite obtained was at 2% multi-walled carbon nanotubes. The 2% nanocomposite also had higher rat bone marrow stem cell adhesion, proliferation, and differentiation characteristics compared to the pure poly(3-hydroxybutyrate-co-3-hydroxyvalerate). The apoptosis in the later stage of rat bone marrow stem cells decreased in the 2% nanocomposites group at different time points. Based on histology and micro-computed tomography tests 6 weeks after in vivo implantation, the 2% multi-walled carbon nanotubes/poly(3-hydroxybutyrate-co-3-hydroxyvalerate) treated animals had a higher volume of bone formation compared to the pure poly(3-hydroxybutyrate-co-3-hydroxyvalerate) group. Thus, the presence of multi-walled carbon nanotubes has an apparent positive effect on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in assisting osteogenesis.
A novel three-dimensional construct was designed to serve as a substitute for the natural meniscus tissue, and tested in vitro. The design consisted of mats of aligned collagen micro/nanofibers, entrapped within a macroporous poly(
Designing degradable hydrogels is complicated by the structural and temporal complexities of the gel and evolving tissue. A major challenge is to create scaffolds with sufficient mechanical properties to restore initial function while simultaneously controlling temporal changes in the gel structure to facilitate tissue formation. Poly(ethylene glycol) was used in this work, to form biodegradable poly(ethylene glycol)-based hydrogels with hydrolyzable poly-
Amino acid pairing peptide-based nanoparticles were recently introduced as promising carriers for hydrophobic anticancer drugs. The AC8 peptide, n-FEFQFNFK-c, is based on the amino acid pairing (AAP) design with 8 amino acids and hence the designated name AAP8. The nanoparticles (NPs) AAP8 have modified either on the C-terminal or on both terminal, by conjugation with diethylene glycol (DEG) . Here, the in vitro biocompatibilities of the NPs and their modified versions were compared and the potential of these NPs as carriers for the hydrophobic anticancer drug pirarubicin was determined as well as the peptide-drug co-assembly complexes. The toxicity of the NPs, DEGylated NPs, and blended mixtures with pirarubicin, was tested against the human adenocarcinoma lung cancer cell line, A549. The amino-end DEGylated NP, (NP-I), had superior biocompatibility over the non-modified NPs or double DEGylated NPs (NP-II). NP-I had very low hemolytic activity (1%) while NP and NP-II had marginal (8%) and acceptable (5%) hemolytic activity, respectively. All three types of NPs did not activate the complement system via the classical and alternative pathways nor did they activate the anaphylotoxin C3a. However, NP-II and its drug complex effectively activate the complement terminal attack complex. The lectin pathway was not activated by NP-I and NP-II, but was to a small extent by the non-modified NPs, with no lectin activation when complexed with drug. These results indicate NP-I is the most promising peptide for use as a drug delivery system, highlighting the importance of proper modification in peptides for drug delivery systems.
A new material was prepared to reduce catheter infection composed of a flocked silicone sheet (AmTiO2NP-F) with TiO2 nanoparticle–immobilized poly(ethylene terephthalate) fibers modified with surface amino groups. This system was used in conjunction with a tissue adhesive composed of disuccinimidyl tartrate and human serum albumin. At a fixed disuccinimidyl tartrate content of 0.2 mmol in human serum albumin solution, AmTiO2NP-F bonded well with collagen-based casing (a model material for skin), with bond strength increasing to a maximum of 38 w/v% human serum albumin. The adhesive bonded AmTiO2NP-F to subcutaneous tissue in mice, and infiltration of the tissue into the AmTiO2NP-F further increased the bond strength for long-term insertions. The material was degraded within 7 days of implantation, and tissue reaction was mild, while infection was completely prevented. These results indicate that the combined use of AmTiO2NP-F and disuccinimidyl tartrate-A for implanted catheters can significantly alleviate the associated risk of infection.
A novel ampicillin prodrug containing two carboxylic acid functionalities was synthesized by reacting ampicillin with acyl chloride in the presence of base. This prodrug was subsequently converted into a poly(anhydride-amide) via solution polymerization. The polymer, which chemically incorporates the ampicillin prodrug into the polymeric backbone, was developed as a film to prevent infections associated with medical devices by controlled, localized release of antimicrobials. The robust polymer coatings exhibiting strong adhesion to stainless steel were produced under elevated temperature and reduced pressure. The in vitro hydrolytic degradation of the polymer into the ampicillin prodrug was measured and the antibacterial activity of polymer-derived coatings was examined using a Gram-positive bacterium, Staphylococcus aureus. Furthermore, the polymer cytotoxicity was screened using fibroblasts. The ampicillin prodrug demonstrated antibacterial activity and the polymer demonstrated no cytotoxic effects on fibroblasts. Based on these results, the biodegradation of the antimicrobial-based poly(anhydride-amide) into the prodrug displays substantial promise as an implant or implant coating to reduce device failure resulting from bacterial infections.
Polymeric fibrous scaffolds based on the biocompatible and biodegradable three-arm-branched star poly(-caprolactone) (Mw = 189,000 g/mol) were prepared by a melt electrospinning technique. The possibility of processing polymers without the use of organic solvents is one of the main advantages over solution electrospinning. Scaffolds were biologically tested for their ability of supporting skin tissue regeneration. For this purpose, mouse embryo fibroblast (BALB/3T3 clone A31) and human keratinocyte (HaCaT) cell lines were selected as models, and seeded onto the polymeric supports both as single and co-culture. Cell viability, proliferation, and collagen production were assessed by WST-1 assay and Direct Red 80 dye, respectively. Cell morphology and colonization of the supports were evaluated by scanning electron microscopy and confocal laser scanning microscopy. Results highlighted that the star poly(-caprolactone) scaffolds were able to promote collagen production by fibroblasts. In co-culture studies, scaffolds supported adhesion, proliferation, and spatial organization of both cell lines. By virtue of the observed results, the developed polymeric scaffolds appeared suitable as biodegradable and biocompatible three-dimensional supports for skin tissue regeneration in wound healing dressing.
Poly[2-ethyl(2-pyrrolidone) methacrylate] and hyaluronic acid hydrogels were synthesized via free-radical polymerization of 2-ethyl(2-pyrrolidone) methacrylate, hyaluronic acid and different crosslinkers. The ability of these hydrogels to induce apatite formation by incubating in simulated body fluid was investigated. The effect of hyaluronic acid content, crosslinkers and immersion time on mineralization behaviour and interface properties as well as the metabolic activity of different cultured cells were also determined. The bioactivity of the poly[2-ethyl(2-pyrrolidone) methacrylate] and hyaluronic acid hydrogels along with cell viability data indicated their potential application in bone tissue engineering.
Appropriate pore structures and mechanical properties are required for scaffolds that are used for tissue engineering and regenerative medicine. In this study, pre-prepared ice particulates were used as a porogen material to prepare collagen porous scaffolds with well-controlled pore structures and improved mechanical properties. Porogen ice particulates initiated the formation of interconnected large spherical pores surrounded by small pores. The large spherical pores were well compacted and increased the elastic modulus of the scaffolds. The unique pore structures facilitated cell penetration, resulting in a homogeneous cell distribution throughout the scaffolds. The excellent mechanical properties protected the scaffolds from deformation during cell culturing and implantation. The collagen porous scaffolds facilitated cartilage regeneration when bovine articular chondrocytes were cultured in these scaffolds. The use of pre-prepared ice particulates as a porogen material proved to be a useful method to control the pore structure and improve the mechanical properties of collagen-based porous scaffolds.
Although poly(L-lactic acid) nanofibers are known to promote osteogenic differentiation of bone marrow stromal cells, their relative hydrophobicity and surface inertia tend to hinder their biomedical application. We explored a feasible and effective technique to improve the bioactivity and biocompatibility of poly(L-lactic acid) fibers for further application in regenerative medicine. A low-temperature atmospheric plasma was used to treat poly(L-lactic acid) nanofibers for 1, 5, and 10 min, and the surface properties and dose-dependent effects on the behavior of bone marrow stromal cells were studied. Both the amino group content and surface hydrophilicity of the nanofibers increased with treatment time, whereas the spreading and proliferation of bone marrow stromal cells were greatest on nanofibers which had been treated for 5 min, followed by samples treated for 1 and 10 min. The quantitative reverse transcription–polymerase chain reaction analysis of the bone marrow stromal cells on the 5-min-treated nanofibers had the highest expression level of osteogenic marker genes including RUNX2, BMP2, ALP, COL1A1, OPN, and OCN. The nanofibers treated for 5 min also promoted the high levels of alkaline phosphatase activity. These results suggest the exertion of dose-dependent effects by atmospheric plasma treatment on the surface of poly(L-lactic acid) nanofibers, and that this treatment is a feasible and effective technique to improve biomaterial biocompatibility and promotion of osteogenic differentiation of bone marrow stromal cells.