Bringing pathology to nanomedicine: a comparison of in vivo toxicity of polymeric nanoparticle carriers with and without chitosan coating.

Over the last years, there has been an increasing number of proposals for the use of nanomaterials in medicine. The safety of novel technologies must be verified, prior to their clinical application. Pathology has much to contribute towards this end. In this study, we compared the in vivo toxicity effects of poly- (lactic-co-glycolic acid) nanoparticles with and without chitosan shell. Both nanoparticle types were loaded with curcumin. The nanoparticles were assessed in vitro for potential cytotoxicity with cell viability studies. For the in vivo test, 36 adult Wistar rats were used, four of which were the control group. The remaining 32 were divided into 2 groups, each of which was administered differentially coated drug carriers: (A) nanoparticles without chitosan coating and (B) nanoparticles with chitosan coating. For both groups, the subcutaneous route was used for administration. Each group was further divided into 2 sub-groups of 8 animals each. The animals of the first sub-groups were sacrificed 24 h after the injection and those of the second on the 7th day. The control group was also divided into 2 subgroups of 2 animals each. At the appointed post-administrative date, the rats were sacrificed, and specimens from the brain, liver, kidneys, heart, stomach, lungs, and from the skin at the injection site were collected and studied histopathologically. The evaluation of both in vitro and in vivo testing shows that nanoparticles with chitosan have significantly less, if any, toxic effects compared to those without chitosan.

Morphological Characteristics of Drug-Eluting Biodegradable Polymeric Thin Films Developed on the Surface of Intraocular Lenses by Three Techniques: A Comparative Study.

Background Cataract surgery is a very popular operation that requires a postoperative period of frequent instillation of antibiotic and anti-inflammatory eye drops. Modified drug-eluting intraocular lenses (IOLs) may eliminate the need for eye drops after surgery. Aim The purpose of this study is to compare the morphological characteristics of dexamethasone eluting biodegradable polymeric thin films developed on the surface of commercially available IOLs by three different methods. Method This experimental study was conducted between May and August of 2021 in the Lab for Thin Films - Nanobiomaterials - Nanosystems & Nanometrology (LTFN) of the Aristotle University of Thessaloniki. A mixture of two organic polymers [Poly (D, L-lactide-co-glycolide)(PLGA), lactide: glycolide (75:25) and Polycaprolactone (PCL)] and dexamethasone was prepared and then deposited on the surface of three-piece IOLs by spin coating, by spray coating, and by gravure printing. The modified IOLs were sterilized with the use of ultraviolet (UV) radiation and plasma treatment. Their structural properties were studied with the use of atomic force microscopy (AFM). Results Spin coating and gravure printing produced uniform thin films on the surface of the IOLs which were not damaged during the sterilization process. Spray coating led to the partial coating of the surface of the IOLs; the thin films underwent alterations following plasma treatment. Conclusions Thin films developed by spin coating and gravure printing on IOLs demonstrate the desired morphological characteristics that make them suitable candidates for further research.

Design, characterisation and drug release study of polymeric, drug-eluting single layer thin films on the surface of intraocular lenses.

To design, develop and study a novel drug delivery system for intraocular applications. The spin coating technique was applied to develop a polymeric, drug-eluting thin film consisting of a blend of organic polymers [poly (D, L lactide coglycolide) lactide: glycolide 75: 25, PLGA and polycaprolactone, PCL] and dexamethasone on the surface of intraocular lenses (IOLs). The initial durability of the IOLs during spinning was assessed. Information about the structural and optical properties of the modified IOLs was extracted using atomic force microscopy, scanning electron microscopy and spectroscopic ellipsometry. A drug release study was conducted for 8 weeks. The IOLs were durable in spinning speeds higher than the ones used to develop thin films. Single-layer thin films were successfully developed on the optics and the haptics of the lenses. The films formed nanopores with encapsulated aggregates of dexamethasone. The spectroscopic ellipsometry showed an acceptable optical transparency of the lenses regardless of the deposition of the drug-eluting films on their surface. The drug release study demonstrated gradual dexamethasone release over the selected period. In conclusion, the novel drug-eluting IOL system exhibited desired properties regarding its transparency and drug release rate. Further research is necessary to assess their suitability as an intraocular drug delivery system.

Biomimetic and biodegradable cellulose acetate scaffolds loaded with dexamethasone for bone implants.

There is, as a matter of fact, an ever increasing number of patients requiring total hip replacement (Pabinger, C.; Geissler, A. Osteoarthritis Cartilage2014,22, 734-741). Implant-associated acute inflammations after an invasive orthopedic surgery are one of the major causes of implant failure. In addition, there are instability, aseptic loosening, infection, metallosis and fracture (Melvin, J. S.; Karthikeyan, T.; Cope, R.; Fehring, T. K. J. Arthroplasty2014,29, 1285-1288). In this work, a drug-delivery nanoplatform system consisting of polymeric celluloce acetate (CA) scaffolds loaded with dexamethasone was fabricated through electrospinning. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) indicated the successful fabrication of these structures. Cytotoxicity studies were performed by using MTT assay, methylene-blue staining and SEM fixation and showed very good cell adhesion and proliferation, indicating the cytocompatibility of these fibrous scaffolds. Drug-release kinetics was measured for the evaluation of a controllable and sustained release of anti-inflammatory drug onto the engineered implants and degradation study was conducted in order to assess the mass loss of polymers. This drug-delivery nanoplatform as coating on titanium implants may be a promising approach not only to alleviate but also to prevent implant-associated acute inflammations along with a simultaneous controlled release of the drug.

Design and fabrication of drug-eluting polymeric thin films for applications in ophthalmology.

To study the development, characterisation, and drug release of one- and two-layered thin films based on organic polymers [poly(D,L-lactide-co-glycolide) lactide:glycolide (65:35), poly(D,L-lactide-co-glycolide) lactide:glycolide (75:25), and polycaprolactone] and dexamethasone. To examine their applicability for intraocular lenses (IOLs) and function in intraocular drug delivery systems. Four series of thin films, single and double-layer, were prepared by the spin-coating method on a silicon substrate. The films were studied using atomic force microscopy and spectroscopic ellipsometry. The release rate of dexamethasone was studied for a period of ten weeks. Series A and C demonstrated the formation of large dexamethasone aggregates. The monolayer films of series C and D formed pores, in agreement with previous findings. The spectroscopic ellipsometry study demonstrated that the samples were transparent. The drug release study demonstrated that dexamethasone was released during the first 6 weeks at a desirable rate. The films exhibited properties suitable for use in intraocular drug delivery systems. The single-layer thin films demonstrated a sufficient encapsulation of dexamethasone and appropriate release of the therapeutic substance. Further studies are necessary to investigate the possibility of developing the films directly on the surface of the IOL.

Surface modification of endovascular stents with rosuvastatin and heparin-loaded biodegradable nanofibers by electrospinning.

This study describes the development of drug-loaded nanofibrous scaffolds as a nanocoating for endovascular stents for the local and sustained delivery of rosuvastatin (Ros) and heparin (Hep) to injured artery walls after endovascular procedures via the electrospinning process. PURPOSE: The proposed hybrid covered stents can promote re-endothelialization; improve endothelial function; reduce inflammatory reaction; inhibit neointimal hyperplasia of the injured artery wall, due to well-known pleiotropic actions of Ros; and prevent adverse events such as in-stent restenosis (ISR) and stent thrombosis (ST), through the antithrombotic action of Hep. METHODS: Biodegradable nanofibers were prepared by dissolving cellulose acetate (AC) and Ros in N,N-dimethylacetamide (DMAc) and acetone-based solvents. The polymeric solution was electrospun (e-spun) into drug-loaded AC nanofibers onto three different commercially available stents (Co-Cr stent, Ni-Ti stent, and stainless steel stent), resulting in nonwoven matrices of submicron-sized fibers. Accordingly, Hep solution was further used for fibrous coating onto the engineered Ros-loaded stent. The functional encapsulation of Ros and Hep drugs into polymeric scaffolds further underwent physicochemical analysis. Morphological characterization took place via scanning electron microscopy (SEM) and atomic force microscopy (AFM) analyses, while scaffolds' wettability properties were obtained by contact angle (CA) measurements. RESULTS: The morphology of the drug-loaded AC nanofibers was smooth, with an average diameter of 200-800 nm, and after CA measurement, we concluded to the superhydrophobic nature of the engineered scaffolds. In vitro release rates of the pharmaceutical drugs were determined using a high-performance liquid chromatography assay, which showed that after the initial burst, drug release was controlled slowly by the degradation of the polymeric materials. CONCLUSION: These results imply that AC nanofibers encapsulated with Ros and Hep drugs have great potential in the development of endovascular grafts with anti-thrombogenic properties that can accelerate the re-endothelialization, reduce the neointimal hyperplasia and inflammatory reaction, and improve the endothelial function.

Insights on the Optical Properties of Poly(3,4-Ethylenedioxythiophene):Poly(styrenesulfonate) Formulations by Optical Metrology.

Poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) is among the most widely used polymers that are used as printed transparent electrodes for flexible Organic Electronic (OE) devices, such as Organic Photovoltaics (OPVs). The understanding of their optical properties and the correlation of the optical properties with their electronic properties and metallic-like behavior can lead to the optimization of their functionality as transparent electrodes in multilayer OE device architectures. In this work, we study the optical properties of different PEDOT:PSS formulations by non-destructive Spectroscopic Ellipsometry (SE), from the infrared to the far ultraviolet spectral regions. The optical response of PEDOT:PSS includes an intense optical absorption originated from the conductive part (PEDOT) at lower photon energies, whereas the electronic transition energies of the non-conductive PSS part have been measured at higher photon energies. Based on the different PEDOT:PSS formulations, the optical investigation revealed significant information on the relative contribution of conductive PEDOT and insulating PSS parts of the PEDOT:PSS formulation in the overall optical response, which can strongly impact the final device functionality and its optical transparency.

The melding of nanomedicine in thrombosis imaging and treatment: a review.

Thromboembolic diseases constitute a plague in our century, wherein an imbalance of hemostasis leads to thrombus formation and vessels constriction reducing blood flow. Hence, the recent rise of nanomedicine gives birth to advanced diagnostic modalities and therapeutic agents for the early diagnosis and treatment of such diseases. Multimodal nanoagents for the detection of intravascular thrombi and nanovehicles for thrombus-targeted fibrinolytic therapy are few paradigms of nanomedicine approaches to overcome current diagnostic treatment roadblocks and persistent clinical needs. This review highlights the nanomedicine strategies to improve the imaging and therapy of acute thrombi by nanoparticles and nanotheranostics, the detailed imaging of thrombogenic proteins and platelets via atomic force microscopy with the knowledge basis of thrombosis pathophysiology and nanotoxicity.

Nanomedicine for Atherosclerosis: Molecular Imaging and Treatment.

Cardiovascular diseases (CVDs) and the underlying process of atherosclerosis are considered to be the most frequent causes of mortality and morbidity in developed societies. Atherosclerosis constitutes a systemic, chronic and progressive inflammatory disease that is initiated by early endothelial dysfunction and is subsequently amplified by oxidative stress, lipid deposition and monocyte recruitment. An interplay occurs among diverse cells, chemoattractants, adhesion molecules and low-density lipoproteins in the subendothelium. Due to the complexity of its pathogenesis, effective therapeutic strategies have not yet been applied in routine clinical practice. With the advent of nanotechnology, nanoparticulate systems with diagnostic and therapeutic moieties for the site-specific targeting of atherosclerotic lesions as well as nanomaterials that are suitable for cardiovascular implants may offer possible solutions to certain shortfalls of current treatment regimens. This article describes the recent advances that involve different types of nanoparticles for the early detection and concurrent therapy of atherosclerotic lesions. Moreover, it provides a state-of-the-art overview of stent technology in the restoration of normal blood flow to ischemic myocardial sites and underscores its drawbacks in light of substantial nanotechnology-based improvements. Emphasis is placed on the contribution of nanomedicine to the development of novel and effective therapies for atherosclerosis, ranging from 'nanotheranostic' strategies for vulnerable plaques to the nanoporous and nanoparticulate drug-delivery platforms that have been applied to stent technology. By striking a balance between the efficacy and the potential toxicity of nanotechnology-enabled systems, new frontiers in atherosclerosis treatment will emerge.

Oxygen-plasma-modified biomimetic nanofibrous scaffolds for enhanced compatibility of cardiovascular implants.

Electrospun nanofibrous scaffolds have been extensively used in several biomedical applications for tissue engineering due to their morphological resemblance to the extracellular matrix (ECM). Especially, there is a need for the cardiovascular implants to exhibit a nanostructured surface that mimics the native endothelium in order to promote endothelialization and to reduce the complications of thrombosis and implant failure. Thus, we herein fabricated poly-ε-caprolactone (PCL) electrospun nanofibrous scaffolds, to serve as coatings for cardiovascular implants and guide tissue regeneration. Oxygen plasma treatment was applied in order to modify the surface chemistry of the scaffold and its effect on cell attachment and growth was evaluated. The conditions of the surface modification were properly adjusted in order to define those conditions of the treatment that result in surfaces favorable for cell growth, while maintaining morphological integrity and mechanical behavior. Goniometry (contact angle measurements), scanning electron microscopy (SEM), atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS) measurements were used to evaluate the morphological and chemical changes induced by the plasma treatment. Moreover, depth-sensing nanoindentation was performed to study the resistance of the plasma-treated scaffolds to plastic deformation. Lastly, the cell studies indicated that all scaffolds were cytocompatible, with the plasma-treated ones expressing a more pronounced cell viability and adhesion. All the above findings demonstrate the great potential of these biomimetic tissue-engineering constructs as efficient coatings for enhanced compatibility of cardiovascular implants.

Measurements of elastic modulus for human anterior lens capsule with atomic force microscopy: the effect of loading force.

The purpose of the study was to appraise the effect of loading force magnitude on the determination of the elastic modulus of the anterior lens capsule through atomic force microscopy. Four human anterior lens capsules taken during phacoemulsification cataract surgery were studied, free of epithelial cells, with atomic force microscopy. For the experiment, five different indentation loading forces were applied to near areas of the specimen. Experimental data was exported and analyzed according to the Hertz model to obtain the Young's modulus with regards to the elastic behavior of the material. Force-distance curves were acquired by applying a load of 2, 5, 10, 20 and 30 nN. When examining the results it was evident that determination of Young's modulus of the anterior lens capsule is dependent on the loading force concerning the examined range. Loading forces of 10 and 20 nN led to results without significant difference (p > 0.05) and more reproducible (coefficients of variation 12.4 and 11.7 %, respectively).

Novel nanostructured biomaterials: implications for coronary stent thrombosis.

BACKGROUND: Nanomedicine has the potential to revolutionize medicine and help clinicians to treat cardiovascular disease through the improvement of stents. Advanced nanomaterials and tools for monitoring cell-material interactions will aid in inhibiting stent thrombosis. Although titanium boron nitride (TiBN), titanium diboride, and carbon nanotube (CNT) thin films are emerging materials in the biomaterial field, the effect of their surface properties on platelet adhesion is relatively unexplored. OBJECTIVE AND METHODS: In this study, novel nanomaterials made of amorphous carbon, CNTs, titanium diboride, and TiBN were grown by vacuum deposition techniques to assess their role as potential stent coatings. Platelet response towards the nanostructured surfaces of the samples was analyzed in line with their physicochemical properties. As the stent skeleton is formed mainly of stainless steel, this material was used as reference material. Platelet adhesion studies were carried out by atomic force microscopy and scanning electron microscopy observations. A cell viability study was performed to assess the cytocompatibility of all thin film groups for 24 hours with a standard immortalized cell line. RESULTS: The nanotopographic features of material surface, stoichiometry, and wetting properties were found to be significant factors in dictating platelet behavior and cell viability. The TiBN films with higher nitrogen contents were less thrombogenic compared with the biased carbon films and control. The carbon hybridization in carbon films and hydrophilicity, which were strongly dependent on the deposition process and its parameters, affected the thrombogenicity potential. The hydrophobic CNT materials with high nanoroughness exhibited less hemocompatibility in comparison with the other classes of materials. All the thin film groups exhibited good cytocompatibility, with the surface roughness and surface free energy influencing the viability of cells.

Development of a nanoporous and multilayer drug-delivery platform for medical implants.

Biodegradable polymers can be applied to a variety of implants for controlled and local drug delivery. The aim of this study is to develop a biodegradable and nanoporous polymeric platform for a wide spectrum of drug-eluting implants with special focus on stent-coating applications. It was synthesized by poly(DL-lactide-co-glycolide) (PLGA 65:35, PLGA 75:25) and polycaprolactone (PCL) in a multilayer configuration by means of a spin-coating technique. The antiplatelet drug dipyridamole was loaded into the surface nanopores of the platform. Surface characterization was made by atomic force microscopy (AFM) and spectroscopic ellipsometry (SE). Platelet adhesion and drug-release kinetic studies were then carried out. The study revealed that the multilayer films are highly nanoporous, whereas the single layers of PLGA are atomically smooth and spherulites are formed in PCL. Their nanoporosity (pore diameter, depth, density, surface roughness) can be tailored by tuning the growth parameters (eg, spinning speed, polymer concentration), essential for drug-delivery performance. The origin of pore formation may be attributed to the phase separation of polymer blends via the spinodal decomposition mechanism. SE studies revealed the structural characteristics, film thickness, and optical properties even of the single layers in the triple-layer construct, providing substantial information for drug loading and complement AFM findings. Platelet adhesion studies showed that the dipyridamole-loaded coatings inhibit platelet aggregation that is a prerequisite for clotting. Finally, the films exhibited sustained release profiles of dipyridamole over 70 days. These results indicate that the current multilayer phase therapeutic approach constitutes an effective drug-delivery platform for drug-eluting implants and especially for cardiovascular stent applications.

Nanomedicine for the reduction of the thrombogenicity of stent coatings.

The treatment of patients with drug-eluting stents (DES) continues to evolve with the current emergence of DES technology that offers a combination of pharmacological and mechanical approaches to prevent arterial restenosis. However, despite the promising short-term and mid-term outcomes of DES, there are valid concerns about adverse clinical effects of late stent thrombosis. In this study, we present an example of how nanomedicine can offer solutions for improving stent coating manufacturing, by producing nanomaterials with tailored and controllable properties. The study is based on the exploitation of human platelets response towards carbon-based nanocoatings via atomic force microscope (AFM). AFM can facilitate the comprehensive analysis of platelets behavior onto stent nanocoatings and enable the study of thrombogenicity. Platelet-rich plasma from healthy donors was used for the real-time study of biointerfacial interactions. The carbon nanomaterials were developed by rf magnetron sputtering technique under controllable deposition conditions to provide favorable surface nanotopography. It was shown that by altering the surface topography of nanocoatings, the activation of platelets can be affected, while the carbon nanocoatings having higher surface roughness were found to be less thrombogenic in terms of platelets adhesion. This is an actual solution for improving the stent coating fabrication.

Atomic force microscopy probing platelet activation behavior on titanium nitride nanocoatings for biomedical applications.

There is increasing interest in developing novel coatings to enhance the biocompatibility of medical implants. A key issue in biocompatibility research is platelet activation and aggregation on the biomaterials' surface. Stoichiometric and nonstoichiometric titanium nitride (TiN(x)) films were developed by sputtering as case study materials, for probing platelet activation behavior onto them. Atomic force microscopy (AFM) facilitates the real-time studies of cells and guarantees cellular viability. In this work a methodology for platelets study by AFM was developed. The morphological, structural, optical, and wettability properties of the TiN(x) films were obtained by AFM, x-ray diffraction, spectroscopic ellipsometry, and contact angle measurements.The properties of TiN(x) films were correlated with their thrombogenicity involving platelets' adhesion, activation and protein clustering mechanisms. It was found that the TiN(x) films stoichiometry and surface roughness affect the platelet response. The stoichiometric and smoother TiN films promote platelets adhesion and activation.

Renal function of patients with a failing Fontan circuit undergoing total cavopulmonary revision surgery.

This report characterizes renal dysfunction after total cavopulmonary (TCPC) revision surgery for atriopulmonary Fontan (APF) circulations, a known risk factor for a poor outcome. The perioperative data for 23 consecutively identified patients were reviewed. The preoperative mean glomerular filtration rate (GFR) was 101 +/- 30 ml/min/1.73 m(2), decreasing to 65 +/- 41 ml/min/1.73 m(2) early in the postoperative period. The preoperative GFR was highly correlated with age at APF (r = -0.5; p = 0.024), age at TCPC (r = -0.5; p = 0.01), and mixed venous saturation (r = 0.6; p = 0.01). Three of four patients requiring renal replacement therapy (RRT) died at a median age of 3 months (range, 18 days to 9 months). Determinants of early GFR and RRT were preoperative GFR (p = 0.016) and creatinine (p = 0.035). Younger age at primary Fontan (p = 0.008), higher preoperative mixed venous saturation (p = 0.019), and higher preoperative blood pressure (p = 0.006) independently predicted better GFRs at the latest follow-up evaluation. Renal function declines acutely after TCPC revision, often necessitating RRT. A requirement for RRT marks greater mortality. Higher preoperative creatinine levels identify those at greatest risk.

Impact of surface electric properties of carbon-based thin films on platelets activation for nano-medical and nano-sensing applications.

Electric surface properties of biomaterials, playing key role to various biointerfacial interactions, were related to hemocompatibility and biosensing phenomena. In this study, the examination of surface electric properties of amorphous hydrogenated carbon thin films (a-C:H) was carried out by means of electrostatic force microscope (EFM) and observation of differences in spatial charge distribution on the surface of the examined films during platelets adhesion was made. The thrombogenic potential of a-C:H thin films developed by magnetron sputtering with approximately 42% sp(3) content and hydrogen partial pressure during deposition was evaluated, by in situ observation with atomic force microscope (AFM) of platelets' activation and their subsequent adhesion. Platelet-rich plasma drawn from healthy donors was used and semi-contact mode of AFM was applied. Platelets behavior and their correlation with the electric surface properties of the examined a-C:H films by EFM was made for hemocompatibility enhancement and sensing platelets that are less electrical negatively charged and with higher tendency to aggregate and form thrombus. The results are discussed in view of the effect of different deposition conditions of hydrogenated carbon films on their structural and morphological characteristics, surface roughness and electrical properties attributing to different hemocompatibility and sensing aspects.