Publications

Graphene and graphene oxide (GO), due to their unique chemical and physical properties, possess biochemical characteristics that can trigger intercellular signals promoting tissue regeneration. Clinical applications of thin GO-derived sheets have inspired the development of various tissue regeneration and repair approaches. In this study, we demonstrate that ultrathin sheets of plasma-functionalized and reduced GO, with the oxygen content ranging from 3.2 % to 22 % and the nitrogen content from 0 % to 8.3 %, retain their essential mechanical and molecular integrity, and exhibit robust potential for regenerating bone tissue and blood vessels across multiple cellular and animal models. Initially, we observed the growth of blood vessels and bone tissue in vitro using these functionalized GO sheets on human adipose-derived mesenchymal stem cells and umbilical vein endothelial cells. Remarkably, our study indicates a 2.5-fold increase in mineralization and two-fold increase in tubule formation even in media lacking osteogenic and angiogenic supplements. Subsequently, we observed the initiation, conduction, and formation of bone and blood vessels in a rat tibial osteotomy model, evident from a marked 4-fold increase in the volume of low radio-opacity bone tissue and a significant elevation in connectivity density, all without the use of stem cells or growth factors. Finally, we validated these findings in a mouse critical-size calvarial defect model (33 % higher healing rate) and a rat skin lesion model (up to 2.5-fold increase in the number of blood vessels, and 35 % increase in blood vessels diameter). This study elucidates the proosteogenic and pro-angiogenic properties of both pristine and plasma-treated GO ultrathin films. These properties suggest their significant potential for clinical applications, and as valuable biomaterials for investigating fundamental aspects of bone and blood vessel regeneration.

JTD Keywords: Adhesion, Angiogenesis, Biocompatibilit, Bone regeneratio, Coatings, Fibronectin, Graphene oxide, Growth, Mesenchymal stem-cells, Osteoblast, Osteogenic differentiation, Plasma treatment, Protein, Tissue regeneration

Polypropylene (PP), thermoplastic polyurethane (TPU), polyethylene terephthalate glycol (PETG) and polylactic acid (PLA) 3D printed specimens, which are intrinsically non-electroresponsive materials, have been converted into electroresponsive electrodes applying a low-pressure oxygen plasma treatment. After complete chemical, morphological and electrochemical characterization, plasma treated samples have been applied as integrated electrochemical sensors for detecting dopamine and serotonin by cyclic voltammetry and chronoamperometry. Results show differences in the sensing behavior, which have been explained on the basis of the chemical structure of the pristine materials. While plasma treated PLA exhibits the highest performance as electrochemical sensor in terms of sensitivity (lowest limits of detection and quantification) and selectivity (against uric acid and ascorbic acid as interfering substances), plasma treated PP displays the poorest behavior due to its low polarity compared to PLA 3D-printed electrodes. Instead, plasma treated TPU and PETG shows a very good response, much closer to PLA, as sensitive electrodes towards neurotransmitter molecules (dopamine and serotonin). Overall, results open a new door for the fabrication of electrochemical conductive sensors using intrinsically insulating materials, without the need of chemical functionalization processes.

JTD Keywords: 3d printing, Amines, Ascorbic acid, Chemical characterization, Cyclic voltammetry, Dopamine, Electrochemical characterizations, Electrochemical sensor s, Electrochemical sensors, Electrode materials, Electroresponsive materials, Low-pressure oxygen-plasma treatments, Morphological characterization, Multiwalled carbon nanotubes (mwcn), Neurophysiology, Oxygen, Oxygen plasmas, Plastic bottles, Polyethylene terephthalate glycol, Polyethylene terephthalate glycols, Polyethylene terephthalates, Polylact i c acid, Polylactic acid, Polylactic acid pla, Polyols, Polypropylene, Polypropylene oxides, Polypropylenes, Polyurethanes, Reinforced plastics, Supercapacitors, Thermoplast i c polyurethane, Thermoplastic polyurethane, Thermoplastic polyurethanes

The incorporation of exogenous lactate into cardiac tissues is a regenerative strategy that is rapidly gaining attention. In this work, two polymeric platforms were designed to achieve a sustained release of lactate, combining immediate and prolonged release profiles. Both platforms contained electrospun poly(lactic acid) (PLA) fibers and an alginate (Alg) hydrogel. In the first platform, named L/K(x)/Alg-PLA, lactate and proteinase K (x mg of enzyme per 1 g of PLA) were directly loaded into the Alg hydrogel, into which PLA fibers were assembled. In the second platform, L/Alg-K(x)/PLA, fibers were produced by electrospinning a proteinase K:PLA solution and, subsequently, assembled within the lactate-loaded hydrogel. After characterizing the chemical, morphological, and mechanical properties of the systems, as well as their cytotoxicity, the release profiles of the two platforms were determined considering different amounts of proteinase K (x = 5.2, 26, and 52 mg of proteinase K per 1 g of PLA), which is known to exhibit a broad cleavage activity. The profiles obtained using L/Alg-K(x)/PLA platforms with x = 26 and 52 were the closest to the criteria that must be met for cardiac tissue regeneration. Finally, the amount of lactate directly loaded in the Alg hydrogel for immediate release and the amount of protein in the electrospinning solution were adapted to achieve a constant lactate release of around 6 mM per day over 1 or 2 weeks. In the optimized bioplatform, in which 6 mM lactate was loaded in the hydrogel, the amount of fibers was increased by a factor of ×3, the amount of enzyme was adjusted to 40 mg per 1 g of PLA, and a daily lactate release of 5.9 ± 2.7 mM over a period of 11 days was achieved. Accordingly, the engineered device fully satisfied the characteristics and requirements for heart tissue regeneration.

JTD Keywords: biodegradable fibers, cardiac tissue regeneration, cell, drug-release, elastic-modulus, electrospinning, heart, nanoindentation, plasma treatment, proteinase, scaffold, stiffness, Alginate, Biodegradable fibers, Cardiac tissue regeneration, Electrospinning, Nanoindentation, Plasma treatment, Proteinase, Skeletal-muscle

Isotactic polypropylene (i-PP) nonabsorbable surgical meshes are modified by incorporating a conducting polymer (CP) layer to detect the adhesion and growth of bacteria by sensing the oxidation of nicotinamide adenine dinucleotide (NADH), a metabolite produced by the respiration reactions of such microorganisms, to NAD+. A three-step process is used for such incorporation: (1) treat pristine meshes with low-pressure O2 plasma; (2) functionalize the surface with CP nanoparticles; and (3) coat with a homogeneous layer of electropolymerized CP using the nanoparticles introduced in (2) as polymerization nuclei. The modified meshes are stable and easy to handle and also show good electrochemical response. The detection by cyclic voltammetry of NADH within the interval of concentrations reported for bacterial cultures is demonstrated for the two modified meshes. Furthermore, Staphylococcus aureus and both biofilm-positive (B+) and biofilm-negative (B-) Escherichia coli cultures are used to prove real-time monitoring of NADH coming from aerobic respiration reactions. The proposed strategy, which offers a simple and innovative process for incorporating a sensor for the electrochemical detection of bacteria metabolism to currently existing surgical meshes, holds considerable promise for the future development of a new generation of smart biomedical devices to fight against post-operative bacterial infections.

JTD Keywords: adhesion, bacteria metabolism, behavior, biocompatibility, conducting polymer, electrochemical sensor, hernia repair, in-vivo, liquid, nadh detection, plasma treatment, prevention, reinforcement, sensor, smart meshes, Bacteria metabolism, Polypropylene mesh, Smart meshes

Electrochemical sensors for real-time detection of several bioanalytes have been prepared by additive manufacturing, shaping non-conductive poly(lactic acid) (PLA) filaments, and applying a physical treatment to create excited species. The latter process, which consists of the application of power discharge of 100 W during 2 min in a chamber at a low pressure of O-2, converts electrochemically inert PLA into an electrochemically responsive material. The electric discharge caused the oxidation of the PLA surface as evidenced by the increment in the quantity of oxygenated species detected by FTIR spectroscopy and X-ray photoelectron spectroscopy (XPS). Indeed, changes in the surface chemical composition became more pronounced with increasing O-2 pressure. After demonstrating the performance of the chemically modified material as individual dopamine and glucose sensors, multiplexed detection has been achieved by measuring simultaneously the two voltammetric signals. This has been performed by collecting the signals in two different regions, a naked chemically modified PLA for dopamine detection and a chemically modified PLA region functionalized with Glucose Oxidase. These outcomes led to define a new paradigm for manufacturing electrodes for electrochemical sensors based on 3D printing without using conducting materials at any stage of the process.

JTD Keywords: Additive manu f a c turing, Carbon, Conductivity, Degradation, Dopamine, Dopamine detection, Glucose detection, Glucose sensors, Immobilization, Multiplexed detect i o n, Oxidase, Plasma treatment

In this work, we present a successful strategy to convert recycled LDPE films, which usually end up in landfills or leak into the environment, into an advanced biomedical product. More specifically, LDPE films for food packaging have been treated with atmosphere corona discharge plasma for electrochemical detection of glucose. Enzyme-functionalized sensors manufactured using such recycled materials, which act as a mediator capable of electrocommunicating with the glucose oxidase (GOx) enzyme, are able to detect glucose concentrations in sweat and are fully compatible with the levels of such bioanalytes in both healthy and diabetic patients. Covalent immobilization of the GOx enzyme on the plasma-treated LDPE films has been successfully performed using the carbodiimide method, as proved by X-ray photoelectron spectroscopy. Then, the electronic communication between the deeply buried active site of the GOx and the reactive excited species formed at the surface of the plasma-treated LDPE has been demonstrated by linear sweep voltammetry. Finally, cyclic voltammetry in artificial sweat has been used to show that the LDPE-functionalized sensor has a linear response in the concentration of range of 50 μM to 1 mM with a limit of detection of 375 μA·μM–1·cm–2. Comparison of the performance of sensors prepared using recycled (i.e. with additives) and pristine (i.e. without additives) LDPE indicates that the utilization of the former does not require any pretreatment to eliminate additives. The present strategy demonstrates a facile approach for recycling LDPE waste into a high value-added product, which will potentially pave the way for the treatment of plastic waste in the future. Noninvasive glucose sensors based on recycled LDPE may play a crucial role in monitoring diabetes in underdeveloped regions.

JTD Keywords: Biosensors, Diabetes monitoring, High-value recycling, Plasma treatment, Sweat sensors