Development and Characterization of Polymer-based Magnetoelectric Nanofibers

With the rapid development of bionics, where biological systems meet electronics, there is an interest in polymer-based electrode systems that are soft, flexible, easily processed and fabricated. In this research area, magnetoelectric (ME) composites bring new and exciting opportunities, including contactless or “wireless” electrical stimulation, less-invasive integration in the form of dispersible, injectable nanoelectrodes, and applications as biodegradable sensors and bioenergy harvesters in the biomedical field. When ME composites are exposed to a magnetic field, a magnetostrictive (MS) component transfers strain to a piezoelectric (PE) component that generates an output voltage. In doing so, ME composites have the ability to enable magnetic-to-electrical conversion and thus can be utilized to power devices or electrically stimulate tissues or cells from a remote magnetic stimulus. To date, ceramic materials have mostly been applied in nanostructured ME composites, however, these may become fragile and cause deleterious reactions at the interface regions, leading to low electrical resistivity and high dielectric losses and ultimately low output voltage. To overcome these shortcomings, polymer-based ME composites offer new solutions to develop softer, contactless electrodes, without electrical connections, for easier and unique fabrication approaches (e.g. incorporation into soft gels). Their strain-mediated ME effect in large scale devices has been thoroughly studied both experimentally and theoretically. Polymer-based ME composites have almost exclusively used the PE polymer, poly (vinylidene fluoride) (PVDF), due to its high PE coefficient and as such developments in exploring other types of PE polymers have not been forthcoming. For example, other PE polymers such as poly (vinylidene fluoride-co-hexafluoropropylene) (P(VDF-HFP)) and poly (lactic acid) (PLA) have yet to be investigated though have the potential to bring added-value and function to polymer-based ME composites. Compared to PVDF and its copolymer P(VDF-TrFE), the piezoelectricity of another copolymer, P(VDF-HFP), is less-well understood. As a biocompatible polymer, PLA has been extensively investigated for applications in drug delivery and tissue engineering. Instead of being used only as a biodegradable and bioactive thermoplastic material, PLA is promising as a PE polymer, which has potential to mimic PE functions of tissues. Thus, in addition to PVDF, the thesis investigates the PE properties of P(VDF-HFP) and PLA and aims to develop ME composite nanofibers based on these polymers

Beyond Tissue replacement: The Emerging role of smart implants in healthcare

Smart implants are increasingly used to treat various diseases, track patient status, and restore tissue and organ function. These devices support internal organs, actively stimulate nerves, and monitor essential functions. With continuous monitoring or stimulation, patient observation quality and subsequent treatment can be improved. Additionally, using biodegradable and entirely excreted implant materials eliminates the need for surgical removal, providing a patient-friendly solution. In this review, we classify smart implants and discuss the latest prototypes, materials, and technologies employed in their creation. Our focus lies in exploring medical devices beyond replacing an organ or tissue and incorporating new functionality through sensors and electronic circuits. We also examine the advantages, opportunities, and challenges of creating implantable devices that preserve all critical functions. By presenting an in-depth overview of the current state-of-the-art smart implants, we shed light on persistent issues and limitations while discussing potential avenues for future advancements in materials used for these devices

Design, characterization and validation of integrated bioelectronics for cellular studies: from inkjet-printed sensors to organic actuators

Mención Internacional en el título de doctorAdvances in bioinspired and biomimetic electronics have enabled coupling engineering devices to biological systems with unprecedented integration levels. Major efforts, however, have been devoted to interface malleable electronic devices externally to the organs and tissues. A promising alternative is embedding electronics into living tissues/organs or, turning the concept inside out, lading electronic devices with soft living matters which may accomplish remote monitoring and control of tissue’s functions from within. This endeavor may unleash the ability to engineer “living electronics” for regenerative medicine and biomedical applications. In this context, it remains a challenge to insert electronic devices efficiently with living cells in a way that there are minimal adverse reactions in the biological host while the electronics maintaining the engineered functionalities. In addition, investigating in real-time and with minimal invasion the long-term responses of biological systems that are brought in contact with such bioelectronic devices is desirable. In this work we introduce the development (design, fabrication and characterization) and validation of sensors and actuators mechanically soft and compliant to cells able to properly operate embedded into a cell culture environment, specifically of a cell line of human epithelial keratinocytes. For the development of the sensors we propose moving from conventional microtechnology approaches to techniques compatible with bioprinting in a way to support the eventual fabrication of tissues and electronic sensors in a single hybrid plataform simultaneously. For the actuators we explore the use of electroactive, organic, printing-compatible polymers to induce cellular responses as a drug-free alternative to the classic chemical route in a way to gain eventual control of biological behaviors electronically. In particular, the presented work introduces inkjet-printed interdigitated electrodes to monitor label-freely and non-invasively cellular migration, proliferation and cell-sensor adhesions of epidermal cells (HaCaT cells) using impedance spectroscopy and the effects of (dynamic) mechanical stimulation on proliferation, migration and morphology of keratinocytes by varying the magnitude, frequency and duration of mechanical stimuli exploiting the developed biocompatible actuator. The results of this thesis contribute to the envision of three-dimensional laboratory-growth tissues with built-in electronics, paving exciting avenues towards the idea of living smart cyborg-skin substitutes.En los útimos años los avances en el desarrollo de dispositivos electrónicos diseñados imitando las propiedades de sistemas vivos han logrado acoplar sistemas electrónicos y órganos/tejidos biológicos con un nivel de integración sin precedentes. Convencionalmente, la forma en que estos sistemas bioelectrónicos son integrados con órganos o tejidos ha sido a través del contacto superficial entre ambos sistemas, es decir acoplando la electrónica externamente al tejido. Lamentablemente estas aproximaciones no contemplan escenarios donde ha habido una pérdida o daño del tejido con el cual interactuar, como es el caso de daños en la piel debido a quemaduras, úlceras u otras lesiones genéticas o producidas. Una alternativa prometedora para ingeniería de tejidos y medicina regenerativa, y en particular para implantes de piel, es embeber la electrónica dentro del tejido, o presentado de otra manera, cargar el sistema electrónico con células vivas y tejidos fabricados por ingeniería de tejidos como parte innata del propio dispositivo. Este concepto permitiría no solo una monitorización remota y un control basado en señalizaciones eléctricas (sin químicos) de tejidos biológicos fabricados mediante técnicas de bioingeniería desde dentro del propio tejido, sino también la fabricación de una “electrónica viva”, biológica y eléctricamente funcional. En este contexto, es un desafío insertar de manera eficiente dispositivos electrónicos con células vivas sin desencadenar reacciones adversas en el sistema biológico receptor ni en el sistema electrónico diseñado. Además, es deseable monitorizar en tiempo real y de manera mínimamente invasiva las respuestas de dichos sistemas biológicos que se han añadido a tales dispositivos bioelectrónicos. En este trabajo presentamos el desarrollo (diseño, fabricación y caracterización) y validación de sensores y actuadores mecánicamente suaves y compatibles con células capaces de funcionar correctamente dentro de un entorno de cultivo celular, específicamente de una línea celular de células epiteliales humanas. Para el desarrollo de los sensores hemos propuesto utilizar técnicas compatibles con la bioimpresión, alejándonos de la micro fabricación tradicionalmente usada para la manufactura de sensores electrónicos, con el objetivo a largo plazo de promover la fabricación de los tejidos y los sensores electrónicos simultáneamente en un mismo sistema de impresión híbrido. Para el desarrollo de los actuadores hemos explorado el uso de polímeros electroactivos y compatibles con impresión y hemos investigado el efecto de estímulos mecánicos dinámicos en respuestas celulares con el objetivo a largo plazo de autoinducir comportamientos biológicos controlados de forma electrónica. En concreto, este trabajo presenta sensores basados en electrodos interdigitados impresos por inyección de tinta para monitorear la migración celular, proliferación y adhesiones célula-sustrato de una línea celular de células epiteliales humanas (HaCaT) en tiempo real y de manera no invasiva mediante espectroscopía de impedancia. Por otro lado, este trabajo presenta actuadores biocompatibles basados en el polímero piezoeléctrico fluoruro de poli vinilideno y ha investigado los efectos de estimular mecánicamente células epiteliales en relación con la proliferación, migración y morfología celular mediante variaciones dinámicas de la magnitud, frecuencia y duración de estímulos mecánicos explotando el actuador biocompatible propuesto. Ambos sistemas presentados como resultado de esta tesis doctoral contribuyen al desarrollo de tejidos 3D con electrónica incorporada, promoviendo una investigación hacia la fabricación de sustitutos equivalentes de piel mitad orgánica mitad electrónica como tejidos funcionales biónicos inteligentes.The main works presented in this thesis have been conducted in the facilities of the Universidad Carlos III de Madrid with support from the program Formación del Profesorado Universitario FPU015/06208 granted by Spanish Ministry of Education, Culture and Sports. Some of the work has been also developed in the facilities of the Fraunhofer-Institut für Zuverlässigkeit und Mikrointegration (IZM) and University of Applied Sciences (HTW) in Berlin, under the supervision of Prof. Dr. Ing. H-D. Ngo during a research visit funded by the Mobility Fellows Program by the Spanish Ministry of Education, Culture, and Sports. This work has been developed in the framework of the projects BIOPIELTEC-CM (P2018/BAA-4480), funded by Comunidad de Madrid, and PARAQUA (TEC2017-86271-R) funded by Ministerio de Ciencia e Innovación.Programa de Doctorado en Ingeniería Eléctrica, Electrónica y Automática por la Universidad Carlos III de MadridPresidente: José Antonio García Souto.- Secretario: Carlos Elvira Pujalte.- Vocal: María Dimak

THE ACOUSTIC WAVE SENSOR AND SOFT LITHOGRAPHY TECHNOLOGIES FOR CELL BIOLOGICAL STUDIES

Recently, cell-based biosensors have attracted many attentions because of their potential applications in fundamental biological research, drug development, and other fields. Acoustic wave biosensors offer powerful tools to probe cell behaviors and properties in a non-invasive, simple, and quantitative manner. Current studies on cell-based acoustic wave sensors are focused on experimental investigation of thickness shear mode (TSM) sensors for monitoring cell attachment and spreading. There are no theoretical models for cell-based TSM biosensors. No studies on other cell biological applications of TSM sensors or on surface acoustic wave cell-based biosensors have been performed. The reliability and sensitivity of current cell-based biosensors are low. Improving them requires studies on engineering cells and understanding the effects of cell morphology on cell function.The overall objective of this dissertation is to develop acoustic wave sensor systems for cell biological studies and to determine the effects of cell shape on cell function. Our study includes three parts: (1) Development of cell-based TSM sensor system; (2) Studies of Love mode devices as cell-based biosensors; (3) Studies of the effects of cell shape on cell function. In the first part, a theoretical model was developed, changes in cell adhesion were monitored and cell viscoelasticity was characterized by TSM sensor systems. The TSM sensor systems were demonstrated to provide a non-invasive, simple, and reliable method to monitor cell adhesion and characterize cell viscoelasticity. In the second part, a theoretical model was developed to determine signal changes in Love mode sensors due to cells attaching on their surface. Experimental results validated the model. In the third part, cell shape was patterned to different aspect ratios. Elongated tendon cells were found to express higher collagen type I than shorter cells. Changes in cell shape induced alterations in cytoskeleton, focal adhesions, and traction forces in cells, which may collectively prompt the observed differential collagen type I expression in cells with different shapes. Overall, our research expanded the applications of acoustic wave cell-based biosensors. Studies on cell shape control and the effects of cell shape on cell function will be useful for increasing the sensitivity of cell-based biosensors in future research

Silk-Based Biopolymers Promise Extensive Biomedical Applications in Tissue Engineering, Drug Delivery, and BioMEMS

As an FDA-approved biopolymer, silk has been contemplated for a wide range of applications based on its unique merits, such as biocompatibility, biodegradability, and piezoelectricity. As silk, in both crystalline structure and amorphous state, exhibits unique physical, mechanical, and biological properties (promoting cell migration, differentiation, growth, and protein-surface interaction), it is fruitful to understand its potential applications. Sensors, actuators, and drug delivery systems are the best in case. As such, the current effort first introduces silk fibroin (SF) and delineates its characteristics. It then explores the extensive use of this biomaterial in tissue engineering approaches, in addition to its biosensor and electro-active wearable bioelectronic application. To this end, the SF application in cardiovascular, skin, cartilage, and drug delivery systems for cancer therapy and wound healing was studied precisely. Compositing any type of other variables to induce a specific application or improve any SF barriers, namely its hydrophobicity, poor electrical conductivity, or tuning its mechanical properties, especially in tissue engineering applications, has also been discussed wherever it is deemed informative.</p

Silk-Based Biopolymers Promise Extensive Biomedical Applications in Tissue Engineering, Drug Delivery, and BioMEMS

As an FDA-approved biopolymer, silk has been contemplated for a wide range of applications based on its unique merits, such as biocompatibility, biodegradability, and piezoelectricity. As silk, in both crystalline structure and amorphous state, exhibits unique physical, mechanical, and biological properties (promoting cell migration, differentiation, growth, and protein-surface interaction), it is fruitful to understand its potential applications. Sensors, actuators, and drug delivery systems are the best in case. As such, the current effort first introduces silk fibroin (SF) and delineates its characteristics. It then explores the extensive use of this biomaterial in tissue engineering approaches, in addition to its biosensor and electro-active wearable bioelectronic application. To this end, the SF application in cardiovascular, skin, cartilage, and drug delivery systems for cancer therapy and wound healing was studied precisely. Compositing any type of other variables to induce a specific application or improve any SF barriers, namely its hydrophobicity, poor electrical conductivity, or tuning its mechanical properties, especially in tissue engineering applications, has also been discussed wherever it is deemed informative.</p

Frontiers in microfluidics, a teaching resource review

This is a literature teaching resource review for biologically inspired microfluidics courses or exploring the diverse applications of microfluidics. The structure is around key papers and model organisms. While courses gradually change over time, a focus remains on understanding how microfluidics has developed as well as what it can and cannot do for researchers. As a primary starting point, we cover micro-fluid mechanics principles and microfabrication of devices. A variety of applications are discussed using model prokaryotic and eukaryotic organisms from the set of bacteria (Escherichia coli), trypanosomes (Trypanosoma brucei), yeast (Saccharomyces cerevisiae), slime molds (Physarum polycephalum), worms (Caenorhabditis elegans), flies (Drosophila melangoster), plants (Arabidopsis thaliana), and mouse immune cells (Mus musculus). Other engineering and biochemical methods discussed include biomimetics, organ on a chip, inkjet, droplet microfluidics, biotic games, and diagnostics. While we have not yet reached the end-all lab on a chip, microfluidics can still be used effectively for specific applications

Wireless Technologies for Implantable Devices

Wireless technologies are incorporated in implantable devices since at least the 1950s. With remote data collection and control of implantable devices, these wireless technologies help researchers and clinicians to better understand diseases and to improve medical treatments. Today, wireless technologies are still more commonly used for research, with limited applications in a number of clinical implantable devices. Recent development and standardization of wireless technologies present a good opportunity for their wider use in other types of implantable devices, which will significantly improve the outcomes of many diseases or injuries. This review briefly describes some common wireless technologies and modern advancements, as well as their strengths and suitability for use in implantable medical devices. The applications of these wireless technologies in treatments of orthopedic and cardiovascular injuries and disorders are described. This review then concludes with a discussion on the technical challenges and potential solutions of implementing wireless technologies in implantable devices