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Biocompatible nanoparticles have been increasingly used in a variety of medical applications, including photodynamic therapy. Although the impact of synthesis parameters and purification methods is reported in previous studies, it is still challenging to produce a reliable protocol for the fabrication, purification, and characterization of nanoparticles in the 200-300 nm range that are highly monodisperse for biomedical applications.We investigated the synthesis of chitosan nanoparticles in the 200-300 nm range by evaluating the chitosan to sodium tripolyphosphate (TPP) mass ratio and acetic acid concentration of the chitosan solution. Chitosan nanoparticles were also crosslinked to rose bengal and incubated with human breast cancer cells (MCF-7) to test photodynamic activity using a green laser (λ = 532 nm, power = 90 mW).We established a simple protocol to fabricate and purify biocompatible nanoparticles with the most frequent size occurring between 200 and 300 nm. This was achieved using a chitosan to TPP mass ratio of 5:1 in 1% v/v acetic acid at a pH of 5.5. The protocol involved the formation of nanoparticle coffee rings that showed the particle shape to be spherical in the first approximation. Photodynamic treatment with rose bengal-nanoparticles killed ~98% of cancer cells.A simple protocol was established to prepare and purify spherical and biocompatible chitosan nanoparticles with a peak size of ~200 nm. These have remarkable antitumor activity when coupled with photodynamic treatment.

Organic electrochemical transistors (OECTs) for bioelectronic applications require the design of conjugated polymers that are stable in aqueous environments and afford high energy efficiency and good performance in OECTs. Polymers based on poly(ethylenedioxythiophene) (PEDOT) are promising in this area due to their low oxidation potential and reversible redox, but they often require cross-linking to prevent dissolution and yield OECTs operating in the less efficient depletion mode. In this work, a new conjugated polymer PEDOT-Phos is presented, which combines a conjugated poly(ethylenedioxythiophene) (PEDOT) backbone with alkyl-protected phosphonate groups. PEDOT-Phos exhibits a low oxidation onset potential (-0.157 V vs Ag/AgCl) and its nanoporous morphology affords it a high volumetric capacitance (282 ± 62 F cm–3). Without any cross-linking, additives, or post-treatment, PEDOT-Phos can be used in aqueous OECTs with efficient accumulation mode operation, long-term stability when immersed in aqueous media, low threshold voltages (-0.161 ± 0.005 V), good transconductances (9.3 ± 1.8 mS), and ON/OFF current ratios (618 ± 54) comparable to other PEDOT-based materials in OECTs. These results highlight the great promise of PEDOT-Phos as a stand-alone channel material for energy-efficient, bioelectronic devices.

Radical polymerization is one of the most widely used methods for the synthesis of polymeric materials for biomedical applications, such as drug delivery, 3D cell culture, and regenerative medicine. Among radical polymerization reactions, thiol-ene click chemistry has shown excellent orthogonality in diverse reaction conditions. However, our preliminary investigations revealed that it fails in cell culture environment. Herein, we investigate the mechanisms by which cell culture media interfere with radical photoreactions. Three different models including free radical linear photopolymerization (N,N-dimethylacrylamide photopolymerization), free radical photohydrogelation (poly(ethylene glycol) diacrylate photohydrogelation), and thiol-ene photohydrogelation (4-arm poly(ethylene glycol)-norbornene thiol-ene photohydrogelation) were investigated. We showed that common cell culture media ingredients can interfere with radical polymerization by two different pathways; namely, radical chain transfer and radical scavenging effects. Thiol-ene photoclick hydrogelation was seriously affected by cell culture media especially under the alkaline conditions of many of them, due to the impact of deprotonation of the thiol reactant. We intend these findings to serve as a reference guide to researchers employing free radical-based molecular synthesis in cell culture settings. The nonbenign impact of media components, pH, and concentration should provide a cue for future studies that aim to prepare well-defined polymeric materials in the presence of cell culture media.

The application of transparent conductive films to flexible biomedical optoelectronics is limited by stringent requirements on the candidate materials' electromechanical and optical properties as well as their biological performance. Thin films of graphene and poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) are sought as mechanically flexible alternatives to traditional indium tin oxide (ITO). However, they require more understanding of their suitability for biomedical optoelectronic devices in terms of transmission behavior and electromechanical stability. This study shows that the relative increase in sheet resistance under cyclic loading for ITO, graphene, and PEDOT:PSS was 3546±3908%,12±2.7%, and 62±68%, respectively. Moreover, graphene and PEDOT:PSS showed a transmission uniformity of 9.3% and 36.3% (380-2000 nm), respectively, compared with ITO film (61%). Understanding the optical, electrical, and mechanical limits of the transparent conductive films facilitates the optimization of flexible optoelectronic designs to fit multiple biomedical research and clinical applications.

Organic bioelectronics based on conjugated polymers as the active electronic material have been shown to operate efficiently at the biointerface. Their translation into a commercial medical device ...

Hydrogels based on cellulose nanofibrils (CNFs) have been widely used as scaffolds for biomedical applications, however, the poor mechanical properties of CNF hydrogels limit their use as ink for 3D bioprinting in order to generate scaffolds for tissue engineering applications. In this study, a dual crosslinkable hydrogel ink composed of a poly(ethylene glycol) (PEG) star polymer and 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO)-oxidized nanocellulose fibers (CNFs) is presented. As the resulting hydrogel had low structural integrity, at first crosslinking of CNFs was introduced by Ca2+. Strong physical interactions between CNFs and Ca2+ cations allowed easy regulation of the viscosity of the inks for extrusion printing raising the solution viscosity by more than 1.5 times depending on the amount of Ca2+ added. The resulting hydrogel had high structural integrity and was further stabilized in a second step by photo crosslinking of PEG under visible light. In only a few seconds, hydrogels with Young's modulus between ∼10 and 30 kPa were obtained just by altering the CNF and Ca2+ content. 3D printed hydrogels supported fibroblasts with excellent cell viability and proliferation. The dual crosslinkable hydrogel ink herein developed is versatile, easy to prepare, and suitable for 3D printing of bioscaffolds with highly tailored viscoelastic and mechanical properties applicable in a wide range of regenerative medicines.

L-3,4-dihydroxyphenylalanine (L-DOPA) is a naturally occurring catechol that is known to increase the adhesive strength of various materials used for tissue repair. With the aim of fortifying a porous and erodible chitosan-based adhesive film, L-DOPA was incorporated in its fabrication for stronger photochemical tissue bonding (PTB), a repair technique that uses light and a photosensitiser to promote tissue adhesion. The results showed that L-DOPA did indeed increase the tissue bonding strength of the films when photoactivated by a green LED, with a maximum strength recorded of approximately 30 kPa, 1.4 times higher than in its absence. The addition of L-DOPA also did not appreciably change the swelling, mechanical and erodible properties of the film. This study showed that strong, porous and erodible adhesive films for PTB made from biocompatible materials can be obtained through a simple inclusion of a natural additive such as L-DOPA, which was simply mixed with chitosan without any chemical modifications. In vitro studies using human fibroblasts showed no negative effect on cell proliferation indicating that these films are biocompatible. The films are convenient for various surgical applications as they can provide strong tissue support and a microporous environment for cellular infusion without the use of sutures. Statement of significance Tissue adhesives are not as strong as sutures on wounds under stress. Our group has previously demonstrated that strong sutureless tissue repair can be realised with chitosan-based adhesive films that photochemically bond to tissue when irradiated with green light. The advantage of this technique is that films are easier to handle than glues and sutures, and their crosslinking reactions can be controlled with light. However, these films are not optimal for high-tension tissue regenerative applications because of their non-porous structure, which cannot facilitate cell and nutrient exchange at the wound site. The present study resolves this issue, as we obtained a strong and porous photoactivated chitosan-based adhesive film, by simply using freeze drying and adding L-DOPA.

Donor-acceptor copolymers featuring electron-deficient isoindigo units and electron-rich 3,4-ethyl-E enedioxy (EDOT) groups are presented as new materials for accumulation mode organic electrochemi ...

Photochemical tissue bonding with chitosan-based adhesive films is an experimental surgical technique that avoids the risk of thermal tissue injuries and the use of sutures to maintain strong tissue connection. This technique is advantageous over other tissue repair methods as it is minimally invasive and does not require mixing of multiple components before or during application. To expand the capability of the film to beyond just a tissue bonding device and promote tissue regeneration, in this study, we designed bioadhesive films that could also support stem cells. The films were modified with oligomeric chitosan to tune their erodibility and made porous through freeze-drying for better tissue integration. Of note, porous adhesive films (pore diameter ∼110 μm), with 10% of the chitosan being oligomeric, could retain similar tissue bonding strengths (13-15 kPa) to that of the nonporous chitosan-based adhesives used in previous studies when photoactivated. When tested in vitro, these films exhibited a mass loss of ∼20% after 7 days, swelling ratios of ∼270-300%, a percentage elongation of ∼90%, and both a tensile strength and Young's modulus of ∼1 MPa. The physical properties of the films were suitable for maintaining the viability and multipotency of bone-marrow-derived human mesenchymal stem cells over the duration of culture. Thus, these biocompatible, photoactivated porous, and erodible adhesive films show promise for applications in controlled cell delivery and regenerative medicine.

In any sustainable electrical systems, various high-voltage (HV) power apparatuses such as electrical cable, transformer etc. play a pivotal role to transfer electrical energy from the generation end to the consumer end. The degradation due to electrical stress is very unlikely to avoid in operation. Prominently, the insulation system is mostly expected to be strong enough, and its degradation always provides a constant threat to the reliable operation. Moreover, the continual deterioration can gradually lead to the complete breakdown of HV equipment. To assess the overall dielectric properties, the regular monitoring/diagnosing of electrical insulation has a paramount importance in power systems. One of the potential tools for diagnostic assessment is the tan-delta (tan δ ) measurement. Considering different aspects at various frequencies, this study elucidates a comprehensive review of tan δ measurement. First, a detailed review and explanation of multiple challenges associated with dielectric loss measurement are presented. Various dielectric losses, their contribution to tan δ , measurement techniques and their comparisons are also discussed. This study further reviews the analogy of different physical dipolar theories, modelling and a brief analysis of commercially available measurement instruments.

The effect of introducing pores in bioelectronic patches was investigated. Using laser ablation, pores of different sizes and distances were introduced in a chitosan film. Polyaniline was then polymerised on the surface to produce bioelectronic patches with tailored large surface areas and pore densities varying from ∼4% to 40%. Conductivity measurements confirmed that the polyaniline formed a connected conductive network with values ranging from 0.22 ± 0.04 S cm−1 (lowest porosity) to 0.08 ± 0.01 S cm−1 (highest porosity). Their mechanical properties also varied as function of porosity. As a potential bioelectronic patch for cardiac applications, electronic fatigue was investigated in response to cyclic stretching mimicking the heart contraction cycle. Patches exhibited either a decrease in resistance (low porosity patch) or a minimal increase in resistance (

Translation into the clinic of organic bioelectronic devices having conjugated polymers as the active material will hinge on their long-term operation in vivo. This will require the device to be subject to clinically approved sterilization techniques without a deterioration in its physical and electronic properties. To date, there remains a gap in the literature addressing the impact of this critical pre-operative procedure on the properties of conjugated polymers. This study aims to address this gap by assessing the physical and electronic properties of a sterilized porous bioelectronic patch having polyaniline as the conjugated polymer. The patch was sterilized by autoclave, ethylene oxide and gamma (γ-) irradiation at 15, 25, and 50 kGy doses. Autoclaving resulted in cracking and macroscopic degradation of the patch, while patches sterilized by γ-irradiation at 50 kGy exhibited reduced mechanical and electronic properties, attributed to chain scission and non-uniform crosslinking caused by the high dose irradiation. Ethylene oxide and γ-irradiation at 15 and 25 kGy sterilization appeared to be the most effective at maintaining the mechanical and electronic properties of the patch, as well as inducing a minimal immune response as revealed by a receding fibrotic capsule after 4 weeks implantation. Our findings pave the way towards closing the gap for the translation of organic bioelectronic devices from acute to long-term in vivo models.

Conducting polymers (CPs) are organic semiconductors that have gained popularity in more recent years as components of bioelectronic devices designed to electrically communicate with biological environments. Synergy between the material and biological tissue, both on a structural and functional level, is paramount for the proper performance of an implantable biomedical device. As such, significant progress has been made on understanding the fundamental impact of the molecular and macro structure of CPs on their functional properties such as conductivity and charge mobility. At the same time, the development of a variety of synthetic approaches has yielded a library of CPs with improved mechanical and electronic properties. Specifically, chemical biofunctionalization of CP films has significantly decreased the foreign body response, the main contributor to device failure. Therefore, this review covers the advances and challenges made in the chemical biofunctionalization of CP films for potential implantable devices. This is achieved by covalently attaching the biocompatible or biofunctional group to the CP backbone via a reactive functional group to create a material with physical and electronic properties that better matches biological tissue. A perspective is presented that this synthetic chemistry approach to biofunctionalization is valuable for the integration of CPs into commercial implantable bioelectronic devices.

Infections caused by drug-resistant pathogens are rapidly increasing in incidence and pose an urgent global health concern. New treatments are needed to address this critical situation while preventing further resistance acquired by the pathogens. One promising approach is antimicrobial photodynamic therapy (PDT), a technique that selectively damages pathogenic cells through reactive oxygen species (ROS) that have been deliberately produced by light-activated chemical reactions via a photosensitiser. There are currently some limitations to its wider deployment, including aggregation, hydrophobicity, and sub-optimal penetration capabilities of the photosensitiser, all of which decrease the production of ROS and lead to reduced therapeutic performance. In combination with nanoparticles, however, these challenges may be overcome. Their small size, functionalisable structure, and large contact surface allow a high degree of internalization by cellular membranes and tissue barriers. In this review, we first summarise the mechanism of PDT action and the interaction between nanoparticles and the cell membrane. We then introduce the categorisation of nanoparticles in PDT, acting as nanocarriers, photosensitising molecules, and transducers, in which we highlight their use against a range of bacterial and fungal pathogens. We also compare the antimicrobial efficiency of nanoparticles to unbound photosensitisers and examine the relevant safety considerations. Finally, we discuss the use of nanoparticulate drug delivery systems in clinical applications of antimicrobial PDT.

Bioelectronic materials based on conjugated polymers are being developed in the hope to interface with electroresponsive tissues. We have recently demonstrated that a polyaniline chitosan patch can efficiently electro-couple with cardiac tissue modulating its electrophysiology. As a promising bioelectronic material that can be tailored to different types of devices, we investigate here the impact of varying the synthesis conditions and time of the in situ polymerization of aniline (An) on the sheet resistance of the bioelectronic patch. The sheet resistance increases significantly for samples that have either the lowest molar ratio of oxidant to monomer or the highest molar ratio of dopant to monomer, while the polymerization time does not have a significant effect on the electrical properties. Conductive atomic force microscopy reveals that the patch with the lowest sheet resistance has a connected network of the conductive phase. In contrast, patches with higher sheet resistances exhibit conductive areas of lower current signals or isolated conductive islands of high current signals. Having identified the formulation that results in patches with optimal electrical properties, we used it to fabricate patches that were implanted in rats for two weeks. It is shown that the patch retains an electroactive nature, and only mild inflammation is observed with fibrous tissue encapsulating the patch.

The clinical use of photodynamic therapy (PDT) with rose bengal (RB) is emerging as an effective t reatment for a range of applications given its non-invasive and localised mode of delivery. In particular, rose bengal PDT has shown promising antifungal action in vitro. While focus has largely been on the physical and chemical impacts of PDT on the cell, an understanding of the role of genetics underpinning the cellular response is still limited. We have, therefore, reported a screen of the entire non-essential gene library of the model organism, Saccharomyces cerevisiae, using rose bengal PDT to ascertain the key genetic pathways affecting fungal tolerance to PDT. We also investigated the dosage of PDT required to eradicate Trichophyton rubrum spores, the main causative organism of onychomycosis infection. Following this, we conducted a pilot patient study of six patients (seven toenails) for the treatment of onychomycosis using rose bengal PDT (140 μM RB and ~763 J/cm2 green light), where the clinical treatment protocol was developed on the basis of the in vitro outcomes. The key biochemical pathways identified by the genetic screen as having altered tolerance to PDT included ergosterol biosynthesis, vacuolar acidification, and purine/S-adenosyl-L-methionine biosynthesis. The subsequent pilot patient study saw the complete cure of onychomycosis for all patients within three to five treatment sessions in the absence of pain or other local side effects. The outcome of the genetic screen for tolerance may thus inform the development of efficient clinical treatments using rose bengal PDT.

Since the permeation of the inflammatory cytokines into hydrogel scaffolds has been shown to cause dysfunction of encapsulated cells, appropriate design strategies to circumvent this are essential. In the present work, it was hypothesized that highly crosslinked PVA-fucoidan and PVA-carrageenan hydrogels can control permeation of the trefoil-shaped inflammatory cytokine IL-1β while allowing the permeation of the globular protein albumin. PVA, fucoidan, and carrageenans were functionalized with methacrylate groups and the functionalized polymers were co-crosslinked by UV photopolymerization. The resultant hydrogels were characterized physicochemically and the release of fucoidan and carrageenans was quantified by developing a colorimetric assay, which was validated by XPS analysis. The permeability characteristics of the hydrogels were evaluated using bovine serum albumin (BSA), IgG, and IL-1β. The results demonstrated an increase in hydrogel swelling through the incorporation of the polysaccharides with minimal overall mass loss. The release studies showed hydrogel stability, where the formulations exhibited ~43% retention of fucoidan and ~60-80% retention of carrageenans consistently up to 7 days. The permeation data revealed very low permeation of IgG and IL-1β through the hydrogels, with1% permeation after 24 h, while allowing6% permeation of BSA. These data indicate that such hydrogels can be used as the basis for cytokine-protective implantable devices for clinical applications.

The organic electrochemical transistor (OECT) with a conjugated polymer as the active material is the elementary unit of organic bioelectronic devices. Improved functionalities, such as low power c ...

Interaction of conjugated polymers with liposomes is an attractive approach that benefits from both systems' characteristics such as electroactivity and enhanced interaction with cells. Conjugated polymer-liposome complexes have been investigated for bioimaging, drug delivery, and photothermal therapy. Their fabrication has largely been achieved by multistep procedures that require first the synthesis and processing of the conjugated polymer. Here, a new one step fabrication approach is reported based on in situ polymerization of a conjugated monomer precursor around liposomes. Polyaniline (PANI) doped with phytic acid is synthesized via oxidative polymerization in the presence of 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) vesicles to produce a conductive aqueous suspension of Liposome-PANI complexes. PANI interacts with liposomes without disrupting the bilayer as shown using differential scanning calorimetry and fluorescence quenching studies of the hydrophobic Nile red probe. The electronic conductivity of the Liposome-PANI complexes, which stems from the doped PANI accessible on the liposome surface, is confirmed using conductive atomic force microscopy and electrochemical impedance spectroscopy. Further, short-term in vitro cell studies show that the complexes colocalize with the cell membrane without reducing cell proliferation. This study presents a novel fabrication route to conductive suspensions of conjugated polymer-liposome complexes suitable for potential applications at the biointerface.

Background Extracellular matrices (ECMs) are often used in reconstructive surgery to enhance tissue regeneration and remodeling. Sutures and staples are currently used to fix ECMs to tissue although they can be invasive devices. Other sutureless and less invasive techniques, such as photochemical tissue bonding, cannot be coupled to ECMs because of their intrinsic opacity to light. Results We succeeded in fabricating a biocompatible and adhesive device that is based on ovine forestomach matrix (OFM) and a chitosan adhesive. The natural opacity of the OFM has been overcome by adding the adhesive into the matrix that allows for the light to effectively penetrate through it. The OFM-chitosan device is semitransparent (attenuation length ~ 106 µm) and can be photoactivated by green light to bond to tissue. This device does not require sutures or staples and guarantees a bonding strength of ~ 23 kPa. Conclusions A new semitransparent and biocompatible bandage has been successfully fabricated and characterized for sutureless tissue bonding.

The stem cell microenvironment or niche plays a critical role in the regulation of survival, differentiation and behavior of stem cells and their progenies. Recapitulating each aspect of the stem cell niche is therefore essential for their optimal use in in vitro studies and in vivo as future therapeutics in humans. Engineering of optimal conditions for three-dimensional stem cell culture includes multiple transient and dynamic physiological stimuli, such as blood flow and tissue stiffness. Bioprinting and microfluidics technologies, including organs-on-a-chip, are among the most recent approaches utilized to replicate the three-dimensional stem cell niche for human tissue fabrication that allow the integration of multiple levels of tissue complexity, including blood flow. This chapter focuses on the physico-chemical and genetic cues utilized to engineer the stem cell niche and provides an overview on how both bioprinting and microfluidics technologies are improving our knowledge in this field for both disease modeling and tissue regeneration, including drug discovery and toxicity high-throughput assays and stem cell-based therapies in humans.

Functionalized poly(ethylene dioxythiophene) (f-PEDOT) was copolymerized with two vinyl monomers of different hydrophilicity, acrylic acid and hydroxyethyl methacrylate, to produce electroconductive hydrogels with a range of physical and electronic properties. These hydrogels not only possessed tailored physical properties, such as swelling ratios and mechanical properties, but also displayed electroactivity dependent on the chemical composition of the network. Raman spectroscopy indicated that the functional PEDOT in the hydrogels is in an oxidized form, most likely accounting for the good electrochemical response of the hydrogels observed in physiological buffer. In vitro cell studies showed that cardiac cells respond differently when seeded on hydrogel substrates with different compositions. This study presents a facile approach for the fabrication of electroconductive hydrogels with a range of properties, paving the way for scaffolds that can meet the requirements of different electroresponsive tissues.

The application of tissue engineered patches made of conductive polymer scaffolds combined with cardiomyocytes (CMs) could provide a dual method of improving the damaged myocardium after an infarction: firstly by introducing functional CMs to the area; secondly the conductive polymer could modulate electrical transmission across the scar tissue. Polyaniline (PANI) scaffolds are one such example, however, the consequences of growing CMs on conductive PANI scaffolds with regards to CM electrophysiology are unknown. In this study we assess the hypothesis that conductive PANI scaffolds affect CM calcium transients and action potential morphology in culture. Neonatal rat ventricular myocytes (NRVMs) and neonatal rat fibroblasts were co-cultured on conductive and non-conductive (sodium hydroxide treated) PANI scaffolds and remained viable after four days of culture, covering the surface of the construct. Compared to those cultured on non-conductive PANI scaffolds, NRVM cultured on conductive PANI scaffolds show faster calcium transients, measured using Fluo-4AM and field stimulated at 1 Hz, with a decrease in the time to peak (t p non-conductive=105±6 ms, t p conductive= 85±5 ms, p50 non-conductive=212±12 ms, t 50 conductive= 116±7 ms, p90 non-conductive=404±24 ms, t 90 conductive= 266±15 ms, p

Innovative approaches to the control of immune response to tissue engineering scaffolds is of high priority. IL-10, an anti-inflammatory cytokine, has traditionally been conjugated to synthetic polymers for local immunomodulation. Marine-sulfated polysaccharides have been reported to possess anti-inflammatory properties. In the present work, it was hypothesized that photopolymerizable fucoidan and carrageenan play similar roles as the IL-10. The polysaccharides were functionalized with methacrylate groups. Their immunomodulatory properties were evaluated and compared relative to IL-10. The polysaccharides were characterized by NMR spectroscopy, revealing 12-13 % functionalization. The data revealed that fucoidan had the same activity as the IL-10 in decreasing LPS- and IFN-γ-stimulated CD86 expression. In addition, fucoidan had a protective role against LPS- and IFN-γ-induced cell growth inhibition. All polysaccharides demonstrated ∼90 % superoxide radical scavenging and they considerably decreased LPS-stimulated nitric oxide production. These results suggest that photopolymerizable fucoidan can be an alternative to IL-10 in the design of immunomodulatory biomaterials.

Conjugated polymers are the material of choice for organic bioelectronic interfaces as they combine mechanical flexibility with electric and ionic conductivity. Their attractive properties are largely demonstrated in vitro, while the in vivo applications are limited to the coating of inorganic electrodes, where they are used to improve the intimate electronic contact between the device and the tissue. However, there has not been a commensurate rise in the in vivo applications of entirely organic implantable electronic devices based on conjugated polymers. To date, there is no comprehensive understanding of how these devices will interface with real biological systems. With the push toward increasingly thinner and more flexible next generation medical implants, this limitation remains a major detractor in the translation of conjugated polymers toward biological applications. This research news article examines the few reported in vivo studies and attempts to establish why there is such a dearth in the literature.

A range of chitosan–based biomaterials have recently been used to perform sutureless, laser-activated tissue repair. Laser-activation has the advantage of bonding to tissue through a non-contact, aseptic mechanism. Chitosan adhesive films have also been shown to adhere to sheep intestine strongly without any chemical modification to chitosan. In this study, we continue to investigate chitosan adhesive films and explore the impact on the tissue repair strength and tensile strength characteristics of four types of adhesive film based on chitosan with different molecular weight and degree of deacetylation. Results showed that adhesives based on chitosan with medium molecular weight achieved the highest bonding strength, tensile strength and E-modulus when compared to the other adhesives. (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A novel chitosan adhesive film that incorporates the dye 'Rose Bengal' (RB) was used in conjunction with a green laser to repair transected rat median nerves in vivo. Histology and electrophysiological recording assessed the impact of the laser-adhesive technique on nerves. One week post-operatively, the sham-control group (laser-adhesive technique applied on un-transected nerves) conserved the average number and size of myelinated fibres in comparison to its contralateral side and electrophysiological recordings demonstrated no significant difference with un-operated nerves. Twelve weeks after the laser-adhesive anastomoses, nerves were in continuity with regenerated axons that crossed the anastomotic site.

The stem cell microenvironment or niche plays a critical role in the regulation of survival, differentiation and behavior of stem cells and their progenies. Recapitulating each aspect of the stem cell niche is therefore essential for their optimal use in in vitro studies and in vivo as future therapeutics in humans. Engineering of optimal conditions for three-dimensional stem cell culture includes multiple transient and dynamic physiological stimuli, such as blood flow and tissue stiffness. Bioprinting and microfluidics technologies, including organs-on-a-chip, are among the most recent approaches utilized to replicate the three-dimensional stem cell niche for human tissue fabrication that allow the integration of multiple levels of tissue complexity, including blood flow. This chapter focuses on the physico-chemical and genetic cues utilized to engineer the stem cell niche and provides an overview on how both bioprinting and microfluidics technologies are improving our knowledge in this field for both disease modeling and tissue regeneration, including drug discovery and toxicity high-throughput assays and stem cell-based therapies in humans.

Researchers develop sutureless conductive patch with enhanced biostability and effect on heart conduction velocity., Electrically active constructs can have a beneficial effect on electroresponsive tissues, such as the brain, heart, and nervous system. Conducting polymers (CPs) are being considered as components of these constructs because of their intrinsic electroactive and flexible nature. However, their clinical application has been largely hampered by their short operational time due to a decrease in their electronic properties. We show that, by immobilizing the dopant in the conductive scaffold, we can prevent its electric deterioration. We grew polyaniline (PANI) doped with phytic acid on the surface of a chitosan film. The strong chelation between phytic acid and chitosan led to a conductive patch with retained electroactivity, low surface resistivity (35.85 ± 9.40 kilohms per square), and oxidized form after 2 weeks of incubation in physiological medium. Ex vivo experiments revealed that the conductive nature of the patch has an immediate effect on the electrophysiology of the heart. Preliminary in vivo experiments showed that the conductive patch does not induce proarrhythmogenic activities in the heart. Our findings set the foundation for the design of electronically stable CP-based scaffolds. This provides a robust conductive system that could be used at the interface with electroresponsive tissue to better understand the interaction and effect of these materials on the electrophysiology of these tissues.

Poly(ethylene dioxythiophene) with functional pendant groups bearing double bonds is synthesized and employed for the fabrication of electroactive hydrogels with advantageous characteristics: covalently cross-linked porous 3D scaffolds with notable swelling ratio, appropriate mechanical properties, electroactivity in physiological conditions, and suitability for proliferation and differentiation of C2C12 cells. This is a new approach for the fabrication of conductive engineered constructs.

Onychomycosis, a fungal infection of the finger or toenails, is predominantly caused by Trichophyton rubrum. Treatment is difficult due to high recurrence rates and problems with treatment compliance. For these reasons, alternative therapies are needed. Here we describe the photoactivation of Rose Bengal (RB) using a green laser (λ = 532 nm) at fluences of 68, 133 and 228 J/cm2, and assess its fungicidal activity on T. rubrum spore suspensions. A 140 µM RB solution was able to induce a fungicidal effect on T. rubrum when photosensitized with the fluence of 228 J/cm2. RB photosensitization using a green laser provides a potential novel treatment for T. rubrum infections. (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A novel approach to induce disassembly of electroactive multilayer films fabricated by the layer by layer assembly technique is reported. Electroactive multilayer films are constructed using water soluble polythiophenes, i.e., negatively charged poly[ammonium (3-thienyl)ethoxypropanesulfonate] (SPT) and positively charged poly[3-(3′-thienyloxy)ethyltriethylammonium] (APT). “Induced” dissolution of the films in response to applied potential is investigated using a quartz crystal microbalance equipped with an electrochemical cell module (EC-QCM-D). Disassembly of the films is observed in response to three different potentials: +650, –650, and ±650 mV; however the time for dissolution varies as a function of the potential with films subject to +650 mV dissolving fully in 19 h compared to 42 h for films subject to –650 mV. These electroactive films and their controlled dissolution under applied potential represent an attractive architectural feature for bionic devices that could benefit from their conductivity and dissolution over time.

Conducting polymers (CPs) have exciting potential as scaffolds for tissue engineering, typically applied in regenerative medicine applications. In particular, the electrical properties of CPs has been shown to enhance nerve and muscle cell growth and regeneration. Hydrogels are particularly suitable candidates as scaffolds for tissue engineering because of their hydrated nature, their biocompatibility, and their tissue-like mechanical properties. This study reports the development of the fi rst single component CP hydrogel that is shown to combine both electro-properties and hydrogel characteristics. Poly(3-thiopheneacetic acid) hydrogels were fabricated by covalently crosslinking the polymer with 1,1 ′ -carbonyldiimidazole (CDI). Their swelling behavior was assessed and shown to display remarkable swelling capabilities (swelling ratios up to 850%). The mechanical properties of the networks were characterized as a function of the crosslinking density and were found to be comparable to those of muscle tissue. Hydrogels were found to be electroactive and conductive at physiological pH. Fibroblast and myoblast cells cultured on the hydrogel substrates were shown to adhere and proliferate. This is the fi rst time that the potential of a single component CP hydrogel has been demonstrated for cell growth, opening the way for the development of new tissue engineering scaffolds.

Injectable hydrogels have potential biomedical applications ranging from tissue fillers to drug delivery vehicles. This study focussed on evaluating the structure of poly(vinyl alcohol) (PVA) hydrogels of variable solid content and high molecular weight model drug release from the networks formed via either conventional photo-polymerization compared with chemical initiation of polymerization using an oxidation-reduction (redox) reaction. Swelling behaviour was characterised in water to assess the structural properties. Model drugs, FITC-Dextran, 20 kDa (FD20) and 4 kDa (FD4) were loaded in the hydrogels prior to curing and drug release studies conducted. Redox-cured hydrogels were more swollen than UV-cured systems, lost approximately 20% of their polymer mass compared to only 5% from UV-cured hydrogels and subsequently exhibited networks of larger mesh sizes. Also, networks of variable solid contents showed different structural properties with systems of higher polymer concentration exhibiting a smaller mesh size. The difference in structural properties of the networks affected release of FD20, being faster in redox-cured than UV-cured hydrogels, and slower from systems of higher solid content. Release of FD4 was faster than FD20 from networks of same solid content. This study suggested that PVA hydrogels can be cured by redox-initiation to function as a controlled delivery system for macromolecular drugs.

Tissue reconstruction and wound closure rely on sutures, staples and clips in current surgical procedures. These traditional devices are nonetheless unable to prevent leakage of fluids from a variety of tissue including blood vessels and dura mater. Furthermore, sutures are usually difficult to apply during minimal invasive surgery and often induce detrimental scarring that may impair healing. To overcome these disadvantages, biocompatible and biodegradable glues based on fibrin, polyethylene glycol (PEG) and cyanoacrylate have recently been used in patients to seal and repair tissue wounds. Cyanoacrylate glues create typically very strong tissue bonds but have mostly been applied externally for skin wound closure because of their residual cytotoxicity. Other adhesive biomaterials are also emerging; these glues and adhesives are usually based on proteins such as albumin and collagen or polysaccharides like chitosan; these are irradiated with coherent or non-coherent light to trigger their adhesion to tissue. These biomaterial based devices offer significant advantages over sutures, such as their sealing or repairing ability, easy application modality and delivery in situ of compounds for accelerating wound healing. This paper reviews different tissue reconstruction strategies employing adhesive biomaterials currently used in surgical and experimental procedures.

Poly (vinyl alcohol) (PVA) hydrogels are highly attractive for biomedical applications, especially for controlled release of drugs and proteins. Recently, degradable PVA hydrogels have been described, having the advantage that the material disappears over time from the implantation site. Herein, we report the synthesis of radiopaque degradable PVA, which gives a further advantage that the position of the hydrogel can precisely be determined by X-ray fluoroscopy. Radiopacity has been introduced by replacing 0.5% of the pendent alcohol groups on the PVA with 4-iodobenzoylchloride. This level of substitution rendered the polymer adequately radiopaque. The subsequent modification of 0.8% of the pendent hydroxyl groups with an ester acrylate functional group allowed for cross-linking of the macromers. The radiopaque hydrogels degraded over a time span of 140 days. Rheology data suggested that the macromer solutions were appropriate for injection.

Biocompatible, degradable hydrogel systems that can cure in situ following injection as a liquid are useful as a base for tissue engineering and drug delivery. In this study, poly (vinyl alcohol) (PVA) polymers were modified with degradable crosslinkers and formulated for either ultraviolet (UV) light initiation or chemical initiation using an oxidation/reduction (redox) curing method. A major objective was to compare the properties of degradable PVA hydrogels formed via two routes of curing. The effect of macromer concentration, degree of hydrolysis and functional group density on the degradation profiles was investigated. Also, since the hydrogels have been designed to be injected as a liquid for in situ curing, the effect of modified macromer solutions and degradation products on cell growth was investigated. Total degradation times ranged from approximately 20 days up to 120 days and increased in direct proportion with percent macromer. Initiation method (UV or redox) did not significantly impact on time for total degradation. While aqueous solutions of the modified macromer induced some cell growth inhibition, mainly associated with oxidative solutions, degradation products showed relatively low cell growth inhibition. Degradable PVA hydrogels tailored to produce networks with various degradation profiles can be cured by redox initiation and have potential as injectable polymers for soft-tissue engineering and drug delivery.

Combining electrical properties with synthetic scaffolds such as hydrogels is an attractive approach for the design of the ideal synthetic soft tissue, one that mimics the architecture of the native extracellular matrix and provides the electronic functionality needed for cell–cell communication. Conducting polymers (CPs) are carbon-based polymers that are electronically active and consequently are being investigated as the structural material for fabrication of electroactive hydrogels. CPs are attractive in that they could be processed in various forms, their chemistry could be modified to introduce different functionalities and most important is their capability to conduct electrons. In this chapter, electroconductive hydrogels (ECHs) fabricated from CP either as a single component or as an additive to conventional hydrogel networks are reviewed.

Sutureless procedures for wound repair and closure have recently integrated nanostructured devices to improve their effectiveness and clinical outcome. This review highlights the major advances in gecko-inspired bioadhesives that relies mostly on van der Waals bonding forces. These are challenged by the moist environment of surgical settings that weaken adherence to tissue. The incorporation of nanoparticles in biomatrices and their role in tissue repair and drug delivery is also reviewed with an emphasis on procedures involving adhesives that are laser-activated. Nanostructured adhesive devices have the advantage of being minimally invasive to tissue, can seal wounds, and deliver drugs in situ. All these tasks are very difficult to accomplish by sutures or staples that are invasive to host organs and often cause scarring.

Nail fungal infections are notoriously persistent and difficult to treat which can lead to severe health impacts, particularly in the immunocompromized. Current antifungal treatments, including systemic and topical drugs, are prolonged and do not effectively provide a complete cure. Severe side effects are also associated with systemic antifungals, such as hepatotoxicity. Light treatments of onychomycosis are an emerging therapy that has localized photodynamic, photothermal or photoablative action. These treatments have shown to be an effective alternative to traditional antifungal remedies with comparable or better cure rates achieved in shorter times and without systemic side effects. This report reviews significant clinical and experimental studies in the field, highlighting mechanisms of action and major effects related to light therapy; in particular, the impact of light on fungal genetics.

Effective tissue bioadhesion of rose bengal-chitosan films can be achieved by photoactivation using a green laser. In this study, lysozyme was incorporated in these films to enhance the rate of depolymerization and assess the laser impact on lysozyme. The lysozyme loaded films exhibited a 21% mass loss after 4 weeks implantation in rats while control films (without lysozyme) had only 7% mass loss. Capillary electrophoresis-mass spectroscopy showed that chitosan degraded into monomers and oligomers of glucosamine and N-acetyl-glucosamine. Irradiation with laser did not affect the depolymerization of adhesive by lysozyme suggesting that the inclusion of lysozyme in the bioadhesive is a viable technique for tailoring the depolymerization.

A range of chitosan-based biomaterials have recently been used to perform sutureless, laser-activated tissue repair. Laser-activation has the advantage of bonding to tissue through a non-contact, aseptic mechanism. Chitosan adhesive films have also been shown to adhere to sheep intestine strongly without any chemical modification to chitosan. In this study, we continue to investigate chitosan adhesive films and explore the impact on the tissue repair strength and tensile strength characteristics of four types of adhesive film based on chitosan with different molecular weight and degree of deacetylation. Results showed that adhesives based on chitosan with medium molecular weight achieved the highest bonding strength, tensile strength and E-modulus when compared to the other adhesives.

A novel chitosan adhesive film that incorporates the dye 'Rose Bengal' (RB) was used in conjunction with a green laser to repair transected rat median nerves in vivo. Histology and electrophysiological recording assessed the impact of the laser-adhesive technique on nerves. One week post-operatively, the sham-control group (laser-adhesive technique applied on un-transected nerves) conserved the average number and size of myelinated fibres in comparison to its contralateral side and electrophysiological recordings demonstrated no significant difference with un-operated nerves. Twelve weeks after the laser-adhesive anastomoses, nerves were in continuity with regenerated axons that crossed the anastomotic site.

This study describes a simple one-step fabrication process of a chitosan film that has nanoscale pillars. The adhesive is laser-bonded to tissue and does not require pillar coating to enhance bonding in water. In comparison with a ‘flat’ adhesive without pillars, the nanostructured adhesive bonded significantly stronger to tissue under either stress or pressure. This adhesive combines two mechanisms for tissue bonding: one based on the nanopatterned surface that increases the adhesion strength over short distances (van der Waals and electrostatic interactions) and another based on elastic interaction with polymer chains over much longer distances.

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two tissue edges(1,2). These are then irradiated by a laser that is selectively absorbed by the RB. The resulting photochemical reactions supposedly crosslink the collagen fibers in the tissue with minimal heat production(3). In this report, RB has been incorporated in thin chitosan films to fabricate a novel tissue adhesive that is laser-activated. Adhesive films, based on chitosan and containing ~0.1 wt% RB, are fabricated and bonded to calf intestine and rat tibial nerves by a solid state laser (λ=532 nm, Fluence~110 J/cm(2), spot size~0.5 cm). A single-column tensiometer, interfaced with a personal computer, is used to test the bonding strength. The RB-chitosan adhesive bonds firmly to the intestine with a strength of 15 ± 6 kPa, (n=30). The adhesion strength drops to 2 ± 2 kPa (n=30) when the laser is not applied to the adhesive. The anastomosis of tibial nerves can be also completed without the use of sutures. A novel chitosan adhesive has been fabricated that bonds photochemically to tissue and does not require sutures.

Onychomycosis, a fungal infection of the finger or toenails, is predominantly caused by Trichophyton rubrum. Treatment is difficult due to high recurrence rates and problems with treatment compliance. For these reasons, alternative therapies are needed. Here we describe the photoactivation of Rose Bengal (RB) using a green laser (λ = 532 nm) at fluences of 68, 133 and 228 J/cm(2) , and assess its fungicidal activity on T. rubrum spore suspensions. A 140 µM RB solution was able to induce a fungicidal effect on T. rubrum when photosensitized with the fluence of 228 J/cm(2) . RB photosensitization using a green laser provides a potential novel treatment for T. rubrum infections.