The Emerging Role of Silk Fibroin for the Development of Novel Drug Delivery Systems.

In order to reduce the toxicological impact on healthy cells and to improve the therapeutic response, many drug delivery systems have been fabricated and analysed, involving the use of different natural and synthetic materials at macro-, micro- and nanoscales. Among the natural materials which have demonstrated a huge potential for the development of effective drug delivery systems, silk fibroin has emerged for its excellent biological properties and for the possibility to be processed in a wide range of forms, which can be compliant with multiple active molecules and pharmaceutical ingredients for the treatment of various diseases. This review aims at presenting silk fibroin as an interesting biopolymer for applications in drug delivery systems, exploring the results obtained in recent works in terms of technological progress and effectiveness in vitro and in vivo.

Novel Approaches and Biomaterials for Bone Tissue Engineering: A Focus on Silk Fibroin.

Bone tissue engineering (BTE) represents a multidisciplinary research field involving many aspects of biology, engineering, material science, clinical medicine and genetics to create biological substitutes to promote bone regeneration. The definition of the most appropriate biomaterials and structures for BTE is still a challenge for researchers, aiming at simultaneously combining different features such as tissue generation properties, biocompatibility, porosity and mechanical strength. In this scenario, among the biomaterials for BTE, silk fibroin represents a valuable option for the development of functional devices because of its unique biological properties and the multiple chances of processing. This review article aims at providing the reader with a general overview of the most recent progresses in bone tissue engineering in terms of approaches and materials with a special focus on silk fibroin and the related mechanisms involved in bone regeneration, and presenting interesting results obtained by different research groups, which assessed the great potential of this protein for bone tissue engineering.

Bioinspired Materials for Wound Healing Application: The Potential of Silk Fibroin.

Nature is an incredible source of inspiration for scientific research due to the multiple examples of sophisticated structures and architectures which have evolved for billions of years in different environments. Numerous biomaterials have evolved toward high level functions and performances, which can be exploited for designing novel biomedical devices. Naturally derived biopolymers, in particular, offer a wide range of chances to design appropriate substrates for tissue regeneration and wound healing applications. Wound management still represents a challenging field which requires continuous efforts in scientific research for definition of novel approaches to facilitate and promote wound healing and tissue regeneration, particularly where the conventional therapies fail. Moreover, big concerns associated to the risk of wound infections and antibiotic resistance have stimulated the scientific research toward the definition of products with simultaneous regenerative and antimicrobial properties. Among the bioinspired materials for wound healing, this review focuses attention on a protein derived from the silkworm cocoon, namely silk fibroin, which is characterized by incredible biological features and wound healing capability. As demonstrated by the increasing number of publications, today fibroin has received great attention for providing valuable options for fabrication of biomedical devices and products for tissue engineering. In combination with antimicrobial agents, particularly with silver nanoparticles, fibroin also allows the development of products with improved wound healing and antibacterial properties. This review aims at providing the reader with a comprehensive analysis of the most recent findings on silk fibroin, presenting studies and results demonstrating its effective role in wound healing and its great potential for wound healing applications.

Photo-assisted green synthesis of silver doped silk fibroin/carboxymethyl cellulose nanocomposite hydrogels for biomedical applications.

Silver nanoparticles (AgNPs) and regenerated silk fibroin (RSF) have recently attracted significant interests for their potential applications in preventing wound-related infections and in tissue engineering. Indeed, nano-silver has long been recognized as one of the most effective antimicrobial agents, and silk fibroin is well known for its capability of stimulating cell activities and facilitating tissue regeneration. In this study, a green synthesis approach was used to create a composite hydrogel (CoHy) of RSF stabilized with CarboxymethylCellulose-Na (CMC-Na) and loaded with AgNPs. Their swelling ratios were up to 59 g/g when tested in different physiologically relevant fluids. Material characterizations by Scanning electron microscopy (SEM) with Energy Dispersive X-ray Spectroscopy (EDS), and X-Ray Diffraction (XRD) confirmed the presence of AgNPs on the surface. Antimicrobial properties of the CoHy samples were evaluated using agar diffusion tests. The results showed distinct inhibition zones against major microorganisms found in wound infections, including Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis), Methicillin Resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa (P. aeruginosa), Candida albicans (C. albicans) and Fluconazole-resistant Candida albicans (FRCA). Cytocompatibility studies with rat bone marrow derived mesenchymal stem cells (BMSCs) in vitro showed that the adhesion density of BMScs on the CoHy loaded with 1 mg/mL was similar to the cell-only control group for the first 24 h of culture; moreover, higher cell proliferation was observed on the CoHy without AgNPs, indicating the regenerative potentials of the RSF/CMC composite hydrogels.

Antimicrobial Silver Nanoparticles for Wound Healing Application: Progress and Future Trends.

Recent data have reported that the burden of infections related to antibiotic-resistant bacteria in the European Union and European Economic Area (EEA) can be estimated as the cumulative burden of tuberculosis, influenza, and human immunodeficiency virus (HIV). In wound management, the control of infections represents a crucial issue and a multi-billion dollar industry worldwide. For diabetic wounds ulcers, in particular, infections are related to the majority of amputations in diabetic patients, which today represent an increasing number of the elderly. The greatest barrier to healing is represented by the biofilm, an organized consortium of bacteria encapsulated in a self-produced extracellular polymeric substance with high resistance to conventional antimicrobial therapies. There is an urgent need for novel anti-biofilm strategies and novel antimicrobial agents and, in this scenario, silver nanotechnology has received tremendous attention in recent years in therapeutically enhanced healthcare. Due to its intrinsic therapeutic properties and the broad-spectrum antimicrobial efficacy, silver nanoparticles have opened new horizons towards novel approaches in the control of infections in wound healing. This review aims at providing the reader with an overview of the most recent progress in silver nanotechnology, with a special focus on the role of silver in the wound healing process.

Development of regenerative and flexible fibroin-based wound dressings.

Skin injuries represent a health problem with consequences in terms of morbidity, disability and life quality. Numerous strategies have been developed for the treatment of wounds, including skin substitutes, biomembranes, scaffolds, and smart dressings. The excellent properties of fibroin can be exploited for the development of advanced wound dressing biomaterials, aiming at promoting the wound healing process. In this work, silk fibroin films modified through the addition of glucose were developed to enhance flexibility of medical device without affecting the biocompatibility, to promote wound healing and to improve the patient well-being. The glucose/fibroin blend was characterized through Fourier transform infrared spectroscopy and differential scanning calorimetry to analyze the protein structure. Absorption capacity, mechanical properties, wettability and bacterial biofilm formation on silk fibroin films were also analyzed to study the effect of the addition of a plasticizer on the properties of the wound dressing. The stability of the films was analyzed through in vitro biodegradability tests. The biocompatibility and regenerative properties were demonstrated through appropriate cellular assays. The results demonstrated that the addition of glucose induced crystallization and provided good flexibility and absorption capacity of silk fibroin films. Glucose modified silk fibroin films were biocompatible and had a positive effect in promoting the wound closure. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 7-18, 2019.

A combined approach for the development of novel sutures with antibacterial and regenerative properties: the role of silver and silk sericin functionalization.

Chronic wounds and related infections cause physical and psychological distress in patients, increased mortality, disability and high health care costs. The healing process can be delayed by several factors and in particular by the risk of infections, which can be further complicated by the increasing number of antibiotic-resistant microorganisms. New approaches in wounds management have been encouraged, aiming at preventing infections and improving wound healing. In this scenario, silver has emerged as an ideal antimicrobial agent due to its recognized efficacy against bacteria, viruses and fungi. Moreover, silk and in particular silk sericin from Bombyx mori has demonstrated excellent biological properties and can be considered a good candidate for skin tissue engineering. In this study absorbable PLGA sutures were functionalized with silk sericin and, then, they were treated with silver through an in situ photochemical deposition technology in order to develop an antibacterial and regenerative biomedical device. Morphological analysis was performed by Scanning Electron Microscopy and Energy Dispersive X-Ray Spectroscopy (SEM-EDX) in order to evaluate the presence and distribution of silver deposited on the sutures. The stability and durability of the sericin/silver coatings were tested and the results were related to both antibacterial properties and sample degradation. The biological analyses also aimed at studying the biocompatibility and wound healing properties of the device, evaluating the synergistic effect between sericin and silver.

Spectroscopic Characterization and Nanosafety of Ag-Modified Antibacterial Leather and Leatherette.

The development of antibacterial coatings is of great interest from both industry and the consumer's point of view. In this study, we characterized tanned leather and polyurethane leatherette, typically employed in the automotive and footwear industries, which were modified by photo-deposition of antibacterial silver nanoparticles (AgNPs). Material surface chemical composition was investigated in detail by X-ray photoelectron spectroscopy (XPS). The material's antibacterial capability was checked against Escherichia coli and Staphylococcus aureus, as representative microorganisms in cross transmissions. Due to the presence of silver in a nanostructured form, nanosafety issues were considered, as well. Ionic release in contact media, as well as whole nanoparticle release from treated materials, were quantitatively evaluated, thus providing specific information on potential product nanotoxicity, which was further investigated through cytocompatibility MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, also after surface abrasion of the materials. The proved negligible nanoparticle release, as well as the controlled release of antibacterial ions, shed light on the materials' potentialities, in terms of both high activity and safety.

Combined Approach for the Development of Efficient and Safe Nanoantimicrobials: The Case of Nanosilver-Modified Polyurethane Foams.

Silver nanophases are routinely used as bioactive additives in commercial products. Besides their antimicrobial activity, nanosafety issues regarding the application of (silver-based) nanoantimicrobials should be considered, as well. In this study, we modified polyurethane foams, typically employed in air filtration and stuffing, by photodeposited silver nanoparticles for preparing hybrid materials (Ag-PU) with antibacterial properties. The composite materials were characterized in terms of morphology, surface chemical composition, ionic release in contact media, bioactivity, as well as whole nanoparticle release. Cytocompatibility was also assessed on 3T3 mouse fibroblasts. The proposed systematic approach allows for defining suitable composite final properties, in terms of bioactivity and safety, by properly tuning the deposition parameters.

Efficacy of silver coated surgical sutures on bacterial contamination, cellular response and wound healing.

The resistance demonstrated by many microorganisms towards conventional antibiotics has stimulated the interest in alternative antimicrobial agents and in novel approaches for prevention of infections. Silver, a natural braod-spectrum antimicrobial agent known since antiquity, has been widely employed in biomedical field due to its recognized antibacterial, antifungal and antiviral properties. In this work, antibacterial silver coatings were deposited on absorbable surgical sutures through the in situ photo-chemical deposition of silver clusters. Scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDX) and thermo-gravimetric analysis (TGA) were performed in order to investigate the presence and distribution of the silver clusters on the substrate. The amounts of silver deposited and released by the silver treated sutures were calculated through Inductively Coupled Plasma-Mass Spectroscopy (ICP-MS), and the results were related to the biodegradation of the material. The microbiological properties and the potential cytotoxicity of the silver-treated sutures were investigated in relation with hydrolysis experiments, in order to determine the effect of the degradation on antibacterial properties and biocompatibility.

The potential of photo-deposited silver coatings on Foley catheters to prevent urinary tract infections.

Catheter-associated urinary tract infection (CAUTI) represents one of the most common causes of morbidity and mortality. The resistance demonstrated by many microorganisms to conventional antibiotic therapies and the increasing health-care costs have recently encouraged the definition of alternative preventive strategies, which can have a positive effect in the management of infections. Antimicrobial urinary catheters have been developed through the photo-chemical deposition of silver coatings on the external and luminal surfaces. The substrates are exposed to ultraviolet radiation after impregnation into a silver-based solution, thus inducing the in situ synthesis of silver particles. The effect of the surface treatment on the material was investigated through scanning electron microscopy (SEM) and silver ion release measurements. The ability of microorganisms commonly associated with urinary tract infections was investigated in terms of bacterial viability, proliferation and biofilm development, using Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis as target organisms. The silver coatings demonstrated good distribution of silver particles to the substrate, and proved an effective antibacterial capability in simulated biological conditions. The low values of silver ion release demonstrated the optimum adhesion of the coating. The results indicated a good potential of silver-based antimicrobial materials for prevention of catheter-associated urinary tract infection.

Development of hybrid cotton/hydrogel yarns with improved absorption properties for biomedical applications.

Hyperhidrosis, or excessive sweating, is an overlooked and potentially disabling symptom, which is often seen in social anxiety disorder. In this work an innovative advanced textile material was developed for application in the management of excessive sweating, preparing a drying yarn providing improved comfort. Hybrid cotton/hydrogel yarns were obtained by combining cotton with superabsorbent hydrogels through an optimization study focused on the achievement of the most promising product in terms of absorption properties and resistance to washings. Swelling and washing tests were performed using different hydrogels, and the effect of an additional crosslinking on the materials was also evaluated by testing different solutions containing Al(3+) and Ca(2+) ions. Scanning electron microscopy and infrared spectroscopy analyses were adopted to characterize morphology and chemical structure of the hydrogels undergoing different production processes. The biocompatibility of the hybrid fabrics was demonstrated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide colorimetric assay (MTT) through the extract method.

In Vitro Assessment of the Antibacterial Potential of Silver Nano-Coatings on Cotton Gauzes for Prevention of Wound Infections.

Multidrug-resistant organisms are increasingly implicated in acute and chronic wound infections, thus compromising the chance of therapeutic options. The resistance to conventional antibiotics demonstrated by some bacterial strains has encouraged new approaches for the prevention of infections in wounds and burns, among them the use of silver compounds and nanocrystalline silver. Recently, silver wound dressings have become widely accepted in wound healing centers and are commercially available. In this work, novel antibacterial wound dressings have been developed through a silver deposition technology based on the photochemical synthesis of silver nanoparticles. The devices obtained are completely natural and the silver coatings are characterized by an excellent adhesion without the use of any binder. The silver-treated cotton gauzes were characterized through scanning electron microscopy (SEM) and thermo-gravimetric analysis (TGA) in order to verify the distribution and the dimension of the silver particles on the cotton fibers. The effectiveness of the silver-treated gauzes in reducing the bacterial growth and biofilm proliferation has been demonstrated through agar diffusion tests, bacterial enumeration test, biofilm quantification tests, fluorescence and SEM microscopy. Moreover, potential cytotoxicity of the silver coating was evaluated through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide colorimetric assay (MTT) and the extract method on fibroblasts and keratinocytes. Inductively coupled plasma mass spectrometry (ICP-MS) was performed in order to determine the silver release in different media and to relate the results to the biological characterization. All the results obtained were compared with plain gauzes as a negative control, as well as gauzes treated with a higher silver percentage as a positive control.

In-situ photo-assisted deposition of silver particles on hydrogel fibers for antibacterial applications.

Silver nanoparticles (AgNPs) have attracted intensive research interest and have been recently incorporated in polymers, medical devices, hydrogels and burn dressings to control the proliferation of microorganisms. In this study a novel silver antibacterial coating was deposited for the first time on hydrogel fibers through an in-situ photo-chemical reaction. Hydrogel blends obtained by mixing different percentages of silver-treated and untreated fibers were characterized by thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Four different fluids, such as phosphate buffered saline (PBS), simulated body fluid (SBF), chemical simulated wound fluid (cSWF), and deionized water (DI water), were used for evaluating the swelling properties. The results obtained confirmed that the presence of silver did not affect the properties of the hydrogel. Moreover, the results obtained through inductively coupled plasma mass spectrometry (ICP-MS) demonstrated very low silver release values, thus indicating the perfect adhesion of the silver coating to the substrate. Good antibacterial capabilities were demonstrated by any hydrogel blend on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) through agar diffusion tests and optical density readings.

Metal-Based Antibacterial Substrates for Biomedical Applications.

The interest in nanotechnology and the growing concern for the antibiotic resistance demonstrated by many microorganisms have recently stimulated many efforts in designing innovative biomaterials and substrates with antibacterial properties. Among the implemented strategies to control the incidence of infections associated with the use of biomedical device and implants, interesting routes are represented by the incorporation of bactericidal agents onto the surface of biomaterials for the prevention of bacterial adhesion and biofilm growth. Natural products and particularly bioactive metals such as silver, copper and zinc represent an interesting alternative for the development of advanced biomaterials with antimicrobial properties. This review presents an overview of recent progress in the modification of biomaterials as well as the most attractive techniques for the deposition of antimicrobial coatings on different substrates for biomedical application. Moreover, some research activities and results achieved by the authors in the development of antibacterial materials are also presented and discussed.

Silver-doped self-assembling di-phenylalanine hydrogels as wound dressing biomaterials.

Chronic and acute wounds can be quickly contaminated and infected by microorganisms such as bacteria, multi-resistant organisms or fungi. The introduction of silver as anti-microbial agent into wound management has widely been demonstrated to be effective and contribute to wound healing. As a consequence, many approaches and different materials have been employed to synthesize antibacterial silver-hydrogels. In this work the introduction of silver particles into the fibrillar structure of self-assembling aromatic di-phenylalanine derivatives modified with aromatic groups such as 9-fluorenylmethoxycarbonyl is proposed to produce antibacterial wound dressings. Hydrogels doped with increasing amounts of silver were tested and adopted to modify flax textiles. The influence of silver on the structure of hydrogels was studied using light and confocal microscopy, while SEM-EDX allowed the characterization of the hydrogel coating on the surface of the textile substrates as well as the identification and distribution of silver nanoparticles. The antibacterial potential of the treated flax was demonstrated through microbiological tests on Staphylococcus aureus. The combination of the physico-chemical and anti-bacterial properties, together with the ease of preparation of these biomaterials, fulfils the requirement of clinically-effective wound dressings.