Utilizing Robust Design to Optimize Composite Bioadhesive for Promoting Dermal Wound Repair.

Catechol-modified bioadhesives generate hydrogen peroxide (H2O2) during the process of curing. A robust design experiment was utilized to tune the H2O2 release profile and adhesive performance of a catechol-modified polyethylene glycol (PEG) containing silica particles (SiP). An L9 orthogonal array was used to determine the relative contributions of four factors (the PEG architecture, PEG concentration, phosphate-buffered saline (PBS) concentration, and SiP concentration) at three factor levels to the performance of the composite adhesive. The PEG architecture and SiP wt% contributed the most to the variation in the results associated with the H2O2 release profile, as both factors affected the crosslinking of the adhesive matrix and SiP actively degraded the H2O2. The predicted values from this robust design experiment were used to select the adhesive formulations that released 40-80 µM of H2O2 and evaluate their ability to promote wound healing in a full-thickness murine dermal wound model. The treatment with the composite adhesive drastically increased the rate of the wound healing when compared to the untreated controls, while minimizing the epidermal hyperplasia. The release of H2O2 from the catechol and soluble silica from the SiP contributed to the recruitment of keratinocytes to the wound site and effectively promoted the wound healing.

Thermomagnetic-Responsive Self-Folding Microgrippers for Improving Minimally Invasive Surgical Techniques and Biopsies.

Traditional open surgery complications are typically due to trauma caused by accessing the procedural site rather than the procedure itself. Minimally invasive surgery allows for fewer complications as microdevices operate through small incisions or natural orifices. However, current minimally invasive tools typically have restricted maneuverability, accessibility, and positional control of microdevices. Thermomagnetic-responsive microgrippers are microscopic multi-fingered devices that respond to temperature changes due to the presence of thermal-responsive polymers. Polymeric devices, made of poly(N-isopropylacrylamide-co-acrylic acid) (pNIPAM-AAc) and polypropylene fumarate (PPF), self-fold due to swelling and contracting of the hydrogel layer. In comparison, soft metallic devices feature a pre-stressed metal bilayer and polymer hinges that soften with increased temperature. Both types of microdevices can self-actuate when exposed to the elevated temperature of a cancerous tumor region, allowing for direct targeting for biopsies. Microgrippers can also be doped to become magnetically responsive, allowing for direction without tethers and the retrieval of microdevices containing excised tissue. The smaller size of stimuli-responsive microgrippers allows for their movement through hard-to-reach areas within the body and the successful extraction of intact cells, RNA and DNA. This review discusses the mechanisms of thermal- and magnetic-responsive microdevices and recent advances in microgripper technology to improve minimally invasive surgical techniques.

Electroactive Polymeric Composites to Mimic the Electromechanical Properties of Myocardium in Cardiac Tissue Repair.

Due to the limited regenerative capabilities of cardiomyocytes, incidents of myocardial infarction can cause permanent damage to native myocardium through the formation of acellular, non-conductive scar tissue during wound repair. The generation of scar tissue in the myocardium compromises the biomechanical and electrical properties of the heart which can lead to further cardiac problems including heart failure. Currently, patients suffering from cardiac failure due to scarring undergo transplantation but limited donor availability and complications (i.e., rejection or infectious pathogens) exclude many individuals from successful transplant. Polymeric tissue engineering scaffolds provide an alternative approach to restore normal myocardium structure and function after damage by acting as a provisional matrix to support cell attachment, infiltration and stem cell delivery. However, issues associated with mechanical property mismatch and the limited electrical conductivity of these constructs when compared to native myocardium reduces their clinical applicability. Therefore, composite polymeric scaffolds with conductive reinforcement components (i.e., metal, carbon, or conductive polymers) provide tunable mechanical and electroactive properties to mimic the structure and function of natural myocardium in force transmission and electrical stimulation. This review summarizes recent advancements in the design, synthesis, and implementation of electroactive polymeric composites to better match the biomechanical and electrical properties of myocardial tissue.

Controlling the Release of Hydrogen Peroxide from Catechol-Based Adhesive Using Silica Nanoparticle.

Catechol-based bioadhesives generate hydrogen peroxide (H2O2) as a byproduct during the curing process. H2O2 can have both beneficial and deleterious effects on biological systems depending on its concentration. To control the amount of H2O2 released from catechol-containing polyethylene glycol-based adhesive (PEG-DA), adhesive was formulated with silica nanoparticles (SiNP) prepared with increased porosity and acid treatment to increase Si-OH surface content. These SiNP demonstrated increased surface area, which promoted interaction with catechol and resulted in increased cure rate, bulk mechanical properties and adhesive properties of PEG-DA. Most importantly, SiNP demonstrated a 50% reduction in the released H2O2 while improving the cell viability and proliferation of three primary cell types, including rat dermal fibroblasts, human epidermal keratinocytes, and human tenocytes. Additionally, SiNP degraded into soluble Si, which also contributed to increased cell proliferation. Incorporation of porous and acid-treated SiNP can be a useful approach to simultaneously modulate the concentration of H2O2 while increasing the adhesive performance of catechol-based adhesives.

Biomimetic hydrogels with spatial- and temporal-controlled chemical cues for tissue engineering.

Biomimetic hydrogels have emerged as the most useful tissue engineering scaffold materials. Their versatile chemistry can recapitulate multiple physical and chemical features to integrate cells, scaffolds, and signaling molecules for tissue regeneration. Due to their highly hydrophilic nature hydrogels can recreate nutrient-rich aqueous environments for cells. Soluble regulatory molecules can be incorporated to guide cell proliferation and differentiation. Importantly, the controlled dynamic parameters and spatial distribution of chemical cues in hydrogel scaffolds are critical for cell-cell communication, cell-scaffold interaction, and morphogenesis. Herein, we review biomimetic hydrogels that provide cells with spatiotemporally controlled chemical cues as tissue engineering scaffolds. Specifically, hydrogels with temporally controlled growth factor-release abilities, spatially controlled conjugated bioactive molecules/motifs, and targeting delivery and reload properties for tissue engineering applications are discussed in detail. Examples of hydrogels that possess clinically favorable properties, such as injectability, self-healing ability, stimulus-responsiveness, and pro-remodeling features, are also covered.

Hydroxyl Radical Generation Through the Fenton-Like Reaction of Hematin- and Catechol-Functionalized Microgels.

Hydroxyl radical (•OH) is a potent reactive oxygen species with the ability to degrade hazardous organic compounds, kill bacteria, and inactivate viruses. However, an off-the-shelf, portable, and easily activated biomaterial for generating •OH does not exist. Here, microgels were functionalized with catechol, an adhesive moiety found in mussel adhesive proteins, and hematin (HEM), a hydroxylated Fe3+ ion-containing porphyrin derivative. When the microgel was hydrated in an aqueous solution with physiological pH, molecular oxygen in the solution oxidized catechol to generate H2O2, which was further converted to •OH by HEM. The generated •OH was able to degrade organic dyes, including orange II and malachite green. Additionally, the generated •OH was antimicrobial against both gram-negative (Escherichia coli) and gram-positive (Staphylococcus epidermidis) bacteria with the initial concentration of 106-107 CFU/mL. These microgels also reduced the infectivity of a non-enveloped porcine parvovirus and an enveloped bovine viral diarrhea virus by 3.5 and 4.5 log reduction values, respectively (99.97-99.997% reduction in infectivity). These microgels were also functionalized with positively charged [2-(methacryloyloxy)ethyl] trimethylammonium chloride (METAC), which significantly enhanced the antibacterial and antiviral activities through electrostatic interaction between the negatively charged pathogens and the microgel. These microgels can potentially serve as a lightweight and portable source of disinfectant, for an on-demand generation of •OH with a wide range of applications.

Development and Characterization of an Antimicrobial Polydopamine Coating for Conservation of Humpback Whales.

Migration patterns of humpback whales have been monitored using 316L stainless steel (SS) satellite telemetry tags. The potential for tissue infection and necrosis is increased if the bacteria, naturally a part of the diverse microbiome on the skin of humpback whales, can adhere to and colonize the surface of the tags. Polydopamine (pDA) has the potential to prevent the adhesion of one of the most prevalent bacterial strains on the surface of the skin of cetaceans (Psychrobacter) through the release of hydrogen peroxide. The release of hydrogen peroxide from the pDA coatings (40-100 µM) has the ability to induce a bacteriostatic response in E. coli, a commonly used bacteria strain in antimicrobial studies and potentially P. cryohalolentis, a common humpback associated bacteria. The H2O2 dose required to induce bacteriostatic conditions in E. coli is approximately 60 µM and in P. cryohalolentis is 100 µM. Bacterial adhesion on the surface of pDA coated SS coupons was measured first using E. coli. The coating successfully prevented adhesion of E. coli on the surface of SS coupons under certain conditions (60% reduction, p < 0.05) but the same was not seen with P. cryohalolentis. When coating conditions were altered (an increase in pH and temperature) the adhesion of P. cryohalolentis was reduced (80% reduction, p < 0.001). Overall, the pDA coatings have the capacity to generate varying amounts of hydrogen peroxide by altering the coating conditions and have the ability to reduce bacterial adhesion on the surface of satellite telemetry tags, and therefore the potential to increase tag functional service lifetime.

Development of an Injectable Nitric Oxide Releasing Poly(ethylene) Glycol-Fibrin Adhesive Hydrogel.

Fibrin microparticles were incorporated into poly(ethylene) glycol (PEG)-fibrinogen hydrogels to create an injectable, composite that could serve as a wound healing support and vehicle to deliver therapeutic factors for tissue engineering. Nitric oxide (NO), a therapeutic agent in wound healing, was loaded into fibrin microparticles by blending S-Nitroso-N-acetyl penicillamine (SNAP) with a fibrinogen solution. The incorporation of microparticles affected swelling behavior and improved tissue adhesivity of composite hydrogels. Controlled NO release was induced via photolytic and thermal activation, and modulated by weight percent of particles incorporated. These NO-releasing composites were non-cytotoxic in culture. Cells maintained morphology, viability, and proliferative character. Fibrin microparticles loaded with SNAP and incorporated into a PEG-fibrinogen matrix, creates a novel injectable composite hydrogel that offers improved tissue adhesivity and inducible NO-release for use as a regenerative support for wound healing and tissue engineering applications.

Multifunctional Biomedical Adhesives.

Currently available biomedical adhesives are mainly engineered to have one function (i.e., providing mechanical support for the repaired tissue). To improve the performance of existing bioadhesives and broaden their applications in medicine, numerous multifunctional bioadhesives are reported in the literature. These adhesives can be categorized as passive or active by design. Passive multifunctional bioadhesives contain inherent compositions and structural designs that can carry out additional functions without added external influences. These adhesives exhibit new functionalities such as antimicrobial properties, self-healing abilities, the ability to promote cellular ingrowth, and the ability to be reshaped. Conversely, active multifunctional bioadhesives respond to environmental changes (e.g., pH, temperature, electricity, light, and biomolecule concentration), which initiate a change in the adhesive to release encapsulated drugs or to activate or deactivate the bioadhesive for interfacial binding. This review article highlights recent advances in multifunctional bioadhesives.

Control of cellular adhesion and myofibroblastic character with sub-micrometer magnetoelastic vibrations.

The effect of sub-cellular mechanical loads on the behavior of fibroblasts was investigated using magnetoelastic (ME) materials, a type of material that produces mechanical vibrations when exposed to an external magnetic AC field. The integration of this functionality into implant surfaces could mitigate excessive fibrotic responses to many biomedical devices. By changing the profiles of the AC magnetic field, the amplitude, duration, and period of the applied vibrations was altered to understand the effect of each parameter on cell behavior. Results indicate fibroblast adhesion depends on the magnitude and total number of applied vibrations, and reductions in proliferative activity, cell spreading, and the expression of myofibroblastic markers occur in response to the vibrations induced by the ME materials. These findings suggest that the subcellular amplitude mechanical loads produced by ME materials could potentially remotely modulate myofibroblastic activity and limit undesirable fibrotic development.

Effect of incorporating clustered silica nanoparticles on the performance and biocompatibility of catechol-containing PEG-based bioadhesive.

A composite adhesive capable of inducing cellular infiltration was prepared by incorporating control clustered silica microparticle (MP) derived from the aggregation of silica nanoparticle (NP) into a catechol-terminated poly(ethylene glycol) bioadhesive (PEG-DA). Incorporation of MP into PEG-DA significantly improved the mechanical and adhesive properties of the bioadhesive. There was no statistical difference between the measured values for NP- and MP-incorporated adhesives, indicating that MP was equally as effective in enhancing the material properties of PEG-DA as NP. Most importantly, MP was significantly less cytotoxic when compared to NP when these particles were directly exposed to L929 fibroblast. When the adhesives were implanted subcutaneously in rats, MP-containing PEG-DA also exhibited reduced inflammatory responses, attracted elevated levels of regenerative M2 macrophage to its interface, and promoted cellular infiltration due to increased porosity within the adhesive network. Control clustered silica MP can be used to improve the performance and biocompatibility of PEG-based adhesive while minimizing undesirable cytotoxicity of silica NP.

Application of Composite Hydrogels to Control Physical Properties in Tissue Engineering and Regenerative Medicine.

The development of biomaterials for the restoration of the normal tissue structure⁻function relationship in pathological conditions as well as acute and chronic injury is an area of intense investigation. More recently, the use of tailored or composite hydrogels for tissue engineering and regenerative medicine has sought to bridge the gap between natural tissues and applied biomaterials more clearly. By applying traditional concepts in engineering composites, these hydrogels represent hierarchical structured materials that translate more closely the key guiding principles required for improved recovery of tissue architecture and functional behavior, including physical, mass transport, and biological properties. For tissue-engineering scaffolds in general, and more specifically in composite hydrogel materials, each of these properties provide unique qualities that are essential for proper augmentation and repair following disease and injury. The broad focus of this review is on physical properties in particular, static and dynamic mechanical properties provided by composite hydrogel materials and their link to native tissue architecture and, ultimately, tissue-specific applications for composite hydrogels.

Bioactive vapor deposited calcium-phosphate silica sol-gel particles for directing osteoblast behavior.

Silica-based materials are being developed and used for a variety of applications in orthopedic tissue engineering. In this work, we characterize the ability of a novel silica sol vapor deposition system to quickly modify biomaterial substrates and modulate surface hydrophobicity, surface topography, and composition. We were able to show that surface hydrophobicity, surface roughness, and composition could be rapidly modified. The compositional modification was directed towards generating apatitic-like surface mineral compositions (Ca/P ratios ∼1.30). Modified substrates were also capable of altering cell proliferation and differentiation behavior of preosteoblasts (MC3T3) and showed potential once optimized to provide a simple means to generate osteo-conductive substrates for tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2135-2148, 2016.

Monitoring the Long-Term Degradation Behavior of Biomimetic Bioadhesive Using Wireless Magnetoelastic Sensor.

The degradation behavior of a tissue adhesive is critical to its ability to repair a wound while minimizing prolonged inflammatory response. Traditional degradation tests can be expensive to perform, as they require large numbers of samples. The potential for using magnetoelastic resonant sensors to track bioadhesive degradation behavior was investigated. Specifically, biomimetic poly (ethylene glycol)- (PEG-) based adhesive was coated onto magnetoelastic (ME) sensor strips. Adhesive-coated samples were submerged in solutions buffered at multiple pH levels (5.7, 7.4 and 10.0) at body temperature (37 °C) and the degradation behavior of the adhesive was tracked wirelessly by monitoring the changes in the resonant amplitude of the sensors for over 80 days. Adhesive incubated at pH 7.4 degraded over 75 days, which matched previously published data for bulk degradation behavior of the adhesive while utilizing significantly less material (∼10(3) times lower). Adhesive incubated at pH 10.0 degraded within 25 days while samples incubated at pH 5.7 did not completely degrade even after 80 days of incubation. As expected, the rate of degradation increased with increasing pH as the rate of ester bond hydrolysis is higher under basic conditions. As a result of requiring a significantly lower amount of samples compared to traditional methods, the ME sensing technology is highly attractive for fully characterizing the degradation behavior of tissue adhesives in a wide range of physiological conditions.

Injectable dopamine-modified poly(ethylene glycol) nanocomposite hydrogel with enhanced adhesive property and bioactivity.

A synthetic mimic of mussel adhesive protein, dopamine-modified four-armed poly(ethylene glycol) (PEG-D4), was combined with a synthetic nanosilicate, Laponite (Na(0.7+)(Mg5.5Li0.3Si8)O20(OH)4)(0.7-)), to form an injectable naoncomposite tissue adhesive hydrogel. Incorporation of up to 2 wt % Laponite significantly reduced the cure time while enhancing the bulk mechanical and adhesive properties of the adhesive due to strong interfacial binding between dopamine and Laponite. The addition of Laponite did not alter the degradation rate and cytocompatibility of PEG-D4 adhesive. On the basis of subcutaneous implantation in rat, PEG-D4 nanocomposite hydrogels elicited minimal inflammatory response and exhibited an enhanced level of cellular infiltration as compared to Laponite-free samples. The addition of Laponite is potentially a simple and effective method for promoting bioactivity in a bioinert, synthetic PEG-based adhesive while simultaneously enhancing its mechanical and adhesive properties.

Application of sub-micrometer vibrations to mitigate bacterial adhesion.

As a prominent concern regarding implantable devices, eliminating the threat of opportunistic bacterial infection represents a significant benefit to both patient health and device function. Current treatment options focus on chemical approaches to negate bacterial adhesion, however, these methods are in some ways limited. The scope of this study was to assess the efficacy of a novel means of modulating bacterial adhesion through the application of vibrations using magnetoelastic materials. Magnetoelastic materials possess unique magnetostrictive property that can convert a magnetic field stimulus into a mechanical deformation. In vitro experiments demonstrated that vibrational loads generated by the magnetoelastic materials significantly reduced the number of adherent bacteria on samples exposed to Escherichia coli, Staphylococcus epidermidis and Staphylococcus aureus suspensions. These experiments demonstrate that vibrational loads from magnetoelastic materials can be used as a post-deployment activated means to deter bacterial adhesion and device infection.

Engineered nanotopography on electrospun PLLA microfibers modifies RAW 264.7 cell response.

In this study, we created a new method of electrospinning capable of controlling the surface structure of individual fibers (fiber nanotopography). The nanotopographical features were created by a phase separation in the fibers as they formed. To control the phase separation, a nonsolvent (a chemical insoluble with the polymer) was added to an electrospinning solution containing poly-l-lactic acid (PLLA) and chloroform. The nanotopography of electrospun fibers in the PLLA/chloroform solution was smooth. However, adding a small weight (<2% of total solution) of a single nonsolvent (water, ethanol, or dimethyl sulfoxide) generated nanoscale depressions on the surface of the fibers unique to the nonsolvent added. Additionally, nanoscale depressions on electrospun fibers were observed to change with dimethyl sulfoxide (DMSO) concentration in the PLLA/chloroform solution. A nonlinear relationship was found between the concentration of DMSO and the number and size of nanotopographical features. The surface depressions did not alter the hydrophobicity of the scaffold or degradation of the scaffold over a two-day period. To determine if fiber nanotopography altered cell behavior, macrophages (RAW 264.7 cells) were cultured on fibers with a smooth nanotopography or fibers with nanoscale depressions. RAW 264.7 cells spread less on fibers with nanoscale depressions than fibers with a smooth topography (p<0.05), but there were no differences between groups with regard to cell metabolism or the number of adherent cells. The results of this study demonstrate the necessity to consider the nanotopography of individual fibers as these features may affect cellular behavior. More importantly, we demonstrate a versatile method of controlling electrospun fiber nanotopography.

S-nitroso-N-acetylpenicillamine (SNAP) derivatization of peptide primary amines to create inducible nitric oxide donor biomaterials.

An S-nitroso-N-acetylpenicillamine (SNAP) derivatization approach was used to modify existing free primary amines found in fibrin (a natural protein-based biomaterial) to generate a controlled nitric oxide (NO) releasing scaffold material. The duration of the derivatization reaction affects the NO release kinetics, the induction of controlled NO-release, hydrophobicity, swelling behavior, elastic moduli, rheometric character, and degradation behavior. These properties were quantified to determine changes in fibrin hydrogels following covalent attachment of SNAP. NO-releasing materials exhibited minimal cytotoxicity when cultured with fibroblasts or osteoblasts. Cells maintained viability and proliferative character on derivatized materials as demonstrated by Live/Dead cell staining and counting. In addition, SNAP-derivatized hydrogels exhibited an antimicrobial character indicative of NO-releasing materials. SNAP derivatization of natural polymeric biomaterials containing free primary amines offers a means to generate inducible NO-releasing biomaterials for use as an antimicrobial and regenerative support for tissue engineering.

Development of vapor deposited silica sol-gel particles for use as a bioactive materials system.

Silica-based sol-gel and bioglass materials are used in a variety of biomedical applications including the surface modification of orthopedic implants and tissue engineering scaffolds. In this work, a simple system for vapor depositing silica sol-gel nano- and micro-particles onto substrates using nebulizer technology has been developed and characterized. Particle morphology, size distribution, and degradation can easily be controlled through key formulation and manufacturing parameters including water:alkoxide molar ratio, pH, deposition time, and substrate character. These particles can be used as a means to rapidly modify substrate surface properties, including surface hydrophobicity (contact angle changes >15°) and roughness (RMS roughness changes of up to 300 nm), creating unique surface topography. Ions (calcium and phosphate) were successfully incorporated into particles, and induced apatitie-like mineral formation upon exposure to simulated body fluid Preosteoblasts (MC3T3) cultured with these particles showed up to twice the adhesivity within 48 h when compared to controls, potentially indicating an increase in cell proliferation, with the effect likely due to both the modified substrate properties as well as the release of silica ions. This novel method has the potential to be used with implants and tissue engineering materials to influence cell behavior including attachment, proliferation, and differentiation via cell-material interactions to promote osteogenesis.

Fabrication of biocompatible, vibrational magnetoelastic materials for controlling cellular adhesion.

This paper describes the functionalization of magnetoelastic (ME) materials with Parylene-C coating to improve the surface reactivity to cellular response. Previous study has demonstrated that vibrating ME materials were capable of modulating cellular adhesion when activated by an externally applied AC magnetic field. However, since ME materials are not inherently biocompatible, surface modifications are needed for their implementation in biological settings. Here, the long-term stability of the ME material in an aqueous and biological environment is achieved by chemical-vapor deposition of a conformal Parylene-C layer, and further functionalized by methods of oxygen plasma etching and protein adsorption. In vitro cytotoxicity measurement and characterization of the vibrational behavior of the ME materials showed that Parylene-C coatings of 10 µm or greater could prevent hydrolytic degradation without sacrificing the vibrational behavior of the ME material. This work allows for long-term durability and functionality of ME materials in an aqueous and biological environment and makes the potential use of this technology in monitoring and modulating cellular behavior at the surface of implantable devices feasible.