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.

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.

Correlating the mass and mechanical property changes during the degradation of PEG-based adhesive.

Change in mechanical property of a degrading adhesive is critical to its performance. However, characterization of degradation behavior is often limited to tracking its mass loss. 4-armed PEG end modified with dopamine (PEG-DA) was used as a model bioadhesive to correlate its change in mass with change in mechanical property. Shear modulus (G) was calculated based on the mass and average molecular weight between crosslinks ( M ¯ c ) of PEG-DA, while the storage modulus (G') was determined by oscillatory rheometry. G decreased slowly within the first week of degradation (10% reduction by week 2), while G' decreased by 60% during the same period. This large discrepancy is due to the partially disconnected and elastically ineffective PEG polymer, which is trapped within the adhesive network. This resulted in minimal mass change and higher calculated G value during the earlier time points. Therefore, tracking mass loss profile alone is inadequate to completely describe the degradation behavior of an adhesive. Additionally, PEG-DA was coated onto magnetoelastic (ME) sensors, and the change in the resonance amplitude of the sensor corresponded well with dry mass loss of PEG-DA. ME sensing provide a non-destructive method to track the mass loss of the coated adhesive.

Catechol-functionalized hydrogels: biomimetic design, adhesion mechanism, and biomedical applications.

Hydrogels are a unique class of polymeric materials that possess an interconnected porous network across various length scales from nano- to macroscopic dimensions and exhibit remarkable structure-derived properties, including high surface area, an accommodating matrix, inherent flexibility, controllable mechanical strength, and excellent biocompatibility. Strong and robust adhesion between hydrogels and substrates is highly desirable for their integration into and subsequent performance in biomedical devices and systems. However, the adhesive behavior of hydrogels is severely weakened by the large amount of water that interacts with the adhesive groups reducing the interfacial interactions. The challenges of developing tough hydrogel-solid interfaces and robust bonding in wet conditions are analogous to the adhesion problems solved by marine organisms. Inspired by mussel adhesion, a variety of catechol-functionalized adhesive hydrogels have been developed, opening a door for the design of multi-functional platforms. This review is structured to give a comprehensive overview of adhesive hydrogels starting with the fundamental challenges of underwater adhesion, followed by synthetic approaches and fabrication techniques, as well as characterization methods, and finally their practical applications in tissue repair and regeneration, antifouling and antimicrobial applications, drug delivery, and cell encapsulation and delivery. Insights on these topics will provide rational guidelines for using nature's blueprints to develop hydrogel materials with advanced functionalities and uncompromised adhesive properties.

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 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.

Antibacterial Properties of Mussel-Inspired Polydopamine Coatings Prepared by a Simple Two-Step Shaking-Assisted Method.

A simple two-step, shaking-assisted polydopamine (PDA) coating technique was used to impart polypropylene (PP) mesh with antimicrobial properties. In this modified method, a relatively large concentration of dopamine (20 mg ml-1) was first used to create a stable PDA primer layer, while the second step utilized a significantly lower concentration of dopamine (2 mg ml-1) to promote the formation and deposition of large aggregates of PDA nanoparticles. Gentle shaking (70 rpm) was employed to increase the deposition of PDA nanoparticle aggregates and the formation of a thicker PDA coating with nano-scaled surface roughness (RMS = 110 nm and Ra = 82 nm). Cyclic voltammetry experiment confirmed that the PDA coating remained redox active, despite extensive oxidative cross-linking. When the PDA-coated mesh was hydrated in phosphate saline buffer (pH 7.4), it was activated to generate 200 µM hydrogen peroxide (H2O2) for over 48 h. The sustained release of low doses of H2O2 was antibacterial against both gram-positive (Staphylococcus epidermidis) and gram-negative (Escherichia coli) bacteria. PDA coating achieved 100% reduction (LRV ~3.15) when incubated against E. coli and 98.9% reduction (LRV ~1.97) against S. epi in 24 h.

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.

Biomimetic recyclable microgels for on-demand generation of hydrogen peroxide and antipathogenic application.

Microgels that can generate antipathogenic levels of hydrogen peroxide (H2O2) through simple rehydration in solutions with physiological pH are described herein. H2O2 is a widely used disinfectant but the oxidant is hazardous to store and transport. Catechol, an adhesive moiety found in mussel adhesive proteins, was incorporated into microgels, which generated 1-5 mM of H2O2 for up to four days as catechol autoxidized. The sustained release of low concentrations of H2O2 was antimicrobial against both gram-positive (Staphylococcus epidermidis) and gram-negative (Escherichia coli) bacteria and antiviral against both non-enveloped porcine parvovirus (PPV) and enveloped bovine viral diarrhea virus (BVDV). The amount of released H2O2 is several orders of magnitude lower than H2O2 concentration previously reported for antipathogenic activity. Most notably, these microgels reduced the infectivity of the more biocide resistant non-envelope virus by 3 log reduction value (99.9% reduction in infectivity). By controlling the oxidation state of catechol, microgels can be repeatedly activated and deactivated for H2O2 generation. These microgels do not contain a reservoir for storing the reactive H2O2 and can potentially function as a lightweight and portable dried powder source for the disinfectant for a wide range of applications. STATEMENT OF SIGNIFICANCE: Researchers have designed bioadhesives and coatings using the adhesive moiety catechol to mimic the strong adhesion capability of mussel adhesive proteins. During catechol autoxidation, hydrogen peroxide (H2O2) is generated as a byproduct. Here, catechol was incorporated into microgels, which can generate millimolar levels of H2O2 by simply hydrating the microgels in a solution with physiological pH. The sustained release of H2O2 was both antimicrobial and antiviral, inactivating even the more biocide resistant non-enveloped virus. These microgels can be repeatedly activated and deactivated for H2O2 generation by incubating them in solutions with different pH. This simplicity and recyclability will enable this biomaterial to function as a lightweight and portable source for the disinfectant for a wide range of applications.

Effect of Ionic Functional Groups on the Oxidation State and Interfacial Binding Property of Catechol-Based Adhesive.

Adhesive hydrogels were prepared by copolymerizing dopamine methacrylamide with either acrylic acid (AAc) or N-(3-aminopropyl)methacrylamide hydrochloride (APMH). The effect of incorporating the anionic and cationic side chains on the oxidation state of catechol was characterized using the FOX assay to track the production of hydrogen peroxide byproduct generated during the autoxidation of catechol, and the interfacial binding property of the adhesive was determined by performing Johnson-Kendall-Roberts contact mechanics tests tested over a wide range of pH values (pH 3.0-9.0). The ionic species contributed to interfacial binding to surfaces with the opposite charge with measured work of adhesion values that were comparable to or in some cases higher than those of catechol. Addition of AAc minimized the oxidation of catechol even at a pH of 8.5 and correspondingly preserved the elevated adhesive property of catechol to both quartz and amine-functionalized surfaces. However, AAc lost its buffering capacity at pH 9.0, and catechol was oxidized at this pH. On the other hand, catechol formed a cohesive covalent bond with the network-bound amine side chain of APMH at basic pH, which interfered with the interfacial binding capability of APMH and the catechol.

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.

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.