Synthesis of Polymerizable Superoxide Dismutase Mimetics to Reduce Reactive Oxygen Species Damage in Transplanted Biomedical Devices

A new polymerizable superoxide dismutase (SOD) mimetic metalloporphyrin macromer was synthesized to minimize inflammatory damage associated with tissue transplantation and biomaterial implantation, such as the use of encapsulated pancreatic islets for the treatment of type I diabetes mellitus (TIDM). This functional SOD mimetic, Mn(III) Tetrakis[1‐(3‐acryloxy‐propyl)‐4‐pyridyl] porphyrin (MnTPPyP‐Acryl), was copolymerized and crosslinked with poly(ethylene glycol) diacrylate (PEGDA) to form hydrogel networks that may actively reduce reactive oxygen species (ROS) damage associated with biomaterial implantation. Solution phase activity assays with MnTPPyP‐Acryl macromers showed comparable SOD activity to MnTMPyP, a non‐polymerizable commercially available SOD mimetic. This work also describes the development of a new, simple, and inexpensive solid phase assay system that was developed to assess the activity of MnTPPyP‐Acryl macromers polymerized within PEGDA hydrogels, which has the potential to fulfill an existing void with the biochemical tools available for testing other immobilized ROS antagonists. With this new assay system, hydrogels containing up to 0.25 mol% MnTPPyP‐Acryl showed significantly higher levels of SOD activity, whereas control hydrogels polymerized with inactive TPPyP‐Acryl macromers showed only background levels of activity. The potential for repeated use of such hydrogel devices to consistently reduce superoxide anion concentrations was demonstrated upon retention of ～100% SOD activity for at least 72 h post‐polymerization. These results demonstrate the potential that polymerizable SOD mimetics may have for integration into medical devices for the minimization of inflammatory damage upon transplantation, such as during the delivery of encapsulated pancreatic islets.     

Facile Stem Cell Delivery to Bone Grafts Enabled by Smart Shape Recovery and Stiffening of Degradable Synthetic Periosteal Membranes

Delivering stem/progenitor cells via a degradable synthetic membrane to devitalized allogenic tissue graft surfaces presents a promising allograft‐mediated tissue regeneration strategy. However, balancing degradability and bioactivity of the synthetic membrane with physical characteristics demanded for successful clinical translation is challenging. Here, well‐integrated composites of hydroxyapatite (HA) and amphiphilic poly(lactide‐co‐glycolide)‐b‐poly(ethylene glycol)‐b‐poly(lactide‐co‐glycolide) (PELGA) with tunable degradation rates are designed that stiffen upon hydration and exhibit excellent shape recovery ability at body temperature for efficiently delivering skeletal progenitor cells around bone grafts. Unlike conventional degradable polymers that weaken upon wetting, these amphiphilic composites stiffen upon hydration as a result of enhanced polyethylene glycol (PEG) crystallization. HA‐PELGA composite membranes support the attachment, proliferation, and osteogenesis of rat periosteum‐derived cells in vitro, as well as the facile transfer of confluent cell sheets of green fluorescent protein‐labeled bone marrow stromal cells. With efficient shape memory behaviors around physiological temperature, the composite membranes can be programmed with a permanent tubular configuration, deformed into a flat temporary shape desired for cell seeding/cell sheet transfer, and triggered to wrap around a femoral bone allograft upon 37 °C saline rinse and subsequently stiffen. These properties combined make electrospun HA‐PELGA promising smart synthetic periosteal membranes for augmenting allograft healing.     

Nanoliter Liquid Packaging in a Bioresorbable Microsystem by Additive Manufacturing and its Application as a Controlled Drug Delivery Device

Precise packaging of nanoliter amounts of liquid in a microsystem is important for many biomedical applications. However, existing liquid encapsulation technologies have limitations in terms of liquid waste, evaporation, trapped bubbles, and liquid degradation. In this study, multiple additive manufacturing techniques for nanoliter liquid packaging in bioresorbable microsystems is used. Two-photon photolithography is used for bioresorbable reservoir fabrication, while inkjet printing (IJP) is used for precise nanoliter liquid packaging. Dual IJP allows for micro-reservoirs to be filled with precise amounts of drug solution and subsequently and rapidly sealed with a layer of lipids mixed with Fe₃O₄ nanoparticles. Combining these two printing techniques can overcome the previous limitations of liquid encapsulation technologies. To demonstrate the relevance of this technique, a wirelessly activated, bioresorbable multi-reservoir microcapsule that can be used for controlled drug delivery is presented. The microcapsules and their content are shown to be stable during fabrication, storage, and operation. Multiple cargo release events are triggered independently by the local melting of the sealing layer, resulting from magnetically induced Fe₃O₄ nanoparticle heating. The operation of the capsule is demonstrated in tissue phantoms and in vitro cell cultures.     

Impact of Reactive Amphiphilic Copolymers on Mechanical Properties and Cell Responses of Fibrin‐Based Hydrogels

Mechanical properties of hydrogels can be modified by the variation of structure and concentration of reactive building blocks. One promising biological source for the synthesis of biocompatible hydrogels is fibrinogen. Fibrinogen is a glycoprotein in blood, which can be transformed enzymatically to fibrin playing an important role in wound healing and clot formation. In the present work, it is demonstrated that hybrid hydrogels with their improved mechanical properties, tunable internal structure, and enhanced resistance to degradation can be synthesized by a combination of fibrinogen and reactive amphiphilic copolymers. Water‐soluble amphiphilic copolymers with tunable molecular weight and controlled amounts of reactive epoxy side groups are used as reactive crosslinkers to reinforce fibrin hydrogels. In the present work, copolymers that can influence the mechanical properties of fibrin‐based hydrogels are used. The reactive copolymers increase the storage modulus of the hydrogels from 600 Pa to 30 kPa. The thickness of fibrin fibers is regulated by the copolymer concentration. It could be demonstrated that the fibrin‐based hydrogels are biocompatible and support cell proliferation. Their degradation rate is considerably slower than that of native fibrin gels. In conclusion, fibrin‐based hydrogels with tunable elasticity and fiber thickness useful to direct cell responses like proliferation and differentiation are produced.     

Localized Multi‐Component Delivery Platform Generates Local and Systemic Anti‐Tumor Immunity

As tumors employ complementary overlapping and/or independent mechanisms to evade immune surveillance, many emerging cancer immunotherapies attempt to target multiple pathways to eradicate malignant cells. Although modulation of independent pathways by simultaneous administration of multiple immune modulators (e.g., checkpoint inhibitors, cytokines, and growth factors) has shown great promise, the clinical impact remains limited due to severe toxicity associated with high systemic levels of many of these drugs. Therefore, novel platforms for efficient delivery of multi‐component therapies at lower effective doses would be enabling. Here, a drug delivery platform called immunomodulatory molecule delivery system (iMods), which provides sustained extracellular delivery of a checkpoint inhibitor (anti‐PD‐L1) and simultaneously, targeted intracellular delivery of a tumor antigen (OVA) along with adjuvant (poly(I:C)), and the indoleamine deoxygenase inhibitor 1‐MT is described. In melanoma tumor‐bearing mice, combinatorial delivery of these factors with iMods leads to regression of both treated and untreated (contralateral) melanoma tumors and 100% survival. These promising therapeutic outcomes are attributed to significantly enhanced ratios of anti‐tumor CD8 T‐cell/tumor‐protective regulatory T‐cell (Treg) in tumors and tumor draining lymph nodes. Overall, the iMods delivery platform described here represents a promising advance in multi‐factor cancer immunotherapy.     

Biomaterial Approaches for Understanding and Overcoming Immunological Barriers to Effective Oral Vaccinations

Oral vaccines have the potential to reduce cost, improve compliance, and induce immunity at mucosal surfaces that are the first line of defense against infection. The gastrointestinal tract is highly evolved to efficiently digest and absorb nutrients without eliciting aberrant inflammation. However, these functions also limit the efficacy of immunization by the oral route. While mechanisms responsible for the development of tolerance to fed antigens are being illuminated, the application of materials with precise physiochemical properties and immunomodulatory potential may accelerate understanding of immunity within the gastrointestinal tract. Insight into these immunological mechanisms can inform the design of next‐generation oral vaccines to harness critical functions to elicit immunity. Here, material strategies for oral vaccines and their dual function in understanding and overcoming immunological barriers associated with oral immunization are discussed.     

Integration of Self‐Assembled Microvascular Networks with Microfabricated PEG‐Based Hydrogels

Despite tremendous efforts, tissue engineered constructs are restricted to thin, simple tissues sustained only by diffusion. The most significant barrier in tissue engineering is insufficient vascularization to deliver nutrients and metabolites during development in vitro and to facilitate rapid vascular integration in vivo. Tissue engineered constructs can be greatly improved by developing perfusable microvascular networks in vitro in order to provide transport that mimics native vascular organization and function. Here a microfluidic hydrogel is integrated with a self‐assembling pro‐vasculogenic co‐culture in a strategy to perfuse microvascular networks in vitro. This approach allows for control over microvascular network self‐assembly and employs an anastomotic interface for integration of self‐assembled microvascular networks with fabricated microchannels. As a result, transport within the system shifts from simple diffusion to vessel supported convective transport and extra‐vessel diffusion, thus improving overall mass transport properties. This work impacts the development of perfusable prevascularized tissues in vitro and ultimately tissue engineering applications in vivo.     

Dextran Microgels for Time‐Controlled Delivery of siRNA

To apply siRNA as a therapeutic agent, appropriate attention should be paid to the optimization of the siRNA gene silencing effect, both in terms of magnitude and duration. Intracellular time‐controlled siRNA delivery could aid in tailoring the kinetics of siRNA gene knockdown. However, materials with easily tunable siRNA release properties have not been subjected to thorough investigation thus far. This report describes cationic biodegradable dextran microgels which can be loaded with siRNA posterior to gel formation. Even though the siRNAs are incorporated in the hydrogel network based on electrostatic interaction, still a time‐controlled release can be achieved by varying the initial network density of the microgels. To demonstrate the biological functionality of the siRNA loaded gels, we studied their cellular internalization and enhanced green fluorescent protein (EGFP) gene silencing potential in HUH7 human hepatoma cells.     

Cationic Hybrid Hydrogels from Amino‐Acid‐Based Poly(ester amide): Fabrication,Characterization, and Biological Properties

A family of biodegradable, biocompatible, water soluble cationic polymer precursor, arginine‐based unsaturated poly (ester amide) (Arg‐UPEA), is reported. Its incorporation into conventional Pluronic diacrylate (Pluronic‐DA) to form hybrid hydrogels for a significant improvement of the biological performance of current synthetic hydrogels is shown. The gel fraction (G_f), equilibrium swelling ratio (Q_eq), compressive modulus, and interior morphology of the hybrid hydrogels as well as their interactions with human fibroblasts and bovine endothelial cells are fully investigated. It is found that the incorporation of Arg‐UPEA into Pluronic‐DA hydrogels significantly changes their Q_eq, mechanical strength, and interior morphology. The structure–property relationship of the newly fabricated hybrid hydrogels is studied in terms of the chemical structure of the Arg‐UPEA precursor, i.e., the number of methylene groups in the Arg‐UPEA repeating unit. The results indicate that increasing methylene groups in the Arg‐UPEA repeating unit increases Q_eq and decreases the compressive modulus of hydrogels. When compared with a pure Pluronic hydrogel, the cationic Arg‐UPEAs/Pluronic hybrid hydrogels greatly improve the attachment and proliferation of human fibroblasts on hydrogel surfaces. A bovine aortic endothelial cells (BAEC) viability test in the interior of the hydrogels shows that the positively charged hybrid hydrogels can significantly improve the viability of the encapsulated endothelial cell over a 2 week study period when compared with a pure Pluronic hydrogel.     

Protein‐Release Behavior of Self‐Assembled PEG–β‐Cyclodextrin/PEG–Cholesterol Hydrogels

This paper reports on the degradation and protein release behavior of a self‐assembled hydrogel system composed of β‐cyclodextrin‐ (βCD) and cholesterol‐derivatized 8‐arm star‐shaped poly(ethylene glycol) (PEG₈). By mixing βCD‐ and cholesterol‐derivatized PEG₈ (molecular weights 10, 20 and 40 kDa) in aqueous solution, hydrogels with different rheological properties are formed. It is shown that hydrogel degradation is mainly the result of surface erosion, which depends on the network swelling stresses and initial crosslink density of the gels. This degradation mechanism, which is hardly observed for other water‐absorbing polymer networks, leads to a quantitative and nearly zero‐order release of entrapped proteins. This system therefore offers great potential for protein delivery.     

Functionally Graded,Bone‐ and Tendon‐Like Polyurethane for Rotator Cuff Repair

Critical considerations in engineering biomaterials for rotator cuff repair include bone‐tendon‐like mechanical properties to support physiological loading and biophysicochemical attributes that stabilize the repair site over the long‐term. In this study, UV‐crosslinkable polyurethane based on quadrol (Q), hexamethylene diisocyante (H), and methacrylic anhydride (M; QHM polymers), which are free of solvent, catalyst, and photoinitiator, is developed. Mechanical characterization studies demonstrate that QHM polymers possesses phototunable bone‐ and tendon‐like tensile and compressive properties (12–74 MPa tensile strength, 0.6–2.7 GPa tensile modulus, 58–121 MPa compressive strength, and 1.5–3.0 GPa compressive modulus), including the capability to withstand 10 000 cycles of physiological tensile loading and reduce stress concentrations via stiffness gradients. Biophysicochemical studies demonstrate that QHM polymers have clinically favorable attributes vital to rotator cuff repair stability, including slow degradation profiles (5–30% mass loss after 8 weeks) with little‐to‐no cytotoxicity in vitro, exceptional suture retention ex vivo (2.79–3.56‐fold less suture migration relative to a clinically available graft), and competent tensile properties (similar ultimate load but higher normalized tensile stiffness relative to a clinically available graft) as well as good biocompatibility for augmenting rat supraspinatus tendon repair in vivo. This work demonstrates functionally graded, bone‐tendon‐like biomaterials for interfacial tissue engineering.     

Recent Advances in Design of Functional Biocompatible Hydrogels for Bone Tissue Engineering

Bone related diseases have caused serious threats to human health owing to their complexity and specificity. Fortunately, owing to the unique 3D network structure with high aqueous content and functional properties, emerging hydrogels are regarded as one of the most promising candidates for bone tissue engineering, such as repairing cartilage injury, skull defect, and arthritis. Herein, various design strategies and synthesis methods (e.g., 3D-printing technology and nanoparticle composite strategy) are introduced to prepare implanted hydrogel scaffolds with tunable mechanical strength, favorable biocompatibility, and excellent bioactivity for applying in bone regeneration. Injectable hydrogels based on biocompatible materials (e.g., collagen, hyaluronic acid, chitosan, polyethylene glycol, etc.) possess many advantages in minimally invasive surgery, including adjustable physicochemical properties, filling irregular shapes of defect sites, and on-demand release drugs or growth factors in response to different stimuli (e.g., pH, temperature, redox, enzyme, light, magnetic, etc.). In addition, drug delivery systems based on micro/nanogels are discussed, and its numerous promising designs used in the application of bone diseases (e.g., rheumatoid arthritis, osteoarthritis, cartilage defect) are also briefed in this review. Particularly, several key factors of hydrogel scaffolds (e.g., mechanical property, pore size, and release behavior of active factors) that can induce bone tissue regeneration are also summarized in this review. It is anticipated that advanced approaches and innovative ideas of bioactive hydrogels will be exploited in the clinical field and increase the life quality of patients with the bone injury.     

Site‐Selectively Coated,Densely‐Packed Microprojection Array Patches for Targeted Delivery of Vaccines to Skin

Densely packed dry‐coated microprojections are shown to deliver vaccines to targeted locations within the skin that are rich in immune cells, thus inducing protective immune responses against a lethal virus challenge. Selectively limiting the antigen coating to the tips of the projections, which penetrate the skin, would significantly reduce the amount of vaccine required in immunization. In this paper a simple technique, dip‐coating the microprojections, is introduced to meet this goal. By increasing the coating solution viscosity, an otherwise strong capillary action is mitigated and the desired controlled coating length on projections is achieved. Following application to the skin, most of the coated vaccine material is rapidly released from the projections (82.6% in mass within 2 min) to the target locations within the skin strata and a potent immune response is induced when a conventional influenza vaccine (Fluvax) is tested in a mouse model. The utility of this coating approach to a variety of molecules representative of vaccines (e.g., chicken egg ovalbumin (OVA) protein, DNA, and fluorescent dyes) is demonstrated. These collective attributes, together with the simplicity of the approach, position the dip‐coating method for practical utility in large vaccination campaigns.     

Theranostic USPIO‐Loaded Microbubbles for Mediating and Monitoring Blood‐Brain Barrier Permeation

Efficient and safe drug delivery across the blood‐brain barrier (BBB) remains one of the major challenges of biomedical and (nano‐) pharmaceutical research. Here, it is demonstrated that poly(butyl cyanoacrylate)‐based microbubbles (MB), carrying ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles within their shell, can be used to mediate and monitor BBB permeation. Upon exposure to transcranial ultrasound pulses, USPIO‐MB are destroyed, resulting in acoustic forces inducing vessel permeability. At the same time, USPIO are released from the MB shell, they extravasate across the permeabilized BBB and they accumulate in extravascular brain tissue, thereby providing non‐invasive R ₂*‐based magnetic resonance imaging information on the extent of BBB opening. Quantitative changes in R ₂* relaxometry are in good agreement with 2D and 3D microscopy results on the extravascular deposition of the macromolecular model drug fluorescein isothiocyanate (FITC)‐dextran into the brain. Such theranostic materials and methods are considered to be useful for mediating and monitoring drug delivery across the BBB and for enabling safe and efficient treatment of CNS disorders.     

Porous Structures: In situ Porous Structures: A Unique Polymer Erosion Mechanism in Biodegradable Dipeptide‐Based Polyphosphazene and Polyester Blends Producing Matrices for Regenerative Engineering (Adv. Funct. Mater. 17/2010)

Synthetic biodegradable polymers serve as temporary substrates that accommodate cell infiltration and tissue in‐growth in regenerative medicine. To allow tissue in‐growth and nutrient transport, traditional three‐dimensional (3D) scaffolds must be prefabricated with an interconnected porous structure. Here we demonstrated for the first time a unique polymer erosion process through which polymer matrices evolve from a solid coherent film to an assemblage of microspheres with an interconnected 3D porous structure. This polymer system was developed on the highly versatile platform of polyphosphazene‐polyester blends. Co‐substituting a polyphosphazene backbone with both hydrophilic glycylglycine dipeptide and hydrophobic 4‐phenylphenoxy group generated a polymer with strong hydrogen bonding capacity. Rapid hydrolysis of the polyester component permitted the formation of 3D void space filled with self‐assembled polyphosphazene spheres. Characterization of such self‐assembled porous structures revealed macropores (10–100 μm) between spheres as well as micro‐ and nanopores on the sphere surface. A similar degradation pattern was confirmed in vivo using a rat subcutaneous implantation model. 12 weeks of implantation resulted in an interconnected porous structure with 82–87% porosity. Cell infiltration and collagen tissue in‐growth between microspheres observed by histology confirmed the formation of an in situ 3D interconnected porous structure. It was determined that the in situ porous structure resulted from unique hydrogen bonding in the blend promoting a three‐stage degradation mechanism. The robust tissue in‐growth of this dynamic pore forming scaffold attests to the utility of this system as a new strategy in regenerative medicine for developing solid matrices that balance degradation with tissue formation.     

Ascidian‐Inspired Fast‐Forming Hydrogel System for Versatile Biomedical Applications: Pyrogallol Chemistry for Dual Modes of Crosslinking Mechanism

Exploitation of unique biochemical and biophysical properties of marine organisms has led to the development of functional biomaterials for various biomedical applications. Recently, ascidians have received great attention, owing to their extraordinary properties such as strong underwater adhesion and rapid self‐regeneration. Specific polypeptides containing 3,4,5‐trihydroxyphenylalanine (TOPA) in the blood cells of ascidians are associated with such intrinsic properties generated through complex oxidative processes. In this study, a bioinspired hydrogel platform is developed, demonstrating versatile applicability for tissue engineering and drug delivery, by conjugating pyrogallol (PG) moiety resembling ascidian TOPA to hyaluronic acid (HA). The HA–PG conjugate can be rapidly crosslinked by dual modes of oxidative mechanisms using an oxidant or pH control, resulting in hydrogels with different mechanical and physical characteristics. The versatile utility of HA–PG hydrogels formed via different crosslinking mechanisms is tested for different biomedical platforms, including microparticles for sustained drug delivery and tissue adhesive for noninvasive cell transplantation. With extraordinarily fast and different routes of PG oxidation, ascidian‐inspired HA–PG hydrogel system may provide a promising biomaterial platform for a wide range of biomedical applications.     

Rational Design of Bioactive Hydrogels toward Periodontal Delivery: From Pathophysiology to Therapeutic Applications

Periodontitis is a biofilm-induced, host-mediated inflammatory disease that results in periodontal tissue destruction. The design of functional biomaterials based on disease pathophysiology is essential for enhancing their therapeutic effects in periodontitis treatment. As promising localized drug delivery systems and tissue engineering scaffolds, hydrogels have gained significant interest for controlled and sustained release of bioactive agents in periodontal applications. The rational design of bioactive hydrogels can facilitate bacterial control and modulate destructive host inflammation, thereby preventing the progression of periodontitis. In this review, the pathophysiological mechanisms underlying periodontitis as fundamental principles for the design of functional hydrogel systems are first introduced. In the following part, an overview is systematically provided of the types and functions of the bioactive hydrogel systems loaded with anti-bacterial and anti-inflammatory agents for periodontal delivery. Finally, the remaining challenges and future perspectives of hydrogel delivery systems for periodontal applications are proposed. It is believed that this review will inspire the rational design and development of innovative functional hydrogel biomaterials toward periodontal therapy.     

Spatiotemporally Controllable Chemical Delivery Utilizing Electroosmotic Flow Generated in Combination of Anionic and Cationic Hydrogels

Spatiotemporally controlled chemical delivery is crucial for various biomedical engineering applications. Here, a novel concept of electrically controllable delivery utilizing electroosmotic flow (EOF) generated in a combination of anionic and cationic hydrogels (A- and C-hydrogels) is reported. The unique advantages of the A/C-hydrogel combination are demonstrated utilizing a flexible sheet-shaped and a thin tubular devices. Since the directions of EOF in the A- and C-hydrogels are opposite to each other, the ionic current for EOF generation flows inside the delivery devices, enabling chemical delivery without accompanying external ionic current that could stimulate target cells and tissues. A thin tubular device, which can be inserted into narrow in vivo structures and be integrated with other flexible devices, exhibits high robustness and repeatability thanks to the flexibility and water retentivity of hydrogels. The EOF devices with A/C-hydrogels combination show high controllability superior to the pumping with a conventional syringe; the volumetric flow rate is able to be controlled proportionally to the current applied, for example, ≈0.4 µL (mA min)⁻¹ for the tubular device. The developed EOF-based devices are versatile for delivery of most chemicals regardless of their charge and size, and have great potential for both biomedical researches and therapeutics.