Qiongyu Guo

Photoactivated Composite Biomaterial for Soft Tissue Restoration in Rodents and in Humans

Photoactivated composite poly(ethylene glycol)–hyaluronic acid biomaterials demonstrate enhanced physicochemical properties for facial soft tissue reconstruction. Photogenic Polymers Can Fix the Flaws Some people just love the spotlight; apparently, some polymers do too. Here, Hillel et al. introduce a class of composite polymers that react favorably to light by crosslinking within minutes. These polymers, composed of synthetic poly(ethylene glycol) (PEG) and natural hyaluronic acid (HA), have been developed for reconstructing facial soft tissue. Deformities in craniofacial soft tissue are a clinical challenge because even small defects can have a major impact on a person’s social behavior and psychological well-being. Hillel and colleagues created a polymeric composite that can be injected into the damaged site, massaged into shape, and then crosslinked in situ with light. A transdermal light exposure method would allow clinicians to inject a liquid polymer, rather than surgically inserting already-polymerized material. First, the authors designed an array of light-emitting diodes to penetrate up to 4 mm of human skin (both light and dark) without any painful side effects. A 2-min exposure to light was enough to crosslink the PEG-HA material under the skin. Next, the polymer was tailored to closely match the elastic properties of native soft tissues, such as human fat. Various amounts of PEG and concentrations of HA were tested, with the authors arriving at an optimal combination of 100 mg PEG and 24 mg/ml HA. When polymerized subcutaneously in rats, the PEG-HA implants were able to maintain near their original volume for up to 491 days, whereas control HA injections were completely resorbed. Notably, these HA-based materials were partially reversible with the addition of the enzyme hyaluronidase. To translate this material to the clinic, Hillel et al. then tested the PEG-HA composites in humans. The polymer was injected into the intradermal space in the abdomen of three patients scheduled to undergo abdominoplasty surgery. Similar to the rodent studies, the PEG-HA material persisted for 12 weeks, whereas the control HA injections lost their shape. An inflammatory response was observed surrounding the injections. It is clear that this new photo-friendly polymer and transdermal crosslinking method will be clinically useful for soft tissue reconstruction—perhaps even encouraging more people to put their best faces forward in the spotlight. Soft tissue reconstruction often requires multiple surgical procedures that can result in scars and disfiguration. Facial soft tissue reconstruction represents a clinical challenge because even subtle deformities can severely affect an individual’s social and psychological function. We therefore developed a biosynthetic soft tissue replacement composed of poly(ethylene glycol) (PEG) and hyaluronic acid (HA) that can be injected and photocrosslinked in situ with transdermal light exposure. Modulating the ratio of synthetic to biological polymer allowed us to tune implant elasticity and volume persistence. In a small-animal model, implanted photocrosslinked PEG-HA showed a dose-dependent relationship between increasing PEG concentration and enhanced implant volume persistence. In direct comparison with commercial HA injections, the PEG-HA implants maintained significantly greater average volumes and heights. Reversibility of the implant volume was achieved with hyaluronidase injection. Pilot clinical testing in human patients confirmed the feasibility of the transdermal photocrosslinking approach for implantation in abdomen soft tissue, although an inflammatory response was observed surrounding some of the materials.

Structure and properties of collagen vitrigel membranes for ocular repair and regeneration applications

The frequency of ocular injuries on the battlefield has been steadily increasing during recent conflicts. Combat-related eye injuries are difficult to treat and solutions requiring donor tissue are not ideal and are often not readily available. Collagen vitrigels have previously been developed for corneal reconstruction, but increased transparency and mechanical strength are desired for improved vision and ease of handling. In this study, by systematically varying vitrification temperature, relative humidity and time, the collagen vitrigel synthesis conditions were optimized to yield the best combination of high transparency and high mechanical strength. Optical, mechanical, and thermal properties were characterized for each set of conditions to evaluate the effects of the vitrification parameters on material properties. Changes in denaturing temperature and collagen fibril morphology were evaluated to correlate properties with structure. Collagen vitrigels with transmittance up to 90%, tensile strength up to 12 MPa, and denaturing temperatures that significantly exceed the eye/body temperature have been synthesized at 40 °C and 40% relative humidity for one week. This optimal set of conditions enabled improvements of 100% in tensile strength and 11% in transmittance, compared to the previously developed collagen vitrigels.

Light activated cell migration in synthetic extracellular matrices

Synthetic extracellular matrices provide a framework in which cells can be exposed to defined physical and biological cues. However no method exists to manipulate single cells within these matrices. It is desirable to develop such methods in order to understand fundamental principles of cell migration and define conditions that support or inhibit cell movement within these matrices. Here, we present a strategy for manipulating individual mammalian stem cells in defined synthetic hydrogels through selective optical activation of Rac, which is an intracellular signaling protein that plays a key role in cell migration. Photoactivated cell migration in synthetic hydrogels depended on mechanical and biological cues in the biomaterial. Real-time hydrogel photodegradation was employed to create geometrically defined channels and spaces in which cells could be photoactivated to migrate. Cell migration speed was significantly higher in the photo-etched channels and cells could easily change direction of movement compared to the bulk hydrogels.

Application of a Collagen-Based Membrane and Chondroitin Sulfate-Based Hydrogel Adhesive for the Potential Repair of Severe Ocular Surface Injuries

This study was performed to evaluate the potential of a chondroitin sulfate-polyethylene glycol (CS-PEG) adhesive and collagen-based membrane (collagen vitrigel, CV) combination as a method to treat penetrating ocular injuries on the battlefield and to improve this method with two technologies: an antibiotic releasing CS-PEG adhesive and a corneal shaped CV. Burst testing using porcine cadaveric eyes, high-performance liquid chromatography, the Kirby-Bauer bacterial inhibition test, and CV implantations on the live and cadaveric rabbit eyes were performed. The ocular burst test showed CS-PEG adhesive could successfully repair 5-mm to 6-mm length wounds in the corneal and corneoscleral regions but would require CS-PEG + CV to treat larger wounds similar to those seen on the battlefield. In addition, high performance liquid chromatography and the Kirby-Bauer bacterial inhibition test presented evidence suggesting the vancomycin incorporated CS-PEG could inhibit Staphylococcus infection for 9 days. Furthermore, the curved CV showed an advantage by matching the corneal contour without any wrinkle formation. Although this pilot study showed a limited range of possible applications, we demonstrated that the combination of CS-PEG adhesive + CV is a promising method and the 2 technologies improve their applicability to the special demands of the battlefield.

Fibre-reinforced hydrogels with high optical transparency

Abstract Fibre-reinforced hydrogels with high optical transparency are an emerging composite material with great promise to enable new applications, such as a transparent wound dressing with custom tuned mechanical properties that provides desirable mechanical and physical properties along with optical clarity for facile wound inspection. Stand-alone hydrogels are an important class of materials comprising a cross-linked polymer network surrounded by a water matrix. However, their mechanical properties are typically very modest compared with other materials. While significant research is going on in parallel in the fields of hydrogels and reinforcement fibres, researchers are only starting to scratch the surface of the possibilities of combining the two. This report provides a review of natural and synthetic reinforcement fibre research with special emphasis placed on nanofibres. These provide the added benefit of transparency by being much smaller than the wavelength of visible light. A review of hydrogel materials is also presented. The mechanical properties, optical properties and biological functionality of hydrogel systems are also described. Ocular, load-bearing tissue, wound management and sensing/device applications are all discussed. Transparent fibre-reinforced hydrogels provide a compelling potential solution to enable advanced functionality, in particular in the wound care and optical application areas.

Moxifloxacin in situ gelling microparticles-bioadhesive delivery system

Antibiotic use for ocular treatments has been largely limited by poor local bioavailability with conventional eyedrops formulations. Here, we developed a controlled delivery system composed of moxifloxacin-loaded poly(lactic-co-glycolic acid) (PLGA) microparticles encapsulated in a chondroitin sulfate-based, two-component bioadhesive hydrogel. Using a simple and fast electrohydrodynamic spray drying (electrospraying) technique, surfactant-free moxifloxacin-loaded microparticles were fabricated with diameters on the order of 1 μm. A mixed solvent system of methanol/dichloromethane (MeOH/DCM) was employed to prepare the microparticles for the electrospraying processing. Extended release of moxifloxacin using a series of MeOH/DCM mixed solvents was accomplished over 10 days with release concentrations higher than the minimum inhibitory concentration (MIC). In contrast, moxifloxacin loaded directly in hydrogels was released rapidly within 24 h. We observed a decrease of the drug release rate from the microparticles when using an increased percentage of methanol in the mixed solvent from 10% to 30% (v/v), which can be explained by the mixed solvent system providing a driving force to form a gradient of the drug concentrations inside the microparticles. In addition, the delivery system developed in this study, which incorporates a bioadhesive to localize drug release by in situ gelling, may potentially integrate antibiotic prophylaxis and wound healing in the eye.

Influence of collagen source on fibrillar architecture and properties of vitrified collagen membranes.

Collagen vitrigel membranes are transparent biomaterials characterized by a densely organized, fibrillar nanostructure that show promise in the treatment of corneal injury and disease. In this study, the influence of different type I collagen sources and processing techniques, including acid-solubilized collagen from bovine dermis (Bov), pepsin-solubilized collagen from human fibroblast cell culture (HuCC), and ficin-solubilized collagen from recombinant human collagen expressed in tobacco leaves (rH), on the properties of the vitrigel membranes was evaluated. Postvitrification carbodiimide crosslinking (CX) was also carried out on the vitrigels from each collagen source, forming crosslinked counterparts BovXL, HuCCXL, and rHXL, respectively. Collagen membrane ultrastructure and biomaterial properties were found to rely heavily on both collagen source and crosslinking. Bov and HuCC samples showed a random fibrillar organization of collagen, whereas rH vitrigels showed remarkable regional fibril alignment. After CX, light transmission was enhanced in all groups. Denaturation temperatures after CX increased in all membranes, of which the highest increase was seen in rH (14.71°C), suggesting improved thermal stability of the collagen fibrils in the membranes. Noncrosslinked rH vitrigels may be reinforced through CX to reach levels of mechanical strength and thermal stability comparable to Bov.

Future perspectives for regenerative medicine in ophthalmology

Repair and reconstruction of the cornea has historically relied on synthetic materials or tissue transplantation. However, the future holds promise for treatments using smart biomaterials and stem cells that direct tissue repair and regeneration to ultimately create new ocular structures that are indistinguishable from the original native tissue. The cornea is a remarkable engineering structure. By understanding the physical structure of the tissue and the resulting impact of the structure on biological function, we can design novel materials for a number of ophthalmic clinical applications. Furthermore, by extending this structure-function approach to characterizing corneal disease processes, new therapies can be engineered.

Entanglement-Based Thermoplastic Shape Memory Polymeric Particles with Photothermal Actuation for Biomedical Applications

Triggering shape-memory functionality under clinical hyperthermia temperatures could enable the control and actuation of shape-memory systems in clinical practice. For this purpose, we developed light-inducible shape-memory microparticles composed of a poly(d,l-lactic acid) (PDLLA) matrix encapsulating gold nanoparticles (Au@PDLLA hybrid microparticles). This shape-memory polymeric system for the first time demonstrates the capability of maintaining an anisotropic shape at body temperature with triggered shape-memory effect back to a spherical shape at a narrow temperature range above body temperature with a proper shape recovery speed (37 < T < 45 °C). We applied a modified film-stretching processing method with carefully controlled stretching temperature to enable shape memory and anisotropy in these micron-sized particles. Accordingly, we achieved purely entanglement-based shape-memory response without chemical cross-links in the miniaturized shape-memory system. Furthermore, these shape-memory microparticles exhibited light-induced spatiotemporal control of their shape recovery using a laser to trigger the photothermal heating of doped gold nanoparticles. This shape-memory system is composed of biocompatible components and exhibits spatiotemporal controllability of its properties, demonstrating a potential for various biomedical applications, such as tuning macrophage phagocytosis as demonstrated in this study.

Developing biomimetic collagen-based matrix using cyclodextrin for corneal repair

The worldwide demand for corneal transplantations cannot match by the supply of donor corneas, especially in most developing countries. There is a large need of developing new corneal substitute mimicking the native cornea in terms of optical, biomechanical and biological properties. The cornea has four main types of proteoglycan that play a critical role in collagen fibrogenesis and transparency. However, obtaining purified proteoglycans has been a challenging task due to their high cost. In this study, we employed cyclodextrins as a proteoglycan substitute to engineer a biomimetic collagen-based matrix. The incorporation of cyclodextrin in collagen membrane increased collagen thermal stability and reduced collagen fibrogenesis. As a result, a thick, transparent and mechanically strong collagen-based membrane was formed. This cyclodextrin-collagen membrane holds a great potential to be used as a therapeutic eye patch for corneal repair.