In Situ Synthesis and Self-Assembly of Peptide-PEG Conjugates: A Facile Method for the Construction of Fibrous Hydrogels.

Peptide-based hydrogels have gained considerable attention as a compelling platform for various biomedical applications in recent years. Their attractiveness stems from their ability to seamlessly integrate diverse properties, such as biocompatibility, biodegradability, easily adjustable hydrophilicity/hydrophobicity, and other functionalities. However, a significant drawback is that most of the functional self-assembling peptides cannot form robust hydrogels suitable for biological applications. In this study, we present the synthesis of novel peptide-PEG conjugates and explore their comprehensive hydrogel properties. The hydrogel comprises double networks, with the first network formed through the self-assembly of peptides to create a ß-sheet secondary structure. The second network is established through covalent bond formation via N-hydroxysuccinimide chemistry between peptides and a 4-arm PEG to form a covalently linked network. Importantly, our findings reveal that this hydrogel formation method can be applied to other peptides containing lysine-rich sequences. Upon encapsulation of the hydrogel with antimicrobial peptides, the hydrogel retained high bacterial killing efficiency while showing minimum cytotoxicity toward mammalian cells. We hope that this method opens new avenues for the development of a novel class of peptide-polymer hydrogel materials with enhanced performance in biomedical contexts, particularly in reducing the potential for infection in applications of tissue regeneration and drug delivery.

Change in the geometry of positive- and negative-powered soft contact lenses during wear.

Contact lens wear causes mutual interactions between the ocular surface and the lens, which may affect comfort as well as vision. The aim of this study was to examine deformations in modern positive- and negative-powered silicone hydrogel soft contact lenses (SiH SCLs) after 7 days of continuous wear. This pre-post interventional study included 64 eyes: 42 eyes with myopia of -3.00 D and 22 eyes with hyperopia of +3.00 D. All patients underwent general ophthalmic examination, corneal topography/tomography, total corneal and epithelial thickness mapping, and specular microscopy before and after the wearing period. SiH SCLs made of senofilcon A were worn continuously for 7 days on all eligible eyes. The geometry of the new and used lenses was measured 3 to 6 minutes after removal in two perpendicular planes using a custom-made swept source optical coherence tomography (SS-OCT) system for in vitro measurements. The anterior and posterior radii of curvature decreased in -3.00 D lenses in two perpendicular planes. This effect correlated significantly with average keratometry of the cornea. Sagittal lens height was lower in +3.00 D lens after wear, which correlated moderately with the corneal sagittal height. A significant decrease in central corneal epithelial thickness was observed after wearing +3.0 D lenses. In conclusion, SiH SCLs made of senofilcon A undergo minor deformations after 7-day continuous wear. Geometry modifications are different for -3.00 D and +3.00 D lenses, and they imitate the shape of the anterior eye surface. These geometric changes are accompanied by a decrease in the central thickness of corneal epithelium after +3.00 D lens wear.

Evaluation of a cell-based osteogenic formulation compliant with good manufacturing practice for use in tissue engineering.

Proper bony tissue regeneration requires mechanical stabilization, an osteogenic biological activity and appropriate scaffolds. The latter two elements can be combined in a hydrogel format for effective delivery, so it can readily adapt to the architecture of the defect. We evaluated a Good Manufacturing Practice-compliant formulation composed of bone marrow-derived mesenchymal stromal cells in combination with bone particles (Ø = 0.25 to 1 µm) and fibrin, which can be readily translated into the clinical setting for the treatment of bone defects, as an alternative to bone tissue autografts. Remarkably, cells survived with unaltered phenotype (CD73+, CD90+, CD105+, CD31-, CD45-) and retained their osteogenic capacity up to 48 h after being combined with hydrogel and bone particles, thus demonstrating the stability of their identity and potency. Moreover, in a subchronic toxicity in vivo study, no toxicity was observed upon subcutaneous administration in athymic mice and signs of osteogenesis and vascularization were detected 2 months after administration. The preclinical data gathered in the present work, in compliance with current quality and regulatory requirements, demonstrated the feasibility of formulating an osteogenic cell-based tissue engineering product with a defined profile including identity, purity and potency (in vitro and in vivo), and the stability of these attributes, which complements the preclinical package required prior to move towards its use of prior to its clinical use.

Manufacturing considerations for producing and assessing decellularized extracellular matrix hydrogels.

Although several decellularized extracellular matrix (ECM) sheets or patches have been commercialized for use in the clinic, only one injectable decellularized ECM hydrogel, a decellularized myocardial matrix, has reached clinical trials. Consequently, very little information is available for established manufacturing standards or assessments of these materials. Here we present detailed methodology for investigating three parameters related to manufacturing optimization for a porcine derived skeletal muscle ECM hydrogel - animal-to-animal variability, bioburden reduction, and harvesting conditions. Results from characterization assays, including residual dsDNA content and sulfated glycosaminoglycan content, did not yield noteworthy differences amongst individual animals or following the addition of a bioburden reducing agent. However, the tissue collected under different harvesting conditions contained varying amounts of fat, and the protein compositions of the decellularized products differed, which could ultimately impact subsequent efficacy in vitro or in vivo. As decellularized ECM hydrogels continue to be evaluated for various applications, the differences between laboratory-scale and manufacturing-scale material batches should be thoroughly considered to avoid costly and timely optimization during scale-up.

A review on mechanical considerations for chronically-implanted neural probes.

This review intends to present a comprehensive analysis of the mechanical considerations for chronically-implanted neural probes. Failure of neural electrical recordings or stimulation over time has shown to arise from foreign body reaction and device material stability. It seems that devices that match most closely with the mechanical properties of the brain would be more likely to reduce the mechanical stress at the probe/tissue interface, thus improving body acceptance. The use of low Young's modulus polymers instead of hard substrates is one way to enhance this mechanical mimetism, though compliance can be achieved through a variety of means. The reduction of probe width and thickness in comparison to a designated length, the use of soft hydrogel coatings and the release in device tethering to the skull, can also improve device compliance. Paradoxically, the more compliant the device, the more likely it will fail during the insertion process in the brain. Strategies have multiplied this past decade to offer partial or temporary stiffness to the device to overcome this buckling effect. A detailed description of the probe insertion mechanisms is provided to analyze potential sources of implantation failure and the need for a mechanically-enhancing structure. This leads us to present an overview of the strategies that have been put in place over the last ten years to overcome buckling issues. Particularly, great emphasis is put on bioresorbable polymers and their assessment for neural applications. Finally, a discussion is provided on some of the key features for the design of mechanically-reliable, polymer-based next generation of chronic neuroprosthetic devices.

The effect of temperature on soft contact lens modulus and diameter.

PURPOSE: To examine the relative changes in diameter and modulus of soft contact lenses when the temperature is raised from room temperature (RT) to eye temperature (ET). METHODS: Thirteen lens types including 9 silicone hydrogel lenses were measured for diameter and elastic modulus at RT (20 ± 1°C) and ET (34 ± 1°C). Lens diameter measurements were undertaken after equilibration in ISO saline in a temperature-controlled lens analyzer (Optimec, Ltd, Malvern, United Kingdom). Measurements of flexural modulus of elasticity were made using an Instron 3343 tensiometer (Instron, Norwood, MA) with the samples suspended in a temperature-controlled saline bath. RESULTS: All lens types reduced in diameter when raised to ET. The largest mean changes with silicone hydrogel and conventional hydrogel lenses were with Biofinity (Δ0.35 mm) and Acuvue 2 (Δ0.28 mm), respectively. All the silicone hydrogels showed a statistically significant reduction in modulus when raised to ET ranging from Δ0.06 MPa with comfilcon A to Δ0.78 MPa with balafilcon A. All the conventional hydrogels showed relatively small changes (<0.05 MPa) in modulus. Two of the four conventional hydrogels showed a statistically significant change in modulus (etafilcon A and ocufilcon A), but these were small and believed to be clinically insignificant. CONCLUSIONS: This study has highlighted some clinically relevant changes in soft contact lens modulus and diameter when raised from RT to ET. It has also shown the importance of standardizing modulus measurement technique.

Cross-linked hydrogels for middle ear packing.

OBJECTIVE: To develop an ideal supportive packing material for ossiculoplasty, tympanoplasty, or other otologic procedures. MATERIALS AND METHODS: Several materials, namely, Carbylan-SX (P-C; Sentrx Surgical, Inc., Salt Lake City, UT), Gelfoam (P-GF; Pharmacia & Upjohn, Kalamazoo, MI), and Merogel (P-MG; Medtronics, Inc., Minneapolis, MN), were prepared and then placed into a Hartley guinea pig's (Elm Hill, Chelmsford, MA) middle ear cavities through a large myringotomy incision. The contralateral ear underwent a large myringotomy without packing material being placed. Preoperative and posteroperative auditory brainstem response studies were performed using Intelligent Hearing system software. The animals were examined weekly. Two weeks after packing placement, the animals were killed, and the temporal bones were harvested. Whole temporal bone sectioning was performed to analyze the presence of implant, surrounding inflammation, presence of osteoneogenesis and fibrosis, or adhesions. RESULTS: All the materials, except the P-MG, were easy to place into the middle ear cavity. The P-MG contains woven strands that are difficult to trim into the small sizes needed for placement. The P-MG group had a smaller average amount of implant present compared with the other groups at 2 weeks. The degree of osteoneogenesis was similar among the P-GF, P-C, and P-MG groups. The P-MG and P-C groups contained the lowest amount of fibrosis between the implant and surrounding middle ear structures. CONCLUSION: This study demonstrates promising results with P-C as a potential supportive packing material for otologic procedures. P-C compares favorably with P-MG and P-GF in a guinea pig model with respect to ease of placement and amount of fibrosis.

Smart hydrogels for in situ generated implants.

The objective of this study was to explore the use of reverse thermo-responsive (RTG) polymers for generating implants at their site of performance, following minimally invasive surgical procedures. Aiming at combining syringability and enhanced mechanical properties, a new family of injectable RTG-displaying polymers that exhibit improved mechanical properties was created, following two different strategies: (1) to synthesize high-molecular-weight polymers by covalenty joining poly(ethylene glycol) and poly(propylene glycol) chains using phosgene as the coupling molecule and (2) to cross-link poly(ethylene oxide) (PEO)-poly(propylene oxide) (PPO)-PEO triblocks after end-capping them with triethoxysilane or methacrylate reactive groups. While the methacrylates cross-linked rapidly, the triethoxysilane groups enabled the system to cross-link gradually over time. The chain-extended PEO/PPO copolymers had molecular weights in the 39 000-54 000 interval and exhibited improved mechanical properties. Reverse thermo-responsive systems displaying gradually increasing mechanical properties were generated by cross-linking triethoxysilane-capped (EO)(99)-(PO)(67)-(EO)(99) (F127) triblocks. Over time, the ethoxysilane groups hydrolyzed and created silanol moieties that subsequently condensated. With the aim of further improving their mechanical behavior, F127 triblocks were reacted with methacryloyl chloride and the resulting dimethacrylate was subsequently cross-linked in an aqueous solution at 37 degrees C. The effect of the concentration of the F127 dimethacrylate on the mechanical properties and the porous structure of the cross-linked matrixes produced was assessed. Rheometric studies revealed that the cross-linked hydrogels attained remarkable mechanical properties and allowed the engineering of robust macroscopic constructs, such as large tubular structures. The microporosity of the matrixes produced was studied by scanning electron microscopy. Monolayered conduits as well as structures comprising two and three layers were engineered in vitro, and their compliance and burst strength were determined.

Three-dimensional tissue fabrication.

In recent years, advances in fabrication technologies have brought a new dimension to the field of tissue engineering. Using manufacturing-based methods and hydrogel chemistries, researchers have been able to fabricate tissue engineering scaffolds with complex 3-D architectures and customized chemistries that mimic the in vivo tissue environment. These techniques may be useful in developing therapies for replacing lost tissue function, as in vitro models of living tissue, and also for further enabling fundamental studies of structure/function relationships in three dimensional contexts. Here, we present an overview of 3-D tissue fabrication techniques based on methods for: scaffold fabrication, cellular assembly, and hybrid hydrogel/cell methods and review their potential utility for tissue engineering.

Metal ion-sensitive holographic sensors.

Holographic sensors for Na+ and K+ have been fabricated from crown ethers incorporated into polymeric hydrogels. The methacrylate esters of a homologous series of hydroxyether crown ethers were synthesized and copolymerized with hydroxyethyl methacrylate and the cross-linker ethylene dimethacrylate (3 mol %) to form stable hydrogel films (approximately 10 m thick) containing covalently bound (0-97 mol %) 12-crown-4, 15-crown-5, and 18-crown-6 pendant functionalities. The films were transformed into silver-based volume holograms using a diffusion method coupled with a holographic recording using a frequency-doubled Nd:YAG laser. The resulting holographic reflection spectrum was used to characterize the shrinkage and swelling behavior of the holograms as a function of polymer composition and the nature and concentration of alkali, alkaline earth, and NH4+ ions in the test media. Optimized film compositions containing 50 mol % crown ether showed substantial responses (< or = 200 nm) within 30 s at ion concentrations of < or = 30 mM, which could be rationalized on the basis of the known complexation behavior of the crown ethers. An 18-crown-6 holographic film was shown to be able to quantitate K+ concentrations over the physiologically relevant range. It was virtually unaffected by variations in the Na+ background concentration within the normal physiological variation (approximately 0.13-0.15 M) and shows promise for developing simple, low-cost K+ sensors for medical applications.

pH-modulated release of insulin entrapped in a spontaneously formed hydrogel system composed of two water-soluble phospholipid polymers.

To develop a polypeptide drug carrier through oral administration, a polymer hydrogel has been found that is very easy to prepare by mixing two water-soluble phospholipid polymers. The polymers having 2-methacryloyloxyethyl phosphorylcholine (MPC) moieties spontaneously formed a hydrogel, which showed controllable dissociation via pH changes. In this study, the MPC polymer hydrogel was prepared from aqueous solutions containing water-soluble poly[MPC-co-methacrylic acid (MA)] (PMA) and poly[MPC-co-n-butyl methacrylate (BMA)] (PMB), and the applicability of the hydrogel as an oral drug delivery carrier was examined. The gelation process from the two MPC polymers is spontaneous, requiring no chemical reactions and/or no physical stimuli. PMB has a hydrophobic domain, which is suitable for loading hydrophobic drugs. Insulin could be very easily loaded to almost 100% in the hydrogel. PMA also has carboxyl groups, which are well known for pH sensitivity. At pH 1.8, the swelling continued for 8 h, with complete dissociation after 16 h. At pH 6.8, the hydrogel completely dissociated within 4 h. The hydrogel remained stable at pH 1.8 and released all the insulin at pH 6.8. The release rate was approximately four times faster at pH 6.8. After release, the insulin did not show any denaturing tendency.