Photopolymerizable Hydrogel Research Articles

Photopolymerizable chitosan-collagen hydrogels for bone tissue engineering.

Photopolymerizable hydrogels derived from naturally occurring polymers have attracted significant interest in tissue-engineering applications due to their excellent biocompatibility, hydrophilic nature favourable for cell ingrowth and ability to be cured in situ through a minimally invasive procedure. In this study, we developed a composite hydrogel consisting of photocrosslinkable methacrylated glycol chitosan (MeGC) and semi-interpenetrating collagen (Col) with a riboflavin photoinitiator under blue light. The incorporation of Col in MeGC hydrogels enhanced the compressive modulus and slowed the degradation rate of the hydrogels. MeGC-Col composite hydrogels significantly enhanced cellular attachment, spreading, proliferation and osteogenic differentiation of mouse bone marrow stromal cells (BMSCs) seeded on the hydrogels compared with pure MeGC hydrogels, as observed by upregulated alkaline phosphatase (ALP) activity as well as increased mineralization. Similarly, when cells were encapsulated within hydrogels, BMSCs exhibited greater proliferation, ALP activity and mineral deposits in the presence of Col. These findings demonstrate that MeGC-Col composite hydrogels may be useful in promoting bone regeneration. Copyright © 2014 John Wiley & Sons, Ltd.

Photopolymerizable hydrogels for implants: Monte-Carlo modeling and experimentalin vitrovalidation

Photopolymerization is commonly used in a broad range of bioapplications, such as drug delivery, tissue engineering, and surgical implants, where liquid materials are injected and then hardened by means of illumination to create a solid polymer network. However, photopolymerization using a probe, e.g., needle guiding both the liquid and the curing illumination, has not been thoroughly investigated. We present a Monte Carlo model that takes into account the dynamic absorption and scattering parameters as well as solid-liquid boundaries of the photopolymer to yield the shape and volume of minimally invasively injected, photopolymerized hydrogels. In the first part of the article, our model is validated using a set of well-known poly(ethylene glycol) dimethacrylate hydrogels showing an excellent agreement between simulated and experimental volume-growth-rates. In the second part, in situ experimental results and simulations for photopolymerization in tissue cavities are presented. It was found that a cavity with a volume of 152 mm3 can be photopolymerized from the output of a 0.28-mm2 fiber by adding scattering lipid particles while only a volume of 38 mm3 (25%) was achieved without particles. The proposed model provides a simple and robust method to solve complex photopolymerization problems, where the dimension of the light source is much smaller than the volume of the photopolymerizable hydrogel.

Additive manufacturing of photosensitive hydrogels for tissue engineering applications

AbstractHydrogels are extensively explored as scaffolding materials for 2D/3D cell culture and tissue engineering. Owing to the substantial complexity of tissues, it is increasingly important to develop 3D biomimetic hydrogels with user-defined architectures and controllable biological functions. To this end, one promising approach is to utilize photolithography-based additive manufacturing technologies (AMTs) in combination with photosensitive hydrogels. We here review recent advances in photolithography-based additive manufacturing of 3D hydrogels for tissue engineering applications. Given the importance of materials selection, we firstly give an overview of water-soluble photoinitiators for single- and two-photon polymerization, photopolymerizable hydrogel precursors and light-triggered chemistries for hydrogel formation. Through the text we discuss the design considerations of hydrogel precursors and synthetic approaches to polymerizable hydrogel precursors of synthetic and natural origins. Next, we shift to how photopolymerizable hydrogels could integrate with photolithography-based AMTs for creating well-defined hydrogel structures. We illustrate the working-principles of both single- and two-photon lithography and case studies of their applications in tissue engineering. In particular, two-photon lithography is highlighted as a powerful tool for 3D functionalization/construction of hydrogel constructs with μm-scale resolution. Within the text we also explain the chemical reactions involved in two-photon-induced biofunctionalization and polymerization. In the end, we summarize the limitations of available hydrogel systems and photolithography-based AMTs as well as a future outlook on potential optimizations.

Controlling uniformity of photopolymerized microscopic hydrogels

This paper studies hydrogels created by photopolymerization with a uniform beam of light. Under some conditions the density profiles of the resulting hydrogels were uniform cylinders, mirroring the illumination profiles. However, under other conditions, gels with hollow cylindrical shapes were formed. We studied the photopolymerization of poly-N-isopropylacrylamide (pNIPAAM), a hydrogel that has been widely used in tissue engineering and microfluidic applications, and examined how the size and uniformity of pNIPAAM microscopic gels can be controlled by varying parameters such as exposure time, exposure area, exposure intensity, monomer concentration, photoinitiator concentration and terminator concentration. A simplified reaction-diffusion model of the polymerization process was developed and was found to describe the experiment for a wide range of parameters. This general framework will guide attempts to establish optimal conditions for the construction of microscopic hydrogels using photolithography, which is a method that has found applications in fields such as microfluidics, drug delivery, cell and tissue culturing, and high resolution 3D printing.

Enzymatic synthesis of hyaluronic acid vinyl esters for two-photon microfabrication of biocompatible and biodegradable hydrogel constructs

A novel class of photopolymerizable hydrogel system based on hyaluronan vinyl esters has been developed for two-photon 3D microfabrication of biocompatible hydrogel constructs.

Cryotemplation for the Rapid Fabrication of Porous, Patternable Photopolymerized Hydrogels.

Download options Please wait... Supplementary files Supplementary information PDF (1797K) Article information DOI https://doi.org/10.1039/C4TB00585F Article type Paper Submitted 12 Apr 2014 Accepted 20 May 2014 First published 28 May 2014 Download Citation J. Mater. Chem. B, 2014,2, 4521-4530 BibTex EndNote MEDLINE ProCite ReferenceManager RefWorks RIS Permissions Request permissions Cryotemplation for the rapid fabrication of porous, patternable photopolymerized hydrogels A. M. Thomas and L. D. Shea, J. Mater. Chem. B, 2014, 2, 4521 DOI: 10.1039/C4TB00585F To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page. If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given. If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page. Read more about how to correctly acknowledge RSC content. Social activity Tweet Share Search articles by author Aline M. Thomas Lonnie D. Shea

Hydrogels for Two‐Photon Polymerization: A Toolbox for Mimicking the Extracellular Matrix

AbstractThe natural extracellular matrix (ECM) represents a complex and dynamic environment. It provides numerous spatio‐temporal signals mediating many cellular functions including morphogenesis, adhesion, proliferation and differentiation. The cell–ECM interaction is bidirectional. Cells dynamically receive and process information from the ECM and remodel it at the same time. Theses complex interactions are still not fully understood. For better understanding, it is indispensable to deconstruct the ECM up to the point of investigating isolated characteristics and cell responses to physical, chemical and topographical cues. Two‐photon polymerization (2PP) allows the exact reconstruction of cell specific sites in 3D at micro‐ and nanometer precision. Processing biocompatible synthetic and naturally‐derived hydrogels, the microenvironment of cells can be designed to specifically investigate their behavior in respect to key chemical, mechanical and topographical attributes. Moreover, 3D manipulation can be performed in the presence of cells, guiding biological tissue formation in all stages of its development. Here, advances in 2PP microfabrication of synthetic and naturally based hydrogels are reviewed. Key components of photopolymerizable hydrogel precursors, their structure–property relationships and their polymerization mechanisms are presented. Furthermore, it is shown how biocompatible 2PP fabricated constructs can act as biologically relevant matrices to study cell functions and tissue development.

Facile preparation of photodegradable hydrogels by photopolymerization

Photodegradable hydrogels have emerged as a powerful material platform for studying and directing cell behaviors, as well as for delivering drugs. The premise of this technique is to use a cytocompatible light source to cleave linkers within a hydrogel, thus causing reduction of matrix stiffness or liberation of matrix-tethered biomolecules in a spatial-temporally controlled manner. The most commonly used photodegradable units are molecules containing nitrobenzyl moieties that absorb light in the ultraviolet (UV) to lower visible wavelengths (∼280–450 nm). Because photodegradable linkers and hydrogels reported in the literature thus far are all sensitive to UV light, highly efficient UV-mediated photopolymerizations are less likely to be used as the method to prepare these hydrogels. As a result, currently available photodegradable hydrogels are formed by redox-mediated radical polymerizations, emulsion polymerizations, Michael-type addition reactions, or orthogonal click chemistries. Here, we report the first photodegradable poly(ethylene glycol)-based hydrogel system prepared by step-growth photopolymerization. The model photolabile peptide cross-linkers, synthesized by conventional solid phase peptide synthesis, contained terminal cysteines for step-growth thiol-ene photo-click reactions and a UV-sensitive 2-nitrophenylalanine residue in the peptide backbone for photo-cleavage. Photolysis of this peptide was achieved through adjusting UV light exposure time and intensity. Photopolymerization of photodegradable hydrogels containing photolabile peptide cross-linkers was made possible via a highly efficient visible light-mediated thiol-ene photo-click reaction using a non-cleavage type photoinitiator eosin-Y. Rapid gelation was confirmed by in situ photo-rheometry. Flood UV irradiation at controlled wavelength and intensity was used to demonstrate the photodegradability of these photopolymerized hydrogels.

Mechanical Properties of a Photopolymerizable Hydrogel for Intervertebral Disc Replacement

We report on the modelling and experimental validation of a photopolymerizable hydrogel for a Nucleus Pulposus replacement.

Structural effects in photopolymerized sodium AMPS hydrogels crosslinked with poly(ethylene glycol) diacrylate for use as burn dressings

Synthetic hydrogel polymers were prepared by free radical photopolymerization in aqueous solution of the sodium salt of 2-acrylamido-2-methylpropane sulfonic acid (Na-AMPS). Poly(ethylene glycol) diacrylate (PEGDA) and 4,4′-azo-bis(4-cyanopentanoic acid) were used as the crosslinker and UV-photoinitiator, respectively. The effects of varying the Na-AMPS monomer concentration within the range of 30–50% w/v and the crosslinker concentration within the range of 0.1–1.0% mol (relative to monomer) were studied in terms of their influence on water absorption properties. The hydrogel sheets exhibited extremely high swelling capacities in aqueous media which were dependent on monomer concentration, crosslink density, and the ionic strength and composition of the immersion medium. The effects of varying the number-average molecular weight of the PEGDA crosslinker from = 250 to 700 were also investigated. Interestingly, it was found that increasing the molecular weight and therefore the crosslink length at constant crosslink density decreased both the rate of water absorption and the equilibrium water content. Cytotoxicity testing by the direct contact method with mouse fibroblast L929 cells indicated that the synthesized hydrogels were nontoxic. On the basis of these results, it is considered that photopolymerized Na-AMPS hydrogels crosslinked with PEGDA show considerable potential for biomedical use as dressings for partial thickness burns. This paper describes some structural effects which are relevant to their design as biomaterials for this particular application.

Incorporation of graphene into photopolymerizable hydrogels of N-acyl glutanamides: Rheological and swelling behavior study of soft nanocomposite materials

Two different respectively N-stearoyl and diacetylenic fatty acyl glutanamide were prepared and employed as both potent low molecular weight hydrogelators (LMWGs). These compounds were able to gelate water at 1.0 wt% of concentration independently of the pH. Due to the presence of two free amino groups on the extremities of the gelator, an aqueous colloidal dispersion of graphene oxide (GO) could be stabilized into the hydrogel matrix simply by ionic bonds. The inclusion of a slightly amount of GO inside the gel networks increased its natural rigidity, confirmed by rheology study. An in-situ reduction of GO afforded after treatment, a semi-transparent hydrogel containing reduced graphene (rGO) that could be easily manipulated without apparent syneresis phenomenon. When the diacetylenic type hydrogel containing GO was irradiated upon UV light, a deep blue coloration change was observed; evidence of an effective photopolymerization. In case of thermal stimulation of the resulted blue polydiacetylene (PDA) hydrogel, the material turned in a red solution precursor of a robust red (PDA) hydrogel at room temperature. This red hydrogel never displayed reversible color change and its swelling behavior was studied at different temperatures. All hydrogels were characterized by different methods such as Field-Emission Scanning Electron Microscopy (FE-SEM) for determining the homogeneity of the gel networks, but also by (FT-IR) Fourier Transform-Infrared spectroscopy and UV–visible measurement.

3D Differentiation of Neural Stem Cells in Macroporous Photopolymerizable Hydrogel Scaffolds

Neural stem/progenitor cells (NSPCs) are the stem cell of the adult central nervous system (CNS). These cells are able to differentiate into the major cell types found in the CNS (neurons, oligodendrocytes, astrocytes), thus NSPCs are the mechanism by which the adult CNS could potentially regenerate after injury or disorder. Microenviromental factors are critical for guiding NSPC differentiation and are thus important for neural tissue engineering. In this study, D-mannitol crystals were mixed with photocrosslinkable methacrylamide chitosan (MAC) as a porogen to enhance pore size during hydrogel formation. D-mannitol was admixed to MAC at 5, 10 and 20 wt% D-mannitol per total initial hydrogel weight. D-mannitol crystals were observed to dissolve and leave the scaffold within 1 hr. Quantification of resulting average pore sizes showed that D-mannitol addition resulted in larger average pore size (5 wt%, 4060±160 µm2, 10 wt%, 6330±1160 µm2, 20 wt%, 7600±1550 µm2) compared with controls (0 wt%, 3150±220 µm2). Oxygen diffusion studies demonstrated that larger average pore area resulted in enhanced oxygen diffusion through scaffolds. Finally, the differentiation responses of NSPCs to phenotypic differentiation conditions were studied for neurons, astrocytes and oligodendrocytes in hydrogels of varied porosity over 14 d. Quantification of total cell numbers at day 7 and 14, showed that cell numbers decreased with increased porosity and over the length of the culture. At day 14 immunohistochemistry quantification for primary cell types demonstrated significant differentiation to the desired cells types, and that total percentages of each cell type was greatest when scaffolds were more porous. These results suggest that larger pore sizes in MAC hydrogels effectively promote NSPC 3D differentiation.

Nanomechanical measurements of polyethylene glycol hydrogels using atomic force microscopy

Poly(ethylene glycol) (PEG)-based hydrogels are among the most widely used synthetic polymers for biomedical applications. Critical parameters of importance for PEG hydrogels are their mechanical properties which can be highly tuned. While properties such as elastic moduli have been measured at the bulk scale, it is often important to measure them at the micro and nanoscales. Further, non-destructive measurements of material properties can enable in situ and high-throughput monitoring for applications including modulating cellular interactions. In this research, the elastic modulus and the stiffness of polyethylene glycol diacrylate (PEG-DA) hydrogel matrices at the nanoscale are determined via nanoindentation using an atomic force microscope (AFM). The effect of varying parameters including monomer molecular weight, initiator concentration and rates of hydration on the mechanical strength of photopolymerized hydrogels were investigated. We present the effects of indentation parameters including loads and indent depths on such measurements. Mechanical characteristics of versatile PEG hydrogels can be adjusted based on polymer chain length and crosslinking, while completely hydrated hydrogels have mechanical properties similar to articular cartilage. A better understanding of these properties can enable tailoring hydrogel based biomaterials for various applications in scaffolds and tissue engineering.

Relative survivability of human osteoblasts is enhanced by 39 °C and ascorbic acid after exposure to photopolymerization ingredients

Photopolymerizable hydrogels offer great potential in cartilage tissue engineering due to their ability to conform to irregular defect shapes and be applied in a potentially minimally invasive manner. An important process requirement in the use of photopolymerizable hydrogels is the ability of the suspended cells to withstand low intensity ultraviolet light (UV) exposure (4-5mW/cm(2)) and photoinitiator concentrations. For cartilage integration with underlying subchondral bone tissue, robust localized osteoblast activity is necessary. Yet, while it is known that osteoblasts do not respond well to UV light, limited work has been conducted to improve their survivability. In this study, we evaluated the cellular cytotoxicity of five different human cell sources at different UV exposure times, with and without a commercially used photoinitiator. We were able to confirm that human osteoblasts were the least tolerant to varying UV exposure times in comparison to bone marrow stem cell, periodontal ligament cell, smooth muscle and endothelial cell lineages. Moreover osteoblasts cultured at 39°C did not deteriorate in terms of alkaline phosphatase expression or calcium deposition within the extracellular matrix (ECM), but did reduce cell proliferation. We believe however that the lower proliferation diminished osteoblast sensitivity to UV and the photoinitiator. In fact, the relative survivability of osteoblasts was found to be augmented by the combination of a biochemical factor and an elevated incubation temperature; specifically, the use of 50mg/l of the anti-oxidant, ascorbic acid significantly (P<0.05) increased the survivability of osteoblasts when cultured at 39°C. We conclude that ascorbic acid at an incubation temperature of 39°C can be included in in vitro protocols used to assess cartilage integration with bone ECM. Such inclusion will enhance conditions of the engineered tissue model system in recapitulating in vivo osteoblast activity.

Synthesis and Photopolymerization of Tween 20 Methacrylate/N-vinyl-2-Pyrrolidone Blends

Poly(oxyethylene 20 sorbitan) monolaurate (Tween® 20) methacrylates were synthesized by coupling methacryloyl chloride (MeOCl) to Tween 20 in the presence of 4-(N,N-dimethylamino) pyridine, using THF as a solvent, in order to investigate their suitability as precursors for photopolymerizable hydrogels in tissue engineering applications. The degree of substitution could be controlled by adjusting the molar ratio of MeOCl and Tween 20, giving three different monomers: Tween 20 monomethacrylate, Tween 20 dimethacrylate and Tween 20 trimethacrylate. Combined 1H NMR and MALDI-TOF MS confirmed these monomers to be of high purity and to have polydispersities less than 1.3. It was shown that aqueous solutions of the monomers were photoactive, all the methacrylate groups reacting within 30 minutes exposure to a UV light intensity of 145mW/cm2. Aqueous Tween 20 trimethacrylate was then combined with N-vinyl-2-pyrrolidone (NVP), giving tough copolymer hydrogels on photopolymerization, whose swelling ratios and swelling rates could be tuned by varying the Tween 20 trimethacrylate content. The use of a flexible spacer with a multifunctional monomer gives a permanent three-dimensional network, whilst maintaining degrees of swelling of between 60 and 85%, with potential for a wide range of biological and non-biological applications.

Controlled hydrogel photopolymerization inside live systems by X-ray irradiation

We successfully demonstrate the in situ formation by X-ray irradiation induced polymerization of a three-dimensional (3D) hydrogel based on poly(ethylene glycol) (PEG) diacrylate (PEG DA), with the option of heparin incorporation to enhance cell adhesion. The polymerization reaction can be completed in tens of seconds, thanks to the fast and uniform X-ray induced process. Eliminating polymerization initiators and the residual unreacted monomers improves the biocompatibility and stability of the hydrogel. The photopolymerization mechanism can be locally controlled by using a trimmed or a focused X-ray beam to allow precise in vivo administration deep in the tissue, without surgery. We also demonstrate that photopolymerized hydrogels containing polyethylenimine (PEI)/nucleic acid (DNA or small interfering RNA, siRNA) nanoparticles (NPs) can deliver such nanoparticles. The hydrogel is biocompatible with mice and is not toxic to epithelial cells.

Fabricating gradient hydrogel scaffolds for 3D cell culture.

Optimizing cell-material interactions is critical for maximizing regeneration in tissue engineering. Combinatorial and high-throughput (CHT) methods can be used to systematically screen tissue scaffolds to identify optimal biomaterial properties. Previous CHT platforms in tissue engineering have involved a two-dimensional (2D) cell culture format where cells were cultured on material surfaces. However, these platforms are inadequate to predict cellular response in a three-dimensional (3D) tissue scaffold. We have developed a simple CHT platform to screen cell-material interactions in 3D culture format that can be applied to screen hydrogel scaffolds. Herein we provide detailed instructions on a method to prepare gradients in elastic modulus of photopolymerizable hydrogels.

Synthesis of a novel photopolymerized nanocomposite hydrogel for treatment of acute mechanical damage to cartilage

Intra-articular fractures initiate a cascade of pathobiological and pathomechanical events that culminate in post-traumatic osteoarthritis (PTOA). Hallmark features of PTOA include destruction of the cartilage matrix in combination with loss of chondrocytes and acute mechanical damage (AMD). Currently, treatment of intra-articular fractures essentially focuses completely on restoration of the macroanatomy of the joint. However, current treatment ignores AMD sustained by cartilage at the time of injury. We are exploring aggressive biomaterial-based interventions designed to treat the primary pathological components of AMD. This study describes the development of a novel injectable co-polymer solution that forms a gel at physiological temperatures that can be photocrosslinked, and can form a nanocomposite gel in situ through mineralization. The injectable co-polymer solution will allow the material to fill cracks in the cartilage after trauma. The mechanical properties of the nanocomposite are similar to those of native cartilage, as measured by compressive and shear testing. It thereby has the potential to mechanically stabilize and restore local structural integrity to acutely injured cartilage. Additionally, in situ mineralization ensures good adhesion between the biomaterial and cartilage at the interface, as measured through tensile and shear testing. Thus we have successfully developed a new injectable co-polymer which forms a nanocomposite in situ with mechanical properties similar to those of native cartilage, and which can bond well to native cartilage. This material has the potential to stabilize injured cartilage and prevent PTOA.

In vivo tissue engineering of functional skeletal muscle by freshly isolated satellite cells embedded in a photopolymerizable hydrogel

The success of skeletal muscle reconstruction depends on finding the most effective, clinically suitable strategy to engineer myogenic cells and biocompatible scaffolds. Satellite cells (SCs), freshly isolated or transplanted within their niche, are presently considered the best source for muscle regeneration. Here, we designed and developed the delivery of either SCs or muscle progenitor cells (MPCs) via an in situ photo-cross-linkable hyaluronan-based hydrogel, hyaluronic acid-photoinitiator (HA-PI) complex. Partially ablated tibialis anterior (TA) of C57BL/6J mice engrafted with freshly isolated satellite cells embedded in hydrogel showed a major improvement in muscle structure and number of new myofibers, compared to muscles receiving hydrogel + MPCs or hydrogel alone. Notably, SCs embedded in HA-PI also promoted functional recovery, as assessed by contractile force measurements. Tissue reconstruction was associated with the formation of both neural and vascular networks and the reconstitution of a functional SC niche. This innovative approach could overcome previous limitations in skeletal muscle tissue engineering.

A photopolymerized antimicrobial hydrogel coating derived from epsilon-poly-l-lysine

Hydrogels made from epsilon-poly-l-lysine-graft-methacrylamide (EPL-MA) have been found to have impressive wide spectrum antimicrobial activity against both bacteria (specifically Escherichia coli, Pseudomonas aeruginosa, Serratia marcescens and Staphylococcus aureus) and fungi (specifically Candida albicans and Fusarium solani). The EPL-MA hydrogel also possesses in vitro biocompatibility and EPL-MA solution is relatively non-hemolytic: the concentration needed for onset of human red blood cell (hRBC) hemolysis is 12,500 μg/mL so that the selectivity for the pathogenic microorganisms over hRBCs is 230–1560. Further, EPL-MA hydrogel can be conveniently ultraviolet-immobilized onto plasma-treated plastic surfaces to form thin highly adherent antimicrobial hydrogel coatings for medical devices and implants.