Surita R. Bhatia

The influence of hydrogel modulus on the proliferation and differentiation of encapsulated neural stem cells

There has been an increasing interest in understanding how the mechanical properties of the microenvironment influence stem cell fate. We describe studies of the proliferation and differentiation of neural stem cells (NSCs) encapsulated within three-dimensional scaffolds--alginate hydrogels--whose elastic moduli were varied over two orders of magnitude. The rate of proliferation of neural stem cells decreased with increase in the modulus of the hydrogels. Moreover, we observed the greatest enhancement in expression of the neuronal marker beta-tubulin III within the softest hydrogels, which had an elastic modulus comparable to that of brain tissues. To our knowledge, this work represents the first demonstration of the influence of modulus on NSC differentiation in three-dimensional scaffolds. Three-dimensional scaffolds that control stem cell fate would be broadly useful for applications in regenerative medicine and tissue engineering.

Synthetically Simple, Highly Resilient Hydrogels

Highly resilient synthetic hydrogels were synthesized by using the efficient thiol-norbornene chemistry to cross-link hydrophilic poly(ethylene glycol) (PEG) and hydrophobic polydimethylsiloxane (PDMS) polymer chains. The swelling and mechanical properties of the hydrogels were controlled by the relative amounts of PEG and PDMS. The fracture toughness (G(c)) was increased to 80 J/m(2) as the water content of the hydrogel decreased from 95% to 82%. In addition, the mechanical energy storage efficiency (resilience) was more than 97% at strains up to 300%. This is comparable with one of the most resilient materials known: natural resilin, an elastic protein found in many insects, such as in the tendons of fleas and the wings of dragonflies. The high resilience of these hydrogels can be attributed to the well-defined network structure provided by the versatile chemistry, low cross-link density, and lack of secondary structure in the polymer chains.

Effect of mammalian cell culture medium on the gelation properties of Pluronic F127.

We investigate the gelation of a poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) copolymer, Pluronic F127, in mammalian cell culture medium for applications in tissue engineering and cell encapsulation. In both minimum essential medium (MEM) and MEM with added fetal bovine serum (MEM-FBS), the gel-phase boundary shifts to lower temperatures and concentrations as compared to pure water. The thermodynamics of gel formation are similar in MEM, MEM-FBS, and pure water, suggesting that the mechanism of gelation is similar in all three solvents. The shift of the sol-gel boundary to lower concentrations is particularly significant for development of cell encapsulation protocols using Pluronics and applications where copolymer concentration must be minimized due to toxicity concerns.

Hydrogel‐Perfluorocarbon Composite Scaffold Promotes Oxygen Transport to Immobilized Cells

Cell encapsulation provides cells a three‐dimensional structure to mimic physiological conditions and improve cell signaling, proliferation, and tissue organization as compared to monolayer culture. Encapsulation devices often encounter poor mass transport, especially for oxygen, where critical dissolved levels must be met to ensure both cell survival and functionality. To enhance oxygen transport, we utilized perfluorocarbon (PFC) oxygen vectors, specifically perfluorooctyl bromide (PFOB) immobilized in an alginate matrix. Metabolic activity of HepG2 liver cells encapsulated in 1% alginate/10% PFOB composite system was 47–104% higher than alginate systems lacking PFOB. A cubic model was developed to understand the oxygen transport mechanism in the alginate/PFOB composite system. The theoretical flux enhancement in alginate systems containing 10% PFOB was 18% higher than in alginate‐only systems. Oxygen uptake rates (OURs) of HepG2 cells were enhanced with 10% PFOB addition under both 20% and 5% O2 boundary conditions, by 8% and 15%, respectively. Model predictions were qualitatively and quantitatively verified with direct experimental OUR measurements using both a perfusion reactor and oxygen sensing plate, demonstrating a greater OUR enhancement under physiological O2 boundary conditions (i.e., 5% O2). Inclusion of PFCs in an encapsulation matrix is a useful strategy for overcoming oxygen limitations and ensuring cell viability and functionality both for large devices (>1 mm) and over extended time periods. Although our results specifically indicate positive enhancements in metabolic activity using the model HepG2 liver system encapsulated in alginate, PFCs could be useful for improving/stabilizing oxygen supply in a wide range of cell types and hydrogels.

Block copolymer assembly to control fluid rheology

Abstract Recent experimental studies on the rheology of block copolymer micelles are reviewed. Where appropriate, we draw analogies between the viscoelastic properties of polymeric micelles and those of colloidal dispersions. We also present some important differences between these two classes of complex fluids, namely the ability to tune self-assembly through solvent–polymer interactions. Finally, new experimental results for attractive micellar solutions of polyelectrolytes are presented.

Nanoparticle-Reinforced Associative Network Hydrogels

ABA triblock copolymers in solvents selective for the midblock are known to form associative micellar gels. We have modified the structure and rheology of ABA triblock copolymer gels comprising poly(lactide)-poly(ethylene oxide)-poly(lactide) (PLA-PEO-PLA) through addition of a clay nanoparticle, laponite. Addition of laponite particles resulted in additional junction points in the gel via adsorption of the PEO corona chains onto the clay surfaces. Rheological measurements showed that this strategy led to a significant enhancement of the gel elastic modulus with small amounts of nanoparticles. Further characterization using small-angle X-ray scattering and dynamic light scattering confirmed that nanoparticles increase the intermicellar attraction and result in aggregation of PLA-PEO-PLA micelles.

New properties from PLA–PEO–PLA hydrogels

Polymeric materials are important in many medical applications. Regenerative medicine offers the potential to repair or replace damaged tissue and polymers are an essential component of many tissue engineering approaches. Hydrogels have many advantageous properties but, generally, lack robust mechanical properties. At the same time, mounting evidence points to the importance of the matrix modulus when constructing devices. In this context, triblock copolymers made from poly(-lactide)-poly(ethylene glycol)-poly(-lactide) have been prepared and formulated into hydrogels. Investigations into their mechanical properties found the elastic modulus to be greater than 10 kPa which is at least one order of magnitude stiffer than previously reported from macromolecules composed of similar monomers. Part of the reason is the presence of crystalline lactide domains. Creating hydrogels with tailored modulus across the kPa range will likely have important ramifications in regenerative medicine.

Poly(lactic acid)-poly(ethylene oxide) block copolymers: new directions in self-assembly and biomedical applications.

In this review, we focus on recent developments in biomaterials of poly(lactic acid)-poly(ethylene oxide)-poly(lactic acid) (PLA-PEO-PLA) triblock copolymers. This system has been widely explored for a number of applications in controlled and sustained release of drugs and in tissue engineering devices. New insights into self-assembly of these materials have resulted in new PLA-PEOPLA solutions and gels with novel structural, mechanical, and drug release properties. Recent innovations include hydrogels with nanoscale crystalline domains, solutions and gels based on PLA stereocomplexes, and nanoparticle-copolymer assemblies. We first briefly review synthetic approaches to these materials. We then describe characterization of the solution properties, formation of micelles, drug release characteristics, and investigation of the sol-gel transition. The properties of PLA-PEO-PLA hydrogels are then discussed, including the effect of crystalline domains on the gel microstructure and efforts to tune the elastic modulus and degredation properties of gels through the addition of chemical crosslinks. In the second half of the review, we discuss the wide variety of biomedical applications currently being pursued for PLA-PEO-PLA triblock copolymer systems. Polymer-nanoparticle complexes have been investigated to facilitate the formation of metal nanoclusters used as biosensors, as well as to enhance the elastic modulus of hydrogels. Thin polymer films have also been investigated for use as tissue engineering scaffolds and as drug-eluding coatings for stents and other medical implants. Finally, we discuss future directions for biomedical applications of this system, including new strategies for improving the specificity and cell affinity of PLA-based biomaterials.

Effect of pharmaceuticals on thermoreversible gelation of PEO-PPO-PEO copolymers

Pluronic F127, a triblock copolymer of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO), has generated considerable interest as a drug delivery vehicle due to its ability to gel at physiological temperatures. This work examines the gelation behavior of Pluronic F127 in the presence of a series of hydrophobic pharmaceuticals, to determine whether there is any correlation between gelation and physicochemical parameters of drug solutes. The study includes the local anesthetics dibucaine, lidocaine, and tetracaine; the pharmaceutical additives methyl paraben, ethyl paraben, and propyl paraben; the anti-cancer agents paclitaxel and baccatin III; and the anti-inflammatory agent sulindac. The results indicate that the presence of local anesthetics and pharmaceutical additives allows F127 solutions to form gels at lower copolymer concentrations; local anesthetics and pharmaceutical additives also shift gelation down to a lower gelation temperature. This behavior is strongly dependent on drug solubility; poorly soluble drugs (paclitaxel, baccatin III, sulindac) do not change the lower gelation temperature or minimum F127 concentration for gelation. An equation relating the decrease in gelation temperature to drug solubility is presented, and the equation fits the data well. The results have significant positive implications on the toxicity and economic issues related to use of Pluronic F127 in drug delivery.

Autostratification in Drying Colloidal Dispersions: Effect of Particle Interactions

When particles differing in size or charge are mixed and cast, vertical segregation is an inevitable phenomenon in the produced films. Apart from the Peclet number, which is the ratio of evaporation to diffusion rates, particle interactions play a crucial role in determining the distribution of particles in the dried films. Trueman et al. (1) developed a model for vertical segregation of particles during drying. Their numerical solution assumed that the chemical potentials were determined entirely by entropy. We report the effect of particle interactions in various systems: (i) charged particles with different Peclet numbers and (ii) charged particles with the same Peclet numbers. An experimental study has also been carried out for particles with Peclet numbers straddling unity; the experimental results conform with the behavior predicted theoretically.