In a first part, the alkoxyamine initiator derived from BlocBuilder® based on the nitroxide SG1 and bearing an N‐succinimidyl ester function was used to synthesize a variety of well‐defined polymers with the activated ester group at the α‐end. The grafting of those polymer chains onto primary‐amine functionalized Stöber silica particles was performed in soft conditions at room temperature and was studied as a function of the polymer chain length, the type of solvent, and the concentrations of both polymer and amine. Polymer grafting densities were mainly in the 0.1–0.2 chain nm−2 range. To increase the grafting density in simple experimental conditions, a direct one‐step method was then proposed: the primary‐amine modified silica, the N‐succinimidyl ester functionalized alkoxyamine, and the monomer were all introduced into the reaction medium at once. This technique allowed grafting and chain growth to take place simultaneously at the polymerization temperature (i.e., 120 °C for styrene), in a single step, and produced hybrid particles with very high grafting density, up to 0.9 chain nm−2, and long polymer chains. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 173–185, 2010

Controlled release drug delivery systems are well established as oral and implantable dosage forms. However, the controlled release paradigm can also be used to present complex soluble signals responsible for cellular organization during development. Endochondral ossification (EO), the developmental process of bone formation from a cartilage matrix is controlled by several soluble signals with distinct functions that vary in structure, molecular weight and stability. This makes delivering them from a single vehicle rather challenging. Herein, a gelatin-based delivery system suitable for the delivery of small molecules as well as recombinant human (rh) proteins (rhWNT3A, rhFGF2, rhVEGF, rhBMP4) is reported. The release behavior and biological activity of the released molecules was validated using analytical and biological assays, including cell reporter systems. The simplicity of fabrication of the gelatin device should foster its adaptation by the diverse scientific community interested in interrogating developmental processes, in vivo.

Soluble signal gradients play an important role in organ patterning, cell migration, and differentiation. Currently, signal gradients in 2D cell culture are realized using microfluidics and here cells are exposed to high and nonphysiological shear stress. Tissue morphogenesis (organogenesis) however occurs in 3D and therefore there is a need for simple and practical systems to impose gradients to cells dispersed in 3D matrix. Herein, a 3D gradient generator based on passive diffusion elements that recapitulates interstitial flow and is capable of imposing predictable gradients over long length scales (6 mm) lasting up to 48 h to cells dispersed in a hydrogel environment is reported. Using recombinant human WNT3A (rhWNT3A), the spatiotemporal activation of the canonical WNT pathway in human epithelial kidney cells and human mesenchymal stems cells expressing a green fluorescence protein reporter on a transcription factor/lymphoid enhancer‐binding factor (TCF/LEF) promoter is demonstrated. By refining computation models based on experimental findings, the diffusion coefficient of rhWNT3A in presence of human cells in 3D is determined. Furthermore, the formation of rhBMP4 gradients is visualized using immunohistochemistry by staining for phospho‐SMAD1/5, the downstream targets of the bone morphogenetic protein (BMP) pathway. The simplicity of the gradient generator is expected to spur its adoption in studying developmental biology paradigms in vitro.