Resilin, a protein found in insect cuticles, is renowned for its outstanding elastomeric properties. The authors' laboratory previously developed a recombinant protein, which consisted of consensus resilin‐like repeats from Anopheles gambiae, and demonstrated its potential in cartilage and vascular engineering. To broaden the versatility of the resilin‐like protein, this study utilizes a cleavable crosslinker, which contains a disulfide bond, to develop smart resilin‐like hydrogels that are redox‐responsive. The hydrogels exhibit a porous structure and a stable storage modulus (G′) of ≈3 kPa. NIH/3T3 fibroblasts cultured on hydrogels for 24 h have a high viability (>95%). In addition, the redox‐responsive hydrogels show significant degradation in a reducing environment (10 mm glutathione (GSH)). The release profiles of fluorescently labeled dextrans encapsulated within the hydrogels are assessed in vitro. For dextran that is estimated to be larger than the mesh size of the gel, faster release is observed in the presence of reducing agents due to degradation of the hydrogel networks. These studies thus demonstrate the potential of using these smart hydrogels in a variety of applications ranging from scaffolds for tissue engineering to drug delivery systems that target the intracellular reductive environments of tumors.

Therapeutic strategies aim to regulate vasculature either by encouraging vessel growth for tissue engineering or inhibiting vascularization around a tumor. Vascular endothelial growth factor (VEGF) is essential to these processes, and there are several strategies that manipulate VEGF signaling. Here we develop a method to control the surface density of VEGF, which is covalently attached to tissue culture polystyrene (TCPS), and explore cellular responses to surfaces with varying VEGF densities. We show that the crosslinker reduces but does not eliminate the biological activity of soluble VEGF as measured by endothelial proliferation. However, endothelial cells cultured on surfaces of covalently bound VEGF did not proliferate in response to surface cues. Interestingly, compared to cells incubated with soluble VEGF (10 ng/ml) and cultured on TCPS, lower cell proliferation was observed when endothelial cells were cultured on high VEGF surface densities (5.9 ng/cm(2)), whereas higher cell proliferation occurred when cells were cultured on low surface densities (0.04 ng/cm(2)). High density surfaces (5.9 ng/cm(2)) also acted in synergy with an inhibitor of VEGF receptors to further suppress endothelial cell proliferation. We also examined the effect of VEGF surfaces on endothelial differentiation of mesenchymal stem cells. No effect was observed when cells were cultured on VEGF surfaces; however, the VEGF surfaces acted in synergy with an inhibitor of VEGF receptors to decrease the ability of differentiated cells to form vascular networks. Together, these results suggest that surface density of bound VEGF can be used to modulate cell behavior and inhibit an angiogenic response.

For clinical applications of engineered vascular replacements, endothelial cells may not be available in sufficient quantities due to limited harvesting sites and slow in vitro expansion rates. Soluble vascular endothelial growth factor (VEGF) is often added to differentiate mesenchymal stem cells (MSCs) into endothelial cells; however, recent studies demonstrate that VEGF is not required to upregulate endothelial markers. In contrast to previous assumptions, this study demonstrates that exogenous VEGF does not enhance or accelerate the upregulation of common endothelial markers during endothelial differentiation of human MSCs. MSCs were cultured at confluence for up to 3 weeks in either basal medium or medium containing VEGF. Cells were examined for gene and protein expression as well as the ability to internalize acetylated low density lipoprotein. With either treatment, endothelial differentiation occurred as evidenced by upregulation of gene and protein expression of typical endothelial markers and the ability to internalize acetylated low density lipoproteins. Interestingly, the addition of VEGF at typical or high concentrations (50 or 100 ng/ml) did not result in differences in gene or protein expression levels of many typical endothelial markers. However, high concentrations of VEGF did significantly increase protein expression of the arterial marker Ephrin-B1. Thus, VEGF did not accelerate or enhance differentiation of human MSCs towards endothelial cells but was vital for specification of arterial fate.

The Formulation Workstream of the BioPhorum Development Group (BPDG), an industry-wide consortium, has identified the increased use of closed system drug-transfer devices (CSTDs) with biologics, without an associated compatibility assessment, to be of significant concern. The use of CSTDs has increased significantly in recent years due to the recommendations by NIOSH and USP that they be used during preparation and administration of hazardous drugs. While CSTDs are valuable in the healthcare setting to reduce occupational exposure to hazardous compounds, these devices may present particular risks that must be adequately assessed prior to use to ensure their compatibility with specific types of drug products, such as biologic drugs, which may be sensitive. The responsibility of ensuring quality of biologic products through preparation and administration to the patient lies with the drug product sponsor. Due to the significant number of marketed CSTD systems, and the large variety of components offered for each system, a strategic, risk-based approach to assessing compatibility is recommended herein. In addition to traditional material compatibility, assessment of CSTD compatibility with biologics should consider additional parameters to address specific CSTD-related risks. The BPDG Formulation Workstream has proposed a systematic risk-based evaluation approach as well as a mitigation strategy for establishing suitability of CSTDs for use.