Biorelevant subcutaneous in vitro test predicts the release of human and fast acting insulin formulations.

The administration of insulins by subcutaneous injection is nowadays widely prevalent. The injection site is located below the dermis and composed of cells and the extracellular matrix formed of a network of macromolecules such as hyaluronic acid and collagen. Following an injection, the insulins from the formulated products are timely released as drug molecules from the injection site into systemic circulation. In this publication, we show the development of an in vitro setup utilizing a hydrogel composed of a special collagen-hyaluronic acid mixture that mimics the extracellular matrix. Another setup was used for differentiation of the commercially available and research insulin formulations by determining the in vitro permeation characteristics with the results that were correlated with the human in vivo data. Significant differentiation was achieved at 90 % confidence level between the permeation curves of insulin glulisine containing formulations (U100 and a concentrated research formulation), while in case of the insulin lispro containing formulations (U100 and U200) the permeation curves showed similarity. These results demonstrated that the in vitro setup may be used as a tool for formulation development and drug candidate profiling as it is able to differentiate or show similarities between the agglomeration states and concentration of the active pharmaceutical ingredients.

Natural Presentation of Glycosaminoglycans in Synthetic Matrices for 3D Angiogenesis Models.

Glycosaminoglycans (GAGs) are long, linear polysaccharides that occur in the extracellular matrix of higher organisms and are either covalently attached to protein cores, as proteoglycans or in free form. Dependent on their chemical composition and structure, GAGs orchestrate a wide range of essential functions in tissue homeostasis. Accordingly, GAG-based biomaterials play a major role in tissue engineering. Current biomaterials exploit crosslinks between chemically modified GAG chains. Due to modifications along the GAG chains, they are limited in their GAG-protein interactions and accessibility to dissect the biochemical and biophysical properties that govern GAG functions. Herein, a natural presentation of GAGs is achieved by a terminal immobilization of GAGs to a polyethylene glycol (PEG) hydrogel. A physicochemical characterization showed that different end-thiolated GAGs can be incorporated within physiological concentration ranges, while the mechanical properties of the hydrogel are exclusively tunable by the PEG polymer concentration. The functional utility of this approach was illustrated in a 3D cell culture application. Immobilization of end-thiolated hyaluronan enhanced the formation of capillary-like sprouts originating from embedded endothelial cell spheroids. Taken together, the presented PEG/GAG hydrogels create a native microenvironment with fine-tunable mechanobiochemical properties and are an effective tool for studying and employing the bioactivity of GAGs.

Small Physical Cross-Linker Facilitates Hyaluronan Hydrogels.

In this study, we demonstrate that small charged molecules (NH4+, GluA+, dHA+) can form physical cross-links between hyaluronan chains, facilitating polymerization reactions between synthetically introduced thiol groups (HA-DTPH). These hybrid hydrogels can be obtained under physiological conditions ideally suited for 3D cell culture systems. The type and concentration of a physical crosslinker can be adjusted to precisely tune mechanical properties as well as degradability of the desired hydrogel system. We analyze the influence of hydrogen bond formation, concentration and additional ionic interactions on the polymerization reaction of HA-DTPH hydrogels and characterize the resulting hydrogels in regard to mechanical and biocompatibility aspects.