Dual-reactive nanogels for orthogonal functionalization of hydrophilic shell and amphiphilic network.

Amphiphilic nanogels (NGs) combine a soft, water-swollen hydrogel matrix with internal hydrophobic domains. While these domains can encapsulate hydrophobic cargoes, the amphiphilic particle surface can reduce colloidal stability and/or limit biological half-life. Therefore, a functional hydrophilic shell is needed to shield the amphiphilic network and tune interactions with biological systems. To adjust core and shell properties independently, we developed a synthetic strategy that uses preformed dual-reactive nanogels. In a first step, emulsion copolymerization of pentafluorophenyl methacrylate (PFPMA) and a reduction-cleavable crosslinker produced precursor particles for subsequent network modification. Orthogonal shell reactivity was installed by using an amphiphilic block copolymer (BCP) surfactant during this particle preparation step. Here, the hydrophilic block poly(polyethylene glycol methyl ether methacrylate) (PPEGMA) contains a reactive alkyne end group for successive functionalization. The hydrophobic block (P(PFPMA-co-MAPMA) contains random methacryl-amido propyl methacrylamide (MAPMA) units to covalently attach the surfactant to the growing PPFPMA network. In the second step, orthogonal modification of the core and shell was demonstrated. Network functionalization with combinations of hydrophilic (acidic, neutral, or basic) and hydrophobic (cholesterol) groups gave a library of pH- and redox-sensitive amphiphilic NGs. Stimuli-responsive properties were demonstrated by pH-dependent swelling and reduction-induced degradation via dynamic light scattering. Subsequently, copper-catalyzed azide-alkyne cycloaddition was used to attach azide-modified rhodamine as model compound to the shell (followed by UV-Vis). Overall, this strategy provides a versatile platform to develop multi-functional amphiphilic nanogels as carriers for hydrophobic cargoes.

Influence of Nanogel Amphiphilicity on Dermal Delivery: Balancing Surface Hydrophobicity and Network Rigidity.

Polymeric nanogels are promising nonirritating nanocarriers for topical delivery applications. However, conventional hydrophilic networks limit encapsulation of hydrophobic therapeutics and hinder tailored interactions with the amphiphilic skin barrier. To address these limitations, we present amphiphilic nanogels containing hydrophilic networks with hydrophobic domains. Two competing factors determine favorable nanogel-skin interactions and need to be balanced through network composition: suitable surface hydrophobicity and low network rigidity (through physical hydrophobic cross-links). To ensure comparability in such investigations, we prepared a library of nanogels with increasing hydrophobic cholesteryl amounts but similar colloidal features. By combining mechanical and surface hydrophobicity tests (atomic force microscopy (AFM)) with dermal delivery experiments on excised human skin, we can correlate an increased delivery efficacy of Nile red to the viable epidermis with a specific network composition, i.e., 20-30 mol % cholesterol. Thus, our nanogel library identifies a specific balance between surface amphiphilicity and network rigidity to guide developments of advanced dermal delivery vehicles.

Amphiphilic Nanogels: Fuzzy Spheres with a Pseudo-Periodic Internal Structure.

Amphiphilic polymer nanogels (NGs) are promising drug delivery vehicles that extend the application of conventional hydrophilic NGs to hydrophobic cargoes. By randomly introducing hydrophobic groups into a hydrophilic polymer network, loading and release profiles as well as surface characteristics of these colloids can be tuned. However, very little is known about the underlying internal structure of such complex colloidal architectures. Of special interest is the question how the amphiphilic network composition influences the internal morphology and the "fuzzy" surface structure. To shine light into the influence of varying network amphiphilicity on these structural features, we investigated a small library of water-swollen amphiphilic NGs using small-angle X-ray scattering (SAXS). It was found that overall hydrophilic NGs, consisting of pure poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA), display a disordered internal structure as indicated by the absence of a SAXS peak. In contrast, a SAXS peak is present for amphiphilic NGs with various amounts of incorporated hydrophobic groups such as cholesteryl (CHOLA) or dodecyl (DODA). The internal composition of the NGs is considered structurally homologous to microgels. Application of the Teubner-Strey model reveals that hydrophilic PHPMA NGs have a disordered internal structure (positive amphiphilicity factor) while CHOLA and DODA samples have an ordered internal structure (negative amphiphilicity factor). From the SAXS data it can be derived that the internal structure of the amphiphilic NGs consists of regularly alternating hydrophilic and hydrophobic domains with repeat distances of 3.45-5.83 nm.

Amphiphilic nanogels: influence of surface hydrophobicity on protein corona, biocompatibility and cellular uptake.

BACKGROUND AND PURPOSE: Nanogels (NGs) are promising drug delivery tools but are typically limited to hydrophilic drugs. Many potential new drugs are hydrophobic. Our study systematically investigates amphiphilic NGs with varying hydrophobicity, but similar colloidal features to ensure comparability. The amphiphilic NGs used in this experiment consist of a hydrophilic polymer network with randomly distributed hydrophobic groups. For the synthesis we used a new synthetic platform approach. Their amphiphilic character allows the encapsulation of hydrophobic drugs. Importantly, the hydrophilic/hydrophobic balance determines drug loading and biological interactions. In particular, protein adsorption to NG surfaces is dependent on hydrophobicity and critically determines circulation time. Our study investigates how network hydrophobicity influences protein binding, biocompatibility and cellular uptake. METHODS: Biocompatibility of the NGs was examined by WST-1 assay in monocytic-like THP-1 cells. Serum protein corona formation was investigated using dynamic light scattering and two-dimensional gel electrophoresis. Proteins were identified by liquid chromatography-tandem mass spectrometry. In addition, cellular uptake was analyzed via flow cytometry. RESULTS: All NGs were highly biocompatible. The protein binding patterns for the two most hydrophobic NGs were very similar to each other but clearly different from the hydrophilic ones. Overall, protein binding was increased with increasing hydrophobicity, resulting in increased cellular uptake. CONCLUSION: Our study supports the establishment of structure-property relationships and contributes to the accurate balance between maximum loading capacity with low protein binding, optimal biological half-life and good biocompatibility. This is an important step to derive design principles of amphiphilic NGs to be applied as drug delivery vehicles.

Metabolomic analysis of resveratrol-induced effects in the human breast cancer cell lines MCF-7 and MDA-MB-231.

Resveratrol is a naturally occurring anticancer compound present in grapes and wine with antiproliferative properties against breast cancer cells and xenografts. Our objective was to investigate the metabolic alterations that characterize the effects of resveratrol in the human breast cancer cell lines MCF-7 and MDA-MB-231 using high-throughput liquid chromatography-based mass spectrometry. In both cell lines, growth inhibition was dose dependent and accompanied by substantial metabolic changes. For all 21 amino acids analyzed levels increased more than 100-fold at a resveratrol dose of 100 µM with far lower concentrations in MDA-MB-231 compared to MCF-7 cells. Among the biogenic amines and modified amino acids (n = 16) resveratrol increased the synthesis of serotonin, kynurenine, and spermindine in both cell lines up to 61-fold indicating that resveratrol strongly interacts with cellular biogenic amine metabolism. Among the eicosanoids and oxidized polyunsaturated fatty acids (n = 17) a pronounced increase in arachidonic acid and its metabolite 12S-HETE was observed in MDA-MB-231 and to a lesser extent in MCF-7 cells, indicating release from cell membrane phospholipids upon activation of phospholipase A2 and subsequent metabolism by 12-lipoxygenase. In conclusion, metabolomic analysis elucidated several small molecules as markers for the response of breast cancer cells to resveratrol.

Perspective: Biotech funding trends: Insights from entrepreneurs and investors.

Based on a qualitative study analyzing a series of interviews with dedicated biotech entrepreneurs and high-level investors, "Biotech Funding Trends" provides a comprehensive overview of current trends in biotech funding by taking a close look behind the scenes of the biotech industry. In particular, it illustrates the tensions between both key players based on their different backgrounds and expectations. Here we outline the various funding opportunities for the biotech industry in Europe and identify ways for both sides, entrepreneurs and investors to overcome their prejudices to successfully thrive in a competitive environment. The results are also discussed in the light of the current financial and economic crisis.