Biomimetic nanostructures for the silicone-biosystem interface: tuning oxygen-plasma treatments of polydimethylsiloxane

Polydimethylsiloxanes (PDMS) have drawn attention because of their applicability in medical implants, soft robotics and microfluidic devices. This article examines the formation of dedicated nanostructures on liquid submicrometer PDMS films when exposed to oxygen-plasma treatment. We show that by using a vinyl-terminated PDMS prepolymer with a molecular weight of 800 g/mol, one can bypass the need of solvent, copolymer, or catalyst to fabricate wrinkled films. The amplitude and periodicity of the wrinkles is tuned varying the thickness of the PDMS film between 150 and 600 nm. The duration of the plasma treatment and the oxygen pressure determine the surface morphology. The amplitude was found between 30 and 300 nm with periodicities ranging from 500 to 2800 nm. Atomic force microscopy was used to measure film thickness, amplitude and wrinkle periodicity. The hydrophobic recovery of the nanostructured PDMS surface, as assessed by dynamic contact angle measurements, scales with nanostructure’s fineness, associated with an improved biocompatibility. The mechanical properties were extracted out of 10,000 nanoindentations on 50×50-μm2 spots. The mechanical mapping with sub-micrometer resolution reveals elastic properties according to the film morphology. Finally, we tailored the mechanical properties of a 590±120-nm-thin silicone film to the elastic modulus of several MPa, as required for dielectric elastomer actuators, to be used as artificial muscles for incontinence treatments

Immunocompatibility of Rad-PC-Rad liposomes in vitro, based on human complement activation and cytokine release

Liposomal drug delivery systems can protect pharmaceutical substances and control their release. Systemic administration of liposomes, however, often activate the innate immune system, resulting in hypersensitivity reactions. These pseudo-allergic reactions can be interpreted as activating the complement system. Complement activation destroys and eliminates foreign substances, either directly through opsonization and the formation of the membrane attack complex (MAC), or by activating leukocytes and initiating inflammatory responses via mediators, such as cytokines. In this study, we investigated the in vitro immune toxicity of the recently synthesized Rad-PC- Rad liposomes, analyzing the liposome-induced complement activation. In five human sera, Rad-PC-Rad liposomes did not induce activation, but in one serum high sensitivity via alternative pathway was detected. Such a behavior in adverse phenomena is characteristic for patient-to-patient variation and, thus, the number of donors should be in the order of hundreds rather than tens – hence the present study based on six donors has preliminary character. In order to further prove the suitability of mechano-responsive Rad-PC-Rad liposomes for clinical trials, the production of pro-inflammatory cytokines was examined by human white blood cells. The concentrations of the pro-inflammatory cytokines, IL-6, IL-12p70, TNF-α, and IL-1β, induced by Rad-PC-Rad liposomal formulations, incubated with whole blood samples, were smaller or comparable to saline (negative control). Because of this favorable in vitro hemo- compatibility, in vivo investigations using these mechano-responsive liposomes should be designed

Immunological response to nitroglycerin-loaded shear-responsive liposomes in vitro and in vivo

Liposomes formulated from the 1,3-diamidophospholipid Pad-PC-Pad are shear- responsive and thus promising nano-containers to specifically release a vasodilator at stenotic arteries. The recommended preclinical safety tests for therapeutic liposomes of nanometer size include the in vitro assessment of complement activation and the evaluation of the associated risk of complement activation-related pseudo-allergy (CARPA) in vivo. For this reason, we measured complement activation by Pad-PC- Pad formulations in human and porcine sera, along with the nanopharmaceutical- mediated cardiopulmonary responses in pigs. The evaluated formulations comprised of Pad-PC-Pad liposomes, with and without polyethylene glycol on the surface of the liposomes, and nitroglycerin as a model vasodilator. The nitroglycerin incorporation efficiency ranged from 25% to 50%. In human sera, liposome formulations with 20 mg/mL phospholipid gave rise to complement activation, mainly via the alternative pathway, as reflected by the rises in SC5b-9 and Bb protein complex concentrations. Formulations having a factor of ten lower phospholipid content did not result in measurable complement activation. The weak complement activation induced by Pad- PC-Pad liposomal formulations was confirmed by the results obtained by performing an in vivo study in a porcine model, where hemodynamic parameters were monitored continuously. Our study suggests that, compared to FDA-approved liposomal drugs, Pad-PC-Pad exhibits less or similar risks of CARPA

CNS border-associated macrophages in the homeostatic and ischaemic brain

CNS border-associated macrophages (BAMs) are a small population of specialised macrophages localised in the choroid plexus, meningeal and perivascular spaces. Until recently, the function of this elusive cell type was poorly understood and largely overlooked, especially in comparison to microglia, the primary brain resident immune cell. However, the recent single cell immunophenotyping or transcriptomic analysis of BAM subsets in the homeostatic brain, coupled with the rapid emergence of new studies exploring BAM functions in various cerebral pathologies, including Alzheimer's disease, hypertension-induced neurovascular and cognitive dysfunction, and ischaemic stroke, has unveiled previously unrecognised heterogeneity and spatial-temporal complexity in BAM populations as well as their contributions to brain homeostasis and disease. In this review, we discuss the implications of this new-found knowledge on our current understanding of BAM function in ischaemic stroke. We first provide a comprehensive overview and discussion of the cell-surface expression profiles, transcriptional signatures and potential functional phenotypes of homeostatic BAM subsets described in recent studies. Evidence for their putative physiological roles is examined, including their involvement in immunological surveillance, waste clearance, and vascular permeability. We discuss the evidence supporting the accumulation and genetic transformation of BAMs in response to ischaemia and appraise the experimental evidence that BAM function might be deleterious in the acute phase of stroke, while considering the mechanisms by which BAMs may influence stroke outcomes in the longer term. Finally, we review the therapeutic potential of immunomodulatory strategies as an approach to stroke management, highlighting current challenges in the field and key issues relating to BAMs, and how BAMs could be harnessed experimentally to support future translational research

Biomimetic nanostructures for the silicone-biosystem interface: tuning oxygen-plasma treatments of polydimethylsiloxane

Polydimethylsiloxanes (PDMS) have drawn attention because of their applicability in medical implants, soft robotics and microfluidic devices. This article examines the formation of dedicated nanostructures on liquid submicrometer PDMS films when exposed to oxygen-plasma treatment. We show that by using a vinyl-terminated PDMS prepolymer with a molecular weight of 800 g/mol, one can bypass the need of solvent, copolymer, or catalyst to fabricate wrinkled films. The amplitude and periodicity of the wrinkles is tuned varying the thickness of the PDMS film between 150 and 600 nm. The duration of the plasma treatment and the oxygen pressure determine the surface morphology. The amplitude was found between 30 and 300 nm with periodicities ranging from 500 to 2800 nm. Atomic force microscopy was used to measure film thickness, amplitude and wrinkle periodicity. The hydrophobic recovery of the nanostructured PDMS surface, as assessed by dynamic contact angle measurements, scales with nanostructure's fineness, associated with an improved biocompatibility. The mechanical properties were extracted out of 10,000 nanoindentations on 50×50-μm2 spots. The mechanical mapping with sub-micrometer resolution reveals elastic properties according to the film morphology. Finally, we tailored the mechanical properties of a 590±120-nm-thin silicone film to the elastic modulus of several MPa, as required for dielectric elastomer actuators, to be used as artificial muscles for incontinence treatments

Liposomes - bio-inspired nano-containers for physically triggered targeted drug delivery

For natural scientists and engineers, learning from nature has tradition and is often driven by bio-inspired processes and materials. For example, engineers have designed multifunctional materials with hierarchical structures. Lipid bilayers, the principal components of cell membranes, can form vesicles, termed liposomes. Such liposomes are usually recognized as foreign by the immune system of a patient, which makes it challenging to use liposomes as containers for targeted drug delivery. There are, however, promising non-spherical, mechano-sensitive, artificial liposomes about 100 nm in diameter, which were recently identified. These bio-inspired containers offer a wide range of applications. In particular, the targeted release at critically stenosed arteries formed as a result of atherosclerosis significantly reduces the undesired side effects such as a drop of blood pressure. It is well known that FDA-approved liposomal drugs, currently on the market, often induce adverse immune responses. Therefore, to exclude the hypersensitivity of the recently discovered mechano-sensitive liposomes, we have performed in vitro complement activation experiments and related animal studies with pigs. Recently, it has been shown that the drug-free Pad-PC-Pad liposomes surprisingly lack any complement activation. In this study, we demonstrate that nitroglycerin-loaded liposomes with relevant human therapeutic dosage exhibit low complement activation compared to the FDA-approved phospholipid drugs, including Abelcet. Furthermore, the liposomal suspensions applied are stable for a period of more than two months. Consequently, the non-spherical liposomes of nanometer size we have developed are promising containers for physically triggered, targeted drug delivery