Nucleic acid vaccines have become a revolutionary technology to give a fast, safe, cost-effective and efficient response against viral infections, such as SARS-CoV-2 or Human papillomavirus (HPV). However, to ensure their effectiveness, the development of adequate methods to protect, carry, and deliver nucleic acids is fundamental. In this work, nanoparticles (NPs) of chitosan (CS)-tripolyphosphate (TPP)-plasmid DNA (pDNA) were thoroughly modulated and characterized, by measuring the charge and size through dynamic light scattering (DLS) and morphology by scanning electron microscopy (SEM). Stability, cytotoxicity and cellular uptake of NPs were also evaluated. Finally, the effect of polyplexes on the expression of HPV E7 antigen in human fibroblast and RAW cells was investigated through polymerase chain reaction (PCR) and real-time PCR. The results showed NPs with a spherical/oval shape, narrow size distribution 20 mV) and good stability after one month of storage at 4 °C in formulation buffer or when incubated in culture medium and trypsin. In vitro studies of NPs cytotoxicity revealed that the elimination of formulation buffers led to an improvement in the rate of cell viability. The E7 antigen transcription was also increased for NPs obtained with high pDNA concentration (60 μg/mL). The analyzed CS-TPP-pDNA polyplexes can offer a promising vehicle for nucleic acid vaccines, not only in the prevention or treatment of viral infections, but also to fight emergent and future pathogens.

Anisotropic, gecko-inspired, microstructured adhesives are one of the most promising solutions for many applications in robotics and biomedical applications that require controllable adhesives to grip flat surfaces. In such adhesives, normal adhesion is negligible when loaded solely in the normal direction, but becomes available when the adhesive is loaded in shear first. However, much remains to be learned regarding the friction and failure mechanisms of microstructures loaded in shear. In response, we analysed the load–displacement profiles of wedge-shaped microstructured adhesives comprised of nine different silicone elastomers and their mixtures loaded in shear. The results show that the friction profile depends on at least three factors related to material properties: interfacial adhesion strength in the normal direction (work of separation), elastic modulus and the sample's imperfections (e.g. contamination, misalignment and moulding errors). Moreover, the work of separation influences the maximum friction load such that for materials with the same elastic modulus, the strongest interfacial adhesion yields the lowest friction force. To explain this, we suggest that strongly adhering materials will lead to a macroscopic frictional sliding of the array rather than previously reported stick-slip behaviour.

Hydrogels are suitable for osteochondral defect regeneration as they mimic the viscoelastic environment of cartilage. However, their biomechanical properties are not sufficient to withstand high mechanical forces. Therefore, we have prepared electrospun poly-ε-caprolactone-chitosan (PCL-chit) and poly(ethylene oxide)-chitosan (PEO-chit) nanofibers, and FTIR analysis confirmed successful blending of chitosan with other polymers. The biocompatibility of PCL-chit and PEO-chit scaffolds was tested; fibrochondrocytes and chondrocytes seeded on PCL-chit showed superior metabolic activity. The PCL-chit nanofibers were cryogenically grinded into microparticles (mean size of about 500 µm) and further modified by polyethylene glycol–biotin in order to bind the anti-CD44 antibody, a glycoprotein interacting with hyaluronic acid (PCL-chit-PEGb-antiCD44). The PCL-chit or PCL-chit-PEGb-antiCD44 microparticles were mixed with a composite gel (collagen/fibrin/platelet rich plasma) to improve its biomechanical properties. The storage modulus was higher in the composite gel with microparticles compared to fibrin. The E_loss of the composite gel and fibrin was higher than that of the composite gel with microparticles. The composite gel either with or without microparticles was further tested in vivo in a model of osteochondral defects in rabbits. PCL-chit-PEGb-antiCD44 significantly enhanced osteogenic regeneration, mainly by desmogenous ossification, but decreased chondrogenic differentiation in the defects. PCL-chit-PEGb showed a more homogeneous distribution of hyaline cartilage and enhanced hyaline cartilage differentiation.

Bisphosphonates (BPs) are compounds resembling the pyrophosphate structure. BPs bind the mineral component of bones. During the bone resorption by osteoclasts, nitrogen-containing BPs are released and internalized, causing an inhibition of the mevalonate pathway. As a consequence, osteoclasts are unable to execute their function. Alendronate (ALN) is a bisphosphonate used to treat osteoporosis. Its administration could be associated with adverse effects. The purpose of this study is to evaluate four different ALN concentrations, ranging from 10⁻⁶ to 10⁻¹⁰ M, in the presence of different combinations of M-CSF and RANKL, to find out the effect of low ALN concentrations on osteoclastogenesis using rat and human peripheral blood mononuclear cells. The cytotoxic effect of ALN was evaluated based on metabolic activity and DNA concentration measurement. The alteration in osteoclastogenesis was assessed by the activity of carbonic anhydrase II (CA II), tartrate-resistant acid phosphatase staining, and actin ring formation. The ALN concentration of 10⁻⁶ M was cytotoxic. Low ALN concentrations of 10⁻⁸ and 10⁻¹⁰ M promoted proliferation, osteoclast-like cell formation, and CA II activity. The results indicated the induction of osteoclastogenesis with low ALN concentrations. However, when high doses of ALN were administered, their cytotoxic effect was demonstrated.

Microstrucutred (sometimes referred to as gecko-like) adhesives have numerous advantages over flat films, especially for practical applications on non-ideal surfaces that may be uneven or contaminated with dust. However, due to interdependence among material surface and bulk properties, the best material to fabricate such adhesives is still unknown. In this work, we analyzed eleven commercially available silicone elastomers to evaluate their use as flat and microstructured adhesives to address multiple material related questions that may impact the choice of the ``best'' material for microstructured dry adhesives. To illustrate the applicability of the measured properties to modeling microstructured surfaces, we use stalk-shaped microstructures, whose contact mechanics are well understood. We demonstrated that there is no correlation between the adhesion strength of flat and microstructured adhesives; while bulk dissipation is the most important factor influencing the adhesion strength of flat elastomers, after microstructurization, interface toughness becomes more important. Therefore, microstructured elastomers loaded with high surface energy additives may demonstrate higher adhesion than their flat counterparts. We also compare the adhesion of flat and microstructured silicone elastomer on rough substrates. In this case, we show that while flat elastomer adhesion decreases with increasing substrate roughness, microstructured silicone adhesion actually increases with increasing roughness up to 0.19 μm. This is the first time an increase in adhesion strength on rough surfaces is reported for materials stiffer than 1 MPa.

In this study, we developed a novel solid lipid nanoparticle (SLN) formulation for drug delivery of small hydrophilic cargos to the retina. The new formulation, based on a gel core and composite shell, allowed up to two-fold increase in the encapsulation efficiency. The type of hydrophobic polyester used in the composite shell mixture affected the particle surface charge, colloidal stability, and cell internalization profile. We validated SLNs as a drug delivery system by performing the encapsulation of a hydrophilic neuroprotective cyclic guanosine monophosphate analog, previously demonstrated to hold retinoprotective properties, and the best formulation resulted in particles with a size of ±250 nm, anionic charge > −20 mV, and an encapsulation efficiency of ±60%, criteria that are suitable for retinal delivery. In vitro studies using the ARPE-19 and 661W retinal cell lines revealed the relatively low toxicity of SLNs, even when a high particle concentration was used. More importantly, SLN could be taken up by the cells and the release of the hydrophilic cargo in the cytoplasm was visually demonstrated. These findings suggest that the newly developed SLN with a gel core and composite polymer/lipid shell holds all the characteristics suitable for the drug delivery of small hydrophilic active molecules into retinal cells.