Synthesis of polyester nanoparticles in miniemulsion obtained by radical ring-opening of BMDO and their potential as biodegradable drug carriers.

5,6-Benzo-2-methylene-1,3-dioxepane (BMDO) is used to obtain degradable polymeric nanoparticles via a statistical free-radical copolymerization with MMA and styrene in direct miniemulsion. The nanoparticles are analyzed by means of IR, NMR, DLS, SEM, and TEM. They show excellent cellular uptake and drug delivery properties. The cellular uptake into HeLa cells of particles resulting from copolymerization of BMDO with styrene is drastically enhanced compared to pure polystyrene. As a model drug system, paclitaxel is incorporated in PBMDO particles and its release and the effect on HeLa cells is studied and compared to commercial drug formulations. It is found that a drug delivery system based on PBMDO shows an excellent pharmacological effect.

BSA adsorption on differently charged polystyrene nanoparticles using isothermal titration calorimetry and the influence on cellular uptake.

BSA adsorption onto negatively and positively charged polystyrene nanoparticles was investigated. The nanoparticles were characterized in terms of particle size, zeta potential, surface group density, and morphology. The adsorption behavior of BSA on the particle surface, as a function of pH and overall charge of the particle, was studied using ITC. Different thermodynamic data such as enthalpy changes upon binding and stoichiometry of the systems were determined and discussed. The degree of surface coverage with BSA was calculated using the thermodynamic data. The cellular uptake of particles before and after BSA adsorption was studied using HeLa cells in the presence and absence of supplemented FCS in the cell culture medium.

Direct and indirect effects of functionalised fluorescence-labelled nanoparticles on human osteoclast formation and activity.

Recently, it was demonstrated that phosphonate-functionalised nanoparticles were successfully taken up by mesenchymal stem cells without influencing their viability and differentiation capacity, suggesting that they may provide a promising basis for the development of nanoparticles for drug delivery or cell labelling. The present study aimed to investigate the effects of these nanoparticles on osteoclast formation and activity as well as on the inflammatory response of osteoclasts and osteoblasts. The intracellular uptake of the particles by human osteoclasts and osteoblasts was demonstrated by confocal laser scanning microscopy, transmission electron microscopy and fluorescence microscopy. The expression of tartrate-resistant acid phosphatase, carboanhydrase II, cathepsin K, calcitonin receptor and osteoclast-specific vacuolar proton pump subunit TCIRG1 as well as actin ring formation were not significantly altered in osteoclasts by particle treatment, as demonstrated by cytochemical staining and immunostaining. Active calcium phosphate resorption by osteoclasts was also not significantly influenced by the particles. The expression and secretion of pro-inflammatory cytokines (IL-6, IL-1ß) by osteoclasts and osteoblasts and the expression of osteoclast-regulating genes (M-CSF, OPG, RANKL) in osteoblasts were similarly not significantly affected. In conclusion, phosphonate-functionalised nanoparticles did not affect osteoclast formation and activity either directly or indirectly, suggesting that they could provide a promising tool for the development of particle-based treatments for anti-resorptive therapies.

Nanostructured coatings by adhesion of phosphonated polystyrene particles onto titanium surface for implant material applications.

Titanium that is covered with a native oxide layer is widely used as an implant material; however, it is only passively incorporated in the human bone. To increase the implant-bone interaction, one can graft multifunctional phosphonic compounds onto the implant material. Phosphonate groups show excellent adhesion properties onto metal oxide surfaces such as titanium dioxide, and therefore, they can be used as anchor groups. Here, we present an alternative coating material composed of phosphonate surface-functionalized polystyrene nanoparticles synthesized via free radical copolymerization in a direct (oil-in-water) miniemulsion process. Two types of functional monomers, namely, vinylphosphonic acid (VPA) and vinylbenzyl phosphonic acid (VBPA), were employed in the copolymerization reaction. Using VBPA as a comonomer leads to particles with a higher density of surface phosphonate groups in comparison to those obtained with VPA. VBPA-functionalized particles were used for the coating formation on the titanium surface. The particles monolayer was investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM) employing titanium and silicium tip with the native OH groups. Force versus distance curves proves the strong adhesion between the phosphonated particles and the titanium (or silicium) surfaces in contrast to the nonfunctionalized polystyrene particles. Finally, as a proof of concept, the particles adhered to the surface were further used to nucleate hydroxyapatite, which has high potential for bioimplants.

Effect of functionalised fluorescence-labelled nanoparticles on mesenchymal stem cell differentiation.

The combined use of nanoparticles and mesenchymal stem cells (MSC) in regenerative medicine requires the incorporation of the particles and, at the same time, undisturbed cell viability and maintenance of the multi-lineage potential of MSC. The aim of this study was to investigate the uptake of novel phosphonate-functionalised polystyrene nanoparticles prepared by miniemulsion polymerisation. After exposition of human MSC to the particles, their uptake and localisation were analysed by flow cytometry, confocal laser scanning microscopy (CLSM), and transmission electron microscopy (TEM). The osteogenic, adipogenic and chondrogenic differentiation potential was examined by analysing representative marker genes by RT-PCR. Flow cytometry revealed that after 5 and 16 days more than 98% of the MSC and of the cells, which underwent osteogenic and adipogenic differentiation were positive for particle association. CLSM and TEM demonstrated the successful intracellular incorporation of the particles without using any transfection agents and their presence over the cultivation period. The cell viability was found to be unaffected. Particle treated MSC maintained their potential for osteogenic, adipogenic and chondrogenic differentiation. It was concluded that the surface functionalisation with phosphonate groups provides a promising basis for the development of nanoparticles with high intracellular uptake rates for drug delivery or cell labelling.