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The very synthesis of functional microparticles is generally deemed the most necessary, but obviously not the only step in successful product development. The behavior of obtained microparticles has to be tested in environments resembling the end use conditions to ensure the desired functionality. During the testing, various problems concerning particles behavior can arise, e.g. unwanted adhesion (before the successful delivering of particles to the region of interest, they will adhere somewhere else, thus hindering the delivery of transported substance), insufficient adhesion (in cases, when the particle adhesion is desired, e.g. specific adhesion for targeted delivery, the end amount of adhered particles might not be sufficient for reaching the expected concentration of released substance, meaning adhesion is not strong enough under given conditions) or particle breakage (some particles are of more fragile structure, which can result in condition limitations, in which they can exist without damage). Furthermore, regarding specific adhesion, the demonstration of such particle functionality should also be performed before testing on living organisms, preferably in conditions resembling the end use.
Breast cancer (BrCa) is the second commonest cause of cancer-related deaths in women. The metastatic breast cancer exhibits a high affinity to bone, leading to debilitating skeletal complications associated with significant morbidity and poor prognosis. Traditional in vitro and in vivo BrCa bone metastasis models contain many inherent limitations with regards to controllability, reproducibility, and flexibility of design. Thus, the objective of this research is to use a 3D bioprinting system and nanomaterials to recreate a biomimetic and tunable bone model suitable for the effective simulation and study of metastatic BrCa invading and colonizing a bone environment. For this purpose, we designed and 3D printed a series of scaffolds, comprised of a bone microstructure and nano hydroxyapatites (nHA, inorganic nano components in bone). The size and geometry of the bone microstructure was varied with 250 and 150 µm pores, in repeating square and hexagon patterns, for a total of four different pore geometries. 3D bioprinted scaffolds were subsequently conjugated with nHA, using an acetylation chemical functionalization process and then characterized by scanning electron microscope (SEM). SEM imaging showed that our designed microfeatures were printable with the predesigned resolutions described above. Imaging further confirmed that acetylation effectively attached nHA to the surface of scaffolds and induced a nanoroughness. Metastatic BrCa cell 4 h adhesion and 1, 3 and 5 day proliferation were investigated in the bone model in vitro. The cell adhesion and proliferation results showed that all scaffolds are cytocompatible for BrCa cell growth; in particular the nHA scaffolds with small hexagonal pores had the highest cell density. Given this data, it can be stipulated that our 3D printed nHA scaffolds may make effective biomimetic environments for studying BrCa bone metastasis.
We developed a tunnel-current based identification method by using nano-gap integrated devices. We performed electrical measurements for mono-nucleotide and oligo-nucleotide during its translocation of molecules between the nano-gap. Based on this determined electrical conductivity for single-nucleotides, we electrically identify the base-type in oligonucleotides, and found that this time-profiles represents the molecular translocation behaviors inside nano-gap. This method could be a promising for an electrical nucleotide sequencing methodology with label-free, high-speed, and low-cost.
A dual experimental and numerical top-down approach is applied to investigate the link between osteocyte morphology and mechanical perception of their environment at the progenitor and mature stages. The numerical model is based on explicit tissue morphology discretization to identify bone diffuse damage at the cellular scale. The in vitro experimental model presents a live allograft bone system where a patient progenitor or mature osteocytes were reseeded in fresh human donor cortical bone tissues subjected to mechanical loading. The live systems behaved mechanically as fresh bone and the cells spatially reorganized in vitro as in vivo. The system under mechanical load also showed an adaptation of the calcium membrane transport rate to the expected in vivo mechanical load detected by bone cells at different stages of differentiation.
A cylindrical-shaped micropillar array embedded microfluidic device was proposed to enhance the dispersion of cell clusters and the efficiency of single cell encapsulation in hydrogel. Different sizes of micropillar arrays act as a sieve to break Escherichia coli (E. coli) aggregates into single cells in polyethylene glycol diacrylate (PEGDA) solution. We applied the external force for the continuous breakup of cell clusters, resulting in the production of more than 70% of single cells into individual hydrogel particles. This proposed strategy and device will be a useful platform to utilize genetically modified microorganisms in practical applications.