RESUMO
Three-dimensional (3D) tumor spheroids are regarded as promising models for utilization as preclinical assessments of chemo-sensitivity. However, the creation of these tumor spheroids presents challenges, given that not all tumor cell lines are able to form consistent and regular spheroids. In this context, we have developed a novel layer-by-layer coating of cellulose nanofibril-polyelectrolyte bilayers for the generation of spheroids. This technique builds bilayers of cellulose nanofibrils and polyelectrolytes and is used here to coat two distinct 96-well plate types: nontreated/non-sterilized and Nunclon Delta. In this work, we optimized the protocol aimed at generating and characterizing spheroids on difficult-to-grow pancreatic tumor cell lines. Here, diverse parameters were explored, encompassing the bilayer count (five and ten) and multiple cell-seeding concentrations (10, 100, 200, 500, and 1000 cells per well), using four pancreatic tumor cell lines-KPCT, PANC-1, MiaPaCa-2, and CFPAC-I. The evaluation includes the quantification (number of spheroids, size, and morphology) and proliferation of the produced spheroids, as well as an assessment of their viability. Notably, our findings reveal a significant influence from both the number of bilayers and the plate type used on the successful formation of spheroids. The novel and simple layer-by-layer-based coating method has the potential to offer the large-scale production of spheroids across a spectrum of tumor cell lines.
RESUMO
In the present study, a series of multifunctional drug delivery systems based on mesostructured hydroxyapatite coating and superparamagnetic nanoparticles with pH-responsive characters was prepared. The structure of each new synthesized nanoscale composite was fully characterized by XRD, FTIR, TEM, VSM and BET. The results showed a good ordered mesostructure having large pores, high pore volume, high surface area, and varied super paramagnetic properties. The mesoporous hydroxyapatite coated super paramagnetic Fe3O4 nanoparticles were applied as a drug delivery carrier loaded with doxorubicin (DOX) as a model drug. The storage/release properties of the developed nonocarriers in phosphate buffer saline (PBS) were studied in two certain pHs: pH=7.4 (the human blood pH) and pH=5.5 (pH of cancer cells). The large pores in the synthesized mesoporous acted as an excellent carrier for DOX molecules with a loading efficiency of ≈93% which is much higher than that of the conventional hydroxyapatite particles. When the pH of the release medium (PBS) was changed from 7.4 to 5.5, the drug release increased significantly from 10% of the adsorbed drug to about 70%. DOX-loaded mesostructure hydroxyapatite reduced the viability of SKBR3 and T47D cells by 54.7 and 57.3%, respectively, which were very similar to 56.8 and 60.4% reduction resulted from free DOX incubation. This new drug delivery system which benefits from both super paramagnetic properties and pH-responsive performances may serve as a suitable platform for developing new biocompatible drug carriers and could have a good potential use in targeted cancer therapy.
