Coaxial Synthesis of PEI-Based Nanocarriers of Encapsulated RNA-Therapeutics to Specifically Target Muscle Cells.

In this work, we performed a methodological comparative analysis to synthesize polyethyleneimine (PEI) nanoparticles using (i) conventional nanoprecipitation (NP), (ii) electrospraying (ES), and (iii) coaxial electrospraying (CA). The nanoparticles transported antisense oligonucleotides (ASOs), either encapsulated (CA nanocomplexes) or electrostatically bound externally (NP and ES nanocomplexes). After synthesis, the PEI/ASO nanoconjugates were functionalized with a muscle-specific RNA aptamer. Using this combinatorial formulation methodology, we obtained nanocomplexes that were further used as nanocarriers for the delivery of RNA therapeutics (ASO), specifically into muscle cells. In particular, we performed a detailed confocal microscopy-based comparative study to analyze the overall transfection efficiency, the cell-to-cell homogeneity, and the mean fluorescence intensity per cell of micron-sized domains enriched with the nanocomplexes. Furthermore, using high-magnification electron microscopy, we were able to describe, in detail, the ultrastructural basis of the cellular uptake and intracellular trafficking of nanocomplexes by the clathrin-independent endocytic pathway. Our results are a clear demonstration that coaxial electrospraying is a promising methodology for the synthesis of therapeutic nanoparticle-based carriers. Some of the principal features that the nanoparticles synthesized by coaxial electrospraying exhibit are efficient RNA-based drug encapsulation, increased nanoparticle surface availability for aptamer functionalization, a high transfection efficiency, and hyperactivation of the endocytosis and early/late endosome route as the main intracellular uptake mechanism.

Effect of strength training and the practice of Alpine skiing on bone mass density, growth, body composition, and the strength and power of the legs of adolescent skiers.

This work examines the influence of practicing strength training and Alpine skiing over 2 years on bone mineral density (BMD), growth, body composition, and the strength and power of the legs of adolescent skiers. The study subjects were 20 adolescent skiers (10 girls and 10 boys) and 19 sedentary adolescents (9 girls and 10 boys), all 13-16 years of age. The BMDs of the lumbar column (L2-L4) and hip (neck of the femur, trochanter, and Ward's triangle) were determined by dual x-ray photon absorptiometry at the beginning and end of the experimental period. The increase in height and the percentage fat and muscular masses of the subjects were also recorded, as was their ability to jump (countermovement jump [CMJ]), their leg strength and power (squat test), and their leg anaerobic power (continuous jump test [CMJ15″]). No significant differences were seen in the increase in height, body weight, or percentage fat mass between the skiers and sedentary subjects, although the boy skiers showed a significant increase in percentage muscular mass (p < 0.05) compared to the sedentary boys. The improvement in the values of the different CMJ variables was significantly greater among the boy skiers than among the sedentary boys (p < 0.001-0.01). The same was true for the girls (p < 0.001), except for CMJ15″. The skiers experienced a significantly greater increase in L2-L4 BMD than the sedentary subjects (boys p < 0.05; girls p < 0.01). These results suggest that Alpine skiing combined with rational strength training involves no special risk for the physical development of young people, has a positive effect on the power and the percentage of muscle mass in the legs, and helps to have a higher bone density in the lumbar spine (L2-L4).