Introduction: Tissue engineering (TE) is defined as the seeding and adhesion of human cells over a scaffold, arising as a viable alternative for reproduction of organs and tissues. Said scaffold must present a surface that promotes cell adhesion, growth and differentiation, while providing a porous structure for cellular migration and tissue formation. Fibrous scaffolds have been considered appropriate for culturing certain cell lines due to its ability to induce morphological differentiation by mimicking the tissue's original extracellular matrix (ECM). The application of the polymeric blend of Poly (Lactic-co-Glycolic Acid) (PLGA) and Polyisoprene (PI) - Cellprene ®[1] - has already been successfully reported for applications in cranioplasty and pneumology. The fibres of PLGA/PI obtained through the dripping technique is proposed as an attractive alternative for soft tissue engineering, as well as presenting an alternative technique for fibres manufacturing processes.
Materials and Methods: The dripping method[2] consists of exposing the diluted blend into an ethanol solution, under rotational agitation, precipitating polymeric fibers. Presenting a mean diameter of 20,8μm, pore size of around 75 μm, porosity of 73% and density around 0,125g/cm³, these fibres have properties relatable to those presented by other fibrous scaffolds successfully applied in TE. Laminar samples of PLGA/PI fibres were sterilized under UV lights and cultured with murine skeletal-muscle cells (line C2C12). Observed after 1, 3, 7 and 14 days of culture, the material's viability was characterised through: Alamar Blue dyeing for indication of cellular population; double stranded DNA readings for indication of cellular proliferation; and blue toluidine dyeing to provide visual observation over cellular morphology. Blank glass coverslips were used as control group and ANOVA was applied as statistic treatment.
Results and Discussion: During all periods, the readings of cell population and DNA amount was in the same level for culture over PLGA/PI fibres and the control group, without significant difference between groups (α=0,025). For both groups, a significant increase in all the readings between 1-14 days was observed. Optical microscope observations confirm a visual similarity in cell population for both groups. Cell morphology observed after 14 days of culture indicates production of ECM by the cells in both groups, which highlights the biocompatibility of PLGA/PI fibres obtained through the dripping process.
Conclusions: These primary results indicate the suitability of PLGA/PI fibres for application in muscle tissue engineering, as well as the viability of the dripping process as an alternative for polymeric fibres production.
CNPq - Brasil; CAPES - Brasil
References:
[1] MARQUES, D. R.; DOS SANTOS, L. A.; SOUSA, V. C.; SANCHES, P. R. S.; MACEDO NETO, A. V. (2011) Blendas Poliméricas de Poli (Ácido Láctico-co-Glicólico) e Poliisopreno. Patent Registration Number: 0000221010682444.
[2] DOS SANTOS, L. A.; SOUSA, V. C.; BERGMANN, C. P.; VASCONCELLOS, L. A. (2011) Processo de Produção de Polímeros em Fibras. Patent Registration Number: 0000221109247952.