STUDY OF CELL VIABILITY OF COMPOSITE POLYMER COATINGS OBTAINED BY EPD METHOD AND POROUS SCAFFOLDS OF PLA MANUFACTURED BY 3D PRINTING

F. LEONARDO REDONDO; DIANA ARGÜELLO; GIAROLI, MARÍA CAROLINA; LAUTARO RIVERA; DIEGO BUSTOS; MARINA UHART; ANDRÉS E. CIOLINO; MARIO D. NINAGO

Mar del Plata

Congreso; XII Latin-American Congress of Artificial Organs and Biomaterials; 2023

Biomaterials play an important role in tissue engineering because they provide new approaches to tissue repair or regeneration. Composites made up of polymers, ceramics and bioactive phosphates are excellent candidates for these uses. Likewise, various methodologies capable of developing these materials have emerged, among which are fused deposition modeling (FDM-3D printing) and electrophoretic deposition (EPD). The aim of this work was to study the adhesion and viability ofmesenchymal stem cells isolated from human periodontal ligament (phMSC) growing on composite coatings based on - caprolactone-block-dimethylsiloxane) (PCL-b-PDMS) with tricalcium phosphate (TCP), and porous scaffolds of poly(lactic acid) (PLA). PLA scaffolds were fabricated by 3D printing, and PCL-b-PDMS/TCP coatings were obtained on stainless steel substrates through EPD technique. The bioactivity was evaluated by immersion in simulated body- fluid (SBF) for 7 and 28 days at 37 °C. Cell viability was studied in cultures using phMSC cells in 12-well plates with Dulbecco´s Modified Eagle Medium (DMEM), 10% of fetal bovine serum and Penicillin and Streptomycin scaffolds and coatings was verified by fluorescence microscopy. The phMSCs were isolated using the explant technique at a density of 100,000 cells/mL. Finally, cell viability assays were performed using the 3-[4,5-dimethylthiazol-2-yl]-2,5diphenyl tetrazolium bromide (MTT) technique. Results and discussion: According to Scanning Electron Microscopy - Energy Dispersive X-ray spectroscopy (SEM- EDX), composite coatings are more compact and uniform at longer immersion times in SBF. Furthermore, EDX analysis revealed Ca/P ratio values between 1.4 and 1.50, close to the stoichiometric values of hydroxyapatite (HA) in human bone. The growth of HA can be attributed to the high diffusion of Ca ionic species relative to P, leading to an increased Ca release at the surface. On the other hand, Fluorescence Microscopy (Nikon TE 2000) analysis on porous scaffolds of PLA revealed 3T3-L1 cells that express the green fluorescent protein (GFP) growing on the porous scaffolds after 3 days of culture in DMEM supplemented with 10 % of FBS. Finally, cell viability results obtained through MTT assay revealed that both materials tested with phMSC cell exhibited a higher absorbance at 540 nm when compared to those without cell growth. Conclusions: The in-vitro assessments evidenced the formation of crystalline structures corresponding to HA in the tested materials. On the other hand, cell viability tests showed the presence of viable phMSC cells in both analyzed materials. The viability of dental mesenchymal stem cells on PCL-b-PDMS/TCP composite materials and porous scaffolds of PLA obtained by 3D printing is demonstrated for the first time, making it of great interest for future applications of regenerative medicine in odontology.