Abstract
Fabricating polymeric scaffolds using cost-effective manufacturing processes is still challenging. Gas foaming techniques using supercritical carbon dioxide (scCO2) have attracted attention for producing synthetic polymer matrices; however, the high-pressure requirements are often a technological barrier for its widespread use. Compressed 1,1,1,2-tetrafluoroethane, known as Freon R134a, offers advantages over CO2 in manufacturing processes in terms of lower pressure and temperature conditions and the use of low-cost equipment. Here, we report for the first time the use of Freon R134a for generating porous polymer matrices, specifically polylactide (PLA). PLA scaffolds processed with Freon R134a exhibited larger pore sizes, and total porosity, and appropriate mechanical properties compared with those achieved by scCO2 processing. PLGA scaffolds processed with Freon R134a were highly porous and showed a relatively fragile structure. Human mesenchymal stem cells (MSCs) attached to PLA scaffolds processed with Freon R134a, and their metabolic activity increased during culturing. In addition, MSCs displayed spread morphology on the PLA scaffolds processed with Freon R134a, with a well-organized actin cytoskeleton and a dense matrix of fibronectin fibrils. Functionalization of Freon R134a-processed PLA scaffolds with protein nanoparticles, used as bioactive factors, enhanced the scaffolds’ cytocompatibility. These findings indicate that gas foaming using compressed Freon R134a could represent a cost-effective and environmentally friendly fabrication technology to produce polymeric scaffolds for tissue engineering approaches.
Highlights
Scaffolds for tissue engineering should be degradable and biocompatible and have an appropriate porous structure and mechanical properties to allow cell colonization and growth [1,2,3,4,5]
We explored the use of compressed Freon R134a for processing PLA matrices at low pressure and compared the results with the same polymeric matrices processed at higher pressure using the well-implemented scCO2
We studied the processing conditions of PLA with CO2 and Freon R134a compressed fluids and their influence on the physicochemical and mechanical properties of the resulting porous matrices
Summary
Scaffolds for tissue engineering should be degradable and biocompatible and have an appropriate porous structure and mechanical properties to allow cell colonization and growth [1,2,3,4,5]. An attractive alternative for overcoming the limitations associated with conventional methods is the “gas foaming” technique, which uses supercritical carbon dioxide (scCO2) to obtain materials with a high degree of porosity (up to 80%) [9,10,11] This organic solvent-free process can occur at physiological temperatures, allowing the incorporation of biological agents [12,13,14,15,16]. We fabricated PLGA matrices using compressed Freon R134a To enhance their bioactivity, pNPs derived from green fluorescence protein (GFP), a commonly used marker for live-cell imaging, were used to functionalize the surface of the resulting porous scaffolds by means of a filtration process. To explore the suitability of scaffolds processed with compressed fluids and functionalized with pNPs for bone tissue engineering applications, we investigated their cytocompatibility using human mesenchymal stem cells (MSCs) as precursors of osteoblasts, the bone-forming cells
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