The biofabrication technique comprises two processes for every volume: first, the fused deposition of the thermoplastic polymer to obtain the scaffold and second, the injection of the bioink and CaCl2 solution to fill the printed scaffold.
Firstly, commercial non-medical grade PLA (1.75 mm filament) was used and it was melted at 210 ºC in a nozzle extruder and deposited, at rate of 1.4 mm/s, in a layerby- layer manner to generate a previously designed matrix (600 micrometer pore size, cylinder-type structure of 10 mm diameter and 10 mm height). After printing four layers of the scaffold (0.35 mm thickness), the bioink (25 microL; 1×10^6 cells mL-1 encapsulated in alginate, 2% w/v) was injected into the scaffold in the selected points (speed: 8 microL/s; 21G nozzle).
Then, cell-loaded alginate solution was chemically cross-linked by following with an injection of 100mM CaCl2 (40 microL; speed: 8 microL/s). All these processes were repeated until the target dimensions of the constructs were fulfilled. Finally, all cell-laden scaffolds were cultured in medium, as previously described, at 37 C in 5% CO2 atmosphere.
References
| Baena, J.M.; Jiménez, G.; López-Ruiz, E.; Antich, C.; Griñán-Lisón, C.; Perán, M.; Gálvez-Martín, P.; Marchal, J.A. Volume-by-volume bioprinting of chondrocytes-alginate bioinks in high temperature thermoplastic scaffolds for cartilage regeneration. Exp. Biol. Med.2019, 244, 13–21. https://doi.org/10.1177/1535370218821128 |
| 1. L opez-Ruiz E, Pera´n M, Cobo-Molinos J, Jime´nez G, Pic on M, Bustamante M, Arrebola F, Herna´ndez-Lamas MC, Delgado-Mart ınez AD, Monta~nez E, Marchal JA. Chondrocytes extract from patients with osteoarthritis induces chondrogenesis in infrapatellar fat pad-derived stem cells. Osteoarthr Cartil.2013;21:246–58 |
| Number | Category | Product | Amount |
|---|---|---|---|
| 1 | Biomaterials | PLA standard-100 gram | 3 |
| 2 | - | Articular chondrocytes | 1 |
| 3 | - | alginate, 2% w/v | 1 |
| 4 | - | 100mM CaCl2 | 1 |