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  • Multi-material 3D bioprinting of porous constructs for cartilage regeneration.

Multi-material 3D bioprinting of porous constructs for cartilage regeneration.

Materials science & engineering. C, Materials for biological applications (2020-04-02)
Laura Ruiz-Cantu, Andrew Gleadall, Callum Faris, Joel Segal, Kevin Shakesheff, Jing Yang
ABSTRACT

The current gold standard for nasal reconstruction after rhinectomy or severe trauma includes transposition of autologous cartilage grafts in conjunction with coverage using an autologous skin flap. Harvesting autologous cartilage requires a major additional procedure that may create donor site morbidity. Major nasal reconstruction also requires sculpting autologous cartilages to form a cartilage framework, which is complex, highly skill-demanding and very time consuming. These limitations have prompted facial reconstructive surgeons to explore different techniques such as tissue engineered cartilage. This work explores the use of multi-material 3D bioprinting with chondrocyte-laden gelatin methacrylate (GelMA) and polycaprolactone (PCL) to fabricate constructs that can potentially be used for nasal reconstruction. In this study, we have investigated the effect of 3D manufacturing parameters including temperature, needle gauge, UV exposure time, and cell carrier formulation (GelMA) on the viability and functionality of chondrocytes in bioprinted constructs. Furthermore, we printed chondrocyte-laden GelMA and PCL into composite constructs to combine biological and mechanical properties. It was found that 20% w/v GelMA was the best concentration for the 3D bioprinting of the chondrocytes without comprising the scaffold's porous structure and cell functionality. In addition, the 3D bioprinted constructs showed neocartilage formation and similar mechanical properties to nasal alar cartilage after a 50-day culture period. Neocartilage formation was also observed in the composite constructs evidenced by the presence of glycosaminoglycans and collagen type II. This study shows the feasibility of manufacturing neocartilage using chondrocytes/GelMA/PCL 3D bioprinted porous constructs which could be applied as a method for fabricating implants for nose reconstruction.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
L-Ascorbic acid, 99%
Sigma-Aldrich
Methacrylic anhydride, contains 2,000 ppm topanol A as inhibitor, ≥94%
Sigma-Aldrich
Transforming Growth Factor-β1 human, TGF-β1, recombinant, expressed in CHO cells, powder, suitable for cell culture
Sigma-Aldrich
Anti-phospho-Histone H2A.X (Ser139) Antibody, clone JBW301, clone JBW301, Upstate®, from mouse
Sigma-Aldrich
Nunc® EasYFlasks, Nunclon® Δ Surface, Nunclon® Δ surface treatment, 75 cm2 angled neck, filter cap, 100/cs
Sigma-Aldrich
Gelatin from porcine skin, gel strength 300, Type A
Sigma-Aldrich
Phosphate buffered saline, tablet
Sigma-Aldrich
Donkey serum
Sigma-Aldrich
MEM Non-essential Amino Acid Solution (100×), without L-glutamine, liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
FGF-2 human, recombinant, expressed in E. coli, ≥95% (SDS-PAGE), ≥95% (HPLC)