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Session: Joints
Session starts: Friday 25 January, 13:00
Presentation starts: 13:30
Room: Lecture room 535

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Ludo van Haasterecht (Department of Biophotonics and Medical Imaging, VU University, Amsterdam Movement Sciences)
Dafydd Visscher (Department of Plastic, Reconstructive & Hand Surgery, VU University Medical Center, Amsterdam Movement Sciences)
Yigitcan Sumbelli (Anadolu University)
M.N. Helder (Department of Oral and Maxillofacial Surgery/Oral Pathology, VU University Medical Center, Amsterdam Movement Sciences)
J.P.J Don Griot (Department of Plastic, Reconstructive & Hand Surgery, VU University Medical Center, Amsterdam Movement Sciences)
M.L. Groot (Department of Biophotonics and Medical Imaging, VU University, Amsterdam Movement Sciences)
P.P.M van Zuijlen (Department of Plastic, Reconstructive & Hand Surgery, VU University Medical Center, Amsterdam Movement Sciences)

Abstract:
Background: Reconstruction of extensively damaged facial cartilage is a challenging task. As autologous and synthetic grafts have significant disadvantages, novel reconstructive strategies are required to provide patients with durable, patient-specific cartilage tissue. Three-dimensional bioprinting has the potential to solve several issues encountered in tissue engineering. However current bioinks, containing both cells and hydrogel, often miss the necessary chondrogenic cues for adequate cartilage formation. The addition of decellularized cartilage powder to the bioink may provide chondrocytes with the necessary signals required for adequate cell proliferation and chondrogenesis. Objective: The objectives were: i) To develop a cartilage-specific hydrogel with adequate bioprinting capabilities, and ii) to support and improve chondrocyte survival and post-printing neocartilage formation. Methods: Native caprine auricular cartilage was decellularized and enzymatically digested to produce extracellular matrix (ECM)-fragments. The effect of ECM addition to gelatin/alginate-based bioink of different concentrations was evaluated. Rheological properties, bioprintability, and swelling capacity, as well as cell survival were evaluated for both the gelatin/alginate bioink and the ECM-containing bioink. Following a culture time of three weeks, the bioprinted scaffolds were subjected to higher harmonic generation microscopy. Second Harmonic Generation (SHG) microscopy has a high specificity for non-centrosymmetric structures and was therefore used in evaluating whether deposition of collagen type I and II took place. Results: The bioink consisting of eight percent gelatin and five percent alginate, showed excellent bioprintability. Alterations in printability due to the addition of ECM fragments were resolved by adjusting the bioink components accordingly. Chondrocytes in bioprinted scaffolds displayed high cell survival, independent of ECM addition. Addition of ECM resulted in increased cell aggregation and collagen formation as shown using SHG microscopy. Conclusion: The addition of ECM fragments to 3D-printed bioink increases chondrocyte proliferation and neo-cartilage formation. Significance: The results in this study constitute a step towards bioprinted elastic cartilage for patient-specific reconstruction. Specifically, the addition of ECM to bioink increases cell proliferation and neocartilage formation necessary for the development of durable cartilage tissue.