WP03 – In vitro assessment

Objectives

The overall objective of WP3 is to evaluate the potential of the developed materials that have been developed in WP1 to support cartilage formation. In addition, these materials must fulfill the criteria in terms of mechanical properties and gelation kinetics for 3D printing. In WP03 materials will be selected for further in vivo evaluation in WP5 and WP6. Accredited testing according to DIN EN ISO/IEC 17025 and DIN ISO 10993-5 will be performed according to the QM/RAM system and product development plan. 

Workpackage Description

In this work package we will evaluate the generated materials WP01 in vitro. Modification of the materials will improve their specific chondrogenic potential. To assess chondrogenic extracellular matrix accumulation and morphology, the performance of all materials will be benchmarked against that of fibrin, a current clinical gold standard for the treatment of cartilage defects.

The chondrogenic potential in vitro of the developed materials (WP01) for the generation of the superficial (top) and middle layers of the HydroZONES construct will be evaluated in vitro in close collaboration by #05 UMCU, #02 UNAV and #03 ORTHO-HD, using both cast and printed hydrogel constructs. Whilst cell culture protocols often differ slightly between institutions, Partners #05 UMCU, #02 UNAV and #03 ORTHO-HD will develop, for the HydroZONES consortium, a standard operating procedure for the isolation and culture of chondrocytes and MSCs to ensure that all culture conditions at the different partners are identical with respect to ensure the exchange and comparability of the acquired data.

The 5 best performing functionalised hydrogels, with respect to cartilage matrix deposition, will be combined with the clinically-approved thermoplast scaffold material of Partner #14 CCT, using #05 UMCU bioprinting technology. Subsequently, biphasic and hybrid constructs will be evaluated for chondrogenic potential. Based on these outcomes the HydroZONES constructs for subsequent in vivo testing (WP05 and 06) will be selected.

Different layers of hard polymer and cell-laden hydrogels
Fig. 1: Different layers of hard (thermoplast) polymer and cell-laden hydrogels can be simultaneously printed to mimic the structure of the native tissue.

WP Leader

University Medical Center Utrecht

Dr. Jos Malda