Additively Manufactured Device for Dynamic Culture of Large Arrays of 3D Tissue Engineered Constructs

Abstract

The ability to test large arrays of cell and biomaterial combinations in 3D environments is still rather limited in the context of tissue engineering and regenerative medicine. This limitation can be generally addressed by employing highly automated and reproducible methodologies. This study reports on the development of a highly versatile and upscalable method based on additive manufacturing for the fabrication of arrays of scaffolds, which are enclosed into individualized perfusion chambers. Devices containing eight scaffolds and their corresponding bioreactor chambers are simultaneously fabricated utilizing a dual extrusion additive manufacturing system. To demonstrate the versatility of the concept, the scaffolds, while enclosed into the device, are subsequently surface-coated with a biomimetic calcium phosphate layer by perfusion with simulated body fluid solution. 96 scaffolds are simultaneously seeded and cultured with human osteoblasts under highly controlled bidirectional perfusion dynamic conditions over 4 weeks. Both coated and noncoated resulting scaffolds show homogeneous cell distribution and high cell viability throughout the 4 weeks culture period and CaP-coated scaffolds result in a significantly increased cell number. The methodology developed in this work exemplifies the applicability of additive manufacturing as a tool for further automation of studies in the field of tissue engineering and regenerative medicine.

In vitro cyclic compressive loads potentiate early osteogenic events in engineered bone tissue

“…Hence, we developed compression bioreactor system to study the effects of cyclic compressive strains on the maturation of engineered bone grafts. Studies have shown the development of array of scaled‐down, multi‐compartment bioreactor systems aid in acceleration of tissue characterization and also help to minimize experimental variations during parametric optimization . Our custom‐built, multi‐chamber compression bioreactor system can apply a range of strains on cellular scaffolds in a semi‐high‐throughput fashion with independent control of loading and medium conditions in each of the chambers.…”

“…Studies have shown the development of array of scaled-down, multi-compartment bioreactor systems aid in acceleration of tissue characterization and also help to minimize experimental variations during parametric optimization. 45,46 Our custom-built, multichamber compression bioreactor system can apply a range of strains on cellular scaffolds in a semi-high-throughput fashion with independent control of loading and medium conditions in each of the chambers. In our study, we seeded MSCs on the scaffolds using fibrin glue which has been proven to facilitate high seeding density, good cellular retention, and robust osteogenesis.…”

A bench‐top molding method for the production of cell‐laden fibrin micro‐fibers with longitudinal topography

“…The method consists of four main steps: (a) the injection of the fibrin components including the cells into the silicone tube, (b) the gelation process, (c) the ejection of the gel after polymerization and (d) the stretching of the resulting fiber. We expect that already existing solutions for automated pipetting, handling, collection, gripping of samples and controlled linear displacement could be assembled to create an ad‐hoc automated system for the production of the fibers in a sterile, closed environment (Bhuthalingam, Lim, Irvine, & Venkatraman, ; Bartolo, Domingos, Gloria, & Ciurana, ; P. F. Costa et al, ; Kikuchi et al, ; Mekhileri et al, ). A GMP conform production is facilitated by the limited number of parts in contact with the materials composing the fibers and the commercial availability of GMP‐conform cell‐work stations.…”

Challenges in engineering large customized bone constructs

“…4). Removal of the scaffolds required breaking apart the outer chamber and would benefit from improved ease of access in future designs; however, a subsequent study did demonstrate the capability for the system to be scaled up with the simultaneous culture of 96 scaffolds (Costa et al, 2015). If fluid paths trace a tortuous, indirect path through the scaffold, the effective distance of mass transport is increased.…”