Shaping collagen for engineering hard tissues: Towards a printomics approach

Hard tissue engineering has evolved over the past decades, with multiple approaches being explored and developed. Despite the rapid development and success of advanced 3D cell culture, 3D printing technologies and material developments, a gold standard approach to engineering and regenerating hard tissue substitutes such as bone, dentin and cementum, has not yet been realised. One such strategy that differs from conventional regenerative medicine approach of other tissues, is the in vitro mineralisation of collagen templates in the absence of cells. Collagen is the most abundant protein within the human body and forms the basis of all hard tissues. Once mineralised, collagen provides important support and protection to humans, for example in the case of bone tissue. Multiple in vitro fabrication strategies and mineralisation approaches have been developed and their success in facilitating mineral deposition on collagen to achieve bone-like scaffolds evaluated. Critical to the success of such fabrication and biomineralisation approaches is the collagen template, and its chemical composition, organisation, and density. The key factors that influence such properties are the collagen processing and fabrication techniques utilised to create the template, and the mineralisation strategy employed to deposit mineral on and throughout the templates. However, despite its importance, relatively little attention has been placed on these two critical factors. Here, we critically examine the processing, fabrication and mineralisation strategies that have been used to mineralise collagen templates, and offer insights and perspectives on the most promising strategies for creating mineralised collagen scaffolds. Statement of significance: In this review, we highlight the critical need to fabricate collagen templates with advanced processing techniques, in a manner that achieves biomimicry of the hierarchical collagen structure, prior to utilising in vitro mineralisation strategies. To this end, we focus on the initial collagen that is selected, the extraction techniques used and the native fibril forming potential retained to create reconstituted collagen scaffolds. This review synthesises current best practises in material sourcing, processing, mineralisation strategies and fabrication techniques, and offers insights into how these can best be exploited in future studies to successfully mineralise collagen templates