Hexagonal pore geometry and the presence of hydroxyapatite enhance deposition of mineralized bone matrix on additively manufactured polylactic acid scaffolds

Additive manufacturing (AM) has revolutionized the design of regenerative scaffolds for orthopaedic applications, enabling customizable geometric designs and material compositions that mimic bone. However, the available evidence is contradictory with respect to which geometric designs and material compositions are optimal. There is a lack of studies that systematically compare different pore sizes and geometries in conjunction with the presence or absence of calcium phosphates. We therefore evaluated the physicochemical and biological properties of additively manufactured scaffolds based on polylactic acid (PLA) in combination with hydroxyapatite (HA). HA was either incorporated in the polymeric matrix or introduced as a coating, yielding 15 and 2% wt., respectively. Pore sizes of the scaffolds varied between 200 and 450 um and were shaped either triangularly or hexagonally. All scaffolds supported the adhesion, proliferation and differentiation of both primary mouse osteoblasts and osteosarcoma cells up to four weeks, with only small differences in the production of alkaline phosphatase (ALP) between cells grown on different pore geometries and material compositions. However, mineralization of the PLA scaffolds was substantially enhanced in the presence of HA, either embedded in the PLA matrix or as a coating at the surface level, and by larger hexagonal pores. In conclusion, customized HA/PLA composite porous scaffolds intended for the repair of critical size bone defects were obtained by a cost-effective AM method. Our findings indicate that the analysis of osteoblast adhesion and differentiation on experimental scaffolds alone is inconclusive without the assessment of mineralization, and the effects of geometry and composition on bone matrix deposition must be carefully considered in order to understand the regenerative potential of experimental scaffolds.