Engineer a pre‐metastatic niched microenvironment to attract breast cancer cells by utilizing a 3D printed polycaprolactone/nano‐hydroxyapatite osteogenic scaffold – An in vitro model system for proof of concept

Abstract Breast cancer bone metastasis is not a random process. It is affected by the local microenvironment which determines the propensity of cancer cells to invade and colonize into the secondary sites. This microenvironment is termed a pre‐metastatic niche. With the flexibility to incorporate different biofactors, tissue‐engineering scaffolds provide an advantageous environment to promote “designed” osteogenesis that may mimic the bony pre‐metastatic niche. In the current study, designed polycaprolactone (PCL) scaffolds enriched with nano‐hydroxyapatite (nHA) were fabricated through three‐dimensional (3D) printing. Subsequently, human mesenchymal stem cells (hMSCs) were seeded onto PCL‐nHA scaffolds for osteogenic differentiation to establish the pre‐metastatic niched microenvironment. Furthermore, transwell migration assay was used to investigate recruitment of MDA‐MB‐231, MCF‐7, and MDA‐MB‐453 breast cancer cells to the osseous PCL‐nHA scaffolds. Our results showed that the mRNA levels of alkaline phosphatase (ALP), runt‐related transcription factor 2 (Runx2), and osteocalcin (OCN) of hMSCs on the PCL‐nHA scaffolds were dramatically increased compared those with the PCL scaffolds (control) at day 7, 14, and 28. Meanwhile, the migration analysis showed that the higher maturation of osteogenesis and bone metabolism collectively contributed to the creation of a more favorable niched site for the cancerous invasion. Moreover, one of the hypothesized key mediators for the promoted migration, CXCL12, was confirmed using an assay of antagonist LIT‐927. This early study demonstrated that a designed tissue engineering scaffold can be utilized to create a bone‐mimicking environment that serves as a novel platform to recapitulate the pre‐metastatic niche and help interrogate the scheme of bone metastasis by breast cancer.