The attributes of implant surfaces are pivotal for successful osseointegration. Among surface engineering strategies, microtopography stands out as a promising approach to promote early cellular interactions. This study aims to design and craft a novel biomimetic osteon-like surface modification and to compare its impact on human mesenchymal stem cells (hMSCs) with four established topographies: blank, inverted pyramids, protrusions, and grooves. Poly-ε-caprolactone samples are fabricated using 2-photon-polymerization and soft lithography, prior to analysis via scanning electron microscopy (SEM), water contact angle (WCA), and protein adsorption assays. Additionally, cellular responses including cell attachment, proliferation, morphology, cytoskeletal organization, and osteogenic differentiation potential are evaluated. SEM confirms the successful fabrication of microtopographies, with minimal effect on WCA and protein adsorption. Cell attachment experiments demonstrate a significant increase on the osteon-like structure, being three times higher than on the blank. Proliferation assays indicate a fourfold increase with osteon-like microtopography compared to the blank, while ALP activity is notably elevated with osteon-like microtopography at days 7 (threefold increase over blank) and 14 (fivefold increase over blank). In conclusion, the novel biomimetic osteon-like structure demonstrates favorable responses from hMSCs, suggesting potential for promoting successful implant integration in vivo.