Hierarchically aligned gradient collagen micropatterns for rapidly screening Schwann cells behavior

Preparation of gradient collagen micropatterns. [Display omitted] •A hierarchically gradient collagen micropattern was successfully fabricated.•The patterns were stable and displayed concentration dependence of collagen on cells.•Micropatterning could effectively up-regulate the key genes of axon and myelination.•The study may be useful for the design and development of artificial implants for nerve regeneration. To penetrate the effect of protein gradient micropattern on peripheral nerve regeneration, the hierarchically aligned gradient collagen micropattern was prepared by micromoulding method and the influence on Schwann cells growth behavior was studied. The morphology, wettability, stability and component variation of the micropatterns were firstly characterized. Then, Schwann cells were cultured and the related mechanism was penetrated. The results showed that the gradient collagen micropattern could be well fabricated. The surface wettability varied with the change of collagen concentration, and the prepared gradient micropattern showed a good stability after PBS immersion for 15 days. The results of Schwann cells culture and morphological index analysis displayed that the prepared gradient collagen micropatten could well regulate the orientation growth of Schwann cells, while a much better cell alignment growth was obtained on the gradient micropattern with higher collagen concentration and wider pattern size. PCR and WB showed that the micropattern structure could effectively up-regulate the key specific genes for axon regeneration and myelination process. Overall, the study provides a systematic and facile method for understanding the effect of various sized micropatterns on cell behavior, which may have a great significance for the development of artificial implants for tissue engineering and regenerative medicine.