Electrospun scaffolds are widely used in tissue engineering; however, a common problem is the poor cell infiltration because of the small pore size and tightly packed structure of these fibrous scaffolds. To address this issue, a novel technique was developed to fabricate electrospun silk fibroin (SF) scaffolds with rather macropores and high porosity using electrospraying-generated PEO microparticles as porogen. The morphology and pore size of MPES scaffolds were evaluated by scanning electron microscopy. It was revealed that MPES scaffold had a relatively loose structure with an increase of mean pore size (i.e., approx. 30 μm of MPES vs. approx. 5 μm of traditional electrospun scaffolds (TES) and porosity (i.e., 95% vs. 84% of TES). Culture of mouse 3T3 fibroblast in TES and MPES scaffold revealed that both scaffolds could support cell attachment, spread and proliferation. Yet, cell inflitration in vitro under the static culture condition only occurred in the MPES scaffold. Subcutaneous implantation of scaffolds in rats further confirmed that the tissue ingrowth was more efficient in the MPES scaffold compared to TES scaffold. Thus, the use of PEO microparticles as porogen was a feasible and effective method for creating macroporous electrospun SF scaffold, which provided an alternative to address the limitation of cell infiltration associated with electrospun fibrous scaffold. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 101A: 3474–3481, 2013.