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Microtia, frequently encountered in plastic surgery practice, is usually corrected by auricular reconstruction with prostheses or autologous cartilages. In recent decades, however, cartilage tissue engineering has been emerging as a promising alternative for its minimal invasion and low immunogenicity. As a critical factor for tissue engineering, scaffolds are expected to be sufficiently porous and stiff to facilitate chondrogenesis. In this work, we introduce novel poly-L-lactic acid (PLLA) porous microsphere-reinforced silk-based hybrid (SBH) scaffolds with a multihierarchical porous structure. The scaffolds are fabricated by embedding PLLA porous microspheres (PMs) into a blending matrix of silk fibroin (SF) and gelatin solution, followed by mixing with a degummed silk fiber mesh and freeze-drying process. Through adjusting the amount of PLLA PMs, the mechanical strength approximates to natural cartilage and also balanced physical properties were realized. Biological evaluations of SBH scaffolds, both in vitro and in vivo, were conducted and PM-free plain silk-based (PSB) scaffolds were applied as control. Overall, it suggests that the incorporation of PLLA PMs remarkably improves mechanical properties and the capability to promote chondrogenesis of SBH scaffolds, and that SBH scaffolds appear to be a promising construct for potential applications in auricular cartilage tissue engineering and relevant fields.

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Tissue engineering has made significant progress as a cartilage repair alternative. It is crucial to promote cell proliferation and migration within three-dimensional (3D) bulk scaffolds for tissue regeneration through either chemical gradients or physical channels. In this study, by developing optimized silk fiber-based composite scaffolds, millimeter-scaled channels were created in the corresponding scaffolds via facile physical percussive drilling and subsequently utilized for auricular cartilage regeneration. We found that by the introduction of poly- l -lactic acid porous microspheres (PLLA PMs), the channels incorporated into the Antheraea pernyi (Ap) silk fiber-based scaffolds were reinforced, and the mechanical features were well maintained. Moreover, Ap silk fiber-based scaffolds reinforced by PLLA PMs containing channels (CMAF) exhibited excellent chondrocyte proliferation, migration, and synthesis of cartilage-specific extracellular matrix (ECM) in vitro . The biological evaluation in vivo revealed that CMAF had a higher chondrogenic capability for an even deposition of the specific ECM component. This study suggested that multihierarchical CMAF may have potential application for auricular cartilage regeneration.