Periosteum plays a pivotal role in bone formation and reconstruction. The ideal repair process for critical-size bone defects with periosteum damage is to induce regeneration of periosteum tissue and the subsequent bone regeneration derived by the periosteum. Inspired by the bilayer structure of the natural periosteum, we develop a periosteum structure/function dual mimicking membrane for the in-situ restoration of periosteum and bone tissue. Among them, the macroporous fluffy guiding layer (TPF) simulates the fibrous layer of the natural periosteum, which is conducive to infiltration and oriented growth of fibroblasts. And the extracellular matrix-like bioactive layer (TN) simulates the cambium layer of the natural periosteum, which significantly enhances the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells. A middle dense layer (PC) connects the above two layers and has the function of preventing the invasion of soft tissues while enhancing the biomimetic periosteum. In vivo restoration results show that the tri-layer biomimetic periosteum (TPF/PC/TN) has an outstanding effect in promoting the regeneration of both vascularized periosteum and bone at the same time. Therefore, the enhanced biomimetic periosteum developed in this research has a great clinical value in the efficient and high-quality reconstruction of critical-size bone defects with periosteum damage.

Piezoelectric stimulation has garnered substantial interest as a promising strategy for tissue regeneration. However, studies investigating its impact on tendon-to-bone healing characterized by fibrocartilage remain scarce. Moreover, there are considerable technical challenges in achieving minimally invasive application of piezoelectric stimulation on the irregular tendon-to-bone interface. Herein, we developed Janus asymmetric piezoelectric adhesives by assembling adhesive hydrogel (GAN) and non-adhesive hydrogel (GM) on each side of piezoelectric poly (L-lactic acid) nanofiber. Piezoelectric adhesives exhibited superior anti-inflammatory effects both in vitro and ex vivo. Notably, the transient receptor potential (TRP) ion channels, a class of versatile signaling molecules, are closely associated with the regulation of inflammation. This study demonstrated that piezoelectric stimulation promoted Ca²⁺ influx through the activation of transient receptor potential vanilloid 1 (TRPV1), further enhancing cAMP signaling pathway in macrophages by RNA sequencing. Additionally, in vivo proteomic analysis revealed Arachidonic acid metabolism and TNF-α signaling pathway downregulation and VEGF signaling pathway upregulation in a rat rotator cuff repair model. Piezoelectric adhesives ultimately achieved inflammation alleviation, angiogenesis enhancement, and fibrocartilage regeneration promotion, improving the biomechanical strength of the enthesis. This study elucidated the mechanism by which piezoelectric stimulation regulated tendon-to-bone healing through multi-omics analysis. The piezoelectric adhesives hold promise as a convenient and effective strategy for enhancing tendon-to-bone healing in clinical practice.