Engineering a complete, physiologically functional, periodontal complex structure remains a great clinical challenge due to the highly hierarchical architecture of the periodontium and coordinated regulation of multiple growth factors required to induce stem cell multilineage differentiation. Using biomimetic self-assembly and microstamping techniques, we construct a hierarchical bilayer architecture consisting of intrafibrillarly mineralized collagen resembling bone and cementum, and unmineralized parallel-aligned fibrils mimicking periodontal ligament. The prepared biphasic scaffold possesses unique micro/nano structure, differential mechanical properties, and growth factor-rich microenvironment between the two phases, realizing a perfect simulation of natural periodontal hard/soft tissue interface. The interconnected porous hard compartment with a Young's modulus of 1409.00 ± 160.83 MPa could induce cross-arrangement and osteogenic differentiation of stem cells in vitro, whereas the micropatterned soft compartment with a Young's modulus of 42.62 ± 4.58 MPa containing abundant endogenous growth factors, could guide parallel arrangement and fibrogenic differentiation of stem cells in vitro. After implantation in critical-sized complete periodontal tissue defect, the biomimetic bilayer architecture potently reconstructs native periodontium with the insertion of periodontal ligament fibers into newly formed cementum and alveolar bone by recruiting host mesenchymal stem cells and activating the transforming growth factor beta 1/Smad3 signaling pathway. Taken together, integration of self-assembly and microstamping strategies could successfully fabricate a hierarchical bilayer architecture, which exhibits great potential for recruiting and regulating host stem cells to promote synergistic regeneration of hard/soft tissues.

由于牙周组织的高度分级结构和诱导干细胞多向分化所需的多种生长因子的协调调节，设计完整的、生理功能的、牙周复合结构仍然是一项巨大的临床挑战。使用仿生自组装和微冲压技术，我们构建了一个分层的双层结构，由类似骨和牙骨质的纤维内矿化胶原蛋白和模拟牙周韧带的未矿化平行排列的纤维组成。制备的双相支架具有独特的微/纳米结构、不同的机械性能和两相间富含生长因子的微环境，实现了对天然牙周硬/软组织界面的完美模拟。体外，而杨氏模量为 42.62 ± 4.58 MPa 的微图案软隔室含有丰富的内源性生长因子，可以指导体外干细胞的平行排列和纤维化分化. 植入临界尺寸的完整牙周组织缺损后，仿生双层结构通过招募宿主间充质干细胞和激活转化生长因子β1/Smad3信号传导，将牙周韧带纤维插入新形成的牙骨质和牙槽骨中，有效地重建天然牙周组织途径。综上所述，自组装和微冲压策略的整合可以成功构建分层双层结构，其在募集和调节宿主干细胞以促进硬/软组织协同再生方面具有巨大潜力。