Spatiotemporally Orchestrated Bone Healing: A Composite Scaffold System for Sequential Hemostasis and Osteogenesis

The instability of hematomas in open bone injuries presents a major barrier to effective healing. To address this issue, we developed a composite scaffold system designed to achieve temporally coordinated hematoma stabilization and osteogenic induction. The system comprises a 3D-printed rigid PCL + CSi-Mg framework integrated with a directionally frozen, highly ordered porous CS + CSi-Mg sponge. The composite scaffold demonstrated outstanding synergistic performance. The internal CS + CSi-Mg sponge, characterized by aligned pore channels and bioactive components (CS and Ca²⁺), facilitated rapid blood absorption and coagulation both in vitro and in vivo, achieving complete blood uptake within 10 s. Meanwhile, the external PCL + CSi-Mg framework provided mechanical strength comparable to that of cancellous bone (∼8.6 MPa). In vitro assays confirmed that the system significantly promoted the proliferation, osteogenic differentiation, and mineralization of bone marrow mesenchymal stem cells (BMSCs), while upregulating key osteogenic markers such as RUNX2, ALP, BSP, and Col1. Importantly, in a rat critical-sized calvarial defect model, the composite scaffold group exhibited markedly increased bone formation (BV/TV) and improved bone quality at 8 weeks compared to single-component controls. In summary, this integrated strategy, combining "hematoma stabilization" with "osteogenic induction", effectively meets the sequential demands of early hemostasis and subsequent bone regeneration, presenting a promising strategy for the development of advanced bone repair materials.