Work

Native hyphal alignment enables spatially anisotropic functions in Pleurotus eryngii-derived membranes for guided bone regeneration

Year: 2026

Type: article

Abstract: Guided bone regeneration (GBR) membranes are required to coordinate mechanical support, osteogenic cell regulation, and soft-tissue exclusion across different spatial directions. Hierarchical architectures with spatial heterogeneity and anisotropy may therefore better accommodate these direction-dependent functional requirements. However, bottom-up construction of such architectures often relies on specialized equipment and complex multistep fabrication. By contrast, biological tissues spontaneously develop intricate heterogeneous and hierarchical structures during growth, providing unique value for the top-down construction of architecture-driven biomaterials. The widely cultivated Pleurotus eryngii has hyphal cell walls composed mainly of chitin and β-glucan, while its fruiting-body stipe exhibits a pronounced longitudinal organization and a robust morphology amenable to sectioning, making it suitable for generating distinct structural presentations simply by changing the cutting orientation. In this study, cross-cut (Tr-C) and longitudinal-cut (Tr-L) membranes were prepared from P. eryngii stipes using the same combined chemical-enzymatic treatment, which substantially reduced cell-associated residues while preserving recognizable tubular hyphal architecture. Tr-C and Tr-L exhibited similar overall chemical characteristics, yet the structural differences introduced by cutting orientation resulted in clearly distinct functional states. Tr-L showed more pronounced directional wet-state mechanical behavior, stronger contact guidance of MC3T3-E1 cells, enhanced osteogenesis-related phenotypes, and greater fibroblast exclusion across the membrane thickness. In contrast, Tr-C exhibited faster liquid infiltration and greater hydration capacity. The two membrane architectures also induced distinct macrophage response patterns under inflammatory stimulation. In a rat calvarial defect model, Tr-L promoted more pronounced centripetal new bone formation. These findings demonstrate that different structural presentations of the same native hyphal network can generate membrane-scale functional anisotropy, highlighting the value of native hyphal organization for constructing GBR membranes with spatially differentiated functions.

Source: Frontiers in Cell and Developmental Biology

DOI: 10.3389/fcell.2026.1997731