Dual siRNA-Loaded Cell Membrane Functionalized Matrix Facilitates Bone Regeneration with Angiogenesis and Neurogenesis.

Vascularization and innervation play irreplaceable roles in bone regeneration and bone defect repair. However, the reconstruction of blood vessels and neural networks is often neglected in material design. This study aims to design a genetically functionalized matrix (GFM) and enable it to regulate angiogenesis and neurogenesis to accelerate the process of bone defect repair. The dual small interfering RNA (siRNA)-polyvinylimide (PEI) (siRP) complexes that locally knocked down soluble vascular endothelial growth factor receptor 1 (sFlt-1) and p75 neurotrophic factor receptor (p75NTR ) are prepared. The hybrid cell membrane (MM) loaded siRP is synthesized as siRNA@MMs to coat on polylactone (PCL) electrospun fibers for mimicking the natural bone matrix. The results indicates that siRNA@MMs could regulate the expression of vascular-related and neuro-related cytokines secreted by mesenchymal stem cells (MSCs). GFMs promote the expression of osteogenic differentiation through paracrine function in vitro. GFMs attenuates inflammation and promotes osseointegration by regulating the coupling of vascularization and innervation in vivo. This study uses the natural hybrid cell membrane to carry genetic material and assist in the vascularization and innervation function of two siRNA. The results present the significance of neuro-vascularized organoid bone and may provide a promising choice for the design of bone tissue engineering scaffold.

Hybrid Cell Membrane-Functionalized Matrixes for Modulating Inflammatory Microenvironment and Improving Bone Defect Repair.

Cell membranes from different sources retain the integrity of the original membrane structure and special membrane protein functions. However, the diversity function of membrane surface proteins is underutilized in bone defect repair. The current study creatively prepared a hybrid membrane (HM) (diameter, 92.25 ± 16.47 nm) and coated it on a poly-ε-caprolactone (PCL) (diameter, 313.79 ± 4.69 nm) electrospinning membrane for cell membrane-functionalized matrixes (MFMs) (diameter, 368.76 ± 5.90 nm) preparation. The effects of the HM and MFMs on inflammatory regulation and bone formation are further explored in vitro and in vivo. The results showed that MFMs can regulate macrophage into anti-inflammatory phenotype and promote alkaline phosphatase secretion and mineralization deposition of mesenchymal stem cells (MSCs) in vitro. The foreign body response is alleviated and bone regeneration is facilitated in rat calvarial critical-size defect in vivo. The study provides an innovative approach to applicate the cell membrane functions from different sources for immunomodulatory and osteogenic differentiation and provides a promising choice for designing bone repair materials.

Matrix stiffness regulates bone repair by modulating 12-lipoxygenase-mediated early inflammation.

Lipid metabolism in macrophages has been increasingly emphasized in exerting an anti-inflammatory effect and accelerating fracture healing. 12-lipoxygenase (12-LOX) is expressed in several cell types, including macrophages, and oxidizes polyunsaturated fatty acids (PUFAs) to generate both pro- and anti-inflammatory lipid mediators, of which the n-3 PUFAs play an important part in tissue homeostasis/fibrosis. Although mechanical factor regulates the lipid metabolic axis of inflammatory cells, specifically matrix stiffness influences macrophages metabolic responses, little is known about how matrix stiffness affects the 12-LOX-mediated early inflammation in bone repair. In the present study, demineralized bone matrix (DBM) scaffolds with different matrix stiffness were constructed by controlling the duration of decalcification (0 h (control), 1 h (high), 12 h (medium), and 5 d (low)) to repair the defected rat skull. The expression of inflammatory cytokines and macrophages polarization were analyzed. The lipid metabolites and lipid mediators' biosynthesis by matrix stiffness-regulated were further detected. The results showed that the low matrix stiffness could polarize macrophages into an anti-inflammatory phenotype, promote the expression of anti-inflammatory cytokines and specialized pro-resolving lipid mediators (SPMs) biosynthesis beneficial for the osteogenesis of mesenchymal stem cells (MSCs). After treated with ML355, the expression of anti-inflammatory cytokines/proteins and SPMs biosynthesis in macrophages cultured on low-matrix stiffness scaffolds were repressed, and there were almost no statistical differences among all groups. Findings from this study support that matrix stiffness regulates bone repair by modulating 12-LOX-mediated early inflammation, which suggest a direct mechanical impact of matrix stiffness on macrophages lipid metabolism and provide a new insight into the clinical application of SPMs for bone regeneration.

Demineralized and decellularized bone extracellular matrix-incorporated electrospun nanofibrous scaffold for bone regeneration.

Extracellular matrix (ECM)-based materials have been employed as scaffolds for bone tissue engineering, providing a suitable microenvironment with biophysical and biochemical cues for cell attachment, proliferation and differentiation. In this study, bone-derived ECM (bECM)-incorporated electrospun poly(ε-caprolactone) (PCL) (bECM/PCL) nanofibrous scaffolds were prepared and their effects on osteogenesis were evaluated in vitro and in vivo. The results showed that the bECM/PCL scaffolds promoted the attachment, spreading, proliferation and osteogenic differentiation of rat mesenchymal stem cells (MSCs), mitigated the foreign-body reaction, and facilitated bone regeneration in a rat calvarial critical size defect model. Thus, this study suggests that bECM can provide a promising option for bone regeneration.