The scaffold showed the highest osteoinduction, and the scaffold with 530 ± 56 μm average pore diameter demonstrated the highest expression of osteodifferentiation marker genes in DPSCs, and the addition of nanoparticles into the polymer matrix led to the decrease in the expression of pro-inflammatory genes in macrophages.
Abstract
We aimed to develop a composite poly (lactic-co-glycolic acid) (PLGA)/calcium phosphate nanoparticles scaffold with the optimal three-dimensional structure to provide an environment for bone tissue regeneration. Composite PLGA-based scaffolds with the inclusion of 15% hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP) nanoparticles, as well as scaffolds with the addition of 15% xenogeneic bone chips, and with different pore diameters were prepared by a solvent casting with particle leaching method. Synthesized HA and β-TCP nanoparticles were characterized using x-ray phase analysis and atomic force microscopy. The scaffold morphology was studied with electron microscopy and energy dispersive x-ray spectroscopy. The scaffold biocompatibility, immunogenicity, inflammatory and osteoinductive properties were investigated in vitro using dental pulp stem cells (DPSCs), human lymphocyte culture, and RAW 264.7 mouse macrophage cells. Among the investigated samples, the PLGA/ β-TCP scaffold showed the highest osteoinduction, and the scaffold with 530 ± 56 μm average pore diameter demonstrated the highest expression of osteodifferentiation marker genes in DPSCs. Furthermore, the addition of nanoparticles into the polymer matrix led to the decrease in the expression of pro-inflammatory genes in macrophages. PLGA with 15% β-TCP and 530 ± 56 μm pore size had the best bioactivity among the tested scaffolds in vitro, and it could be considered as a potential candidate for bone tissue engineering applications.
The persistent inflammatory microenvironment during bone defect repair can inhibit osteogenic differentiation, delay angiogenesis, and reduce the reparative efficacy of implanted materials. Therefore, developing biomaterials that combine immunomodulatory functions with bone-regenerative capacity is of great significance. In this study, composite scaffolds with anti-inflammatory and bone-repair-promoting properties were fabricated through surface functionalization and 3D printing. The structural and physicochemical properties of the scaffolds were systematically characterized by scanning electron microscopy (SEM), elemental mapping, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), contact angle measurement, and mechanical testing. Transmission electron microscopy and related analyses were further used to evaluate their microstructure and surface characteristics. The degradation behavior, pH variation, ion release profile, and antioxidant performance of the scaffolds under inflammatory conditions were then investigated to verify their ability to regulate the local microenvironment. In vitro, cytocompatibility was evaluated using CCK-8 assays, live/dead staining, and immunofluorescence staining. The effects of the scaffolds on macrophage polarization and inflammatory factor expression were further analyzed. Their osteogenic differentiation potential was assessed by alkaline phosphatase (ALP) staining, Alizarin Red staining, and osteogenesis-related gene expression analysis. In addition, Micro-CT, hematoxylin and eosin (HE) staining, and Masson's trichrome staining were performed in a bone defect animal model to evaluate bone regeneration and tissue repair. The results showed that the composite scaffolds effectively improved the inflammatory microenvironment in the defect region, promoted macrophage polarization toward an anti-inflammatory phenotype, enhanced cellular osteogenic activity and mineralization, and facilitated new bone formation and tissue reconstruction. This study provides a theoretical basis and experimental evidence for the design and application of bone repair materials under inflammatory conditions.
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