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Early osteogenic signal expression of rat bone marrow stromal cells is influenced by both hydroxyapatite nanoparticle content and initial cell seeding density in biodegradable nanocomposite scaffolds.

ABSTRACT: Incorporation of hydroxyapatite (HA) within a degradable polymeric scaffold may provide a favorable synthetic microenvironment that more closely mimics natural bone tissue physiology. Both incorporation of HA nanoparticles and alterations of the paracrine cell-cell signaling distance may affect the intercellular signaling mechanism and facilitate enhanced osteogenic signal expression among the implanted cell population. In this study we investigate the effect of the incorporation of HA nanoparticles into poly(propylene fumarate) (PPF) scaffolds on the surface properties of composite scaffolds and early osteogenic growth factor gene expression in relation to initial cell seeding density. The results of surface characterization indicated that HA addition improved the surface properties of PPF/HA composite scaffolds by increasing the roughness, hydrophilicity, protein adsorption, and initial cell attachment. Rat bone marrow stromal cells (BMSCs), which were CD34-, CD45-, CD29+, and CD90+, were cultured on three-dimensional (3-D) macroporous PPF/HA scaffolds at two different initial cell seeding densities (0.33 and 1.00 million cells per scaffold) for 8 days. The results demonstrated that endogenous osteogenic signal expression profiles, including bone morphogenetic protein-2, fibroblast growth factor-2, and transforming growth factor-?1, as well as the transcriptional factor Runx2, were affected by both HA amount and initial cell seeding density. Up-regulated expression of osteogenic growth factor genes was related to subsequent osteoblastic differentiation of rat BMSCs on 3-D scaffolds, as characterized by alkaline phosphatase activity, osteocalcin mRNA expression, and calcium deposition. Thus, the PPF/HA composite scaffold construction parameters, including amount of HA incorporated and initial cell seeding density, may be utilized to induce the osteoblastic differentiation of transplanted rat BMSCs.

SUBMITTER: Kim K

PROVIDER: S-EPMC3031657 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature

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Early osteogenic signal expression of rat bone marrow stromal cells is influenced by both hydroxyapatite nanoparticle content and initial cell seeding density in biodegradable nanocomposite scaffolds.

Kim Kyobum K Dean David D Lu Anqi A Mikos Antonios G AG Fisher John P JP

Acta biomaterialia 20101111 3

Incorporation of hydroxyapatite (HA) within a degradable polymeric scaffold may provide a favorable synthetic microenvironment that more closely mimics natural bone tissue physiology. Both incorporation of HA nanoparticles and alterations of the paracrine cell-cell signaling distance may affect the intercellular signaling mechanism and facilitate enhanced osteogenic signal expression among the implanted cell population. In this study we investigate the effect of the incorporation of HA nanoparti ...[more]

PMID: 21074640

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Project description:Polylactic acid (PLA) has been widely used as filaments for material extrusion additive manufacturing (AM) to develop patient-specific scaffolds in bone tissue engineering. Hydroxyapatite (HA), a major component of natural bone, has been extensively recognized as an osteoconductive biomolecule. Here, inspired by the mussel-adhesive phenomenon, in this study, polydopamine (PDA) coating was applied to the surface of 3D printed PLA scaffolds (PLA@PDA), acting as a versatile adhesive platform for immobilizing HA nanoparticles (nHA). Comprehensive analyses were performed to understand the physicochemical properties of the 3D-printed PLA scaffold functionalized with nHA and PDA for their potent clinical application as a bone regenerative substitute. Scanning electron microscopy (SEM) and element dispersive X-ray (EDX) confirmed a successful loading of nHA particles on the surface of PLA@PDA after 3 and 7 days of coating (PLA@PDA-HA3 and PLA@PDA-HA7), while the surface micromorphology and porosity remain unchanged after surface modification. The thermogravimetric analysis (TGA) showed that 7.7 % and 12.3% mass ratio of nHA were loaded on the PLA scaffold surface, respectively. The wettability test indicated that the hydrophilicity of nHA-coated scaffolds was greatly enhanced, while the mechanical properties remained uncompromised. The 3D laser scanning confocal microscope (3DLS) images revealed that the surface roughness was significantly increased, reaching Sa (arithmetic mean height) of 0.402 μm in PLA@PDA-HA7. Twenty-eight days of in-vitro degradation results showed that the introduction of nHA to the PLA surface enhances its degradation properties, as evidenced by the SEM images and weight loss test. Furthermore, a sustainable release of Ca2+ from PLA@PDA-HA3 and PLA@PDA-HA7 was recorded, during the degradation process. In contrast, the released hydroxyl group of nHA tends to neutralize the local acidic environments, which was more conducive to osteoblastic differentiation and extracellular mineralization. Taken together, this facile surface modification provides 3D printed PLA scaffolds with effective bone regenerative properties by depositing Ca2+ contents, improving surface hydrophilicity, and enhancing the in-vitro degradation rate.

Dataset Information

Early osteogenic signal expression of rat bone marrow stromal cells is influenced by both hydroxyapatite nanoparticle content and initial cell seeding density in biodegradable nanocomposite scaffolds.

Publications

Early osteogenic signal expression of rat bone marrow stromal cells is influenced by both hydroxyapatite nanoparticle content and initial cell seeding density in biodegradable nanocomposite scaffolds.

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