Although biodegradable polymer/ceramic composite scaffolds can overcome the limitations of conventional ceramic bone substitutes, the osteogenic potential of these scaffolds needs to be further enhanced for efficient bone tissue engineering. In this study, bonelike apatite was efficiently coated onto the scaffold surface by using polymer/ceramic composite scaffolds instead of polymer scaffolds and by using an accelerated biomimetic process to enhance the osteogenic potential of the scaffold. The creation of bonelike, apatite-coated polymer scaffold was achieved by incubating the scaffolds in simulated body fluid (SBF). The apatite growth on porous poly(D,L-lactic-co-glycolic acid)/nanohydroxyapatite (PLGA/ HA) composite scaffolds was significantly faster than on porous PLGA scaffolds. In addition, the distribution of coated apatite was more uniform on PLGA/HA scaffolds than on PLGA scaffolds. After a 5-day incubation period, the mass of apatite coated onto PLGA/HA scaffolds incubated in 5 x SBF was 2.3-fold higher than PLGA/HA scaffolds incubated in 1 x SBF. Furthermore, when the scaffolds were incubated in 5 x SBF for 5 days, the mass of apatite coated onto PLGA/HA scaffolds was 4.5-fold higher than PLGA scaffolds. These results indicate that the biomimetic apatite coating can be accelerated by using a polymer/ceramic composite scaffold and concentrated SBF. When seeded with osteoblasts, the apatite-coated PLGA/HA scaffolds exhibited significantly higher cell growth, alkaline phosphatase activity, and mineralization in vitro compared to the apatite-coated PLGA scaffolds. Therefore, the apatite-coated PLGA/HA scaffolds may provide enhanced osteogenic potential when used as scaffold for bone tissue engineering.
Biodegradable polymer/ceramic scaffolds can overcome the limitations of conventional ceramic bone substitutes. However, the conventional methods of polymer/ceramic scaffold fabrication often use organic solvents, which might be harmful to cells or tissues. Moreover, scaffolds fabricated with the conventional methods have limited ceramic exposure on the scaffold surface since the polymer solution envelopes the ceramic particles during the fabrication process. In this study, we developed a novel fabrication method for the efficient exposure of ceramic onto the scaffold surface, which would enhance the osteoconductivity and wettability of the scaffold. Poly(D,Llactide-co-glycolide)/nanohydroxyapatite (PLGA/HA) scaffolds were fabricated by the gas foaming and particulate leaching (GF/PL) method without the use of organic solvents. Selective staining of ceramic particles indicated that HA nanoparticles exposed to the scaffold surface were observed more abundantly in the GF/PL scaffold than in the conventional solvent casting and particulate leaching (SC/PL) scaffold. Both types of scaffolds were implanted to critical size defects in rat skulls for 8 weeks. The GF/PL scaffolds exhibited significantly enhanced bone regeneration when compared with the SC/PL scaffolds. Histological analyses and microcomputed tomography of the regenerated tissues showed that bone formation was more extensive on the GF/PL scaffolds than on the SC/PL scaffolds. Compared with the SC/PL scaffolds, the enhanced bone formation on the GF/PL scaffolds may result from the higher exposure of HA nanoparticles to the scaffold surface. These results show that the biodegradable polymer/ceramic composite scaffolds fabricated with the novel GF/PL method can enhance bone regeneration compared with those fabricated with the conventional SC/PL method.
Previously, the sustained delivery of basic fibroblast growth factor (bFGF) has been demonstrated to promote bone regeneration in bone defects that had not been treated with osteogenic cell transplantation. In this study, we tested the hypothesis that the sustained delivery of bFGF could enhance osteoblast transplantation-mediated ectopic bone formation. Rat osteoblasts and bFGF were mixed with an injectable fibrin matrix and subcutaneously transplanted to rats (cell + bFGF group). The fibrin matrix played roles in both the cell transplantation matrix and the bFGF sustained delivery matrix. The transplantation of osteoblasts suspended in a fibrin matrix without bFGF served as a control. Twelve weeks after transplantation, histological analyses of retrieved transplants showed that new bone formation was more abundant and mature in the cell + bFGF group than in the control group. The bone formation area and the calcium content in the cell + bFGF group were two- and nine-fold higher, respectively, than those in the control group. Enhanced bone formation by the sustained delivery of bFGF may be attributed to the enhanced osteogenic gene expression of the transplanted cells and neovascularization of the transplants, as both mRNA expression of various osteogenic markers and arteriole density in the cell + bFGF group were significantly higher than those in the control group. This study demonstrates that the sustained delivery of bFGF can potentiate ectopic bone regeneration by osteoblast transplantation. This combination therapy may have effective implications for bone regeneration in large bone defects in which extensive osteogenic cell migration and angiogenesis are required.
Nanoparticles (NPs) are being employed for various industrial purposes with increasing frequency, yet the adverse health effects associated with the prolonged exposure of humans and the environment to NPs has not been well-established. Particularly, the effects of the extrinsic (or dynamic) physicochemical properties of NPs in aqueous cell culture media (e.g., hydrodynamic size, aggregation, agglomeration, sedimentation, and dissolution of nanoparticles) on the cytotoxicities of the NPs are barely understood. In this study, to investigate the effects of two important extrinsic properties of Ag NPs, namely the sedimentation and dissolution of Ag NPs, we performed MTT cell viability tests for HeLa cells exposed to Ag NPs with varying extrinsic properties. Ag NPs with different hydrodynamic sizes, sedimentation rates, and dissolution rates were prepared via exposure to NaCl and FBS. Sedimentation of aggregated/agglomerated Ag NPs was found to contribute more significantly to the cytotoxicity of Ag NPs during early periods of exposure, whereas the cytotoxicity was more greatly enhanced later during the exposure period due to the increase in silver ions. Therefore, it is offered that any assessment of NP cytotoxicity should consider the extrinsic properties of NPs, and their time-dependent properties, because the dominant processes affecting NP cytotoxicity may change over time and lead to a misunderstanding or poor prediction of NP cytotoxicity.
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