The development of an artificial bone graft which can promote the regeneration of fractures or diseased bones is currently the most challenging aspect in bone tissue engineering. To achieve the purpose of promoting bone proliferation and differentiation, the artificial graft needs have a similar structure and composition of extracellular matrix. One-step electrospinning method of biocomposite nanofibers containing hydroxyapatite (HA) nanoparticles and collagen (Coll) were developed for potential application in bone tissue engineering. Nanocomposite scaffolds of poly(L-lactide) (PLLA), PLLA/HA, PLLA/Coll, and PLLA/Coll/HA were fabricated by electrospinning. The morphology, diameter, elements, hydrophilicity, and biodegradability of the composite scaffolds have been investigated. The biocompatibility of different nanocomposite scaffolds was assessed using mouse osteoblasts MC3T3-E1 in vitro, and the proliferation, differentiation, and mineralization of cells on different nanofibrous scaffolds were investigated. The results showed that PLLA/Coll/HA nanofiber scaffolds enhanced cell adhesion, spreading, proliferation, differentiation, mineralization, and gene expression of osteogenic markers compared to other scaffolds. In addition, the nanofibrous scaffolds maintained a stable composition at the beginning of the degradation period and morphology wastage and weight loss were observed when incubated for up to 80 days in physiological simulated conditions. The PLLA/Coll/HA composite nanofibrous scaffolds could be a potential material for guided bone regeneration.

Yttrium oxide nanoparticles are an excellent host material for the rare earth metals and have high luminescence efficiency providing a potential application in photodynamic therapy and biological imaging. In this study, the effects of yttrium oxide nanoparticles with four different sizes were investigated using primary osteoblasts in vitro. The results demonstrated that the cytotoxicity generated by yttrium oxide nanoparticles depended on the particle size, and smaller particles possessed higher toxicological effects. For the purpose to elucidate the relationship between reactive oxygen species generation and cell damage, cytomembrane integrity, intracellular reactive oxygen species level, mitochondrial membrane potential, cell apoptosis rate, and activity of caspase-3 in cells were then measured. Increased reactive oxygen species level was also observed in a size-dependent way. Thus, our data demonstrated that exposure to yttrium oxide nanoparticles resulted in a size-dependent cytotoxicity in cultured primary osteoblasts, and reactive oxygen species generation should be one possible damage pathway for the toxicological effects produced by yttrium oxide particles. The results may provide useful information for more rational applications of yttrium oxide nanoparticles in the future.

Two-dimensional materials provide a secluded space for bone formation and preserve the growth of surrounding tissues, thus playing a crucial role in guided bone regeneration (GBR). Graphene oxide (GO) has been widely employed in GBR due to its good mechanical and hydrophilic properties. A single GO membrane, however, does not provide a friendly environment for osteogenic cell adhesion. With their adjustable mechanical properties and excellent biocompatibility, composite membranes can simulate the multicomponent structure of an extracellular matrix for cell adhesion. To obtain two-dimensional membranes with appropriate mechanical strength and sufficient biocompatibility, GO-based composite membranes simultaneously containing chitosan (CS) and hydroxyapatite (HAP) were first prepared using one-step vacuum filtration and a biomimetic mineralization method. CS and HAP improved the mechanical strength and surface hydrophilicity of the membranes. In addition, moderate addition of HAP enhanced the adhesion, differentiation, and mineralization of osteoblasts. The prepared composite membranes were then implanted into a calvarial defect model to evaluate their osteogenic induction effects in vivo. Microcomputed tomography observation and histological analysis indicate that GO/CS/HAP composite membranes can accelerate bone regeneration without the contribution of endogenous cytokines. GO/CS/HAP composite membranes with unique biomimetic porous structures, superior mechanical properties, and excellent bone regeneration capacity are potential materials for application in GBR.

ABSTRACT With the fast development of nanotechnology, the nanomaterials have raised questions concerning its potential toxic effects on human health. In this study, the effects of hydroxyapatite nanoparticles on the primary osteoblasts were investigated. As an indicator of membrane damage, lactate dehydrogenase was quantitatively assessed. The quantitative analysis on cellular uptake of hydroxyapatite nanoparticles could be detected by flow cytometer and inductively coupled plasma mass spectrometry, respectively. The results demonstrated that hydroxyapatite nanoparticles can enter cells through cell membrane and the nanoparticles taken up in the cells followed dose and time dependent manner. The methods could be used for the initial screening of the uptake potential of nanoparticles as an index of nanotoxicity.