Indications exist that paramagnetic calcium phosphates may be able to promote regeneration of bone faster than their regular, diamagnetic counterparts. In this study, analyzed was the influence of paramagnetic cobalt-substituted hydroxyapatite nanoparticles on osteoporotic alveolar bone regeneration in rats. Simultaneously, biocompatibility of the material was tested in vitro, on osteoblastic MC3T3-E1 and epithelial Caco-2 cells in culture. The material was shown to be biocompatible and nontoxic when added to epithelial monolayers in vitro, while it caused a substantial decrease in the cell viability as well as deformation of the cytoskeleton and cell morphology when incubated with the osteoblastic cells. In the course of six months after the implantation of the material containing different amounts of cobalt, ranging from 5 – 12 wt%, in the osteoporotic alveolar bone of the lower jaw, the following parameters were investigated: histopathological parameters, alkaline phosphatase and alveolar bone density. The best result in terms of osteoporotic bone tissue regeneration was observed for hydroxyapatite nanoparticles with the largest content of cobalt ions. The histological analysis showed a high level of reparatory ability of the nanoparticulate material implanted in the bone defect, paralleled by a corresponding increase in the alveolar bone density. The combined effect of growth factors from autologous plasma admixed to cobalt-substituted hydroxyapatite was furthermore shown to have a crucial effect on the augmented osteoporotic bone regeneration upon the implantation of the biomaterial investigated in this study.
Composite biomaterials comprising nanostructured hydroxyapatite (HAp) have an enormous potential for natural bone tissue reparation, filling and augmentation. Chitosan (Ch) as a naturally derived polymer has many physicochemical and biological properties that make it an attractive material for use in bone tissue engineering. On the other hand, poly-D,L-lactide-co-glycolide (PLGA) is a synthetic polymer with a long history of use in sustained drug delivery and tissue engineering. However, while chitosan can disrupt the cell membrane integrity and may induce blood thrombosis, PLGA releases acidic byproducts that may cause tissue inflammation and interfere with the healing process. One of the strategies to improve the biocompatibility of Ch and PLGA is to combine them with compounds that exhibit complementary properties. In this study we present the synthesis and characterization, as well as in vitro and in vivo analyses of a nanoparticulate form of HAp coated with two different polymeric systems: (a) Ch and (b) a Ch-PLGA polymer blend. Solvent/non-solvent precipitation and freeze-drying were used for synthesis and processing, respectively, whereas thermogravimetry coupled with mass spectrometry was used for phase identification purposes in the coating process. HAp/Ch composite particles exhibited the highest antimicrobial activity against all four microbial strains tested in this work, but after the reconstruction of the bone defect they also caused inflammatory reactions in the newly formed tissue where the defect had lain. Coating HAp with a polymeric blend composed of Ch and PLGA led to a decrease in the reactivity and antimicrobial activity of the composite particles, but also to an increase in the quality of the newly formed bone tissue in the reconstructed defect area.
Biphasic calcium phosphate-poly-DL-lactide-co-glycolide composite biomaterial with and without biostimulative agents (protein-rich plasma or fibrin) was synthesised in the form suitable for reconstruction of bone defects. The composite used as filler was obtained by precipitation in solvent-non-solvent systems. The material, calcium phosphate granules covered by polymer, was characterised by wide-angle X-ray structural analysis, scanning electron microscopy, infrared spectroscopy and differential scanning calorimetry. Reparation of bone tissue damaged by osteoporosis was investigated in vivo on rats. The method applied enabled production of granules of calcium phosphate-poly-DL-lactide-co- glycolide composite biomaterial of average diameter 150-200 mum. Histological analysis confirmed recuperation of the alveolar bone, which osteoporosis-induced defects were repaired using composite biomaterial. By addition of biostimulative agents, intensity of osteogenesis increases accompanied by the formation of regular, new bone structure.
In this article, synthesis and application of calcium phosphate/poly-DL-lactide-co-glycolide (CP/PLGA) composite biomaterial in particulate form, in which each CP granule/particle is coated with PLGA, are described. Two types of the particulate material having different particle sizes were synthesized: one with an average particle diameter between 150 and 250 mum (micron-sized particles, MPs) and the other with an average particle diameter smaller than 50 nm (nanoparticles, NPs). A comparative in vivo analysis was done by reconstructing defects in osteoporotic alveolar bones using both composites. The material, CP granules/particles covered with polymer, was characterized using X-ray structural analysis, scanning electron microscopy, and atomic force microscopy. Changes in reparatory functions of tissues affected by osteoporosis were examined in mice in vivo, using these two kinds of composite materials, with and without autologous plasma. Having defined the target segment, histomorphometric parameters-bone area fraction, area, and mean density-were determined. The best results in the regeneration and recuperation of alveolar bone damaged by osteoporosis were achieved with the implantation of a mixture of nanoparticulate CP/PLGA composite and autologous plasma. After the implantation of microparticulate CP/PLGA, in the form of granules, mixed with autologous plasma, into an artificial defect in alveolar bone, new bone formation was also observed, although its formation rate was slower.
Cholecalciferol, vitamin D3, plays an important role in bone metabolism by regulating extracellular levels of calcium. Presented here is a study on the effects of the local delivery of cholecalciferol (D3) using nanoparticulate carriers composed of hydroxyapatite (HAp) and poly(D,L-lactide-co-glycolide) (PLGA). Multifunctional nanoparticulate HAp-based powders were prepared for the purpose of: (a) either fast or sustained, local delivery of cholecalciferol, and (b) the secondary, osteoconductive and defect-filling effect of the carrier itself. Two types of HAp-based powders with particles of narrowly dispersed sizes in the nano range were prepared and tested in this study: HAp nanoparticles as direct cholecalciferol delivery agents and HAp nanoparticles coated with cholecalciferol-loaded poly(D,L)-lactide-co-glycolide (HAp/D3/PLGA). Satisfying biocompatibility of particulate systems, when incubated in contact with MC3T3-E1 osteoblastic cells in vitro, was observed for HAp/D3/PLGA and pure HAp. In contrast, an extensively fast release of cholecalciferol from the system comprising HAp nanoparticles coated with cholecalciferol (HAp/D3) triggered necrosis of the osteoblastic cells in vitro. Artificial defects induced in the osteoporotic bone of the rat mandible were successfully reconstructed following implantation of cholecalciferol-coated HAp nanoparticles as well as those comprising HAp nanoparticles coated with cholecalciferol-loaded PLGA (HAp/D3/PLGA). The greatest levels of enhanced angiogenesis, vascularization, osteogenesis and bone structure differentiation were achieved upon the implementation of HAp/D3/PLGA systems.
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