ABSTRACT
Low temperature setting calcium phosphate cements (CPC) formed from reactive calcium phosphate precursors are receiving great attention in the fields of orthopaedics and tissue engineering. The purpose of this study was to evaluate the mechanical properties and osteocompatibility of a novel calcium deficient hydroxyapatite (CDSHA) with a Ca/P ratio of 1.6 developed in our laboratories and compare it to a previously developed calcium deficient hydroxyapatite (CDHA) with a Ca/P ratio of 1.5. The results demonstrated that the calcium-deficient hydroxyapatites (HA) formed from the CPCs were similar to biological HA at physiological temperature and the elastic moduli of CDHA and CDSHA were found to be 174.42 +/- 20.41 MPa (p < 0.05) and 115.86 +/- 24.8 MPa (p < 0.05), respectively. The surface morphologies of the two calcium deficient HA's formed were identical with a micro/nano porous structure as evidenced from SEM. The cellular proliferation on CDHA, and CDSHA, was comparable to the control, tissue culture polystyrene (TCPS) (p < 0.05). Alkaline phosphatase activity was significantly elevated on CDHA and CDSHA matrices at early time points when compared with the control (TCPS) (p < 0.05). Osteoblast cells gene expression on CDHA, and CDSHA showed type I collagen, alkaline phosphatase, osteocalcin, and osteopontin activity at both 7 and 14 days of culture. Thus, novel calcium-deficient HAs, CDHA, and CDSHA formed at low temperature are promising candidates for orthopaedic applications based on their ability to promote osteoblast cell adhesion and gene expression in vitro.
Subject(s)
Bone Cements , Calcium Phosphates , Nanoparticles , Osteoblasts/cytology , Osteoblasts/metabolism , 3T3 Cells , Alkaline Phosphatase/genetics , Animals , Base Sequence , Bone Cements/chemistry , Calcium Phosphates/chemistry , Cell Adhesion , Collagen Type I/genetics , DNA Primers/genetics , Durapatite/chemistry , Gene Expression , Materials Testing , Mice , Microscopy, Electron, Scanning , Nanoparticles/chemistry , Osteocalcin/genetics , Osteopontin/genetics , Reverse Transcriptase Polymerase Chain Reaction , Surface Properties , Temperature , Tissue EngineeringABSTRACT
This study deals with the synthesis and in vitro osteocompatibility evaluation of two novel alanine-containing biodegradable ester polyphosphazenes as candidates to form self-setting composites with hydroxyapatite (HAp) precursors. The two novel biodegradable polyphosphazenes synthesized were poly[(ethyl alanato)1.0(ethyl oxybenzoate)1.0 phosphazene] (PN-EA/EOB) and poly[(ethyl alanato)1.0(propyl oxybenzoate)1.0 phosphazene] (PN-EA/POB). The polymers were characterized by multinuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), and gel permeation chromatography (GPC). Biodegradability and percentage water absorption of the polymers were evaluated by following the mass change in phosphate buffer (pH 7.4) at 37 degrees C. PN-EA/POB underwent faster degradation and showed higher water absorption compared to PN-EA/EOB. Both polymers became insoluble in common organic solvents following hydrolysis presumably due to crosslinking reactions accompanying the degradation process. Osteoblast cell adhesion and proliferation on PN-EA/EOB and PN-EA/POB was followed by scanning electron microscopy (SEM) and by using a biochemical assay. Both PN-EA/EOB and PN-EA/POB supported the adhesion and proliferation of primary rat osteoblast cells in vitro. Furthermore, the enzymatic activity of the osteoblast cells cultured on the polymers was confirmed by the alkaline phosphatase activity. Thus, these biodegradable amino-acid-based polyphosphazenes are promising new materials for forming self-setting bone cements.
Subject(s)
Biocompatible Materials/chemical synthesis , Bone and Bones , Organophosphorus Compounds/chemical synthesis , Polymers/chemical synthesis , Tissue Engineering , Alanine/chemistry , Alkaline Phosphatase/metabolism , Animals , Biocompatible Materials/chemistry , Biodegradation, Environmental , Cell Adhesion , Cell Proliferation , Cells, Cultured , Materials Testing , Microscopy, Electron, Scanning , Organophosphorus Compounds/chemistry , Osteoblasts/cytology , Osteoblasts/enzymology , Polymers/chemistry , RatsABSTRACT
Electrospinning has developed as a unique and versatile process to fabricate ultrathin fibers in the form of nonwoven meshes or as oriented arrays from a variety of polymers. The very small dimension of these fibers can generate a high surface area, which makes them potential candidates for various biomedical and industrial applications. The objective of the present study was to develop nanofibers from polyphosphazenes, a class of inorganic-organic polymers known for high biocompatibility, high-temperature stability, and low-temperature flexibility. Specifically, we evaluated the feasibility of developing bead-free nonwoven nanofiber mesh from poly[bis(p-methylphenoxy)phosphazene] (PNmPh) by electrospinning. The effect of process parameters such as nature of solvent, concentration of the polymer solution, effect of needle diameter, and applied potential on the diameter and morphology (beaded or bead-free) of resulting nanofibers were investigated. It was found that solution of PNmPh in chloroform at a concentration range of 7% (wt/v) to 9% (wt/v) can be readily electrospun to form bead-free fibers at room temperature. The mean diameter of the fibers obtained under optimized spinning condition was found to be approximately 1.2 microm. The bead-free, cylindrical nanofibers formed under the optimized condition showed a slightly irregular surface topography with indentations of a few nanometer scale. Further, the electrospun nanofiber mats supported the adhesion of bovine coronary artery endothelial cells (BCAEC) as well as promoted the adhesion and proliferation of osteoblast like MC3T3-E1 cells.
Subject(s)
Organophosphorus Compounds/chemistry , Polymers/chemistry , 3T3 Cells , Animals , Biocompatible Materials/chemistry , Cattle , Cell Adhesion , Cell Proliferation , Chloroform , Dose-Response Relationship, Drug , Electrophysiology , Endothelium, Vascular/cytology , Macromolecular Substances/chemistry , Magnetic Resonance Spectroscopy , Mice , Microscopy, Electron, Scanning , Models, Chemical , Nanotechnology , Osteoblasts/cytology , Solvents , Surface Properties , Temperature , Time Factors , Tissue Engineering/methods , Ultraviolet RaysABSTRACT
Amphiphilic diblock copolymers with varying compositions of hydrophilic poly(ethylene oxide) (PEO) and hydrophobic poly[bis(ethyl glycinat-N-yl)phosphazene] (PNgly) were synthesized via the controlled cationic-induced polymerization of a phosphoranimine (Cl(3)P=NSiMe(3)) at ambient temperature using a PEO-phosphoranimine macroinitiator. The aqueous-phase transition behavior of PEO-PNgly-3 (M(n) = 10,000) and micelle formation of both PEO-PNgly-3 and PEO-PNgly-4 (M(n) = 8,500) were investigated using fluorescence techniques and dynamic light scattering. The critical micelle concentrations (cmc's) of PEO-PNgly-3 and PEO-PNgly-4 were determined to be 3 and 12 mg/L with the mean diameters of micelles being 120 and 130 nm, respectively. The hydrolytic degradation of these diblock copolymers was also studied in solution. These studies coupled with the biodegradability of the poly[bis(ethyl glycinat-N-yl)phosphazene] block to give benign products make PEO-PNgly copolymers well-suited for a wide variety of biomedical applications including novel biodegradable drug-delivery systems.
