This study evaluates the cytotoxic and mutagenic effect of synthetic hydroxyapatite granules (source: School of Material and Mineral Resources Engineering, Universiti Sains Malaysia) in the bone marrow cells of mice. Mice are exposed to synthetic hydroxyapatite granules, the bone marrow cells are collected and observed for chromosome aberrations. No chromosome aberrations were noticed in the animals exposed to distilled water (negative control) and to the test substance, synthetic hydroxyapatite granules (treatment) groups. Chromosome aberrations were observed in the animals exposed to Mitomycin C (positive control group). There was no indication of cytotoxicity due to synthetic hydroxyapatite granules in the animals as revealed by the mitotic index. Hence, synthetic hydroxyapatite granules are considered non-mutagenic under the prevailing test conditions.
Bone Marrow Cells/*drug effects ; Bone Substitutes/*toxicity ; *Chromosome Aberrations ; Durapatite/*toxicity ; *Mutagenicity Tests

The present study is aimed at finding the mutagenicity and cytotoxicity of dense form of synthetic hydroxyapatite (Source: School of Materials and Mineral Resources Engineering, Universiti Sains Malaysia) in the blood of sheep. The biomaterial was implanted in the tibia of Malin, an indigenous sheep breed of Malaysia. Blood was collected from the sheep before implantation of the biomaterial, cultured and a karyological study was made. Six weeks after implantation, blood was collected from the same animal, cultured and screened for chromosome aberrations. The mitotic indices and karyological analysis indicated that the implantation of synthetic hydroxyapatite (dense form) did not produce any cytotoxicity or chromosome aberrations in the blood of sheep.
Biocompatible Materials/*toxicity ; Bone Substitutes/*toxicity ; Bone and Bones/pathology ; Cell Survival/drug effects ; *Chromosome Aberrations ; Hydroxyapatites/*toxicity ; Karyotyping ; *Mutagenicity Tests ; *Prostheses and Implants ; Sheep

Mutagenicity of CORAGRAF (natural coral) and REKAGRAF (hydroxyapatite) was tested in Ames test with and without an external metabolic activation system (S9). The test revealed no mutagenic activity of both locally produced osseous substitutes.
Base Pair Mismatch/drug effects ; Biotransformation/physiology ; Bone Substitutes/*toxicity ; Calcium Carbonate/*toxicity ; Chromosome Aberrations ; Escherichia coli/genetics ; Hydroxyapatites/*toxicity ; *Materials Testing ; *Mutagenicity Tests ; Salmonella typhimurium/genetics

Hydroxyapatite is the main component of the bone which is a potential biomaterial substance that can be applied in orthopaedics. In this study, the biocompatibility of this biomaterial was assessed using an in vitro technique. The cytotoxicity and genotoxicity effect of HA2 and HA3 against L929 fibroblast cell was evaluated using the MTT Assay and Alkaline Comet Assay respectively. Both HA2 and HA3 compound showed low cytotoxicity effect as determined using MTT Assay. Cells viability following 72 hours incubation at maximum concentration of both HA2 and HA3 (200 mg/ml) were 75.3 +/- 8.8% and 86.7 +/- 13.1% respectively. However, the cytotoxicity effect of ZnSO4.7H2O as a positive control showed an IC50 values of 46 mg/ml (160 microM). On the other hand, both HA2 and HA3 compound showed a slight genotoxicity effect as determined using the Alkaline Comet Assay following incubation at the concentration 200 mg/ml for 72 hours. This assay has been widely used in genetic toxicology to detect DNA strand breaks and alkali-labile site. The percentage of the cells with DNA damage for both substance was 27.7 +/- 1.3% and 15.6 +/- 1.0% for HA2 and HA3 respectively. Incubation of the cells for 24 hours with 38 microg/ml (IC25) of positive control showed an increase in percentage of cells with DNA damage (67.5 +/- 0.7%). In conclusion, our study indicated that both hydroxyapatite compounds showed a good biocompatibility in fibroblast cells.
Biocompatible Materials/*toxicity ; Bone Substitutes/*toxicity ; Cell Survival/drug effects ; *DNA Damage ; Hydroxyapatites/*toxicity ; L Cells (Cell Line) ; *Mutagenicity Tests ; *Prostheses and Implants

Biomaterials intended for end-use application as bone-graft substitutes have to undergo safety evaluation. In this study, we investigated the in vitro cytotoxic effects especially to determine the mode of death of two hydroxyapatite compounds (HA2, HA3) which were synthesized locally. The methods used for cytotoxicity was the standard MTT assay whereas AO/PI staining was performed to determine the mode of cell death in HA treated L929 fibroblasts. Our results demonstrated that both HA2 and HA3 were not significantly cytotoxic as more than 75% cells after 72 hours treatment were viable. Furthermore, we found that the major mode of cell death in HA treated cells was apoptosis. In conclusion, our results demonstrated that these hydroxyapatite compounds are not cytotoxic where the mode of death was primarily via apoptosis.
Apoptosis/drug effects ; Biocompatible Materials/*toxicity ; Bone Substitutes/*toxicity ; Cell Death/*drug effects ; Durapatite/*toxicity ; L Cells (Cell Line) ; *Prostheses and Implants

Porous nano-hydroxyapatite/polyamide 66 (n-HA/PA66) composite was developed by injection molding method. Uniformly distributed and interconnected pores with an average size of about 500 microm in matrix were obtained. The evaluation of biological safety of the porous composite including cell cytoxicity test, sensitivity test, pyrogen test, haemolysis test was carried out according to GB/T16886 and GB/T16175. The results showed that the porous n-HA/PA66 composite was of no cytotoxicity, no allergen and pyrogen reactions as well as no hemolytic effect.
Animals ; Bone Substitutes ; chemical synthesis ; toxicity ; Durapatite ; chemistry ; Guinea Pigs ; Implants, Experimental ; Materials Testing ; Mice ; Nanocomposites ; chemistry ; Nanoparticles ; chemistry ; Nylons ; chemistry ; Porosity ; Rabbits ; Tissue Engineering

AIMThe piezoelectric properties and cytotoxicity of a porous lead-free piezoelectric ceramic for use as a direct bone substitute were investigated.

METHODOLOGYCold isostatic pressing (CIP) was applied to fabricate porous lithium sodium potassium niobate (Li0.06Na0.5K0.44) NbO3 specimens using a pore-forming method. The morphologies of the CIP-processed specimens were characterized and compared to those of specimens made by from conventional pressing procedures. The effects of the ceramic on the attachment and proliferation of osteoblasts isolated from the cranium of 1-day-old Sprague-Dawley rats were examined by a scanning electron microscopy (SEM) and methylthiazol tetrazolium (MTT) assay.

RESULTSThe results showed that CIP enhanced piezoelectricity and biological performance of the niobate specimen, and also promoted an extracellular matrix-like topography of it. In vitro studies showed that the CIP-enhanced material had positive effects on the attachment and proliferation of osteoblasts.

CONCLUSIONNiobate ceramic generated by CIP shows a promise for being a piezoelectric composite bone substitute.

Animals ; Biocompatible Materials ; chemistry ; toxicity ; Bone Substitutes ; chemical synthesis ; toxicity ; Cell Adhesion ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Ceramics ; chemical synthesis ; toxicity ; Coloring Agents ; Electrochemistry ; Materials Testing ; Microscopy, Electron, Scanning ; Niobium ; toxicity ; Osteoblasts ; drug effects ; Oxides ; chemical synthesis ; toxicity ; Porosity ; Potassium ; toxicity ; Pressure ; Rats ; Rats, Sprague-Dawley ; Skull ; cytology ; Stress, Mechanical ; Surface Properties ; Tetrazolium Salts ; Thiazoles

The cuttlebones, harvested from cuttles, undergo the chemical reaction in high temperature and high pressure for a certain time. The products are qualitatively analysed, and spacial structure observation and cytocompatibility are tested. The results show that the chemical component of the cuttlebone is CaCO3 and the crystal type is aragonite. Cuttlebones undergo a hydro-thermal reaction, and thus transform into hydroxyapatite-that is, the cuttlebone-transformed hydroxyapatite(CBHA). The CBHA materials have the interconnected microporous network structures. Under the high magnification, CBHAs appear to have many micro-spheres, thus construct a new self-assembled nano-material system. The marrow stromal osteoblasts can adhere to and proliferate well on the surface of the CBHAs. These results show that CBHAs have good biocompatibility. Therefore, it can be a potential candidate scaffold for bone tissue engineering.
Animals ; Bone Substitutes ; chemical synthesis ; chemistry ; toxicity ; Cells, Cultured ; Durapatite ; chemical synthesis ; chemistry ; toxicity ; Materials Testing ; Osteoblasts ; cytology ; drug effects ; Rabbits ; Sepia ; anatomy & histology ; Spine ; anatomy & histology ; chemistry ; Tissue Engineering

We examined the biocompatibility and the safety of a-calcium sulfate hemihydrate (CSH)/multi-walled carbon nanotube (MWCNT) composites for bone reconstruction application. The biocompatibility of the CSH/MWCNT composites was evaluated by the measures which taking L929 fibroblast cells cultured in the extracted liquid of the composite soaking solution and putting bone marrow stromal cells planted on the composite pellets in vitro, respectively. The cell proliferation was evaluated by MTT test and further observed using an inverted optical microscope and a scanning electric microscope. The toxicity of the composites was evaluated by acute and subacute systemic toxicity test. Long-term muscle and bone implantation in vivo tests were also conducted. L929 fibroblast cells grew well in the extracted liquid, as well as bone marrow stromal cells that could adhere on the surface of sample pellets and proliferated rapidly. MTT test showed that there were no significant differences between the experimental and control groups (P > 0.05). In vivo test manifested that the composites were no toxicity, no irritation to skin and good for bone defect reconstruction. It was proved that a-calcium sulfate hemihydrate (CSH)/multi-walled carbon nanotube (MWCNT) composites exhibited excellent biocompatibility for the potential application in bone tissue engineering.
Animals ; Biocompatible Materials ; chemistry ; Bone Marrow Cells ; cytology ; Bone Substitutes ; chemical synthesis ; chemistry ; Calcium Sulfate ; chemistry ; Cell Line ; Cell Proliferation ; Fibroblasts ; cytology ; Materials Testing ; Nanotubes, Carbon ; chemistry ; Rabbits ; Stromal Cells ; cytology ; Tissue Engineering ; methods ; Toxicity Tests