The study was carried out to evaluate macroscopically the ability of coral to repair a large size bone defect. A total 12 adult, male sheep were used in the study. The large bone defect (2.5cm x 0.5cm x 0.5cm) was created surgically on the left proximal femur and replaced by a block of coral (Porites sp.). Radiographs were obtained immediately after surgery and at 2, 4, 8 and 12 weeks post-implantation. Ultrasonographic examinations were carried out every 2 weeks after implantation up to 12 weeks using ultrasound machine (TOSHIBA Capasee II) connected with 7MHz frequency transducer. The sheep were euthanased at 2, 4, 8, and 12 weeks post-implantation and the bone examined grossly. Both ultrasonographs and radiographs taken at 8 and 12 weeks showed that the implants had been resorbed and left the space that much reduced in size. There was no sign of implant rejection observed in all animals. The results showed that processed coral has potential to become bone substitute for reconstructive bone surgery.
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.
The aim of this study was to evaluate the in vitro cytotoxicity of biomaterials; Hydroxyapatite (HA), Natural coral (NC) and Polyhydroxybutarate (PHB). Three different materials used in this study; HA (Ca10(PO4)6(OH)2), NC (CaCO3) and PHB (Polymer) were locally produced by the groups of researcher from Universiti Sains Malaysia. The materials were separately extracted in the complete culture medium (100mg/ml) for 72h and introduced to the osteoblast cells CRL-1543. The viability of osteoblast CRL-1543 cultivated with these extraction materials after 72h incubation period was compared to negative control with neutral red assay by using spectrophotometer at 540nm. The results showed the non-cytotoxicity of the materials. After 72h of incubation period, HA showed 123% viable cells, NC was 99.43% and PHB was 176.75%. In this study, cytotoxicity test dealt mainly with the substances that leached out from the biomaterial. The results obtained showed that the materials were not toxic and also promoted cells growth in the sense of biofunctionality.
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.
This in vivo study revealed that porous hydroxyapatite (PHA) and dense hydroxyapatite (DHA) are good implant materials that can accelerate bone healing and resorbed in acceptable time. But there were differences in the mechanism of the resorption of DHA and PHA due to variability in the physical properties and osteogenicity.
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.
There has been, and is still, concern about the high elastic modulus of Ti alloys compared to bone. Any reduction in the Young's modulus value of the implant is expected to enhance stress redistribution to the adjacent bone tissues, minimize stress shielding and eventually prolong device lifetime. Dynamic Monte Carlo simulation is used to predict the gradual reduction in Young's modulus values between the bulk of Ti alloys and the modified surface layers due to Ca ion implantation. The simulation can be used as a screening step when applying new alloys and/or coatings.
In this work, nanometer HA crystals have been synthesized via wet chemical precipitation and characterized. This research studies how key synthesis parameters affect the size and phase purity of the produced HA. Characterization work was carried out using X-ray powder diffraction method and scanning electron microscopy for phase identification and particle sizing, respectively.
Matched MeSH terms: Bone Substitutes/analysis; Bone Substitutes/chemical synthesis*
Bone is unique in its ability to adapt structure to functional requirements, but as is all too obvious in an ever-ageing population it is susceptible to a number of degenerative diseases. Therefore there is an increasing need for materials for bone replacement. Clearly, the ideal material with which to replace bone, would be bone itself, but the major problem now facing us is that there is an insufficient supply of the natural bone to satisfy the clinical requirements. Hence, there is a need for the development of chemically synthesised bone graft substitutes
The study was carried out with the aim to evaluate natural coral (Porites spp.) implanted in sheep femur microscopically. Twelve adult, male sheep were used in this study. The defect area was implanted with coral and monitored for up to 12 weeks. The sheep were euthanased at 2,4,8, and 12 weeks post-implantation. Microscopically, natural coral implanted into bone tissue have shown gradual resorption and progressively replaced by new bone. At 12 weeks post-implantation, the implanted site was almost completely surrounded by mature bone. The results showed that natural coral was found to be a biodegradable and osteo-conductive biomaterial, which acted as a scaffold for a direct osteoblastic apposition.
Avian demineralized bone matrix (ADBM) powder prepared from chicken, pigeon, and turkey sources induced bone formation via endochondral and intramembranous processes, as in mammalian studies. There were no significant differences in percentage of new bone, percentage of cartilage, surface-forming osteoblast area, or osteoclast count between gaps treated with chicken, pigeon, and turkey DBM. However, there was a significantly (p<0.05) higher percentage of inflammatory area in gaps treated with chicken DBM than in gaps treated with pigeon DBM.
Hydroxyapatite (HA; Ca10(PO4)6(OH)2), is one of the significant implant materials used in Orthopaedics and Dental applications. However, synthetically produced HA may not be stable under ionic environment, which it will unavoidably encounter during its applications. In this paper, the in vitro effects of three HA materials derived from different resources, i.e. commercial HA (HAC), synthesised HA from pure chemicals (HAS) and synthesised HA from kapur sireh; derived traditionally from natural limestone (HAK), were studied. The HA disc samples were prepared and immersed in simulated body fluid (SBF) for 31-day period. The evaluation conducted focuses on the changes of the pH and the Calcium ion (Ca-ion) and Phosphate ion (P-ion) concentrations in the SBF solution, as well as the XRD and SEM data representing the reactions on the HA materials. From the XRD, it was found that HAK has the smallest crystallite sizes, which in turn affect the pH of the SBF during immersion. The Ca and P-ion concentrations generally decrease over time at different rates for different HA. Upon 1-day immersion in SBF, apatite growth was observed onto all three surfaces, which became more pronounced after 3-day immersion. However, the appetites formed were observed to be different in shapes and sizes. The reasons for the difference in the apatite-crystals and their subsequent effects on cells are still being investigated.
Matched MeSH terms: Bone Substitutes/chemical synthesis*
Defects were created in the mandible of a rabbit model whereby the right side was implanted with hydroxyapatite (HA) while the left side was left empty to act as control. Both the implant and control sites were evaluated clinically and histologically at 4,12,20,22 weeks. Decalcified sections were studied under confocal laser scanning microscope. No reactive cells were evident microscopically in all sections. There was bone ingrowth as early as 4 weeks when viewed by the topographic method. Enhancement of osteoconduction was evident by the presence of abundant capillaries, perivascular tissue and osteoprogenitor cells of the host. At 22 weeks, the implanted defect showed mature bone formation filling almost the whole field. This study demonstrated that the dense HA exhibits excellent biocompatibility as noted by the complete absence of reactive cells. It also promotes osteoconduction.
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.
This study is to qualitatively evaluate a locally produced hydroxyapatite (HA), made by AMREC-SIRIM in an experimental animal bone defect using New Zealand White (NZW) rabbits. HA cylindrical blocks measuring 2.5 mm (D) x 1.0 mm (H) were implanted in the rabbits' left tibia. The tibias were harvested within one to three weeks post-implantation. The implantion site was cut into thin undecalcified sections of about 30 microm to 60 microm and stained with Toluidine Blue and Goldner's Masson Trichrome. Microscopic examinations using standard light microscopy of these slides were performed.
There was a significant increased in Absolute Contact Length measurements of endosteal bone growth along the Nickel-Titanium (NiTi) implant coated with the natural coral powder and Hydroxyapatite (HA) compared to the non-calcium coated implants. This study demonstrated that coated implants seemed to show earlier and higher osseointergration phenomena compared to non coated ones. Furthermore, there was significantly greater bone-to-implant contact at the apical 1/3rd of the coated implants.
The main objective of the study was to determine the biodegradability, resorption and osteoconductivity potency of coral implant. Coral blocks (CORAGRAF) were prepared from sea coral Porites species. The blocks were implanted in the right mandible of rabbit model. Implants were harvested at 2 and 4 weeks intervals and subjected for light and scanning electron microscopy. Dense hydroxyapatite (DHA) was implanted in the left mandible as a control. The results of this study demonstrated that CORAGRAF is a good implant material that can accelerates bone healing and be resorbed in an acceptable time. The mechanisms of the resorption seemed to be the same (crumbling process), a first step where the edge of the coral become powdery then a second step which could be phagocytosis and dissolution in extracellular fluid.
The present in vitro evaluation indicated that the value added hydroxyapatite (HA) was more toxic than pure HA but the toxicity of value added HA was slight compared to the positive control. In this testing, the conclusion can be made that value added HA is less biocompatible than commercialized pure HA. This toxicity may be caused by both the particle size and degradation (leaching). Further studies should be carried out to determine whether there is particle size effect or leaching effect when using powder as compared to the block materials. The in vivo evaluation should be done to assess the reaction to this value added HA as compared to the pure HA.
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.