Quick setting and poor injectability due to liquid-solid phase separation have limited the clinical use of brushite and monetite cements. The presence of certain ions in the cement during the setting reaction moderate the setting time and properties of the cement. This study reports the preparation of injectable bone cement by using biphasic calcium phosphate (BCP) extracted from femur lamb bone by calcination at 1450 °C. EDX analysis infers the presence of Mg and Na ions as trace elements in BCP. X-ray diffraction patterns of the prepared cement confirmed the formation of brushite (DCPD) along with monetite (DCPA) as a minor phase. DCPA phase diminished gradually with a decrease in powder to liquid ratio (PLR). Initial and final setting time of 5.3 ± 0.5 and 14.67 ± 0.5 min respectively are obtained and within the acceptable recommended range for orthopedic applications. Exceptional injectability of ≈90% is achieved for all prepared bone cement samples. A decrease in compressive strength was observed with increase in the liquid phase of the cement, which is attributed to the higher degree of porosity in the set cement. Immersion of bone cement in simulated body fluid (SBF) for up to 7 days resulted in the formation of apatite layer on the surface of cement with Ca/P ratio 1.71, which enhanced the compressive strength from 2.88 to 9.15 MPa. The results demonstrate that bone cement produced from BCP extracted from femur lamb bone can be considered as potential bone substitute for regeneration and repair of bone defects.
Improvement of the mechanical properties of hydroxyapatite (HA) can be achieved by the incorporation of metal. In addition, incorporation of strontium ion into HA crystal structures has been proved effective to enhance biochemical properties of bone implant. In this research, strontium-doped HA powder was developed via a sol-gel method to produce extraordinarily fine strontium-doped HA (Sr-doped HA) powder. XRD measurement had shown that the powder contained hydroxyapatite phase only for all doping concentration except for 2%, showing that Sr atoms have suppressed the appearance of beta-TCP as the secondary phase. Morphological evaluation by FESEM measurement shows that the particles of the Sr-doped HA agglomerates are globular in shape with an average size of 1-2 microm in diameter while the primary particles have a diameter of 30-150 nm in average.
This paper provides a comprehensive analysis of the dielectric and physicochemical properties of the porous hydroxyapatite/cornstarch (HAp/Cs) composites in a new perspective. The porous composites have been characterized via SEM, FTIR, XRD and dielectric spectroscopy. The dielectric permittivity spectra were obtained in Ku-band (12.4-18.0 GHz) and it was correlated with the physicochemical properties of the porous HAp/Cs. Porous HAp/Cs composites exhibits low ε' and negative ε″, which influenced by the microstructural morphology, interaction between Hap and Cs, as well as crystalline features due to the various proportion of the HAp/Cs. The physicochemical effect of the composites results in the dielectric polarization and energy loss. This phenomenon indicates the presence of the three obvious relaxation responses in the ε' spectrum (13.2-14.0, 15.2-16.0, and 16.6-17.4 GHz) and the negative behaviours in the ε″ spectrum. The relationships between physicochemical and dielectric properties of the porous composite facilitate the development of the non-destructive microwave evaluation test for the porous composite.
Phase composition of calcium phosphate ceramic is a characteristic directly related to the biological response of implants due to the differences in mechanical and biochemical properties of these compounds. The biodegradation rate of biphasic calcium phosphate (BCP) can be controlled by altering the HA to β-TCP ratios. In this study the crystalline phase evolution of BCP synthesized via precipitation from aqueous solution of (NH4)2PO4 titrated into heated solution of Ca (NO3)2 was evaluated. The resulting powder was fabricated into porous scaffold using polyurethane foam method. Bulk powders were sintered from 700 - 1400°C to determine the most significant sintering temperature to obtain a stable and well crystallize BCP phases. The porous scaffolds were then sintered at selected temperature and the effects of various sintering times from 5,7,9,11,13 and 15 h were investigated. Bulk powders were characterized by dilatometer, IR analysis and XRD and porous scaffolds were analyzed by XRD and SEMEdx. RIR method was performed to show that the HA to β-TCP ratios were increased with increasing of sintering time and reached the maximum HA value at 11h. It is found that, the possibilities to manipulate the HA to β-TCP ratios in BCP porous scaffold by just controlling the sintering time of the scaffold without controlling the starting powder characteristics.
Hydroxyapatite powder was mechanochemically synthesized from calcium pyrophosphate (Ca2P2O7) and calcium carbonate (CaCO3) using a solid-state reaction. The two powders were mixed in distilled water, milled for 8 hours, dried and calcined at 1100 degrees C for 1 hour. The phase(s) formed was analyzed by x-ray diffraction (XRD). It was found that hydroxyapatite was not the only one formed. This result will be used as the starting point to produce a single-phase hydroxyapatite in terms of excess hydroxyl group in a mechanochemical reaction.
The present paper reports on the influence of sintering temperature on the porosity and strength of porous hydroxyapatite (HA). HA powder was first prepared by the sol-gel precipitation method using calcium hydroxide and ortho-phosporic acid. The fine HA powder, measuring <50 microm was then mixed into a slurry with the addition of binder agent, being a mixture of sago and PVA. A small amount of sodium dodecyl sulphate was also used as a foaming agent. Porous HA samples were then prepared via slip casting technique. The surface morphology of the sintered samples was observed under scanning electron microscopy at 20 kV and the compositions were determined via SEM-EDX. A universal testing machine was used to determine the compaction strength of the sintered samples.
In Malaysia, the field of genomics in toxicology is still in infancy. The purpose of this study is to focus on the use of toxicogenomics for determination of gene expressions changes in cultured human fibroblast cells treated with genotoxicology free biomaterial (using Ames test), a locally produced hyroxyapatite. Dose and time response is similar to Ames test with time interval up to 21 days. mRNA is extracted, followed with RT-PCR and polyacrilamide gel electrophoresis. Changes of the gene expressions compared to the non-treated fibroblast mRNA would suggest some gene interactions in the molecule level associated with the exposure of the fibroblast cell line to the biomaterials. Further analysis (cloning & sequencing) shall be carried out to investigate the genes involved as simple changes might not signified toxicity.
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
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.
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.
There is a great demand of Hydroxyapatite (HA) material in Orthopaedics and Dental applications due to its similarity to human bone. However, the lack of availability and due to high import cost of this material in Malaysia, research in producing synthetic HA locally is therefore timely. The use of local resources as the raw materials for the production of HA is also desirable in reducing the overall cost of HA. In this study, two HA materials were synthesised from different starting precursors, i.e. commercial pure Ca(OH)2 (HAS) and Ca(OH)2 directly from a local natural limestone deposit (HAL). Whereas a commercially available HA "Captal 60" (HAC) was used as reference. The synthesised powders obtained were fired at 1000 degrees C and at 1250 degrees C. Characterisation evaluations on bulk properties were carried out using XRD, SEM-EDX, ICP and FTIR. The results indicate that both HAS and HAL are comparable to HAC even at 1000 degrees C. Thus, the local natural limestone can be used to form HA. However, the overall appearance of these materials are quite different (HAC - blue, HAS - greenish and HAL - light green). The reasons for this and the subsequent mechanical and bioactive effects of these materials are currently being investigated.
The paper discusses the influence of sintering temperature on the microstructure and strength of hydroxyapatite ceramics prepared using the extrusion process. The average pore diameters observed were in the range of approximately 150mm to 300mm whereas the compaction strength was found to be around 120-160 MPa.
Hydroxyapatite (HA) powder was synthesized via wet method using calcium nitrate hydrate (Ca(NO3)2.H2O) and diammonium hydrogen phosphate ((NH4)2HPO4) as raw materials. Powder obtained was milled using various milling speed ranging from 250 to 400 r.p.m. and sintered at 1300°C for 2hrs. Due to the nature of HA powder that decomposed at high temperature, XRD technique have been used in this work to determine the phase composition of the HA powder and also the crystallite size. The unmilled sample was used as the control group. Results show that sufficient heat supply generated from the milling process, initiates the decomposition of HA phase into ȕ-tricalcium phosphate (ȕ-TCP). Decomposition of HA starts to occur at the milling speed of 300 rpm, i.e the formation of ȕ-TCP was occurred at lower sintering temperature. It was believed that the decomposition of HA was associated with the formation of an intermediate phase, oxyapatite. Moreover, the crystallinity and particle size of the produced powder is very much affected by the milling speed and the stability of the HA. All milled powders possess spherical shape particle.
Biphasic Calcium Phosphate (BCP) with a ratio of 20/80 Hydroxyapatite (HA)/Beta-tricalcium phosphate (β-TCP) promotes the differentiation of human dental pulp cells (HDPCs). In the current study, the genotoxicity of locally produced BCP of modified porosity (65%) with a mean pore size of 300micrometer (μm) was assessed using Comet and Ames assays. HDPCs were treated with BCP extract at three different inhibitory concentrations which were obtained based on cytotoxicity test conducted with concurrent negative and positive controls. The tail moment of HDPCs treated with BCP extract at all three concentrations showed no significant difference compared to negative control (p>0.05), indicating that BCP did not induce DNA damage to HDPCs. The BCP was evaluated using five tester strains of Salmonella typhimurium TA98, TA100, TA102, TA1537 and TA1538. Each strain was incubated with BCP extract with five different concentrations in the presence and absence of metabolic activation system (S9) mix. Concurrently, negative and positive controls were included. The average number of revertant colonies per plate treated with the BCP extract was less than double as compared to the number of revertant colonies in negative control plate and no dose-related increase was observed. Results from both assays suggested that the BCP of modified porosity did not exhibit any genotoxic effect under the present test conditions.
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.
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.
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 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.