The aim is to evaluate the development of an experimental multi-mode/Universal resin-based dentin adhesive modified with synthetic Mg2+ doped hydroxyapatite crystals (HAp) having self-remineralization and antibiofilm properties. HAp doped with Mg2+ was prepared by the precipitation method. Experimental adhesives were subjected to degree of conversion and X-ray diffraction test for size and crystal structure. Bond strength was tested, and electron microscopy (SEM/TEM) imaging of resin-dentin interface was done along with nanoleakage, nanoindentation, confocal and Raman analyses. S. mutans was analysed using CLSM images against modified adhesive specimens. Nucleating abilities within the resin-dentin specimens are determined by measuring Ca2+. Alkaline phosphatase, Runx2, and Ocn transcripts are amplified using quantitative polymerase chain reaction (q-PCR). A calcium assay is performed to quantify level of mineralisation. When compared to control adhesives, the 0.5% Hap/Mg2+ containing experimental dentin adhesive demonstrated improved interaction with dentin. The preservation of uniform intact hybrid layer with the absence of nanoleakage indicated dentin bond integrity with 0.5% HAP/Mg2+ modified adhesive. Self-remineralization and antibiofilm potentials are supported.
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.
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 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.
The effect of the addition of an ionic dopant to calcium phosphates for biomedical applications requires specific research due to the essential roles played in such processes. In the present study, the mechanical and biological properties of Ni-doped hydroxyapatite (HA) and Ni-doped HA mixed with graphene nanoplatelets (GNPs) were evaluated. Ni (3wt.% and 6wt.%)-doped HA was synthesized using a continuous precipitation method and calcined at 900°C for 1h. The GNP (0.5-2wt.%)-reinforced 6% Ni-doped HA (Ni6) composite was prepared using rotary ball milling for 15h. The sintering process was performed using hot isostatic pressing at processing conditions of 1150°C and 160MPa with a 1-h holding time. The results indicated that the phase compositions and structural features of the products were noticeably affected by the Ni and GNPs. The mechanical properties of Ni6 and 1.5Ni6 were increased by 55% and 75% in hardness, 59% and 163% in fracture toughness and 120% and 85% in elastic modulus compared with monolithic HA, respectively. The in-vitro biological behavior was investigated using h-FOB osteoblast cells in 1, 3 and 5days of culture. Based on the osteoblast results, the cytotoxicity of the products was indeed affected by the Ni doping. In addition, the effect of GNPs on the growth and proliferation of osteoblast cells was investigated in Ni6 composites containing different ratios of GNPs, where 1.5wt.% was the optimum value.
Autologous bone grafting remains the gold standard for almost all bone void-filling orthopedic surgery. However, autologous bone grafting has several limitations, thus scientists are trying to identify an ideal synthetic material as an alternative bone graft substitute. Magnesium-doped biphasic calcium phosphate (Mg-BCP) has recently been in the spotlight and is considered to be a potential bone substitute. The Mg-BCP is a mixture of two bioceramics, that is, hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), doped with Mg2+ , and can be synthesized through chemical wet-precipitation, sol-gel, single diffusion gel, and solid state reactions. Regardless of the synthesis routes, it is found that the Mg2+ preferentially accommodates in β-TCP lattice instead of the HA lattice. The addition of Mg2+ to BCP leads to desirable physicochemical properties and is found to enhance the apatite-forming ability as compared to pristine BCP. In vitro results suggest that the Mg-BCP is bioactive and not toxic to cells. Implantation of Mg-BCP in in vivo models further affirmed its biocompatibility and efficacy as a bone substitute. However, like the other bioceramics, the optimum physicochemical properties of the Mg-BCP scaffold have yet to be determined. Further investigations are required regarding Mg-BCP applications in bone tissue engineering.
Surface reactivity of bioactive ceramics contributes in accelerating bone healing by anchoring osteoblast cells and the connection of the surrounding bone tissues. The presence of silicon (Si) in many biocompatible and bioactive materials has been shown to improve osteoblast cell adhesion, proliferation and bone regeneration due to its role in the mineralisation process around implants. In this study, the effects of Si-biphasic calcium phosphate (Si-BCP) on bioactivity and adhesion of human osteoblast (hFOB) as an in vitro model have been investigated. Si-BCP was synthesised using calcium hydroxide (Ca(OH)2) and phosphoric acid (H3PO4) via wet synthesis technique at Ca/P ratio 1.60 of material precursors. SiO2 at 3 wt% based on total precursors was added into apatite slurry before proceeding with the spray drying process. Apatite powder derived from the spray drying process was pressed into discs with Ø 10 mm. Finally, the discs were sintered at atmospheric condition to obtain biphasic hydroxyapatite (HA) and tricalcium phosphate (TCP) peaks simultaneously and examined by XRD, AFM and SEM for its bioactivity evaluation. In vitro cell viability of L929 fibroblast and adhesion of hFOB cell were investigated via AlamarBlue® (AB) assay and SEM respectively. All results were compared with BCP without Si substitution. Results showed that the presence of Si affected the material's surface and morphology, cell proliferation and cell adhesion. AFM and SEM of Si-BCP revealed a rougher surface compared to BCP. Bioactivity in simulated body fluid (SBF) was characterised by pH, weight gain and apatite mineralisation on the sample surface whereby the changes in surface morphology were evaluated using SEM. Immersion in SBF up to 21 days indicated significant changes in pH, weight gain and apatite formation. Cell viability has demonstrated no cytotoxic effect and denoted that Si-BCP promoted good initial cell adhesion and proliferation. These results suggest that Si-BCP's surface roughness (164 nm) was significantly higher than BCP (88 nm), thus enhancing the adhesion and proliferation of the osteoblast.
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.
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.
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