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In vitro cytotoxicity testing on valued added hydroxyapatite as bone replacement material

Shaari R, Samsudin AR

Med J Malaysia, 2004 May;59 Suppl B:109-10.
PMID: 15468842

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.

Matched MeSH terms: Hydroxyapatites/toxicity*
Fulltext Bioactivity studies and adhesion of human osteoblast (hFOB) on silicon-biphasic calcium phosphate material

Ibrahim S, Sabudin S, Sahid S, Marzuke MA, Hussin ZH, Kader Bashah NS, et al.

Saudi J Biol Sci, 2016 Jan;23(1):S56-63.
PMID: 26858566 DOI: 10.1016/j.sjbs.2015.10.024

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.

Matched MeSH terms: Hydroxyapatites
In-vitro evaluation of microleakage of bioceramic root-end filling materials: A spectrophotometric study

Gupta R, Kewalramani R

J Oral Biol Craniofac Res, 2021 03 10;11(2):330-333.
PMID: 33786296 DOI: 10.1016/j.jobcr.2021.03.001

Aim: To evaluate the microleakage of newer bioceramic root-end filling materials.
Material and method: Sixty freshly extracted human single-rooted mandibular premolar teeth were selected for the study. Teeth with fractured root, cracks, anddilacerations were rejected. All teeth were cleaned with ultrasonic scalers. Standard access opening was done and root canal treatment was performed with rotary files followed by obturation. After storing in saline for a week apical 3 mm of the root was resected at 900 angles to the long axis of the root. Retro cavity preparation was done with ultrasonic tips. The teeth were divided into four groups of 15 specimens each. Group I - Biodentin, GroupII-Bioaggregate, Group III - MTA Plus, and Group IV - MTA. After the restoration of retro cavities of all the teeth as per manufacture instructions, two coats of nail varnish were applied to leave apical 3 mm. All teeth were stored in 2% methylene blue for 72 h followed by emersion in 65% nitric acid for the next 72 h for Dye extraction. The obtained supernatant solution was then centrifuged and optical density or absorbance was measured with a UV spectrophotometer.
Result: Microleakage was found to be increasing in this order: Biodentin 0.01).
Conclusion: All materials exhibit some amount of microleakage. Biodentin shows the least microleakage among all the bioceramic material groups. Hence, Biodentin and bioaggregate are better material of choice for the retrograde filling to prevent microleakage.

Matched MeSH terms: Hydroxyapatites
Effect of space holders on fabrication of porous titanium alloy-hydroxyapatite composite through powder injection molding

Farrahshaida Mohd Salleh, Abu Bakar Sulong, Muhammad Rafi Raza, Norhamidi Muhamad, Lim TF

Sains Malaysiana, 2017;46:1651-1657.

owder injection molding (PIM) is able to produce porous titanium alloy/hydroxyapatite composite through the space holder technique. Thermal debinding and sintering processes were the main challenges due to different properties of metal and ceramic in producing such composite. This study focused on the effect of different space holders on the physical and mechanical properties of debound and sintered porous titanium aloi/hydroxyapatite composite. The feedstock is containing of 80 wt. % of titanium alloy/hydroxyapatite with 20 wt. % of space holders such as sodium chloride (NaCl) and polymethylmethacrylate (PMMA), respectively. The binders were then removed from the injected samples by two stages of debinding; solvent and thermal debinding. The sintering was performed at three different temperatures 1100oC, 1200oC and 1300oC at a heating rate of 10oC /min and holding time of 5 h. It was found that the samples containing PMMA space holder was fractured after sintering. While, the samples containing NaCl space holder successfully formed pores and not fractured. At sintering temperature of 1300oC, the density, compressive strength and porosity volume percentages for the sintered sample containing NaCl space holder were 3.05 g/cm3, 91.7 MPa. and 11.9 vol%, respectively.

Matched MeSH terms: Hydroxyapatites
Characterization of nickel-doped biphasic calcium phosphate/graphene nanoplatelet composites for biomedical application

Baradaran S, Moghaddam E, Nasiri-Tabrizi B, Basirun WJ, Mehrali M, Sookhakian M, et al.

Mater Sci Eng C Mater Biol Appl, 2015 Apr;49:656-668.
PMID: 25686995 DOI: 10.1016/j.msec.2015.01.050

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.

Matched MeSH terms: Hydroxyapatites/chemistry
Mutagenicity of CORAGRAF and REKAGRAF in the Ames test

Suzina AH, Azlina A, Shamsuria O, Samsudin AR

Med J Malaysia, 2004 May;59 Suppl B:105-6.
PMID: 15468840

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.

Matched MeSH terms: Hydroxyapatites/toxicity*
Biocompatible magnesium-doped biphasic calcium phosphate for bone regeneration

Ballouze R, Marahat MH, Mohamad S, Saidin NA, Kasim SR, Ooi JP

J Biomed Mater Res B Appl Biomater, 2021 Oct;109(10):1426-1435.
PMID: 33484103 DOI: 10.1002/jbm.b.34802

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.

Matched MeSH terms: Hydroxyapatites/chemistry*
Fulltext Fabrication of Hydroxyapatite with Bioglass Nanocomposite for Human Wharton's-Jelly-Derived Mesenchymal Stem Cell Growing Substrate

Ebrahimi S, Hanim YU, Sipaut CS, Jan NBA, Arshad SE, How SE

Int J Mol Sci, 2021 Sep 06;22(17).
PMID: 34502544 DOI: 10.3390/ijms22179637

Recently, composite scaffolding has found many applications in hard tissue engineering due to a number of desirable features. In this present study, hydroxyapatite/bioglass (HAp/BG) nanocomposite scaffolds were prepared in different ratios using a hydrothermal approach. The aim of this research was to evaluate the adhesion, growth, viability, and osteoblast differentiation behavior of human Wharton's-jelly-derived mesenchymal stem cells (hWJMSCs) on HAp/BG in vitro as a scaffold for application in bone tissue engineering. Particle size and morphology were investigated by TEM and bioactivity was assessed and proven using SEM analysis with hWJMSCs in contact with the HAp/BG nanocomposite. Viability was evaluated using PrestoBlueTM assay and early osteoblast differentiation and mineralization behaviors were investigated by ALP activity and EDX analysis simultaneously. TEM results showed that the prepared HAp/BG nanocomposite had dimensions of less than 40 nm. The morphology of hWJMSCs showed a fibroblast-like shape, with a clear filopodia structure. The viability of hWJMSCs was highest for the HAp/BG nanocomposite with a 70:30 ratio of HAp to BG (HAp70/BG30). The in vitro biological results confirmed that HAp/BG composite was not cytotoxic. It was also observed that the biological performance of HAp70/BG30 was higher than HAp scaffold alone. In summary, HAp/BG scaffold combined with mesenchymal stem cells showed significant potential for bone repair applications in tissue engineering.

Matched MeSH terms: Hydroxyapatites/chemistry*
Fulltext Incorporation of Hydroxyapatite into Glass Ionomer Cement (GIC) Formulated Based on Alumino-Silicate-Fluoride Glass Ceramics from Waste Materials

Wan Jusoh WN, Matori KA, Mohd Zaid MH, Zainuddin N, Ahmad Khiri MZ, Abdul Rahman NA, et al.

Materials (Basel), 2021 Feb 18;14(4).
PMID: 33670465 DOI: 10.3390/ma14040954

Glass ionomer cement (GIC) is a well-known restorative material applied in dentistry. The present work aims to study the effect of hydroxyapatite (HA) addition into GIC based on physical, mechanical and structural properties. The utilization of waste materials namely clam shell (CS) and soda lime silica (SLS) glass as replacements for the respective CaO and SiO2 sources in the fabrication of alumino-silicate-fluoride (ASF) glass ceramics powder. GIC was formulated based on ASF glass ceramics, polyacrylic acid (PAA) and deionized water, while 1 wt.% of HA powder was added to enhance the properties of the cement samples. The cement samples were subjected to four different ageing times before being analyzed. In this study, the addition of HA caused an increment in density and compressive strength results along with ageing time. Besides, X-ray Diffraction (XRD) revealed the formation of fluorohydroxyapatite (FHA) phase in HA-added GIC samples and it was confirmed by Fourier Transform Infrared (FTIR) analysis which detected OH‒F vibration mode. In addition, needle-like and agglomeration of spherical shapes owned by apatite crystals were observed from Field Emission Scanning Electron Microscopy (FESEM). Based on Energy Dispersive X-ray (EDX) analysis, the detection of chemical elements in the cement samples were originated from chemical compounds used in the preparation of glass ceramics powder and also the polyacid utilized in initiating the reaction of GIC.

Matched MeSH terms: Hydroxyapatites
Development of a bioactive dentin adhesive resin modified with magnesium-doped synthetic hydroxyapatite crystals

Daood U, Fawzy A

J Mech Behav Biomed Mater, 2023 Apr;140:105737.
PMID: 36827934 DOI: 10.1016/j.jmbbm.2023.105737

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.

Matched MeSH terms: Hydroxyapatites