The porous hydroxyapatite bioceramics were prepared through combination of sacrificial templet and direct foaming techniques utilizing PMMA granules ( varied from 5 to 50wt % in content ) as a templet and H2O2 solution ( varied from 5 to 30wt % in concentration ) as a foaming agent, severally. The effects of PMMA content and H2O2 concentration on concluding porousness, microstructure and mechanical strengths were studied. The porous samples utilizing PMMA provided the porousness runing from 52 % to 75 % , the samples utilizing H2O2 had the porousness runing from 82 % to 85 % , and the sample utilizing both pore formers provided the porousness runing between 84 % and 90 % . The higher content of PMMA and concentration of H2O2 led the porousness increased, taking to a lessening in the compressive and flexural strengths. Furthermore, this combination technique allowed interrelated pores holding two degrees of pore size, which were come from PMMA and H2O2. The PMMA formed the little pores with the diameter runing between 100 and 300 millimeters, while H2O2 provided the larger pores with the diameter runing from 100 to 1,000 millimeters depending on concentration.
Presents, porous bioceramics have an progressively of import function in biomedical application, such as bone filler, orbital implant and drug bringing bearers. [ 1 ] One of the most popular subject focal points on porous hydroxyapatite ( HA ) . Since it has a chemical composing ( Ca10 ( PO4 ) 6 ( OH ) 2 ) identical to human bone and an first-class biocompatibility. Furthermore, porous construction provides outstanding bone ingrowths, vascularisation and an increased interfacial country between the implant and the tissues ensuing in the stronger fond regard. [ 1,2,3 ]
A figure of pore organizing techniques on bioceramics have been proposed, which can be classified into 3 chief techniques. [ 4 ] There are ( 1 ) reproduction technique, e.g. replamineform and PU impregnation ; [ 5,6 ] ( 2 ) sacrificial templet technique, e.g. amylum consolidation, dual-phase commixture and camphene-based freezing casting ; [ 7,8,9 ] and ( 3 ) direct foaming technique, e.g. frothing method and gelcasting. [ 10,11 ] However, about old surveies were based on utilizing a individual technique. A few plants focused on porous fiction utilizing combination of these techniques. Padilla et Al. studied on porous HA utilizing combination of PU impregnation ( reproduction ) and gelcasting ( direct foaming ) techniques ; and Batulli et Al. studied on porous zirconium oxide utilizing polythene sphere as a sacrificial templet and gelcasting techniques. [ 12,13 ] In malice of these, it seems no survey on fiction of porous HA utilizing combination of sacrificial templet and direct foaming techniques.
The intent of this survey was to qualify the porous HA samples fabricated through combination of sacrificial templet and direct foaming techniques utilizing PMMA granules and H2O2 solution as a pore templet and a foaming agent, severally. Additionally, the effects of PMMA content and H2O2 concentration on the concluding porousness, microstructure, compressive strength and flexural strength of the samples were besides investigated.
2. Materials and methods
2.1 Preparation of hydroxyapatite pulverization
A stoichiometric HA was prepared utilizing the precipitation reaction between 0.5 moles calcium hydrated oxide ( Riedel-de-Haen, Germany ) and 0.3 moles orthophosphoric acid ( Merck, Germany ) . The precipitation reaction was performed at room temperature and the pH was controlled at 10.5 by the add-on of ammonium hydrated oxide solution ( APS Finechem, Australia ) . [ 14 ] During blending procedure, the acerb solution was easy dropped into the smartly stirring suspension, utilizing a peristaltic pump. [ 15 ] After complete commixture of the reactants, the suspension was aged overnight. The precipitate was filtered, dried at 80oC overnight and so land to a pulverization by a stamp and howitzer. Subsequently, the pulverization was sieved with the size of 100 millimeter.
2.2 Fabrication of porous hydroxyapatite
The poly-methyl-methacrylate granules ( PMMA ) being commercial class was applied as a pore former for sacrificial templet technique, while H peroxide solution, ( H2O2 ) was used as a foaming agent for direct foaming technique.
The porous hydroxyapatite ceramics were prepared through three techniques, including sacrificial templet, direct foaming and combination between sacrificial templet and direct foaming. To manufacture the porous samples, the HA pulverization was homogeneously assorted with PMMA granule at assorted content ( 5, 10, 20, 30, 40 and 50wt % ) and H2O2 solution at different concentration ( 5, 10, 20 and 30wt % ) , with the liquid to pulverize ratio ( L/P ratio ) of 1.3 ml/g. A series of sample name were listed in Table 1. To measure the consequence of PMMA content and H2O2 concentration, the mixtures were prepared without add-on of binder, deflocculant and other additives.
After blending, the paste was placed into removable casts, and kept at 60oC nightlong. At this temperature, the decomposition of H2O2 produced the foaming of the paste. Afterwards, the green samples were removed from the casts and so heated at 400oC for 1 hr with the slowly incline rate of 1oC/min for firing out PMMA granules and to avoid the snap. Finally, the samples were sintered at 1100oC for 2 hours with the incline rate of 5oC/min and so furnace cooled.
2.3 Word pictures
The Morphology of the synthesized HA pulverization and the as-received PMMA granule was characterized utilizing optical maser atom size distribution analysis ( LPD ) and scanning negatron microscope ( SEM ) . In add-on to the morphological survey, SEM was besides applied to detect the microstructure of the porous HA samples.
The stage pureness of the sintered HA pulverization was analyzed utilizing X-ray diffraction ( XRD ) with CuK? radiation. The scanning scope of 2q was between 20o and 50o at the scan velocity of 0.5o/min.
The decomposition temperature of the PMMA granules was determined by thermo-gravimetric analysis ( TGA ) utilizing coincident thermic analyser ( STA ) with the heating rate of 10oC/min.
The porousnesss of the sintered samples were calculated from the majority denseness of the sample ( ?bulk ) and the theoretical denseness of hydroxyapatite ( ?HA = 3.156 g/cm3 ) , by the Equation ( 1 ) .
Equation ( 1 )
To analyze the consequence of PMMA content and H2O2 concentration on the porousness, the one-way analysis of discrepancy ( ANOVA ) was performed at the important degree ( a ) of 0.05. In add-on, such consequences were used to develop arrested development theoretical accounts to foretell porousness.
The compressive strength and three-point flexural strength of porous HA samples were characterized harmonizing to ASTM C773-88 and C1161-02c specification, severally.
3. Consequences and treatment
3.1 Features of the HA pulverization and PMMA granule
Figure 1 showed atom size distribution of the HA pulverization and the PMMA granule. The distribution of the HA pulverization was a bimodal distribution with the extremums of about 4 millimeters and 20 millimeter in atom size, while that of the PMMA granules was an unimodal distribution with a extremum of about 150 millimeter. The norm and standard divergence of average atom sizes of the both were summarized in Table 2. The mean average atom sizes of the HA pulverization and PMMA granules were 4.97±0.03 millimeters and 150.75±0.15 millimeter, severally.
Figure 2 exhibited SEM micrographs of the HA pulverization and PMMA granules. In figure 2 ( a ) , the HA pulverization had angular atoms with assorted sizes. This may be due to the manually milling procedure through a stamp and howitzer. This is similar to the consequences from old probe by Gibson et Al. [ 14 ] The larger atoms appeared to be the agglomeration of smaller atoms during sintering procedure. In Figure 2 ( B ) , the PMMA granules evidently had a spherical form with assorted sizes, ranged from 10 to 200 millimeters.
Figure 3 displayed XRD spectra of the HA pulverization sintered at 1100oC. The spectra showed the individual stage of HA, matching to the ICDD standard extremum of stoichiometric hydroxyapatite ( standard No.09-0432 ) . Furthermore, this consequence agrees with a old survey in that sintering at high temperature resulted in the crisp and narrow diffraction extremums. [ 16 ]
Figure 4 illustrated TGA curve of the as-received PMMA granules. A stable weight was attained at approximately 400oC, which indicates that the PMMA granules have been wholly decomposed at approximately 400oC. This consequences is similar to the old study by Yao et Al. [ 17 ] In order to allow PMMA break up wholly and avoid the clefts in HA ceramics, the heating rate of the green HA organic structure should be really slow at sintering temperature below 400oC. This is why the warming rate of the samples was set at 1oC/min.
3.2 Effect of PMMA and H2O2 on porousness
Table 1 showed the executable production scope of the porous HA samples in this survey. The samples prepared utilizing merely PMMA at the content of over 40wt % were handless, while the samples assorted with H2O2 solution at the concentration of over 20wt % were really brickle. To fix the samples utilizing both PMMA and H2O2, a balance between the PMMA content and the H2O2 concentration must be considered as shown in Table 1.
Figure 5 presented consequence of PMMA content and H2O2 concentration on porousness of the HA samples performed by assorted techniques: ( a ) sacrificial templet ; ( B ) direct foaming ; and ( degree Celsius ) combination between sacrificial and direct foaming. Table 3 summarized P-value from analysis of discrepancy ( ANOVA ) for porousness of the porous HA samples prepared by assorted organizing methods ( at a = 0.05 ) . Table 4 listed the arrested development theoretical accounts and R-square values from arrested development analysis for porousness of the porous HA samples prepared by assorted methods.
Figure 5 ( a ) showed the concluding porousness of the specimens utilizing PMMA granules as pore templet. The porousness of the HA sample ( H00-P00 ) was about 52 % . Obviously, the porousness increased with an increasing content of PMMA used. This consequence was agree with a old survey by Yao et Al. [ 17 ] By this technique, the porousness reached about 75 % at the PMMA content of 40wt % . The relation between the porousness ( P ) and the content of PMMA ( CP ) was explained by the arrested development theoretical account ( 1 ) in Table 4
Figure 5 ( B ) exhibited the porousness of the samples utilizing H2O2 solution as pore former. The porousness of the samples increased somewhat from ~82 % up to ~85 % , when the concentration of H2O2 increased from 5wt % to 30wt % . To corroborate the consequence of H2O2 concentration on porousness, ANOVA trial was performed. This statistical analysis confirmed that the concentration of H peroxide had a important consequence on porousness, with the P-value of 0.022, at 95 % confident interval, as shown in Table 3. Furthermore, the porousness of the samples utilizing H2O2 less than 5wt % could be predicted by the arrested development theoretical account ( 2 ) in Table 4. However, compared to the sample H00-P00, adding H2O2 ( 5wt % ) resulted in a dramatic addition in porousness of approximately 30 % . This was because the add-on of H2O2 into ceramic slurry produced gas nothingnesss when it was stored at 60oC, although a little sum of H2O2 was applied. [ 10 ] At elevated temperature, H2O2 could bring forth H2 and O2 gases, taking to the nothingnesss in ceramic organic structures. The gas nothingnesss were driven out by heating during the liquid stage. After drying procedure, the gas nothingnesss staying in the paste became the pores in porous dry organic structures.
Figure 5 ( degree Celsius ) illustrated the porousness of the porous samples prepared through combination technique utilizing PMMA granule and H2O2 solution. It seemed a minor alteration in porousness was found, when both PMMA content and H2O2 concentration increased. From P-values in Table 3, the content of PMMA had a important consequence on porousness when the H2O2 concentration was at 5wt % and 10wt % , with the P-value of 0.001 and 0.003, severally ( at 95 % confident interval ) . However, at 20wt % and 30wt % H2O2, the PMMA content showed an undistinguished consequence on porousness with the P-value of 0.066 and 0.061, severally ( at 95 % confident interval ) . As expected, the samples utilizing H2O2 with the concentration of over 20 wt % were difficult to be prepared. So, blending PMMA into these samples had hardly affected on porousness. The porousness of the samples utilizing both PMMA and H2O2 could be estimated by the arrested development theoretical account ( 3 ) in Table 4.
From above consequences, it seemed that the porousness derived from PMMA granules was dominated by H2O2 solution. This was because PMMA granule formed pores by decomposition at above 400oC in solid province while H2O2 produced pores by enlargement of gas nothingnesss in liquid stage, which provide the higher porousness than PMMA.
3.3 Effect of PMMA and H2O2 on microstructure
Figure 6 illustrated SEM micrographs of the HA samples prepared with different conditions, after sintering at 1100oC for 2 hours. The sintered HA or the sample of H00-P00 had a few little pores with less than 50 millimeter in diameter, as shown in Figure 6 ( a ) . These pores were derived from driving out of H2O during drying procedure.
The porous samples prepared by 10wt % and 30wt % PMMA content were presented in Figure 6 ( B ) and 6 ( degree Celsius ) , severally. From the samples, they were composed of two groups of pore size scope. The big pore group had the diameter scope of 100-300 millimeter, originating from PMMA granules, which conformed to the atom size measured and corresponded to Yao et Al. [ 17 ] However, it seemed that the little pore group had a similar pore size as seen in the H00-P00 sample. Furthermore, the more content of PMMA the higher sum of pores.
The porous samples prepared utilizing 10wt % and 20wt % H2O2 concentration were shown in Figure 6 ( vitamin D ) and Figure 6 ( vitamin E ) , severally. By direct foaming technique, the samples had a figure of pores with the diameter larger than 100 millimeter. Comparison between the sample H10-P00 and H20-P00 indicated that the sum of pores and pore size were increased when utilizing a higher concentration of H2O2. However, it seemed there were a few pores derived from H2O. This consequence is agree with a old survey by Almirall et Al. [ 10 ]
The HA sample prepared through combination technique utilizing 10wt % PMMA content and 10wt % H2O2 concentration was displayed in Figure 6 ( degree Fahrenheit ) . It seemed that the sample consisted of the big pores in the ceramic organic structure and the little pores in the ceramic wall. The big pore size was up to 1000 millimeter in diameter, while little pore size ranged between 100 and 300 millimeter in diameter. The big pores were perchance resulted from H2O2, while little pores might be from PMMA granules. Furthermore, the interrelated pores were observed. The interrelated pores, tracts between pores, behavior cells and vass between pores and therefore prefer bone ingrowth inside ceramics. [ 18 ]
Based on old surveies, the minimal demand for pore size is considered to be ~ 100 millimeters due to cell size, migration demands and conveyance. However, pore sizes over 300 millimeter was favourable to heighten new bone formation and cell regeneration. [ 18,19 ] Therefore, the sample prepared by combination technique utilizing both PMMA and H2O2 can supply the pores achieving these demands.
3.4 Effect of PMMA and H2O2 on mechanical strength
Figure 7 showed the consequence of PMMA content and H2O2 concentration on compressive strength of the HA samples performed by sacrificial templet, direct foaming and combination between sacrificial templet and direct foaming. In Figure 7 ( a ) , the compressive strength of the samples prepared utilizing PMMA granules dropped from about 25 MPa to 0.6 MPa when the templet content used increased from 5wt % to 40wt % , while the HA cement ( H00-P00 ) had the compressive strength of around 35 MPa. In Figure 7 ( B ) , the compressive strength of the samples prepared utilizing H2O2 solution decreased from about 0.3 MPa to 0.15 MPa when concentration of the foaming agent was increased from 5wt % to 30wt % . Compared to the sample H00-P00, an add-on of H2O2 could impact on falling in compressive strength of the HA samples due to a dramatic addition in porousness. [ 10 ] Besides, it seemed that H2O2 has a more influence on impairing the strength than PMMA, because H2O2 could do higher porousness than PMMA. In Figure 7 ( degree Celsius ) , the samples prepared through combination technique showed a lessening in compressive strength when utilizing higher PMMA content and higher concentration of H2O2. In other words, the compressive strength decreases with an addition of concluding porousness.
Figure 8 exhibited the consequence of PMMA content and H2O2 concentration on flexural strength of the HA samples performed by sacrificial templet, direct foaming and combination between both techniques. The flexural strength of the specimens prepared utilizing PMMA decreased from around 5.5 MPa to 0.7 MPa when the content of the pore templet was raised from 5wt % to 40wt % , as shown in Figure 8 ( a ) . The flexural strength of the specimens prepared utilizing H2O2 was diminished from about 0.5 MPa to 0.2 MPa when concentration of the bubbling agent increased from 5wt % to 30wt % , as illustrated in Figure 8 ( B ) . As expect, this consequence is consistent with the compressive strength in that H2O2 had a more consequence than PMMA on the strength. Finally, the specimens prepared through combination technique had a lessening in flexural strength when higher content of PMMA and higher concentration of H2O2 were added.
Figure 9 and Figure 10 presented the consequence of porousness on compressive strength and flexural strength, severally, of the HA samples divided by organizing technique, including sacrificial templet utilizing PMMA, direct frothing utilizing H2O2, combination technique utilizing PMMA and H2O2, and overall techniques.
The samples prepared through sacrificial templet utilizing PMMA provided the widest scope of mechanical strengths ( 35 to ~1 MPa in compressive strength and 6 to ~1 MPa in flexural strength ) , as shown in Figure 9 ( a ) and Figure 10 ( a ) .
The samples prepared by direct frothing utilizing H2O2 showed a lessening in both strengths ( from 0.3 to ~0.1 MPa in compressive strength and from ~0.5 to 0.2 MPa in flexural strength ) when porousness increased, as illustrated in Figure 9 ( B ) and Figure 10 ( B ) .
The samples utilizing both PMMA and H2O2 besides had a lessening in mechanical strengths ( ~0.2-0.05 MPa in compressive strength and 0.4-0.1 MPa in flexural strength ) when porousness rose, as exhibited in Figure 9 ( degree Celsius ) and Figure 10 ( degree Celsius ) .
It seemed that both compressive strength and flexural strength decreased with an increasing porousness for all forming techniques, as summarized in Figure 9 ( vitamin D ) and Figure 10 ( vitamin D ) . These consequences agreed with old research that the mechanical strength of the sample was conversely comparative to the sum of pore former used and concluding porousness. [ 9,10,17 ] Furthermore, the samples with the porousness higher than 80 % showed a fluctuation in strength, peculiarly flexural strength. On the other manus, the samples utilizing H2O2 ( i.e. direct foaming and combination technique ) provided high fluctuation in strength more than those utilizing PMMA. This was because the porousness obtained from H2O2 could non be efficaciously controlled.
However, the mechanical strengths of the samples in this survey were lower than the minimal strength of cancellate bone. The minimal compressive strength and flexural strength of human cancellate bone were about 2 MPa and 10 MPa, severally. [ 20 ] The pore size favourable to cram formation and cell regeneration was over 300 millimeters. [ 19 ] Equally good as the interrelated pores favours to cell ingrowth inside the pore. [ 18 ] But, there are no study on suited porousness for these applications. As a consequence, such samples were non applicable for a scaffold in bone Restoration due to the deficiency of strength. However, these stuffs could be applied for fiction of an oculus ball in orbital implant being porous bioceramics.
Furthermore, the low mechanical strengths of porous hydroxyapatite prepared by this combination technique utilizing PMMA and H2O2 could be developed by adding some additives ; such as binder, deflocculant or wetting agent. Additionally, utilizing H peroxide less than 5wt % concentration might be another manner to better the strength of the porous samples.
This survey investigated the features of the porous HA prepared by three different methods and the consequence of pore former on their mechanical belongingss, summarized as follows:
An increasing porousness of the hydroxyapatite samples resulted in a lessening of both compressive strength and flexural strength.
For sacrificial templet, the more PMMA content had a important consequence on the higher porousness of the HA samples whereas the lower compressive strength and flexural strength. These samples had the pore size ranged between 100 and 300 millimeter.
For direct foaming, the higher concentration of H2O2 resulted in an addition in porousness and pore size, while a lessening in the both mechanical strength. The pore size of these samples ranged from 100 to 1000 millimeter.
There were difficult to fix the porous HA utilizing PMMA content over 40wt % for sacrificial templet and H2O2 concentration over 20wt % for direct foaming when L/P ratio was controlled at 1.3 ml/g.
For combination between sacrificial templet and direct foaming, an increasing usage of PMMA and H2O2 caused a rise in porousness and a bead in both compressive and flexural strengths.
The HA samples prepared through sacrificial templet provided the porousness ranged between 52 % and 75 % , while the samples produced by direct foaming and combination technique provided the porousnesss ranged from 82 % to 85 % and from 84 % to 90 % , severally.