1. Microstructure of Hydroxyapatite from Waste Eggshell Synthesized under Different Temperature
Aekgaran Sangmala
Thank you for introduce myself. Good morning all participant. my name is Aekgaran Sangmala. Today I am going to present on the topic that is Microstructure of Hydroxyapatite from Waste Eggshell Synthesized under Different Temperature.
Department of Physics
King Mongkut’s University of Technology Thonburi

2. Introduction Objectives References
Experimental details
Results & Discussion
Conclusions
In my presentation, there are consist of six main part.
The first part is introduction.
Next part are objective, experimental details, result and discussions, conclusions and references
References

3. BONE Introduction Protection Support Movement Blood Cell Production
The first, the normal human body has about 200 bones which are needed for a variety of functions within the body. There are many functions which bones are in the body.
The first is support. The bone is providing support and structure. Bones are the hardest and most rigid structure in our body. Bones are strong but light, which gives the body support and shape without weighing is down
Protection, the nervous system is the most important in our body. Because the nervous system controls the rest of our body and is very fragile. The bones can protect this system such as skull and backbone. Moreover, the rib bones also protect the vital organs in the chest, such as the lungs and heart.
Movement, the skeletal system also closely interacts with the muscle system in our body. Muscles are attached to bones by bands of tissue called tendons. Bone are necessary to attach to in order to contract and cause motion.
The last one, Blood cell production. The bones also are important as a center for the productions of blood cells. The inside of bones is filled with a jelly-like material called the bone marrow, which is where red blood cells are made.
Blood Cell Production

4. approximately 60-70% is bone mineral
Introduction
Bone structure
approximately % is bone mineral
Calcium phosphate
The bone structure. The bone structure consists of compact bone and spongy bone where inside contain bone marrow. The compact bone is very dense and in the outside. Inside the compact bone is spongy bone which is high porosity. The compositions of bone are inorganic and collagen fiber. The most of inorganic is calcium phosphate that contains approximately 60 to 70 percent as bone mineral. The bone mineral is hydroxyapatites which is one of calcium phosphate phase.
Hydroxyapatite

5. Introduction Broken bone loss bone Accident Porous/Brittle bone
Although human bones are the strongest in the body but the bone can be destructed or damage by some cause
For example, when we have an accident. The bones may be damage, sometimes just incomplete fracture, sometimes the bones are cracked.
And in the older persons, their body system lack of the calcium concentration. So the density of the bones will be decreased called porous or brittle bone, because their body dose not have enough some mineral for make the new bone.
Porous/Brittle bone

6. Bone replacement material
Introduction
Bioactive
Bioinert
Bone replacement material
Bioresorbable
The bioceramic for bone replacement has the main three types.
The first is the bioinert, it is the material that use in the human body, the bioinert most use for dental and load bearing applications.
The second is the bioactive, the bioactive is the biomaterial similarity to the human bone, so it most use for the joint replacement and some application that want to use special properties of its.
So in this work have been use the bioinert and bioactive material to study.
The last one is bioresorbable material, this is material that has special properties for some application. Their properties is the restorable in the body quickly, so this effect can improve the function of using the bone grafting.

7. Human bone contains 60%-70%
Introduction
Bioactive
Alveolar ridge augmentation
Maxillofacial reconstruction
Orbital implants
Spine fusion
Repair of bones defects
Applications
Inorganic Mineral
Ca10(PO4)6(OH)2
Bone and Teeth
Biocompatibility
Bioactive ceramic
Hydroxyapatite
For the bioactive material, Hydroxyapatite is the one of this types. Hydroxyapatite is the inorganic mineral most found in bones and teeth. So it has high biocompatibility and bioactive. It can use several application such as Alveolar ridge augmentation, maxillofacial reconstruction, Orbital implants, spine fusion and repair of bones defects. The most of applications of HA relate the bioactive properties of it.
Human bone contains 60%-70%

8. Hydroxyapatite synthesizing
Introduction
Hydroxyapatite synthesizing
Hydroxyapatite
Calcium phosphate
Sea shell
Calcium ions
Coral reef
How to synthesis hydroxyapatite for bone replacement.
The hydroxyapatite is the one of calcium phosphate phase, calcium phosphate consists of phosphate ions and calcium ions.
In the practical, the researcher try to reduced the cost of hydroxyapatite synthesizing. The reducing of cost is using the calcium source from waste. The calcium source waste have many type such as sea shell, coral reef, bovine bone and eggshell. Thereby there are many waste that can be use for the hydroxyapatite synthesizing. However, some waste should not use. Because they are in the ecology cycle. For example, sea shell may be habitat of small sea animal. And the coral reef are necessary for fish and some animal in the deep sea. If this waste are used, the environmental or ecology may be damage.
Although, sea shell and coral reef should not use for hydroxyapatite synthesizing. But bovine one and eggshell also can be used. They are the waste from household. So they are not profit for another. They are very cheap and not destruct when using in hydroxyapatite synthesizing
So, for this work choose the chicken eggshell to synthesis hydroxyapatite. Because they are abundant in Thailand. And they are full of calcium.
Phosphate ions
Eggshell
Bovine bone

9. Why the eggshell can be used for synthesizing hydroxyapatite?
Introduction
Why the eggshell can be used for synthesizing hydroxyapatite?
Calcium 95%
Membrane
CaCO3 94%
Collagen 35%
Glucosamine 10%
Chondroitin 9%
Hyaluronic acid 5-10%
Ca(PO4)2 1%
Magnesium 1%
Shell
From my work. Chicken eggshell had been used in the hydroxyapatite synthesizing as the calcium source.
The chicken consist of two main part
The inside of shell is shell membrane which contain collagen, and other. So, this part of shell will be removed before use.
The outside part of shell is the shell which are the most of calcium compound about 95%. The calcium compound is the calcium carbonate phase about 94%. So the chicken eggshell is full of calcium and can be used for hydroxyapatite synthesizing
Organic 4%

10. Introduction Some method to synthesis the hydroxyapatite Sol gel
Solid state
Lack of purity
Very low cost
large particle
Sol gel
Small particle
20 – 50 nm
High cost of precursors
Wet chemical
Small particle
low cost
Easy to control
According to previous research, hydroxyapatite can be synthesized by several method
For example, solid state method is the very simple method that is the reaction of calcium group powder with phosphate group powder in the furnace or ball milling machine. This method can produce the hydroxyapatite powder easily but the particle will be large and lack of purity. The purity come from by product and some surface of ball from ball milling method.
The sol-gel is the most method to synthesis hydroxyapatite. This method produce dense hydroxyapatites in small particle. But disadvantages of this method are the high cost of precursors and low throughput.
And the last interesting method is the precipitation. This method is similar to the sol-gel method. But this method is unnecessary the formation of gel so the time of synthesis may be shorter. The hydroxyapatite powder are form into solid and precipitate in the solution. In addition, the condition of synthesizing can be easy to control and by product are easy to remove.
Thereby, this work had been use the precipitation method to synthesis the hydroxyapatite.

11. Objectives To synthesis hydroxyapatite from waste eggshells.
To study the effect of hydroxyapatite synthesizing under different temperature.
To study the temperature effect on hydroxyapatite from waste eggshell.
So, the aim of this work are synthesis hydroxyapatite added zirconia from waste eggshells.
study the effect of adding the zirconia in hydroxyapatite from waste eggshells.
improvement the mechanical properties of hydroxyapatite via zirconia adding

12. Experimental details Chicken eggshell Chemical precursors
In my research, the chicken eggshell has been used as the calcium source. Inner protein membranes of chicken eggshell have been removed before dying process because some protein is in the shell membrane may be a contaminations in the sample. After remove the shell membrane, the chicken eggshell were dried and grounded to the powder.
Next chemical precursor, di ammonium hydrogen orthophosphate was used as a phosphate source. In the other research, by products after hydroxyapatite synthesizing when use di ammonium hydrogen orthophosphate as a phosphate source are ammonium solution and water. So this by products are easy to remove from the sample.
In my experiment, hydroxyapatites has been doped by Zirconium Oxide or Zirconia, this Zirconia were stabilized by amount 3%mol of yttrium adding. Because in the room temperature, un-stabilized Zirconia is the monoclinic phase that is the low mechanical properties phase. Monoclinic Zirconia can be transformed to tetragonal phase that is high strange phase by sintered at very high temperature. So this method is low effective and high cost. However, tetragonal phase can be produced by doped some material such as yttrium, after adding the yttrium in low content, Zirconia were transformed to tetragonal phase suddenly.
Chicken eggshell

13. Hydroxyapatite synthesizing Di-ammonium hydrogen orthophosphate
Experimental details
Hydroxyapatite synthesizing
Di-ammonium hydrogen orthophosphate
Heat at
1300˚c
Di water
PO43- Solution
The first step is about hydroxyapatite synthesizing by precipitation method, chicken eggshells were grounded into the powder. The chicken eggshell powder has been heated at 1300 degree Celsius. In addition, the other research showed that the phase of chicken eggshell is calcite or calcium carbonate. After heat treatment around at 1000 degree Celsius, the phase of chicken eggshell is transform to the calcium oxide. Moreover, calcium oxide phase from heated chicken eggshell is the high crystalinity after 1300 degree Celsius of heat treatment.
The next step, calcium oxide was mixt in the nitric acid. The calcium oxide powder was dissolve and transform to the calcium nitrate solution. So, calcium nitrate solution was used as a calcium solution for hydroxyapatite synthesizing.
Next step, a diammonium orthophosphate was dissolved into the DI water. The diammonium orthophosphate solution was used as a phosphate solution for hydroxyapatite synthesizing.
The phosphate solution was dropped into the calcium solution and mixt together by magnetic stirrer. And the time of stirring is about 24 hours.
Furthermore, the both of calcium and phosphate solution were must adjusted pH value. From the other research, hydroxyapatite is a one of calcium phosphate. But the hydroxyapatite can be formed completely under high pH value. And the suitable pH value is around 10 to 11
The last step, the precipitate of hydroxyapatite were filtered by vacuum filtration. However, before the filtration, the products of synthesizing have been washed by DI water until by products were removed clearly. And the hydroxyapatite powder was dried at 80 degree Celsius for 48 hours.
Keep at oc
4 H
Dried at 80 oc
48 H
CaO
Hydroxyapatite
DI water
Ca(OH)2 Solution
Stir 30 min

14. Sample characterization
Experimental details
Sample characterization
XRD
FTIR
All of sample
The sample were heated in the furnace for study the temperature effect on the microstructure.
And then, all of sample were investigated by XRD for the phase and structure , SEM for the morphology and particle dispersion, and FTIR for the chemical functional groups.
In addition, for the future, Some sample will be study the mechanical properties by UTM tester. The UTM tester will be confirmed the changing of mechanical properties of Zirconia adding.
Heat at ˚c
FESEM
Furnace

15. Hydroxyapatite synthesizing Hydroxyapatite decomposition
Results & Discussions -
2theta (degree)
Intensity (a.u.)
Hydroxyapatite
Ca3(PO4)2 ;TCP
Ca2P2O7
Hydroxyapatite synthesizing
10Ca(OH)2+6(NH4)2HPO4
Ca10(PO4)6(OH)2+12NH3+18H2O
Hydroxyapatite decomposition
Ca10(PO4)6(OH)2
CaO+3Ca3(PO4)2+H2O
700 oC
600 oC
700 oC
500 oC
600 oC
Transmittance (%)
500 oC
400 oC
400 oC
300 oC
300 oC
200 oC
The first result is the structure characterization as shown in the figure 1, in the picture a) show that the XRD pattern of hydroxyapatite synthesized from chicken eggshell by precipitation method. From the result indicated that the sample is only a hydroxyapatite phase. In addition, this method can be explained by this chemical reaction as show in this slide.
In the picture b) show that the XRD pattern of Zirconia that is used in this experiment. The XRD pattern show that Zirconia consist of 2 phase that are major and minor phase. The major phase is a tetragonal phase which is shown in the blue point. And the minor phase is a monoclinic phase which is shown in the orange point. From this result show that Zirconia added in the hydroxyapatite is stabilized to the tetragonal by add content of the yttrium.
In the picture c) show that the XRD pattern of hydroxyapatite various content of Zirconia. The results show that all of the sample have 2 phase consist of hydroxyapatite and Zirconia phase. This result indicate that hydroxyapatite dose not react with Zirconia which confirm that all of the sample can be applied to bone replacement material. Because other phase dose not appear in the XRD pattern. Furthermore, intensity of sample on 30 degree which is Zirconia peak increase with increasing the content of Zirconia. So, it is indicate that Zirconia dose not react with another and were remove in the washing process.
200 oC
Wave number (cm-1)
Function Group
(PO43-)
631
(OH-)
Wavenumber (cm-1)
Figure 2. FTIR spectra of hydroxyapatite were synthesized at various temperature.
Figure 1. XRD patterns of hydroxyapatite were synthesized at various temperature.

16. FESEM images of hydroxyapatite various synthesizing temperature.
Results & Discussions
FESEM images of hydroxyapatite various synthesizing temperature.
a) 200 oC
b) 600 oC
c) 700 oC
50 nm
100 nm
The first result is the structure characterization as shown in the figure 1, in the picture a) show that the XRD pattern of hydroxyapatite synthesized from chicken eggshell by precipitation method. From the result indicated that the sample is only a hydroxyapatite phase. In addition, this method can be explained by this chemical reaction as show in this slide.
In the picture b) show that the XRD pattern of Zirconia that is used in this experiment. The XRD pattern show that Zirconia consist of 2 phase that are major and minor phase. The major phase is a tetragonal phase which is shown in the blue point. And the minor phase is a monoclinic phase which is shown in the orange point. From this result show that Zirconia added in the hydroxyapatite is stabilized to the tetragonal by add content of the yttrium.
In the picture c) show that the XRD pattern of hydroxyapatite various content of Zirconia. The results show that all of the sample have 2 phase consist of hydroxyapatite and Zirconia phase. This result indicate that hydroxyapatite dose not react with Zirconia which confirm that all of the sample can be applied to bone replacement material. Because other phase dose not appear in the XRD pattern. Furthermore, intensity of sample on 30 degree which is Zirconia peak increase with increasing the content of Zirconia. So, it is indicate that Zirconia dose not react with another and were remove in the washing process.
Figure 3. The FESEM images of sample powder were synthesized at various temperature. .

17. Temperature effect of Hydroxyapatite from chicken egg shell
Results & Discussions
Temperature effect of Hydroxyapatite from chicken egg shell
𝐷= 0.9𝜆 𝛽 cos 𝜃
Hydroxyapatite
Ca3(PO4)2 ;TCP
Ca2P2O7
800 oC
700 oC
600 oC
Intensity (a.u.)
Crystalline size (Å)
500 oC
400 oC
The first result is the structure characterization as shown in the figure 1, in the picture a) show that the XRD pattern of hydroxyapatite synthesized from chicken eggshell by precipitation method. From the result indicated that the sample is only a hydroxyapatite phase. In addition, this method can be explained by this chemical reaction as show in this slide.
In the picture b) show that the XRD pattern of Zirconia that is used in this experiment. The XRD pattern show that Zirconia consist of 2 phase that are major and minor phase. The major phase is a tetragonal phase which is shown in the blue point. And the minor phase is a monoclinic phase which is shown in the orange point. From this result show that Zirconia added in the hydroxyapatite is stabilized to the tetragonal by add content of the yttrium.
In the picture c) show that the XRD pattern of hydroxyapatite various content of Zirconia. The results show that all of the sample have 2 phase consist of hydroxyapatite and Zirconia phase. This result indicate that hydroxyapatite dose not react with Zirconia which confirm that all of the sample can be applied to bone replacement material. Because other phase dose not appear in the XRD pattern. Furthermore, intensity of sample on 30 degree which is Zirconia peak increase with increasing the content of Zirconia. So, it is indicate that Zirconia dose not react with another and were remove in the washing process.
300 oC
Heating temperature (oC)
200 oC
Figure 5. The crystal size of hydroxyapatite were heated at various temperature.
Figure 4. XRD patterns of hydroxyapatite were heated at various temperature.
2theta (degree)

18. Conclusions
Hydroxyapatite can be synthesized from waste eggshells by reaction of Ca(OH)2 and (NH4)2(HPO)4
Hydroxyapatite transform to tri-calcium phosphate (Ca3(PO4)2) at 700oc of synthesizing temperature.
The particle size of hydroxyapatite synthesized from waste eggshells is around 50 nm.
The crystalline size of hydroxyapatite increase with increasing the heating temperature and hydroxyapatite transform to Ca3(PO4)2 after heat over 700oc.

19. References
Ghazanfari, S.M.H. and Zamanian, A., 2013, “Phase Transformation Microstructural and Mechanical Properties of Hydroxyapatite/Alumina Nanocomposite Scaffolds Produced by Freeze Casting”, Ceramics International, Vol.39, pp
Bigi, A., Foresti, E., Gandolfi, M., Gazzano, M. and Roveri, N., 1995, “Inhibiting Effect of Zinc on Hydroxyapatite Crystallization”, Journal of Inorganic Biochemistry, Vol.58, pp
Elliott, J.C., Wilson, R.M. and Dowker, S.E.P., 2002, “Apatite Structures”, Advances in X-ray Analysis, Vol. 1, pp
LeGeros, R.Z. and LeGeros, J.P., 1993, “Dense Hydroxyapatite”, in An Introduction to Bioceramics, Hench, L.L. and Wilson, J., 2nd ed., World Scientific, Singapore, pp
Seo, H. J., Cho, Y.E., Kim, T., Shin, H.I. and Kwun, I.S., 2010, “Zinc May Increase Bone Formation Through Stimulating Cell Proliferation, Alkaline Phosphatase Activity and Collagen Synthesis in Osteoblastic MC3T3-E1 Cells”, Nutrition Research and Practice, Vol.4, No.5, pp