Development of Porous AluminaAlumina-Hydroxyapatite Composites using Protein FoamingFoamingConsolidation Method

Prepared by:

Dr. Ahmad Fadli, MT

Jurusan Teknik Kimia Universitas Riau

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Problem Statement

Porous alumina
- Strong

Porous hydroxyapatite
-Weak mechanically -Bone bonding ability -Good bone ingrowth

- Stable & Compatible - Biological inertness

Porous alumina-HA composites

- Strong

- Biocompatible - Bioactive

Problem Statement
The usage of protein such as ovalbumine, Bovine Serum Albumin as foaming agents has been tried by some researcher groups [Eckert, et.al (2000), Garnn, et.al (2004), Bhattacharjee, et.al. (2007)]. However, due to its high foaming capacity, it was hardly to be suitable to make porous ceramics with controlled porosity. In this research, egg yolk is used as a pore creating agent. The main components of egg yolk are - 50% water - 33% lipids - 17% proteins. The lipids phase in yolk would reduce the foaming capacity of protein in making pores. Set up of proper composition between yolk and alumina powder will be able to make porous ceramics with controlled pore size distribution accordingly.
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Novelties and Its significances

Development of a new method called protein foamingconsolidation method The use of yolk as the pore creating agent Yolk functions as foaming and consolidating agent Pore creation can be done at very low temperature (110180C) compared conventional methods (500-600C) Control of properties of porous ceramics by regulating the slurry composition and the foaming condition. The significances are simplicity, quick process and the lowcost processing/materials needed to producing porous ceramics.
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Research philosophy

The foaming ability of yolk on heating of slurry was used for creating pores in ceramic bodies. The properties of ceramics can be determined by manage slurry composition, drying and sintering processes .
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The main objectives

(1) To fabricate porous alumina and porous aluminahydroxyapatite for biomedical applications through protein foaming consolidation method (2) To evaluate the effect of slurry compositions and operating conditions on the rheological behavior and foaming capacity as well as the physical properties of porous bodies such as shrinkage, density, porosity, pore size distribution, interconnectivity and compressive strength (3) To investigate the biocompatibility of the porous aluminahydroxyapatite composites obtained as microcarriers in a spinner vessel bioreactor for Vero cell culture.
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Materials
Commercial alumina Commercial HA Commercial TCP Egg yolk

Sol-gel HA powder

Starch powder

Darvan 821 A 7 as dispersant

Preparation of porous alumina & alumina-HA

Biocompatibility test

TGA analysis of yolk

~100 C a weight loss of 55 % water evaporation. 100 375 C - 22.5 % weight loss removal of lipids. 375 525C, a weight loss of 22.5%. decomposition of proteins
Burned out : 600 C
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The properties of porous alumina

Mass ratio of alumina to yolk (w/w) Dispersant (% w) Drying Temp. (C) Shrinkage (Vol.%) Porosity (%) Density (g/cm3) Strength (MPa)

0.65 0.75 0.83 1.00 1.00 1.00 1.00 1.00 1.00

0 0 0 0 0 0 0.01 0.04 0.05

180 180 180 180 150 110 180 180 180

39.9 36.4 32.5 29.3 36.8 7.8 31.5

Un-measurable Un-measurable

70.6 52.8 50.5 50.4 53.0 43.6 78.4 79.6 82.4

0.9 1.9 2.0 2.0 1.9 2.2 0.85 0.81 0.70

1.1 1.7 3.1 4.6 4.7 5.7 1.43

Unmeasurable Unmeasurable 11

Sintering temperature = 1550C

Porous alumina bodies

CT-Scan

- Cylindrical shape - No cracks

Floating Density < 1 g/cm3

Non Floating 12

Effect of drying time on pore size

Al2O3-to-yol mass ratio = 1.0 min, drying temp. = 180C, Sintering temp. = 1550C

Drying time = 8 min

12 min

60 min - Bigger pore size - 100 800 m - Higher foaming capacity

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Effect of dispersant content on pore size

Dispersant conc. = 0.01% 0.02%

0.04%

0.05%

- Low viscosity of the slurry resulted in high foaming capacity, hence increased the pore generation.

- Bigger pore size (200 to 1000 m) - More interconnected - Less dense 14 - Thinner pore walls.

The effect of drying temperature on foaming capacity and pore size

110C 180C

Foaming in 3 stages; - Pre-heating - Foaming stage - Stabilizing stage.

- Bigger pore size - More interconnected

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Compressive strength models

Porosity have a significant impact on comp. strength. Comp. strength was increased 1.1 to 5.7 MPa by decreasing porosity 71 to 40 vol.%. A fine correlation was obtained with error = 18 25%

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Porous alumina-HA composites

The objective : to combine mechanical strength of alumina and bioactivity of HA.

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Effect of Yolk on rheology of slurries

Yolk addition changed rheological properties from pseudoplastic behavior to Newtonian fluid.
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Effect of HA loading on viscosity

9 8 7 Viscoaity (Pa.s) 6 5 4 3 2 1 0 0,3 0,5 0,7 HA-to-alumina mass ratio (w/w) 1

As HA loading increased, the water amount adsorbed by HA powder increases, thus leading to higher viscosity.
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Effect of sintering temperature on physical properties

Bioactive ceramic Sol-gel derived HA Sintering temp. (C) Shrinkage (Vol.%) Porosity (%) Comp. strength (MPa)

1200 1350 1400 1550

26.2 32.4 31.9 64.2 24.4 28.6 33.0 43.3 18.3 26.1 30.6 52.7

52.1 50.0 51.2 46.5 41.8 48.5 39.0 44.6 45.0 41.0 44.2 38.9

0.1 0.3 0.6 4.9 0.8 1.8 2.1 7.5 0.3 1.1 1.9 10.6
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High sintering temperature: - Melted HA particles - large grain sizes - Higher contact area - improvement of strength.
Commercial HA

1200 1350 1400 1550

Commercial TCP

1200 1350 1400 1550

Effect sintering temperature on pore size

1350C 1550C

- Bigger

- Smaller - In the range 100-700 m - Large grain sizes

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Effect of sintering temp. on grain size

1350C 1550C
XRD patterns

- Larger grain sizes - Improvement of the strength. - Phases in body did not alter during sintering process

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Effect of yolk amount on foaming capacity & properties

- viscosity decreased - increasing foaming capacity - bigger pore size

Yolk amount (g) 34 44 64

Shrinkage (Vol.%) 46.9 57.4 70.3

Porosity (%) 47.8 60.1 63.3

Comp. strength (MPa) 6.4 1.5 1.1

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Effect of HA & TCP loadings

As commercial HA and TCP content increased, strength increased. In contrast, the compressive strength decreased when higher amount of sol-gel HA was used.
Sol-gel HA loading (g) Shrinkage (Vol.%) Porosity (%) Comp. strength (MPa)

3 5 7 10
Comm. HA loading (g)

73.1 57.4 53.1 48.6

Shrinkage (Vol.%)

29.0 48.7 46.9 45.2

Porosity (%)

2.2 0.6 0.2 0.1

Comp. strength (MPa)

4 5
Commercial HA Sol-gel HA

76.7 76.5 83.5

Shrinkage (Vol.%)

45.8 39.7 36.8

Porosity (%)

2.9 4.2 20.4

Comp. strength (MPa)

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TCP loading (g)

5 10

80.0 62.7

56.1 61.6

3.1 4.2 24

Effect of HA content on microstructure

HA = 5 gram HA = 8 gram XRD patterns
HA = 8 gram

HA = 5 gram

- Thicker walls - Larger grain sizes. - Solid bonding - Higher contact area, - lead to the improvement of the strength.

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CT-scan images
3D Images of Thickness of HA with Color Coding HA-to-Al2O3 ratio = 0.3 HA HA ratio = 1.0 HA HA

- HA layer in middle part is thicker than the upper part

- HA particles tend to precipitate in surface part

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Pore size distribution

HA-to-Al2O3 ratio = 0.3

- Bigger pore size - 10 460 m,

ratio = 1.0

- Smaller pore size - 10 350 m,

Addition of HA loading would decrease pore size.

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Biocompatibility test
The objective is to evaluate biocompatibility of porous alumina and alumina-HA composites using culture of Vero cell in vessel bioreactor. Vero cell : Mammalian cells of the organ relevant to a specific application can be used to determine cell function on materials tested
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Effect of HA loading on cell number

Number of cells increases with HA content.

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Effect of HA loading on cell number

Porous alumina

Porous alumina

Porous alumina-HA

- Time = 12 hours - 40000 cells/mL

- Time = 120 hours

- 55000 cells/mL

- HA-to-Alumina ratio = 1.0 - 120 hours - 100000 cells/mL

Number of cells increases with HA content.

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Effect of HA loading on cell growth rate

The growth rate was influenced by: - Composition of microcarriers - Surface morphology.

Pore size of 0.3 w/w microcarrier was bigger than 1.0 w/w microcarrier. Cell metabolism is supported by: - bigger pore size, thus the attached cell on surface of microcarrier pores increased.
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EDX analysis
EDX analysis

Carbon content

Carbon indicated the presence of cells

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Conclusion
1. A novel method for preparation of porous alumina and alumina-hydroxyapatite composites using egg yolk both as the consolidating and foaming agent was successfully developed. The properties of porous ceramics can be controlled by varying ceramic slurry composition, drying and sintering processes. 2. The addition of yolk into the slurries shifted the rheological properties from shear thinning behavior to a Newtonian fluid. 3. The pore size of alumina bodies increased with drying time and found in the range of 100 800 m. 4. Depending on dispersant concentration (0.010.05 wt%) in the alumina slurry, the density ranged from 0.7 to 0.85 g/cm3 and the mean pore size varied from 200 to 1000 m.
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Conclusion
5. The foaming capacity of slurries increased with drying temperature. As the drying temperature increased, the pores became interconnected with less dense and thinner pore. The apparent density of sintered porous alumina decreased from 2.2 to 2.0 g cm-3 when the drying temperature increased from 110 to 180C. 6. The compressive strength of the porous alumina bodies increased from 1.1 MPa at 71.0% porosity to 5.7 MPa at 40.0% porosity, showing that the compressive strength is strongly dependent on porosity. 7. The viscosity of slurry increased with HA loading. The addition of commercial HA and TCP in the bodies was found to increase the compressive strength, whereas the case reverse for the addition of sol-gel derived HA.

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Conclusion
8. At 1550C sintering temperature, the porous alumina-HA bodies prepared using sol-gel derived HA has a compressive strength of 4.9 MPa at 46.5% porosity, whereas the porous alumina-HA bodies with 7.5 MPa compressive strength and 44.6% porosity were obtained using commercial HA. 9. When the sintering temperatures increased from 1200 to 1550C, the shrinkage of porous alumina bodies compositized with sol-gel derived HA, commercial HA and TCP increased from 26.2 to 64.2 vol.%, 24.4 to 43.3 vol.% and 18.3 to 52.7 vol.%, respectively. The grain size of samples enlarged and fused with the increasing temperature. Phases in porous body did not alter with increasing sintering temperatures.

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Conclusion
10. The HA addition in samples would increase cells number attached on surface samples. 11. The cell growth rates of microcarrier containing 0.0, 0.3 and 1.0 w/w were 0.015, 0.019 and 0.017 hour-1, respectively. 12. Carbon content on porous alumina bodies without HA addition was 36.03%, it significantly increases 46.14% when HA added 1.0 w/w.

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Thank You

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