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EFFECT OF GREEN TEA EXTRACT TO ANGIOGENESIS IN BONE HEALING EXPERIMENTAL STUDY ERI KRISWANTO.

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Presentation on theme: "EFFECT OF GREEN TEA EXTRACT TO ANGIOGENESIS IN BONE HEALING EXPERIMENTAL STUDY ERI KRISWANTO."— Presentation transcript:

1 EFFECT OF GREEN TEA EXTRACT TO ANGIOGENESIS IN BONE HEALING EXPERIMENTAL STUDY ERI KRISWANTO

2 Background: Non-union is a big problem in bone healing Angiogenesis plays important role in bone healing Green tea is one of the strong anti- angiogenesis agent There are no any research that study the effect of green tea to angiogenesis in bone healing

3 Green Tea (Camelia sinensis) poliphenols in green tea : • flavanol • flavandiol • flavonol • phenolic acid, • catechin ( 70%) :Epigallocatehchin-3-gallate (EGCG) about 67%, epigallocatehchin (EGC), epicatechin gallate (ECG), epicatechin (EC), dan gallocatechin ( Bokuchava,..et al. 1980), (Kim, S,et,al.2000), (Fasina, G, et,al. 2004).

4 Bone Healing

5 Bone Healing in Mice Inflamation Soft callus (cartilage)  day 7 th Chondrocytes presents VEGF  Day 10 th Hard callus (bone)  day 14 th Remodelling ( Street, J,. et. al, 2002)

6 Effect of EGCG to Angiogenesis

7 fracture Inflamation Soft callus Chondrocyte apoptosis Cartilaginous matrix degradation and removal Angiogenesis Rekrutmen osteogenic cell Matrix production Differentiation Hard callus Remodeling Chondrocyte & osteoblast (Hypoxia) HIF VEGF, FGF, dan TGF Endotel Pericyte Survival, vascular permeability, migration, prolifera-tion, vascular guidance PDGF VEGF PDGFR, ERK 1/2 FGFR, PI3K, ERK 1/2 Green tea ( EGCG) Neo-vessels ERK ½, PI3 K

8 Objective 1.To know the effect of green tea extract to amount of VEGF presenting cells in the callus 2.To know the effect of green tea extract to amount of vessels in the callus 3.To know the effect of green tea extract to cross- sectional-area of vessels in the callus

9 Materials and Method

10 DESIGN Post Test Control Group Design. S1 : sample R : randomization K1 : pure water P1 : 15 mg green tea extract P2 : 20 mg green tea extract P3 : 25 mg green tea extract P4 : 30 mg green tea extract

11 Aclitimation ( 1 week) P3 35 mices with rigth femur fracture P2 P1 20 mg 15 mg 25 mg Day 10 th evaluation Callus harversting IHC staining Analyzation K1 P4 30 mg Pure water

12 Aclitimation ( 1 week) P7 35 mices with rigth femur fracture P6 P5 20 mg 15 mg 25 mg Day 14 th evaluation Callus harversting HE staining Analyzation K2 P8 30 mg Pure water

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15 RESULTS

16 IHC staining VEGF presenting cells 0 mg25 mg30 mg15 mg20 mg

17 DoseNAmount of VEGF presenting cells/15625 µ 2 MeanSDMinMax Pure water54.8260 b,c 1.294753.306.13 15 mg54.7000 b,c.902773.505.80 20 mg53.6600 b.630873.004.40 25 mg52.0200 a.311451.502.30 30 mg42.3250 a.427201.802.80 a,b,c the different superscript shows there is significant difference ( p < 0.05 ) between groups ( Games – Howell)

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19 HE staining - amount of vessels - cross-sectional area of vessels 0 mg25 mg30 mg15 mg20 mg

20 DoseNAmount of vessels / field MeanSDMinMax Pure water548.8000 c,d 32.2133514.0098.00 15 mg537.0000 a,b,c,d 23.1840517.0074.00 20 mg526.4000 a,b,c 18.0776115.0058.00 25 mg512.4000 a 7.092256.0023.00 30 mg516.6000 a,b 6.426516.0023.00 a,b,c the different superscript shows there is significant difference ( p<0.05 ) between groups ( List Significant Data)

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22 DoseNCross-sectional area of vessels (µ 2 ) MeanSDMinMax Pure water5139.8000 d 22.06978115.00167.50 15 mg5101.1000 b,c,d 37.5306575.00165.50 20 mg558.5000 b,c 7.2024350.0067.50 25 mg548.5000 a,b 8.0234037.5060.00 30 mg538.0000 a 7.5828825.0045.00 a,b,c the different superscript shows there is significant difference ( p<0.05 ) between groups ( Games – Howell)

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24 DISCUSSION • Green tea or EGCG per orally shows significant inhibition to angiogenesis (Fassina, G, et, al. 2004). • 1.25 % (4,69 mg/ml) of green tea contains 708 µg/ml EGCG per orally will produce 0,1-0,3 µM EGCG in the blood, this dose is equal with concentration of EGCG in human after drinking 2 or 3 cups of green tea (Wang, Z.Y,et,al. 1992). • Drinking 1.25% of green tea significantly inhibit neo- vascularization in mice’s cornea ( Cao Y, Cao R, 1999 ).

25 Discussion Drinking 1.25% (4.69 mg/ml) of green tea ad libitum in mice is equal with 15 mg of green tea extract single dose. Results : • Significant decrease of cross-sectional area of vessels in callus was found at minimal dose of 20 mg green tea extract. • Significant decrease amount of VEGF presenting cells in callus was found at minimal dose of 25 mg green tea extract • Significant decreases amount of vessels in callus was found at minimal dose of 25 mg green tea extract

26 Conclusion • Drinking of green tea > 15 mg in mice equal with drinking > 2 or 3 cups of green tea in human will decrease VEGF presenting cells, amount of vessels, and amount of cross-sectional area of vessels in bone healing. • Green tea significantly inhibit angiogenesis in bone healing.

27 THANK YOU


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