*Sagitha Indrayana **I G N Wien Aryana

1. Comparation of IGF-1 LEVELBetween Femoral Fracture in White Rat Treated with and without Intrahematoma Rat Recombinant Leptin Injection *Sagitha Indrayana **I G N Wien Aryana Department of Orthopaedic and Traumatology Faculty of Medicine Udayana University Sanglah General Hospital, Bali-Indonesia

2. INTRODUCTION • Many factors can inhibit the healing of bone fracture. • Systemic problem  ↗ time for bone healing DM. • Insulin stimulates synthesis osteoblasts, osteoblast proliferation, and insulin-like growth factor-I ( IGF-1 ) • DM  ↘levels of insulin and IGF-1. • IGF I stimulate collagen synthesis and proliferation sel .

3. Leptin: a small protein, secreted by adipocytes • Affects regulation of bone mass. • ↘ leptin level ↘bone mass &adipocytes accumulation in the bone marrow (Mice) • Diabetic mice low levels of serum leptin • Complications of diabetes : • ↘bone mass • ↗risk of fractures • impaired fracture healing

4. Bone remodeling controlled by hormones and local factors • IGF-1anabolic regulator of bone cells • lower collagen degradation • ↗bone matrix • ↗proliferation of osteoblasts • The understanding of the leptin role in fracture healing used as one of therapy for healing of bone fractures

5. Problem Base Is introduce leptin peripherally can lead to better fracture healing characterise by the increase of IGF-1 level?

6. Study Objective • General Objective Understanding of the effect of introducing leptin peripherally for the fracture healing. • Specific Objective Introducing leptin peripherally can lead to better fracture healing in the rat with femoral fracture characterise by the increase of IGF-1

7. Experimental Benefit • Scientific Benefit: Understanding about the role of introduce leptin peripherally in the better fracture healing in white rat femoral fracture characterise by the increase of IGF 1 level and as the scientific base for larger study in human. • Clinical Benefit: As one of the promising alternative for better fracture healing in orthopaedic practice • Sociobenefit: as a contribution for the society to have an alternative for better fracture healing, decrease of the delay union, nonunion and general expend

8. The Study Concept White Rat with Femoral fracture (IM. Wire) Leptin • INTERNAL FACTORS • Age • Imunity • Sex • genetic • EXTERNAL FACTORS • Environment • Nutrition • Life style • Fracture type • Immobilisation IGF -1 level

9. Study Hypothesis Introduce leptin peripherally can lead to better fracture healing characteristic by the increase of IGF-1 level.

10. Study Design • Experimental study group 1 (Control)IGF-1 level PS R group 2 (treatment)IGF-1 level

11. Place and Time The study take place in the Farmacology laboratory department, medical faculty Udayana University, Denpasa Bali. In March 2013

12. Population and Sample • Variabilitas Population Study populationare white rat eligible with sampe criteria • Sample Criteria Sampel : White rat (Sprague Dawley Rats) with inclusion and exclusion criteria Inclusion Criteria : White rat (Sprague Dawley Rats) 6 months old, 22-25 grams (Smith, 1988). Exclusion Criteria : Sick white rat ( don’t want to eat) Drop out Criteria : Die rats. • Sample Size : is 19,8 ~ 20 sample each group (Pocock, 1983)

13. Study Variable Variable Classification a) Independent variable : Leptin b) Dependent variable : IGF-1 level. c) Control variable : rat species, sex, age, body weight, nutrition, fracture type, immobilisation procedure.

14. Study Variable Definition • Leptin : intrahematoma recombinant rat leptin injection as much as 100 ng • IGF-1 level: blood test to evaluate the level of IGF-1 with Elisa technique • Rat species: Sprague Dawley Rats, Male, 6months old, 22-25 grams • Fracture and immobilisation procedure: iatrogenic fracture at the diafisis femur shaft and treat use an intramedullary wire

15. Materials and Instruments • Materials: -White rat (Sprague Dawley Rats), -6 months old, weight 22-25grams -Rat nutrition -wire 2.0 -sterile bandage -Recombinant rat leptin injection -Elisa reagent • Instrumen : -rat cages -basic set orthopaedic -weight scales -SPSS software

16. RESULTS Table Deskriptif of the IGF-1 level in The Treatment and Control Group

17. Control group: • the average of IGF-1 level :695.7596 • Range from 327.79 –1163.72. • Treatment group: • the average of IGF-1 level 2211.9347 • Range from 290.08 – 4813.07. • Normality test (Shapiro-Wilk)normal distribution. • Independent t-test the mean of IGF-1 level in the treatment group compared with control group is Higher significantly.

18. DISCUSSION • Bone healing after break in continuity Complex mechanism • Involves local and systemic factors. • Systemic problem DM ( well research). • Diabetes mellitus : • ↗time of fracture healing ( 2-3 longer) • ↗risk of delay union and non- union. • Insulin : • Stimulates synthesis osteoblasts • Osteoblast proliferation • ↗insulin-like growth factor-I ( IGF-1 )  stimulate collagen synthesis and proliferation sel

19. In this study : • Calculate IGF-1 levels (ELISA method) • Compared IGF-1 levels  group of rats receiving leptin peripherally Vs did not receive leptin. • Data analysis Independent t test  increased levels of IGF-1 significantly in mice given leptin peripheral compared with mice that did not receive leptin. (p <0:05).

20. IGF-1 potent Bone morphogenic proteins and increase Osteoprogenitor cell differentiation into osteoblast cells osteocytes. • ↗ cell osteocytes ↗bone density matrix &strength. • Osteoblast cell activity can be seen from the activity of IGF-1. • this study found  Administration of leptin peripherally increase IGF-1 levels significantly.  This study as one of refferal source for greater numbers of future experimental in human, for administration of leptin peripheral to increase IGF-1 level and decrease complication of delay union and malunion, especially in diabetic patients.

21. THANK YOU