Antiosteoporotic activity

1. Anti osteoporotic activity of Salvadora persica sticks
extract in an estrogen deficient model of osteoporosis
Presented By:- Shouvik Kumar Nandy (1991402006)
Priyanka Sagar Negi (1991402012)

2. Introduction:-
 Definition of Osteoporosis:- A medical condition in
which the bones brittle and fragile from loss of tissue,
typically as a results of hormonal changes or deficiency
of calcium or vitamin D.
 Low bone density.
 Increase the risk of fracture.
Bones in osteoporosis Bones in normally

3. Molecular signaling of osteoporosis:-

4. Risk factors:-

5. Objectives:-
 The effect of Salvadora persica sticks on prevention of tooth decay is well
established
 But the effect of S. persica stick extract (SPE) on the prevention/treatment of
osteoporosis has not been studied

6. Methods:-
 SPE was administered at 50, 150, and 300 mg/d orally to OVX rats for 16 weeks
 Serum osteocalcin, alkaline phosphatase, calcium, and phosphorus, and urinary
deoxypyridinoline, calcium, and phosphorus were measured
 Bone mineral density (BMD), three point bending test, and histomorphometric
characteristics of the femoral bone were also examined

7. Method:-
Preparation of the extract:-
200gm of ground stems was soaked in the 1000 mL of distilled water
At 4o C for 48 hours, pH will be 6.8
After filtration, the resultant extract was concentrated and lyophilized under reduced
pressure below 4o C to give a crude extract
Administered by morning gavage at doses of 50, 150, and 300 mg/kg dissolved in
0.5-mL distilled water/l00 g of body weight
The lyophilized S. persica extract (SPE) was kept frozen

8. Method:-
Animals and Treatments:-
 Forty-eight, 3-month-old female Sprague-Dawley rats (200-220gm) obtained from
the animal house in a naturally controlled lab upon arrival and acclimatized for 5
days.
 Surgical procedures were done under sodium pentobarbital anesthesia
 4 weeks after surgery, the OVX rats were randomly divided into 5 groups
 8 rats per group:-
• OVX vehicle control
• OVX þ 17bestradiol (E2; 25 mg/kg/d)
• OVX þ SPE (SPE50; 50 mg/kg/d)
• OVX þ SPE (SPE150; 150 mg/kg/d)
• OVX þ SPE (SPE300; 300 mg/kg/d)

9.  Biochemical markers of bone turnover:-
Assay for serum calcium, serum phosphorus and bone-specific alkaline
phosphatase activity was done using commercially available kits
 Assessment of bone density:-
To measure bone changes due to OVX and treatments
 Biomechanical testing:-
It was assessed in the left femurs by the three point bending test using a
materials testing machine
 Digital histomorphometric analysis:-
Microscopic examination of the stained femoral bone sections showed normal
bony architecture of the sham control rats
 Statistical analysis:-
Data were presented as mean ± standard error of the mean
Method:-

10. Method:-
Treatment procedure :-
Animals in the OVX and sham control groups were given the same volume of vehicle
Vehicle, E2, and SPE were given every morning by oral gavage starting from the 4th
week after surgery and lasted for 16 week
Diets were similar in macronutrients, calcium (0.9%), and phosphate (0.7%) contents
The body weights were measured every one week
Urine samples were collected and acidify with 6 mol/L HCL (0.03 vol/vol)
After anesthetized blood collected and centrifuge it
Stored urine sample at -20o C & Weighted the dissected off uterus of each rat
Femurs were dissected, wrapped, and stored at -20o C until biomechanical and
structural analysis

11. Results:-
 Body and uterus weights:-
In this study all rat groups had a similar mean basal body weight.
Fig:- The body weight of the animals was recorded weekly during the experimental
period

12.  Body and uterus weights:-
Fig:- The uterus index was represented as uterus weight divided by body weight before
sacrifice. Values are presented as mean ± standard error of the mean
Result:-

13. Results:-
 Biochemical markers of bone turnover:-
Measurement of drug effect

14. Results:-
 Bone density assessment:-
Effect of 16-weeks treatment with SPE on bone mineral density of OVX rats as
measured dual-energy X-ray absorptiometry(Fig- 2)
Fig:- 2

15. Results:-
 Biomechanical testing:-
Effect of 16- weeks treatments with E2 and SPE on biomechanical
parameter in the femoral diaphysis of OVX rats (table:- 2)

16. Results:-
 Digital histomorphometric analysis:-
Microscopic examination of the stained femoral bone sections showed normal
bony architecture of the sham control rats.

17. Discussion:-
 The dissolution of active ingredients present in S. persica sticks in saliva during
tooth brushing might have effect on skeletal bone turnover.
 It has same pathological characteristics of postmenopausal osteoporosis in human.
 We chose the oral route for SPE administration
 The dose-related effect of SPE was confirmed by measuring serum & urine bone
turnover markers & mechanical properties

18. Conclusion:-
 The results presented confirm at least, partial, dose-dependent protective action of
SPE on rat OVX model of osteoporosis.
 When more than ingredient is responsible for this effect, at this point we cannot
decide which molecule is responsible for the effect, but unlikely E2 have the bone
protective effect.
 We believe that SPE has ability for further advancement as a natural alternative for
the prevention of post- menopausal osteoporosis

19. Reference:-
1. A.-M. Fouda, A.R. Youssef / Osteoporosis and Sarcopenia xxx (2017) 1e6