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العنوان
The Effect of Vitamin D3 on the Osteogenic Potential of Adipose Derived Stem Cells in Treatment of Induced Osteoporosis in Albino Rats:
المؤلف
Ibrahim, Shaymaa Mamdouh Dessouky.
هيئة الاعداد
باحث / Shaymaa Mamdouh Dessouky Ibrahim
مشرف / Souzi Farid Shinaishin
مشرف / Dina Mohamed Abd elkhalik
مناقش / Dina Mohamed Abd elkhalik
تاريخ النشر
2018.
عدد الصفحات
236 P. :
اللغة
الإنجليزية
الدرجة
ماجستير
التخصص
Dentistry (miscellaneous)
تاريخ الإجازة
1/1/2018
مكان الإجازة
جامعة عين شمس - كلية طب الأسنان - قسم بيولوجيا الفم
الفهرس
Only 14 pages are availabe for public view

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from 236

Abstract

Osteoporosis is considered one of the most prevalent diseases. Prolonged use of GC has a detrimental effect on bones resulting in increased incidence of osteoporosis. Whereas, ADSCs are considered a recent treatment for male and female osteoporosis especially if its differentiation is guided toward the osteogenic lineage.
So, this study aimed to investigate the effect of Vit.D3 on the osteogenic differentiation potentiality of ADSCs in treatment of induced osteoporosis in albino rats in mandibular molar area. This will help to improve the osteogenic differentiation potentiality of ADSCs using a potent, low-cost and available agent such as Vit.D3.
Our results were evaluated histologically, immune-histochemically and histo-morphometrically.
Materials and Methods:
This study was carried out on 60 male albino rats with an average weight of 200-250 gm. The rats were divided into main four groups:
 group 1: (control groups):
This group consisted of 20 rats and was divided into:
Subgroup 1A:This subgroup consisted of (10 rats) which were injected subcutaneously with 0.5ml water mixed with benzyl alcohol (corresponding to the solvent of Mpsl) daily for one month then were killed corresponding to experimental groups (Osp. Subgroup 2A) to assess induction of osteoporosis.
Subgroup 1B: This subgroup consisted of (10 rats) which were given 0.5ml water daily (corresponding to the solvent of Vit.D3) orally for the following month, (After one month of benzyl alcohol injection), and then were killed corresponding to experimental groups (Osp. Subgroup 2B, Ex.G.3 & ExG.4).
group 2: (osp.gp):
This group consisted of 20 rats and was divided into:
Subgroup 2A: This subgroup consisted of (10 rats) which were given (Methyl prednisolone) for induction of osteoporosis by daily subcutaneous injection of a dose of 20mg/kg for one month according to (Yujiang et al., 2002), then, the rats were killed to confirm induction of osteoporosis.
Subgroup 2B: After one month, half the number of the Osp.gp (10 rats) was left for the following month to assess normal regeneration of bone after induction of osteoporosis corresponding to experimental groups (3& 4).
group 3: ((osp.gp) treated with ADSCs): This group consisted of 10 osteoporotic rats, ADSCs (2 × 106 cells/rat) were injected once into via tail vein after one month from the beginning of induction of osteoporosis, according to (Li et al., 2012).
group 4: ((osp.gp) treated with ADSCs & Vit.D3): This group consisted of 10 osteoporotic rats, Vit.D3 was administered orally through a gastric tube as a suspension in medium chain Tri-glyceride (MCT) according to (Norman A.W. et al., 1988) at a dose of 1µg/kg BW/day (40 IU/Kg BW) for 4 weeks starting with the injection of the ADSCs after induction of osteoporosis, according to (Massaki et al., 2006).
- Blood samples: Blood samples were collected from each group to ensure induction of osteoporosis- before administration of ADSCs or Vit.D3- by checking the alkaline phosphatase activity according to (Delmas et al., 2009; Jehoon et al., 2012).
-After the experimental period of each group, rats were killed by cervical dislocation. The mandibular molar area was dissected free, kept in formalin then decalcified and prepared for paraffin embedding.
Staining Techniques:
-Sections, 5µm thick each, were examined by L.M using:
1) H&E for routine histological study.
2) Masson Trichrome for histochemical study.
3) Osteonectin monoclonal antibodies for immune-histochemical study.
- The morphometric examination was done using an image analyser (Leica 500 Mc program) for morphometric study regarding:
1) The number of osteoblasts, osteocytes, osteoclasts per unit area with a high power x400.
2) The thickness of buccal and cortical plates at the molar area with a low power field x100.
3) The surface area percentage of spongy bone trabeculae per unit area with a low power field x100.
- All data was collected and tabulated.
Results:
- The results of this study revealed evident histological alternations in the alveolar bone of Osp. Subgroups (2A&2B). This was confirmed first of all by the blood sample which revealed higher ALP in osteoporotic groups compared to the control group indicating higher activity of bone formation to compensate for bone loss resulted from the induced osteoporosis. The histological alternations included alveolar bone resorption associated with increased osteoclastic activity, decreased osteoblastic activity, hardly detectable osteoid and appearance of many osteocytes with shrunken nucleus and empty lacunae. In addition, the resorptive changes were further confirmed by a significant decrease in the number of osteoblasts, osteocytes, thickness of the cortical plates and surface area percentage of spongy bone trabeculae accompanied administration of GC. Also, a significant increase in the number of osteoclasts was observed compared to all groups except Exp.G.3.
- Although rats in Exp.G.3, showed improvement of the osteoporotic features to some extent, rats in Exp.G.4 showed superior results with much more obvious osteoid layer indicating higher bone formation activity. This was confirmed by a significant increase in number of osteoblasts, osteocytes, thickness of cortical plates and surface area percentage of spongy bone trabeculae of Exp.G.4 over Exp.G.3. Also, a significant decrease in number of osteoclasts in Exp.G.4 was clearly observed. These results were also supported by Masson trichrome which revealed predominance of blue color in Exp.G.4 over Exp.G.3, indicating higher collagen synthesis activity. Higher osteoblastic activity also was confirmed in Exp.G.4 immuno-histologically.
Regarding comparison between the control group and Exp.G.4, there were no significant difference in the number of osteoblasts, osteocytes, osteoclasts, thickness of the cortical plates and surface area percentage of spongy bone trabeculae. These results were supported by the histological picture of this group which resembled the control group, with high collagen synthesis confirmed by Masson trichrome and high osteoblastic activity confirmed by high immunoreactivity of this group.