Pattern of bone resorption after extraction

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4 Teeth loss

5 Pattern of bone resorption after extraction 50% in 1st year 2/ 3 in first 3 months Reich KM, Huber CD, Lippnig WR, Um C, Watzek G, Tangl S. (2011, 17). Atrophy of Residual Alveolar Ridge following tooth loss in an historical population, Oral Diseases, 17,

6 Le, B., Rohrer, M. D., & Prassad, H. S. (2010, 02). Screw Tent-Pole Grafting Technique for Reconstruction of Large Vertical Alveolar Ridge Defects Using Human Mineralized Allograft for Implant Site Preparation. Journal of Oral and Maxillofacial Surgery, 68(2), Vertical Alveolar deficiency 3 dimensional Defect (3D bone reconstruction) The larger the defect the greater is the tension Soft tissue tension increase resorption of graft Wound Dehiescence (infection, loss of graft)

7 Management of vertical alveolar ridge deficiency Donor site morbidity Higher rate of bone resorption Wound Dehiscence Zhang, Z. (2011, 12). Bone regeneration by stem cell and tissue engineering in oral and maxillofacial region. Front. Med. Frontiers of Medicine, 5(4),

8 Stem Cell Based Alveolar Regeneration Expanded MSCs

9 Stem Cell Based Alveolar Regeneration Cells Cells Scaffold Scaffold Growth Factor Growth Factor Mechanical Environment Nie e Nie, H., Lee, C. H., Tan, J., Lu, C., Mendelson, A., Chen, M.,... Mao, J. J. (2012, 03). Musculoskeletal tissue engineering by endogenous stem/progenitor cells. Cell Tissue Res Cell and Tissue Research, 347(3), t al.,

10 Stem Cell Based Alveolar Regeneration

11 Cells Rapid expansion Stable with extended passaging Multipotency Ease of isolation Egusa, H., Sonoyama, W., Nishimura, M., Atsuta, I., & Akiyama, K. (2012, 10). Stem cells in dentistry Part II: Clinical applications. Journal of Prosthodontic Research, 56(4),

12 Cells - Proliferation & differentiation - Migration & chemotaxix -Provide cell attachment - Maintain phenotye Tissue -Biocompatible -Biodegradable - Mechanical strength before degradation -Angiogenesis & vasculogenesis - Hydrophillic Surgeon -cost effective - easily handled -available in relevant quantities Bose, S., Roy, M., & Bandyopadhyay, A. (2012, 10). Recent advances in bone tissue engineering scaffolds. Trends in Biotechnology, 30(10),

13 Dohan, D. M., Choukroun, J., Diss, A., Dohan, S. L., Dohan, A. J., Mouhyi, J., & Gogly, B. (2006, 03). Platelet-rich fibrin (PRF): A secondgeneration platelet concentrate. Part II: Platelet-related biologic features. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 101(3). Fibrin Scaffold in Bone tissue Engineering FG PRP PRF BM-MSCs in treatment of articular cartilage defects. BM- MSCs in treatment of alveolar bone defects in rats.

14 PRF in Bone Tissue Engineering Stimulation of cellular adhesion and Proliferation of human gingival fibroblast, dermal prekeratinocyte, preadipocyte and maxillofacial osteoblast and also osteoblastic differentiation Dose dependant stimulation of Proliferation & differentiation of BM-MSCs Stimulate cellular adhesion and proleferation of rat osteoblasts, human osteoblasts, human periodontal ligament fibroblasts, and human pulp fibroblasts. - Ehrenfest, D. M., Diss, A., Odin, G., Doglioli, P., Hippolyte, M., & Charrier, J. (2009, 09). In vitro effects of Choukroun's PRF (platelet-rich fibrin) on human gingival fibroblasts, dermal prekeratinocytes, preadipocytes, and maxillofacial osteoblasts in primary cultures. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 108(3), Ehrenfest, D. M., Doglioli, P., Peppo, G. M., Corso, M. D., & Charrier, J. (2010, 03). Choukroun's platelet-rich fibrin (PRF) stimulates in vitro proliferation and differentiation of human oral bone mesenchymal stem cell in a dose-dependent way. Archives of Oral Biology, 55(3), Wu, C., Lee, S., Tsai, C., Lu, K., Zhao, J., & Chang, Y. (2012, 05). Platelet-rich fibrin increases cell attachment, proliferation and collagenrelated protein expression of human osteoblasts. Australian Dental Journal, 57(2),

15

16 Aim Of The Study (PRF+ MSCs) (PRF+ MSCs +Bi- Phasic Ca- Ph) Pattern of Bone Formation

17 Materials & Methods

18 3 vertical alveolar ridge defects 10 dogs months Kg Weight

19 Stage I Defect induction Gingival harvesting A- In-Vivo Stage II Augmentation Procedures Animal Scarify B- In-Vitro Stem Cell Isolation, Characterization and expansion

20 A- In- Vivo Stage I (Defect Induction )

21 Stage I (Defect Induction )

22 Stage I (Defect Induction )

23 Stage I (Defect Induction ) 4 Weeks

24 Stage I Gingival Harvesting

25 Tissue Engineering Laboratory

26 Stage II Augmentation Procedures Group I (PRF + MSCs) Group II (PRF + MSCs + Bi-phasic Ca- Ph block) Group III (PRF + Bi- phasic Ca- Ph block)

27 PRF Scaffold Preparation

28 Augmentation Procedures

29 Mesh Fixation

30 Group I (PRF+ MSCs)

31 Group II (PRF+ Bi- Phasic Ca- Ph + MSCs) Group III (PRF+ Bi- Phasic Ca- Ph)

32 Group II (PRF+ Bi- Phasic Ca- Ph + MSCs) Group III (PRF+ Bi- Phasic Ca- Ph)

33

34 Results

35 MSCs in Culture In-vitro Cell Surface Marker Expression Osteogenic Multi-lineage Differentiation Chondrogenic

36 MSCs in Culture Day 0 Day 7 Day 14

37 Cell Surface Marker Expression

38 Multi-lineage Differentiation Osteoblastic Differentiation Chondrogenic Differentiation

39 Histology H & E In-vivo Histochemical (Masson Trichrome) Mature Bone Histomorphometry Bone Matrix

40 Histological Results H & E Bone Interface Bone formation in center Bone Trabeculae Reversal Lines Osteocytes Osteoclasts

41 Group I Bone interface Bone formation in center

42 Group I Bone Trabeculae Reversal Lines Osteocytes Osteoclasts

43 Group II Bone interface Bone formation in center

44 Group II Bone Trabeculae Reversal Lines Osteocytes Osteoclasts

45 Group III Bone interface Bone formation in center

46 Group III Bone Trabeculae Reversal Lines Osteocytes Osteoclasts

47 Histomorphometric Results Area percent of Mature Bone

48 Mean area percent of Mature Bone Gp 1 Gp2 Gp3

49 Histochemical Results Group I

50 Group II

51 Group III

52 Histomorphometric Results Area percent of Bone Matrix

53 Percent Mean area percent of Bone Matrix Gp 1 Gp2 Gp3

54 Conclusion

55 PRF membranes loaded with MSCs could induce early bone formation and maturation in vertical alveolar defects. PRF couldn't support osteogensis process in vertical alveolar ridge defects till complete bone formation and restoration of the whole bone volume. The combined use of Bi-phasic calcium phosphate block together with PRF loaded with MSCs is superior to single use of PRF loaded with MSCs in treatment of vertical alveolar ridge defects. The combined use of Bi-phasic calcium phosphate block together with MSCs loaded PRF yields promising results in treatment of vertical alveolar ridge defects

56 Recommendation Larger sample size with longer period of follow up. Radiographic assessment.

57 Thank You

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