The Use of Expanded Mesenchymal Stem Cells (MSCs ...scholar.cu.edu.eg/ashrafseida/files/ph.d._presentation.pdfManagement of vertical alveolar ridge deficiency Donor site morbidity

Teeth loss

Pattern of bone resorption after extraction

2/ 3 in

first 3

months

50% in

1st

year

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, 33-44.

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)

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), 428-435.

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), 401-413.

Stem Cell Based Alveolar

Regeneration

Expanded

MSCs

Stem Cell Based Alveolar Regeneration

Cells

Scaffold

Growth Factor

Cells Scaffold

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), 665-676.

t al., 2012 9

Stem Cell Based Alveolar Regeneration

Cells

Stable with extended

passaging

Ease of isolationMultipotency

Rapid expansion

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), 229-248.

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), 546-554.

Fibrin Scaffold in Bone tissue Engineering

BM-MSCs in treatment of articular cartilage defects.

BM- MSCs in treatment of alveolar bone defects in

rats.

FG PRP PRF

Dohan, D. M., Choukroun, J., Diss, A., Dohan, S. L., Dohan, A. J., Mouhyi, J., & Gogly, B. (2006, 03). Platelet-rich fibrin (PRF): A second-

generation platelet concentrate. Part II: Platelet-related biologic features. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and

Endodontology, 101(3).

PRF in Bone Tissue Engineering

- 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), 341-352.

- 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), 185-194.

- Wu, C., Lee, S., Tsai, C., Lu, K., Zhao, J., & Chang, Y. (2012, 05). Platelet-rich fibrin increases cell attachment, proliferation and collagen-

related protein expression of human osteoblasts. Australian Dental Journal, 57(2), 207-212.

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.

Aim Of The Study

(PRF+ MSCs)(PRF+ MSCs +Bi- Phasic

Ca- Ph)

Pattern of Bone Formation

Materials & Methods

12 – 24 months25- 40 Kg Weight

3 vertical alveolar ridge

defects

10 dogs

Stem Cell Isolation, Characterization

and expansion

Stage II

Animal Scarify

Stage I

A- In-Vivo

B- In-Vitro

Defect induction

Gingival harvesting

Augmentation

Procedures

A- In- Vivo

Stage I

(Defect Induction )

Stage I

(Defect Induction )

Stage I

(Defect Induction )

4 Weeks

Stage I

(Defect Induction )

Stage I

Gingival Harvesting

Tissue Engineering Laboratory

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)

PRF

Scaffold Preparation

Augmentation Procedures

Mesh Fixation

Group I

(PRF+ MSCs)

Group II

(PRF+ Bi- Phasic Ca- Ph +

MSCs)

Group III

(PRF+ Bi- Phasic Ca- Ph)

Group II

(PRF+ Bi- Phasic Ca- Ph +

MSCs)

Group III

(PRF+ Bi- Phasic Ca- Ph)

Results

In-vitro

MSCs in Culture

Multi-lineage

Differentiation

Cell Surface Marker

Expression

Osteogenic

Chondrogenic

MSCs in Culture

Day 0Day 14

Day 7

Cell Surface Marker Expression

Multi-lineage Differentiation

OsteoblasticDifferentiation

ChondrogenicDifferentiation

In-vivo

Histology

H & E

Histomorphometry

Bone

Matrix

Mature

Bone

Histochemical

(Masson Trichrome)

Bone formation

in center

Bone Interface

Bone

Trabeculae

Reversal

LinesOsteocytes Osteoclasts

Histological Results

H & E

Group I

Bone formation in

center Bone interface

Bone

Trabeculae

Reversal

LinesOsteocytes Osteoclasts

Group I

Group II

Bone formation in

center Bone interface

Bone

Trabeculae

Reversal

LinesOsteocytes Osteoclasts

Group II

Group III

Bone formation in

center Bone interface

Bone

Trabeculae

Reversal

LinesOsteocytes Osteoclasts

Group III

Histomorphometric Results

Area percent of Mature Bone

0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

90.00

Gp 1 Gp2 Gp3

75.98

81.91

35.35

Mean area percent of Mature Bone

Histochemical Results

Group I

Group II

Group III

Histomorphometric Results

Area percent of Bone Matrix

0.00

10.00

20.00

30.00

40.00

50.00

60.00

Gp 1 Gp2 Gp3

12.56

50.17

10.60

Perc

ent

Mean area percent of Bone Matrix

Conclusion

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

Recommendation

Larger sample size with longer period of follow up.

Radiographic assessment.

Thank You