1 Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran Stem Cells and Regenerative Medicine Research Group, Academic Center for Education, Culture, and Research (ACECR), Khorasan Razavi Branch, Mashhad, Iran Novel Diagnostics and Therapeutics Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

3 Oral and Maxillofacial Diseases Research Center, School of Dentistry, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Microanatomy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

6 Department of Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

7 Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran Stem Cells and Regenerative Medicine Research Group, Academic Center for Education, Culture, and Research (ACECR), Khorasan Razavi Branch, Mashhad, Iran


Background: Due to the known disadvantages of autologous bone grafting, tissue engineering approaches have become an attractive method for ridge augmentation in dentistry. To the best of our knowledge, this is the first study conducted to evaluate the potential therapeutic capacity of PRP-assisted hADSCs seeded on HA/TCP granules on regenerative healing response of canine alveolar surgical bone defects. This could offer a great advantage to alternative approaches of bone tissue healing-induced therapies at clinically chair-side procedures. Methods: Cylindrical through-and-through defects were drilled in the mandibular plate of 5 mongrel dogs and filled randomly as following: I- autologous crushed mandibular bone, II- no filling material, III- HA/TCP granules in combination with PRP, and IV- PRP-enriched hADSCs seeded on HA/TCP granules. After the completion of an 8-week period of healing, radiographic, histological and histomorphometrical analysis of osteocyte number, newly-formed vessels and marrow spaces were used for evaluation and comparison of the mentioned groups. Furthermore, the buccal side of mandibular alveolar bone of every individual animal was drilled as normal control samples (n=5). Results: Our results revealed that hADSCs subcultured on HA/TCP granules in combination with PRP significantly promoted bone tissue regeneration as compared with those defects treated only with PRP and HA/TCP granules (P<0.05). Conclusion: In conclusion, our results indicated that application of PRP-assisted hADSCs could induce bone tissue regeneration in canine alveolar bone defects and thus, present a helpful alternative in bone tissue regeneration.


Main Subjects

1. Esposito M, Grusovin MG, Felice P, Karatzopoulos
G, Worthington HV, Coulthard P. Interventions for
replacing missing teeth: horizontal and vertical
bone augmentation techniques for dental implant
treatment. Cochrane Database Syst Rev. 2009;
2. Erbe EM, Marx GJ, Clineff TD, Bellincampi LD. Potential
of an ultraporous β-tricalcium phosphate synthetic
cancellous bone void filler and bone marrow aspirate
composite graft. Eur Spine J. 2001; 10(Suppl 2):S141-6.
3. Sulaiman SB, Keong TK, Cheng CH, Saim AB, Idrus
RB. Tricalcium phosphate/hydroxyapatite (TCP-HA)
bone scaffold as potential candidate for the formation
of tissue engineered bone. Indian J Med Res. 2013;
4. Malgieri A, Kantzari E, Patrizi MP, Gambardella S.
Bone marrow and umbilical cord blood human
mesenchymal stem cells: state of the art. Int J Clin Exp
Med. 2010; 3(4):248-69.
5. Naderi‐Meshkin H, Matin MM, Heirani‐Tabasi A,
Mirahmadi M, Irfan‐Maqsood M, Edalatmanesh MA, et
al. Injectable hydrogel delivery plus preconditioning
of mesenchymal stem cells: exploitation of SDF‐1/
CXCR4 axis toward enhancing the efficacy of stem
cells’ homing. Cell Biol Int. 2016; 40(7):730-41.
6. Lin CS, Lin G, Lue TF. Allogeneic and xenogeneic
transplantation of adipose-derived stem cells
in immunocompetent recipients without
immunosuppressants. Stem Cell Dev. 2012;
7. Puissant B, Barreau C, Bourin P, Clavel C, Corre J,
Bousquet C, et al. Immunomodulatory effect of human
adipose tissue‐derived adult stem cells: comparison
with bone marrow mesenchymal stem cells. Br J
Haematol. 2005; 129(1):118-29.
8. Tahami M, Haddad B, Abtahian A, Hashemi A,
Aminian A, Konan S. Potential role of local estrogen in
enhancement of fracture healing: preclinical study in
rabbits. Arch Bone Jt Surg. 2016; 4(4):323-9.
9. Kim ES, Kim JJ, Park EJ. Angiogenic factor-enriched
platelet-rich plasma enhances in vivo bone formation
around alloplastic graft material. J Adv Prosthod.
2010; 2(1):7-13.
10. Dhurat R, Sukesh M. Principles and methods of
preparation of platelet-rich plasma: a review and
author’s perspective. J Cutan Aesthet Surg. 2014;
11. Butterfield KJ, Bennett J, Gronowicz G, Adams D. Effect
of platelet-rich plasma with autogenous bone graft for
maxillary sinus augmentation in a rabbit model. J Oral
Maxillofac Surg. 2005; 63(3):370-6.
12. Roldán JC, Jepsen S, Miller J, Freitag S, Rueger DC, Açil
Y, et al. Bone formation in the presence of plateletrich
plasma vs. bone morphogenetic protein-7. Bone.
2004; 34(1):80-90.
13. Ghavimi SA, Ebrahimzadeh MH, Shokrgozar MA,
Solati-Hashjin M, Osman NA. Effect of starch content
on the biodegradation of polycaprolactone/starch
composite for fabricating in situ pore-forming
scaffolds. Polymer Test. 2015; 43(2):94-102.
14. Ali Akbari Ghavimi S, Ebrahimzadeh MH, Solati-
Hashjin M, Abu Osman NA. Polycaprolactone/
starch composite: fabrication, structure, properties,
and applications. J Biomed Mater Res A. 2015;
15. Gamblin AL, Brennan MA, Renaud A, Yagita H,
Lézot F, Heymann D, et al. Bone tissue formation
with human mesenchymal stem cells and biphasic
calcium phosphate ceramics: the local implication of
osteoclasts and macrophages. Biomaterials. 2014;
16. Houmard M, Fu Q, Genet M, Saiz E, Tomsia AP. On the
structural, mechanical, and biodegradation properties
of HA/β‐TCP robocast scaffolds. J Biomed Mater Res B
Appl Biomater. 2013; 101(7):1233-42.
17. Hahn BD, Park DS, Choi JJ, Ryu J, Yoon WH, Lee BK,
et al. Effect of the HA/β‐TCP ratio on the biological
performance of calcium phosphate ceramic coatings
fabricated by a room‐temperature powder spray in
vacuum. J Am Ceram Soc. 2009; 92(4):793-9.
18. Ripamonti U, Crooks J, Kirkbride A. Sintered porous
hydroxyapatites with intrinsic osteoinductive activity:
geometric induction of bone formation. South Afr J
Sci. 1999; 95(8):335-43.
19. Lobo SE, Livingston Arinzeh T. Biphasic calcium
phosphate ceramics for bone regeneration and tissue
engineering applications. Materials. 2010; 3(2):815-26.
20. Hörner K, Loeffler K, Holtzmann M. Comparison of
the histologic structure of the compact bone of the
long hollow bones of mouse, hamster, rat, guinea pig,
rabbit, cat, and dog during development. Anat Histol
Embryol. 1997; 26(4):289-95.
21. Pearce SG. Animal models for bone repair. Eur Cell
Mater. 2007; 14(1):42-9.
22. Kang Y, Kim S, Khademhosseini A, Yang Y. Creation
of bony microenvironment with CaP and cellderived
ECM to enhance human bone-marrow MSC
behavior and delivery of BMP-2. Biomaterials. 2011;
23. Xie X, Wang Y, Zhao C, Guo S, Liu S, Jia W, et al.
Comparative evaluation of MSCs from bone marrow
and adipose tissue seeded in PRP-derived scaffold
for cartilage regeneration. Biomaterials. 2012;
24. Miloro M, Haralson DJ, Desa V. Bone healing in a rabbit
mandibular defect using platelet-rich plasma. J Oral
Maxillofac Surg. 2010; 68(6):1225-30.
25. Jafarian M, Eslaminejad MB, Khojasteh A, Mashhadi
Abbas F, Dehghan MM, Hassanizadeh R, et al. Marrowderived
mesenchymal stem cells-directed bone
regeneration in the dog mandible: a comparison
between biphasic calcium phosphate and natural
bone mineral. Oral Surg Oral Med Oral Pathol Oral
Radiol Endod. 2008; 105(5):e14-24.

26. Mohammadipour A, Fazel A, Haghir H, Motejaded F,
Rafatpanah H, Zabihi H, et al. Maternal exposure to
titanium dioxide nanoparticles during pregnancy;
impaired memory and decreased hippocampal
cell proliferation in rat offspring. Environ Toxicol
Pharmacol. 2014; 37(2):617-25.
27. Levi B, James AW, Nelson ER, Vistnes D, Wu B, Lee
M, et al. Human adipose derived stromal cells heal
critical size mouse calvarial defects. PloS One. 2010;
28. Tollemar V, Collier ZJ, Mohammed MK, Lee MJ, Ameer
GA, Reid RR. Stem cells, growth factors and scaffolds
in craniofacial regenerative medicine. Genes Dis.
2016; 3(1):56-71.
29. van Bergen CJ, Kerkhoffs GM, Ö􀇆 zdemir M, Korstjens
CM, Everts V, van Ruijven LJ, et al. Demineralized
bone matrix and platelet-rich plasma do not improve
healing of osteochondral defects of the talus: an
experimental goat study. Osteoarth Cartilage. 2013;
30. Wolf BR. An injection of platelet-rich plasma was
not more effective than placebo for rotator cuff
tendinopathy. J Bone Jt Surg. 2014; 96(10):871-8.
31. Chen FM, Wu LA, Zhang M, Zhang R, Sun HH. Homing
of endogenous stem/progenitor cells for in situ tissue
regeneration: promises, strategies, and translational
perspectives. Biomaterials. 2011; 32(12):3189-209.
32. Kovacevic M, Tamarut T, Zoricic S, Bešlic S. A
method for histological, enzyme histochemical
andimmunohistochemical analysis of periapical
diseases on undecalcified bone with teeth. Acta
Stomatol Croat. 2003; 37(3):261-73.