In vitro Assay of Human Gingival Scaffold in Differentiation of Rat’s Bone Marrow Mesenchymal Stem Cells to Keratinocystes

Document Type : Original Article


1 Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran.

3 Dental Research Centre, Faculty of Dentistry, Mashhad University of Medical Sciences, Mashhad, Iran

4 Oral and Maxillofacial Diseases Research Centre, Faculty of Dentistry, Mashhad University of Medical Sciences, Mashhad, Iran


Tissue engineering is an attractive science because it promises new therapeutic strategies for repairing organs that have lost functions due to damage. The purpose of this study was to evaluate induction effect of human gingival scaffold in tissue engineering for skin regeneration.
Materials and Methods
Tissue samples were obtained from crown-lengthening procedures and wisdom teeth removal. The samples were decellularized and used as a scaffold for loading of rat BM-MSCs. The human gingival scaffolds loaded by bone marrow mesenchymal stem cells were derived from Wistar rat. Finally, it was evaluated via electron micrographs, as well as immunohistochemical techniques at day 7, 14, and 28 after initial seeding.
The histologic sections of human gingival scaffold –loaded rat BM-MSCs demonstrated formation of epithelial like layers at days 7, 14 and 28 after initial seeding. Scanning electron microscope (SEM) of the scaffolds indicated formed desmosomal adhesions, which revealed a degree of differentiation toward keratinocytes. The results of immunohistochemical staining were strongly positive for multi cytokeratin (CK) 14 days after initial seeding in epithelial differentiation. Rat BM-MSCs which loaded on human gingival scaffold is capable of differentiating toward keratinocytes.
Gingival tissues were presented as a natural scaffold for attachment and differentiation of bone marrow mesenchymal stem cells towards keratinocytes, and might be used as suitable scaffold for reconstruction of the skin.


1. Bakopoulou A, Leyhausen G, Volk J, Tsiftsoglou A, Garefis P, Koidis P, Geurtsen W. Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells (DPSCs) and stem cells from the apical papilla (SCAP). Arch Oral Biol 2011; 56:709-721.
2. Musina RA, Bekchanova ES, Belyavskii AV, Sukhikh GT. Differentiation potential of mesenchymal stem cells of different origin. Bull Exp Biol Med 2006; 141:147-151.
3. Hwang NS, Zhang C, Hwang YS, Varghese S. Mesenchymal stem cell differentiation and roles in regenerative medicine. Wiley Interdiscip Rev Syst Biol Med 2009; 1:97-106.
4. Markowicz M, Koellensperger E, Neuss S, Koenigschulte S, Bindler C, Pallua N. Human bone marrow mesenchymal stem cells seeded on modified collagen improved dermal regeneration in vivo. Cell Transplant 2006; 15:723-732.
5. Tonello C, Vindigni V, Zavan B, Abatangelo S, Abatangelo G, Brun P, et al. In vitro reconstruction of an endothelialized skin substitute provided with a microcapillary network using biopolymer scaffolds. FASEB J 2005; 19:1546-1548.
6. Roessner ED, Thier S, Hohenberger P, Schwarz M, Pott P, Dinter D, et al. Acellular dermal matrix seeded with autologous fibroblasts improves wound breaking strength in a rodent soft tissue damage model in neoadjuvant settings. J Biomater Appl 2011; 25:413-427.
7. Stark HJ, Boehnke K, Mirancea N, Willhauck MJ, Pavesio A, Fusenig NE, Boukamp P. Epidermal homeostasis in long-term scaffold-enforced skin equivalents. J Investig Dermatol Symp Proc 2006; 11:93-105.
8. Tran CT, Huynh DT, Gargiulo C, Nguyen PT, Tran TT, Huynh MT, et al. In vitro culture of keratinocytes from human umbilical cord blood mesenchymal stem cells: the Saigonese culture. Cell Tissue Bank 2011; 12:125-133.
9. Wu Y, Chen L, Scott PG, Tredget EE. Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells 2007; 25:2648-2659.
10. O'Toole E A. Extracellular matrix and keratinocyte migration. Experimental dermatology. 2001; 26:525-30
11. Nanci A.Ten cate, s oral histology.7th ed. Philadelphia: Mosby Elsevier; 2008.p.328.
12. Eckert RL, Rorket EA. Molecular biology of keratinocyte differentiation. Environ Health Perspect 1989; 80:109-116.
13. McKeown ST, Hyland PL, Locke M, Mackenzie IC, Irwin CR. Keratinocyte growth factor and scatter factor expression by regionally defined oral fibroblasts. Eur J Oral Sci 2003; 111:42-50.
14. Gilbert TW, Sellaro TL, Badylak SF. Biomaterials.Decellularization of tissues and organs. Biomaterials 2006; 27:3675-3683.
15. Elder BD, Eleswarapu SV, Athanasiou KA. Extraction techniques for the decellularization of tissue engineered articular cartilage constructs. Biomaterials 2009; 30:3749-3756.
16. Fröhlich M, Grayson WL, Marolt D, Gimble JM, Kregar-Velikonja N, Vunjak-Novakovic G. Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. Tissue Eng Part A 2010; 16:179-189.
17. Narita Y, Kagami H, Matsunuma H, Murase Y, Ueda M, Ueda Y. Decellularized ureter for tissue-engineered small-caliber vascular graft. J Artif Organs 2008; 11:91-99.
18. Abousleiman RI, Reyes Y, McFetridge P, Sikavitsas V. The human umbilical vein: a novel scaffold for musculoskeletal soft tissue regeneration. Artif Organs 2008; 32:735-742.
19. Clark JM, Saffold SH, Israel JM. Decellularized dermal grafting in cleft palate repair. Arch Facial Plast Surg 2003; 5:40-44.
20. Bannasch H, Stark GB, Knam F, Horch RE, Föhn M. Decellularized dermis in combination with cultivated keratinocytes in a short- and long-term animal experimental investigation. J Eur Acad Dermatol Venereol 2008; 22:41-49.
21. Tamizi M, Amouian B, Mahdavie N. Clinical and histological evaluation of the effect of storage time on healing and success of free gingival graft stored in normal saline. J M Dental S? 1996; 20:38-49.
22. Miettinen M. Immunohistochemistry in tumor diagnosis. Ann Med 1993; 25:221-233.