Human dental pulp cell culture and cell transplantation with an alginate scaffold.
Many studies on tissue stem cells have been conducted in the field of regenerative medicine, and some studies have indicated that cultured dental pulp mesenchymal cells secrete dentin matrix. In the present study we used alginate as a scaffold to transplant subcultured human dental pulp cells subcutaneously into the backs of nude mice. We found that when beta-glycerophosphate was added to the culture medium, dentin sialophosphoprotein mRNA coding dentin sialoprotein (DSP) was expressed. An increase in alkaline phosphatase, which is an early marker for odontoblast differentiation, was also demonstrated. At 6 weeks after implantation the subcutaneous formation of radio-opaque calcified bodies was observed in situ. Immunohistochemical and fine structure studies identified expression of type I collagen, type III collagen, and DSP in the mineralizing transplants. Isolated odontoblast-like cells initiated dentin-like hard tissue formation and scattered autolyzing apoptotic cells were also observed in the transplants. The study showed that subcultured dental pulp cells actively differentiate into odontoblast-like cells and induce calcification in an alginate scaffold.
Kumabe
Shunji
S
Department of Oral Anatomy, Osaka Dental University, 8-1, Kuzuhahanazono-cho, Hirakata-shi, Osaka 573-1121, Japan. kumabe@cc.osaka-dent.ac.jp
Nakatsuka
Michiko
M
Kim
Gi-Seup
GS
Jue
Seong-Suk
SS
Aikawa
Fumiko
F
Shin
Je-Won
JW
Iwai
Yasutomo
Y
D000464:Alginates; D000469:Alkaline Phosphatase / Q000032:analysis / Q000235:genetics; D001672:Biocompatible Materials; D002454:Cell Differentiation; D017690:Cell Transplantation / Q000379:methods*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000032:analysis / Q000235:genetics; D024061:Collagen Type III / Q000032:analysis / Q000235:genetics; D046508:Culture Techniques; D003782:Dental Pulp / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D003784:Dental Pulp Calcification / Q000473:pathology / Q000503:physiopathology; D016326:Extracellular Matrix Proteins; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009804:Odontoblasts / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D010006:Osteoblasts / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D010750:Phosphoproteins; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000032:analysis / Q000235:genetics; D000099311:Dentin Sialophosphoprotein
D000464:Alginates; D000469:Alkaline Phosphatase; D001672:Biocompatible Materials; D024042:Collagen Type I; D024061:Collagen Type III; D016326:Extracellular Matrix Proteins; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D010750:Phosphoproteins; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D000099311:Dentin Sialophosphoprotein