Tooth loss due to periodontal disease, dental caries, trauma, or a variety of genetic disorders continues to affect most adults adversely at some time in their lives. A biological tooth substitute that could replace lost teeth would provide a vital alternative to currently available clinical treatments. To pursue this goal, we dissociated porcine third molar tooth buds into single-cell suspensions and seeded them onto biodegradable polymers. After growing in rat hosts for 20 to 30 weeks, recognizable tooth structures formed that contained dentin, odontoblasts, a well-defined pulp chamber, putative Hertwig’s root sheath epithelia, putative cementoblasts, and a morphologically correct enamel organ containing fully formed enamel. Our results demonstrate the first successful generation of tooth crowns from dissociated tooth tissues that contain both dentin and enamel, and suggest the presence of epithelial and mesenchymal dental stem cells in porcine third molar tissues.
Bohl KS, Shon J, Rutherford B, Mooney DJ -1998-. Role of synthetic extracellular matrix in development of engineered dental pulp. J Biomater Sci-Polym Ed9:749–764.
3.
Bronckers AL, D’Souza RN, Butler WT, Lyaruu DM, van Dijk S, Gay S, et al. -1993-. Dentin sialoprotein: biosynthesis and developmental appearance in rat tooth germs in comparison with amelogenins, osteocalcin and collagen type-1. Cell Tissue Res272:237–247.
4.
Chen JK, McCulloch CA, Sodek J -1993-. Bone sialoprotein in developing porcine dental tissues: cellular expression and comparison of tissue localization with osteopontin and osteonectin. Arch Oral Biol38:241–249.
5.
Choi RS, Vacanti JP -1997-. Preliminary studies of tissue-engineered intestine using isolated epithelial organoid units on tubular synthetic biodegradable scaffolds. Transplant Proc29:848–851.
6.
Deakins M, Volker JF -1941-. Amount of organic matter in enamel from several types of human teeth. J Dent Res20:117–121.
7.
Diekwisch TG -2001-. The developmental biology of cementum. Int J Dev Biol45:695–706.
8.
Glasstone S -1967-. Development of teeth in tissue culture. J Dent Res46:858–861.
9.
Gronthos S, Mankani M, Brahim J, Robey PG, Shi S -2000-. Postnatal human dental pulp stem cells -DPSCs- in vitro and in vivo. Proc Natl Acad Sci USA97:13625–13630.
10.
Grzesik WJ, Cheng H, Oh JS, Kuznetsov SA, Mankani MH, Uzawa K, et al. -2000-. Cementum-forming cells are phenotypically distinct from bone-forming cells. J Bone Miner Res15:52–59.
11.
Jernvall J, Thesleff I -2000-. Reiterative signaling and patterning during mammalian tooth morphogenesis. Mech Dev92:19–29.
12.
Kim SS, Vacanti JP -1999-. The current status of tissue engineering as potential therapy. Semin Pediatr Surg8:119–123.
13.
Laine M, Thesleff I -1986-. Development of mouse embryonic molars in vitro: an attempt to design defined culture conditions allowing mineralization. J Biol Buccale14:15–23.
14.
LeFevre ML, Manly RS -1932-. Moisture, inorganic and organic contents of enamel and dentin from carious teeth. J Am Dent Assoc24:233–242.
15.
Linde A, Goldberg M -1993-. Dentinogenesis. Crit Rev Oral Biol Med4:679–728.
16.
MacDougall M, Simmons D, Luan X, Nydegger J, Feng J, Gu TT -1997-. Dentin phosphoprotein and dentin sialoprotein are cleavage products expressed from a single transcript coded by a gene on human chromosome 4. Dentin phosphoprotein DNA sequence determination. J Biol Chem272:835–842.
17.
Mikos AG, Bao Y, Cima LG, Ingber DE, Vacanti JP, Langer R -1993-. Preparation of poly-glycolic acid- bonded fiber structures for cell attachment and transplantation. J Biomed Mater Res27:183–189.
18.
Mina M, Kollar EJ -1987-. The induction of odontogenesis in non-dental mesenchyme combined with early murine mandibular arch epithelium. Arch Oral Biol32:123–127.
19.
Mooney DJ, Powell C, Piana J, Rutherford B -1996-. Engineering dental pulp-like tissue in vitro. Biotechnol Prog12:865–868.
20.
Peters H, Balling R -1999-. Teeth. Where and how to make them. Trends Genet15:59–65.
Saito M, Iwase M, Maslan S, Nozaki N, Yamauchi M, Handa K, et al. -2001-. Expression of cementum-derived attachment protein in bovine tooth germ during cementogenesis. Bone29:242–248.
23.
Saygin NE, Giannobile WV, Somerman MJ -2000-. Molecular and cell biology of cementum. Periodontology24:73–98.
24.
Slavkin HC, Bringas P, Cameron J, LeBaron R, Bavetta LA -1969-. Epithelial and mesenchymal cell interactions with extracellular matrix material in vitro. J Embryol Exp Morphol22:395–405.
25.
Slavkin HC, Bringas P Jr, Bessem C, Santos V, Nakamura M, Hsu MY, et al. -1989-. Hertwig’s epithelial root sheath differentiation and initial cementum and bone formation during long-term organ culture of mouse mandibular first molars using serumless, chemically-defined medium. J Periodontal Res24:28–40.
26.
Stevens A, Lowe L, Bancroft JD -1990-. Theory and practice of histological techniques. New York: Churchill Livingstone.
27.
Thesleff I -1976-. Differentiation of odontogenic tissues in organ culture. Scand J Dent Res84:353–356.
28.
Thomas HF, Kollar EJ -1989-. Differentiation of odontoblasts in grafted recombinants of murine epithelial root sheath and dental mesenchyme. Arch Oral Biol34:27–35.
29.
Wu D, Ikezawa K, Parker T, Saito M, Narayanan AS -1996-. Characterization of a collagenous cementum-derived attachment protein. J Bone Miner Res11:686–692.
30.
Yamada M, Bringas P Jr, Grodin M, MacDougall M, Cummings E, Grimmett J, et al. -1980-. Chemically-defined organ culture of embryonic mouse tooth organs: morphogenesis, dentinogenesis and amelogenesis. J Biol Buccale8:127–139.
31.
Yoshikawa DK, Kollar EJ -1981-. Recombination experiments on the odontogenic roles of mouse dental papilla and dental sac tissues in ocular grafts. Arch Oral Biol26:303–307.
32.
Zeichner-David M, Diekwisch T, Fincham A, Lau E, MacDougall M, Moradian-Oldak J, et al. -1995-. Control of ameloblast differentiation. Int J Dev Biol39:69–92.
