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<p><strong>INTRODUCTION </strong></p> <p><strong>T</strong>he incidence of children born with missing primary and/or adult teeth is significant (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#NUNN-ETAL-2003">Nunn <em>et al.</em>, 2003</a>), and tooth loss in aged populations is also a prevalent health problem. Current replacement tooth methods use synthetic materials that can elicit an immune induced host rejection response. The ability to generate biological tooth substitutes from autologous human tissues would be a valuable clinical tool. Tissue engineering—a relatively new science integrating developmental, molecular/cellular biology, and genetics with the field of engineering—holds promise in this area (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#LANGER-AND-VACANTI-1993">Langer and Vacanti, 1993</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#SITTINGER-ETAL-1996">Sittinger <em>et al.</em>, 1996</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#CHOI-AND-VACANTI-1997">Choi and Vacanti, 1997</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#BOHL-ETAL-1998">Bohl <em>et al.</em>, 1998</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#KIM-AND-VACANTI-1999">Kim and Vacanti, 1999</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#MOONEY-AND-MIKOS-1999">Mooney and Mikos, 1999</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#STOCK-AND-VACANTI-2001">Stock and Vacanti, 2001</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#VACANTI-ETAL-2001">Vacanti <em>et al.</em>, 2001</a>). In the fields of maxillofacial surgery and periodontics, tissue engineering has been used to generate alveolar bone (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#ABUKAWA-ETAL-2003">Abukawa <em>et al.</em>, 2003</a>) and to regenerate oral tissues lost to cancer, decay, and periodontitis (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#LYNCH-ETAL-1999">Lynch <em>et al.</em>, 1999</a>). Cultured human dental pulp and gingival fibroblasts adhere to biodegradable scaffolds, and proliferate and differentiate <em>in vitro</em> (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#MOONEY-ETAL-1996">Mooney <em>et al.</em>, 1996</a>; <a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#MURPHY-AND-MOONEY-1999">Murphy and Mooney, 1999</a>) and <em>in vivo</em> (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#BUURMA-ETAL-1999">Buurma <em>et al.</em>, 1999</a>). More recently, we have reported the successful bioengineering of whole tooth crowns composed of accurately formed enamel, dentin, and pulp tissues (<a href="http://www.biology-online.org/articles/references-bioengineered_teeth_from_cultured_rat_tooth_bud_cells.htm#YOUNG-ETAL-2002">Young <em>et al.</em>, 2002</a>). </p> <p>The objective of this study was to improve upon our tooth-tissue-engineering methods by optimizing the age of tooth bud cells. In addition, we further define the progenitor cell populations giving rise to bioengineered tooth structures by exclusively using single-cell suspensions of rat tooth bud cells that were first cultured <em>in vitro</em> for 6 days. Finally, we compared the use of PGA and PLGA scaffold materials. The results of this study show that, as previously demonstrated for pig tooth buds, rat tooth bud cells can be used to bioengineer complex tooth crowns, suggesting a common application for mammalian tooth tissue engineering. Furthermore, our demonstrated ability to use 4-dpn tooth bud cells that were first cultured <em>in vitro</em> for 6 days suggests that epithelial and mesenchymal dental stem cells (DSCs) giving rise to bioengineered tooth structures can be maintained in culture. The results of this study significantly advance current tooth-tissue-engineering efforts by demonstrating a general application to mammals, and suggest a potential means to propagate and expand DSCs in culture. </p>
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