In the absence of a basal lamina, ameloblasts absorb enamel in a serumless and chemically defined organ culture system.

OBJECTIVES: Tooth organ development was examined in a serumless, chemically defined organ culture system to determine whether morphological and functional development was identical to that in in vivo and serum-supplemented organ cultures. METHODS: Mouse mandibular first molar tooth organs at 16 days of gestation were cultured for up to 28 days in a Tronwell culture system using a serum-supplemented or serumless, chemically defined medium. After culture, specimens were processed for assessing tooth development using ultrastructural, immunohistochemical, and mRNA expression analyses. RESULTS: In serum-supplemented conditions, inner enamel epithelial cells differentiated into secretory-stage ameloblasts, which formed enamel and reached the maturation stage after 14 and 21 days of culture, respectively. Ameloblasts deposited a basal lamina on immature enamel. Conversely, in serumless conditions, ameloblasts formed enamel on mineralized dentin after 21 days. Moreover, maturation-stage ameloblasts did not form basal lamina and directly absorbed mineralized enamel after 28 days of culture. RT-PCR analysis indicated that tooth organs, cultured in serumless conditions for 28 days, had significantly reduced expression levels of ODAM, amelotin, and laminin-322. CONCLUSIONS: These results indicate that several differences were detected compared to the development in serum-supplemented conditions, such as delayed enamel and dentin formation and the failure of maturation-stage ameloblasts to form basal laminae. Therefore, our results suggest that some factors might be required for the steady formation of mineralized dentin, enamel, and a basal lamina. Additionally, our results indicate that a basal lamina is necessary for enamel maturation.

Resorption analysis of deproteinized cancellous bovine bone.

Commercially available deproteinized cancellous bovine bone (DPBB) has been indicated as non-absorbable bone filling materials. Stoichiometric hydroxyapatite (HA) was not resorbed by osteoclasts while calcium-deficient and carbonate-rich apatite converted from octacalcium (OCP hydrolysate) was resorbed by osteoclasts. We analyzed the chemical composition of DPBB and compared the tissue reactions around two materials after implanting into mouse bone marrow. X-ray diffraction analysis and Fourier transform infrared spectroscopy showed that DPBB was a carbonate-rich apatite. Micro-CT analysis indicated the massive bone formation on both materials at 2 weeks, then gradually resorbed. At 12 weeks, osteoclasts were directly attached to both materials. The ultrastructure of ruffled borders on DPBB was identical to osteoclasts resorbing normal bone while ruffled border on OCP hydrolysate showed irregular shape. These results indicated that DPBB was the absorbed material and that the structure of ruffled border of osteoclasts might be regulated by the size or orientation of HA.