Abstract：
OBJECTIVE: The development of bio-three-dimensional (bio-3D) printers has led to significant advances in regenerative medicine. Three-dimensional constructs, including spheroids, are maintained by extracellular matrix proteins secreted by cells so that the cells can be cultured in conditions closer to the physiological environment. This study aimed to create a useful 3D construct as a model of the dentin-pulp complex.
METHODS: We examined the expression patterns of extracellular matrix proteins and cell proliferation areas in a 3D construct created using O9-1 cells derived from cranial neural crest cells of mice. The 3D construct was created by sticking the spheroid cultures onto a needle array using a bio-3D printer.
RESULTS: Cell proliferation areas along with characteristic expression of tenascin C and DMP1 were evaluated. The expression of tenascin C and DMP1 was significantly enhanced in the spheroids compared to that in two-dimensional cultures. Moreover, cell proliferation regions and tenascin C expression were confirmed in the outer layer of spheroids in the embryonic stem cell medium, with insignificant DMP1 expression being observed. Interestingly, in a 3D construct cultured in calcification-induction medium, DMP1 expression was promoted, and DMP1-positive cells existed in the outermost layer without overlapping with tenascin C expression.
CONCLUSIONS: The extracellular matrix proteins, tenascin C and DMP1, were expressed in a polarized manner in spheroids and 3D constructs, similar to the findings in the dental papilla. Therefore, these 3D constructs show potential as artificial models for studying odontogenesis.
METHODS: We examined the expression patterns of extracellular matrix proteins and cell proliferation areas in a 3D construct created using O9-1 cells derived from cranial neural crest cells of mice. The 3D construct was created by sticking the spheroid cultures onto a needle array using a bio-3D printer.
RESULTS: Cell proliferation areas along with characteristic expression of tenascin C and DMP1 were evaluated. The expression of tenascin C and DMP1 was significantly enhanced in the spheroids compared to that in two-dimensional cultures. Moreover, cell proliferation regions and tenascin C expression were confirmed in the outer layer of spheroids in the embryonic stem cell medium, with insignificant DMP1 expression being observed. Interestingly, in a 3D construct cultured in calcification-induction medium, DMP1 expression was promoted, and DMP1-positive cells existed in the outermost layer without overlapping with tenascin C expression.
CONCLUSIONS: The extracellular matrix proteins, tenascin C and DMP1, were expressed in a polarized manner in spheroids and 3D constructs, similar to the findings in the dental papilla. Therefore, these 3D constructs show potential as artificial models for studying odontogenesis.
摘要：
目的：生物三维（bio-3D）打印机的发展导致了再生医学的重大进展。三维结构，包括球体，由细胞分泌的细胞外基质蛋白维持，因此细胞可以在更接近生理环境的条件下培养。这项研究旨在创建一个有用的3D构建体作为牙本质-牙髓复合物的模型。方法：我们检查了使用源自小鼠颅神经c细胞的O9-1细胞创建的3D构建体中细胞外基质蛋白和细胞增殖区域的表达模式。通过使用生物3D打印机将球状体培养物粘贴到针阵列上来创建3D构建体。
结果：评估细胞增殖面积以及生腱蛋白C和DMP1的特征性表达。与二维培养相比，生腱蛋白C和DMP1在球状体中的表达显着增强。此外，在胚胎干细胞培养基中的球状体外层证实了细胞增殖区和生腱蛋白C的表达，观察到不显著的DMP1表达。有趣的是,在钙化诱导培养基中培养的3D构建体中，DMP1表达被促进，DMP1阳性细胞存在于最外层，与生腱蛋白C表达不重叠。
结论：细胞外基质蛋白，生腱蛋白C和DMP1在球状体和3D结构中以极化方式表达，类似于牙齿乳头的发现。因此,这些3D结构显示出作为研究牙本质发生的人工模型的潜力。
结果：评估细胞增殖面积以及生腱蛋白C和DMP1的特征性表达。与二维培养相比，生腱蛋白C和DMP1在球状体中的表达显着增强。此外，在胚胎干细胞培养基中的球状体外层证实了细胞增殖区和生腱蛋白C的表达，观察到不显著的DMP1表达。有趣的是,在钙化诱导培养基中培养的3D构建体中，DMP1表达被促进，DMP1阳性细胞存在于最外层，与生腱蛋白C表达不重叠。
结论：细胞外基质蛋白，生腱蛋白C和DMP1在球状体和3D结构中以极化方式表达，类似于牙齿乳头的发现。因此,这些3D结构显示出作为研究牙本质发生的人工模型的潜力。
