Stem Cells From the Apical Papilla Cultured in Spheroids Spontaneously Differentiate Into Odontoblasts via N-Cadherin...

Traditional monolayer cultures of stem cells from the apical papilla (SCAP) fall short in their ability to replicate the anatomical organization of SCAP and often fail to provide high-quality SCAP suitable for dental root regeneration. This study aimed to employ agarose-based low-adhesion plates to induce the spontaneous formation of SCAP spheroids and disclose the biological impacts of SCAP in forming papilla-like structures in vivo. Compared with monolayer-cultured SCAP, spheroid-cultured SCAP exhibited enhanced in vitro and in vivo odontoblastic differentiation potentials. RNA sequencing identified the importance of N-cadherin downregulation in enhancing the odontoblastic differentiation potential of SCAP in spheroids compared with monolayer-cultured SCAP. Furthermore, N-cadherin knockdown in vitro significantly upregulated the β-catenin signaling pathway, further promoting SCAP odontoblastic differentiation. SCAP cultured on agarose-based plates spontaneously form spheroids and undergo odontoblastic differentiation through N-cadherin downregulation, offering new insights into SCAP-mediated dentin regeneration.