Establishment of in vitro threedimensional cementocyte differentiation scaffolds to study orthodontic root resorption

Orthodontic-induced root resorption is a severe side effect that can lead to tooth root shortening and loss. Compressive force induces tissue stress in the cementum that covers the tooth root, which is associated with activation of bone metabolism and cementum resorption. To investigate the role of cementocytes in mechanotransduction and osteoclast differentiation, the present study established an in vitro threedimensional (3D) model replicating cellular cementum and observed the effects of static compression on the cellular behavior of the cementocytes. Cell Counting Kit8 assay, alkaline phosphatase staining and dentin matrix protein 1 quantification were used to evaluate the cementocyte differentiation in the 3D scaffolds. Cellular viability under static compression was evaluated using live/dead staining, and expression of mineral metabolismrelated genes were analyzed via reverse transcriptionquantitative PCR. The results suggested that the cementocytes maintained their phenotype and increased the expression of osteoprotegerin (OPG), receptor activator of NFκB ligand (RANKL) and sclerostin (SOST) in the 3D model compared with cells cultured in two dimensions. Compression force increased cell death and induced osteoclastic differentiation via the upregulation of SOST and RANKL/OPG ratio, and the downregulation of osteocalcin. The effect of compression showed a force magnitudedependent pattern. The present study established an in vitro model of cellular cementum to study the biology of cementocytes. The results indicated that cementocytes are sensitive to mechanical loading and may serve potential roles in the metabolic regulation of minerals during orthodontic root resorption. These findings provide a novel tool to study biological processes in the field of orthodontics and expand knowledge of the biological function of cementocytes.