Three-dimensional cultured periodontal ligament stem cells promote osteoclastic differentiation of RAW264.7 under static compression

doi:10.12307/2023.698

Abstract

Abstract: BACKGROUND: The osteogenic potential of periodontal ligament stem cells (as seed cells) has become a research hotspot. The regulation of periodontal ligament stem cells on osteoclast activity is less studied and deserves further exploration.
OBJECTIVE: To investigate the effect of three-dimensional cultured periodontal ligament stem cells under continuous static compression on the osteoclastic differentiation of RAW264.7 cells.
METHODS: Periodontal ligament stem cells at passages 3 and 4 were taken for three-dimensional culture and randomly divided into 13 groups. Continuous static compression of 5 kPa (2, 6, and 12 hours), 25 kPa (2, 6, and 12 hours), 45 kPa (2, 6, and 12 hours), and 65 kPa (2, 6, and 12 hours) was applied by FLEXCELL 5000C, and group 13 with no compression. The cell supernatant was used as a conditioned medium to act on RAW264.7 cells. The expression of osteoclast differentiation genes tartrate resistant acid phosphatase, cathepsin K, and matrix metalloproteinase 9 was detected by qPCR and tartrate resistant acid phosphatase staining after 5 days.
RESULTS AND CONCLUSION: (1) The scaffold morphology was stable and the cell status was good in the compression experimental group and the non-compression control group under three-dimensional culture. (2) The conditioned medium of periodontal ligament stem cells in the compression loading group promoted the osteoclastic differentiation of RAW264.7 cells. (3) Within a certain range, the pro-osteoclastic effect was positively correlated with the force conditions. At 45 kPa and 12 hours, RAW264.7 cells had the strongest osteoclastic differentiation effect. The osteoclast-promoting ability of 65 kPa and 12-hour group was weakened. (4) These findings indicated that the osteoclastic differentiation of RAW264.7 cells was promoted by three-dimensional cultured periodontal ligament stem cells under static compression through the paracrine pathway, and the force conditions were correlated with the osteoclast effect.

Key words: static compression, three-dimensional culture, periodontal ligament stem cell, RAW264.7, osteoclastic differentiation

CLC Number:

Han Xing, Li Wenwen, Ma Wensheng. Three-dimensional cultured periodontal ligament stem cells promote osteoclastic differentiation of RAW264.7 under static compression[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(24): 3810-3817.

Figures/Tables 4

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