Effects of hair keratin on cell cycle regulation of human periodontal ligament cells in histidine-tryptophan-ketoglutarate-based storage media

In this study, the effect of a storage medium (hK-HTCM) in which hair keratin was dissolved in a 1:1 mixed solution of Histidine-Tryptophan-Ketoglutarate and Culture media solution (HTCM) was evaluated on the viability and proliferation of human periodontal ligament cells. There was no difference in cytotoxicity between 0.1% and 0.25% hK-HTCM against 0% hK-HTCM and human periodontal ligament cells. Human periodontal ligament cells were cultured in 0.1% and 0.25% hK-HTCM for 48 hours, and after removing hair keratin from the culture medium, the cells resumed proliferation. When exposed to 0.25% hK-HTCM, human periodontal ligament cells showed differential expression of genes related to cell cycle and cell division regulation. On the other hand, differential expression of genes related to phosphorylation and ubiquitination related to cell cycle resumption was observed in human periodontal ligament cells after removal of 0.25% hK-HTCM. 0.25% hK-HTCM showed the ability to regulate the cell cycle of human periodontal ligament cells without showing cytotoxicity, and its potential to be used as a long-term storage medium for avulsed teeth was confirmed. Quantseq 3' mRNA sequencing was performed on cells before exposure to optimal hK-HTCM, 48 hours after exposure, and 48 hours after washing and medium exchange to identify differentially expressed genes and use bioinformatics methods to identify differentially expressed genes. were analyzed to interpret the biological changes that occurred.