Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (31): 4976-4980.doi: 10.12307/2021.139

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Effect of hydrostatic pressure on odontogenic/osteogenic differentiation of dental pulp stem cells

Li Junqing1, 2, He Wenxi2, Guo Qian2, Wu Jiayuan1, 3   

  1. 1Zunyi Medical University, Special Key Laboratory of Oral Diseases Research of Universities in Guizhou Province, Zunyi 563000, Guizhou Province, China; 2State Key Laboratory of Military Stomatology, Key Laboratory of Shaanxi Province Stomatology, Department of Dental Pulp Diseases, The Third Affiliated Hospital of Air Force Military Medical University, Xi’an 710032, Shaanxi Province, China; 3Stomatological Hospital Affiliated to Zunyi Medical University, Zunyi 563000, Guizhou Province, China
  • Received:2020-10-26 Revised:2020-10-29 Accepted:2020-12-07 Online:2021-11-08 Published:2021-04-25
  • Contact: Wu Jiayuan, MD, Professor, Zunyi Medical University, Special Key Laboratory of Oral Diseases Research of Universities in Guizhou Province, Zunyi 563000, Guizhou Province, China; Stomatological Hospital Affiliated to Zunyi Medical University, Zunyi 563000, Guizhou Province, China
  • About author:Li Junqing, Master candidate, Zunyi Medical University, Special Key Laboratory of Oral Diseases Research of Universities in Guizhou Province, Zunyi 563000,
  • Supported by:
    the National Natural Science Foundation of China, No. 81771060 (to HWX), No. 81970932 (to HWX)

Abstract:

BACKGROUND: Besides biochemical factors, cellular mechanics is increasingly recognized as the key factor affecting the behavior and function of stem cells. 

OBJECTIVE: To explore the effect of hydrostatic pressure on the odontogenic/osteogenic differentiation potential of human dental pulp stem cells. 
METHODS:  Human dental pulp stem cells were isolated and extracted by enzyme digestion. The expression of cell surface markers was detected by flow cytometry. After osteogenic induction for 14 days or adipogenic induction for 21 days, alizarin red or oil red O staining was used to identify the mutidirectional differentiation of human dental pulp stem cells. The human dental pulp stem cells were loaded with hydrostatic pressure of different pressure values (0, 30, 60, 90, 120, and 150 kPa) by using the self-developed mechanical loading device, for 1 hour. At 1, 3, 5, and 7 days, the culture solution was discarded. CCK-8 assay was used to observe the proliferation of human dental pulp stem cells, 1 hour per day, for 14 consecutive days. Alizarin red staining was applied to observe the osteogenic and mineralized abilities. Western blot assay was used to detect the protein expression levels of odontogenic/osteogenic differentiation related genes.   
RESULTS AND CONCLUSION: Dental pulp stem cells expressed high levels of CD29, CD90, CD146, CD105 and low levels of CD45 and CD34, and had the ability of osteogenic and adipogenic differentiation. Hydrostatic pressure could promote the proliferation of human dental pulp stem cells at 90 and 120 kPa, but the proliferation ability could be significantly inhibited when the pressure value was 150 kPa. After 14 days of hydrostatic pressure stimulation, the protein expression levels of dentin sialophosphoprotein, collagen type I, and osteocalcin in dental pulp stem cells were significantly lower than those in the control group (P < 0.05), and their ability to inhibit differentiation became more obvious with the increase of hydrostatic pressure. The results show that hydrostatic pressure is involved in the process of dental pulp stem cell odontogenesis/osteogenesis and has a negative regulatory effect.

Key words: stem cells, dental pulp stem cells , hydrostatic pressure, static pressure, cellular mechanics, cell differentiation

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