Short Pretreatment with Calcitriol Is Far Superior to Continuous
Treatment in Stimulating Proliferation and Osteogenic
Differentiation of Human Adipose Stem Cells
The last two authors equally contributed to this work.
Mokhtari-Jafari F, Amoabediny G, Dehghan MM, Helder MN, Zandieh-Doulabi B, Klein-Nulend J. Short pretreatment with calcitriol Is far superior to continuous treatment in stimulating proliferation and osteogenic differentiation of human adipose stem cells. Cell J. 2020; 22(3): 293-301. doi: 10.22074/ cellj.2020.6773.
This study investigated whether short stimulation (30 minutes) of human adipose stem cells (hASCs) with 1,25-dihydroxyvitamin D3(calcitriol or 1,25-(OH)2VitD3), fitting within the surgical procedure time frame, suffices to induce osteogenic differentiation, and compared this with continuous treatment with 1,25-(OH)2VitD3.
Materials and Methods
In this experimental study, hASCs were pretreated with/without 10 nM calcitriol for 30 minutes, seeded on biphasic calcium phosphate (BCP), and cultured for 3 weeks with/without 1,25-(OH)2VitD3. Cell attachment was determined 30 minutes after cell seeding. AlamarBlue assay, alkaline phosphatase (ALP) assay, ALP staining, real-time polymerase chain reaction (PCR), and protein assay were used to evaluate the effect of short calcitriol pretreatment on proliferation and osteogenic differentiation of hASCs up to 3 weeks.
Pretreatment with 1,25-(OH)2VitD3enhanced the attachment of hASCs to BCP by 1.5-fold compared to non-
treated cells and increased the proliferation by 3.5-fold at day 14, and 2.6-fold at day 21. In contrast, continuous
treatment increased the proliferation by 1.7-fold only at day 14. After 2 weeks, ALP activity was increased by 18.5-fold
when hASCs were pretreated with 1,25-(OH)2VitD3for 30 minutes but increased only 2.6-fold when compared with its
continuous counterpart. Moreover, after 14 days, pretreatment resulted in significant upregulation of the osteogenic
This study is the first research showing that 30 minutes pretreatment of hASCs with 1,25-(OH)2VitD3, not only enhanced cell attachment to the scaffold at seeding time, but also promoted the proliferation and osteogenic differentiation of hASCs more strongly than continuous treatment, suggesting that short pre-treatment with 1,25-(OH)2VitD3is a promising approach for the regeneration of bones in a one-step surgical procedure.