Application of Green Tea Catechin for Inducing the Osteogenic Differentiation of Human Dedifferentiated Fat Cells in Vitro

Abstract

Despite advances in stem cell biology, there are few effective techniques to promote the osteogenic differentiation of human primary dedifferentiated fat (DFAT) cells. We attempted to investigate whether epigallocatechin-3-gallate (EGCG), the main component of green tea catechin, facilitates early osteogenic differentiation and mineralization on DFAT cells in vitro. DFAT cells were treated with EGCG (1.25–10 μM) in osteogenic medium (OM) with or without 100 nM dexamethasone (Dex) for 12 days (hereafter two osteogenic media were designated as OM(Dex) and OM). Supplementation of 1.25 μM EGCG to both the media effectively increased the mRNA expression of collagen 1 (COL1A1) and runt-related transcription factor 2 (RUNX2) and also increased proliferation and mineralization. Compared to OM(Dex) with EGCG, OM with EGCG induced earlier expression for COL1A1 and RUNX2 at day 1 and higher mineralization level at day 12. OM(Dex) with 10 μM EGCG remarkably hampered the proliferation of the DFAT cells. These results suggest that OM(without Dex) with EGCG might be a preferable medium to promote proliferation and to induce osteoblast differentiation of DFAT cells. Our findings provide an insight for the combinatory use of EGCG and DFAT cells for bone regeneration and stem cell-based therapy.

Dedifferentiated Fat (DFAT) cells: A cell source for oral and maxillofacial tissue engineering

“…After 7 days, fat cells have attached to the ceiling; the medium is now removed and the flask is re‐inverted (i.e., the ceiling now becomes the floor of the flask). Fresh medium is added to barely cover the bottom of the flask to allow cells to continue their expansion (Kaida, Honda, Hashimoto, Tanaka, & Baba, ; Kishimoto et al., ; Matsumoto et al., ; Okita et al., ; Sakamoto, Hashimoto, Kishimoto, Honda, & Matsumoto, ; Sakuma et al., ). Adipocytes that have attached to the inner surface of the flask during the ceiling culture divide asymmetrically and produce fibroblast‐like cells.…”

“…Jumabay and colleagues isolated human DFAT cells without using the ceiling culture method. Human DFAT cells differentiated into adipocytes (Matsumoto et al, 2008;Poloni et al, 2012;Sakuma et al, 2009;Watson et al, 2014), chondrocytes (Azumi, Honda, Kishimoto, Hashimoto, & Matsumoto, 2016;Jumabay et al, 2014;Matsumoto et al, 2008;Okita et al, 2015;Poloni et al, 2012;Sakuma et al, 2009), osteoblasts (Hao et al, 2017;Jumabay et al, 2014;Kaida et al, 2015;Matsumoto et al, 2008;Poloni et al, 2012;Sakamoto et al, 2015;Watson et al, 2014), smooth muscle-like cells (Sakuma et al, 2009), and neurogeniclineage cells (Poloni et al, 2012) under appropriate culture conditions in vitro.…”

Sphingosine-1-phosphate/S1PR2-mediated signaling triggers Smad1/5/8 phosphorylation and thereby induces Runx2 expression in osteoblasts

“…Cells were fixed with 3.7% paraformaldehyde in phosphate-buffered saline for 10 min, followed by a treatment with 50% acetone (Wako Pure Chemicals) in ethanol for 1 min. ALP activities were detected by ALP staining kit (Wako Pure chemicals) as described previously [41]. ALP stained areas were quantified and represented as the ratio to the control.…”

Osteogenesis of Multipotent Progenitor Cells using the Epigallocatechin Gallate-Modified Gelatin Sponge Scaffold in Rat Congenital Cleft-Jaw Model

“…DFAT cells differentiate into osteoblasts much earlier than bone marrowderived mesenchymal stem cells or ADSC [41,42]. Furthermore, our previous study proved that EGCG can induce the differentiation of human DFAT cells into osteoblastic cells in vitro [40]. Thus, taken together, these findings indicate that gelatin chemically modified with EGCG (vhEGCG-GS) holds the potential to promote bone formation, even in congenital bone defects, when used with ADSC and DFAT cells in vivo.…”

“…DFAT cells have augmented bone formation in created bone defect in alveolar bone [7,39]. These two cell types have been proved to differentiate into osteoblastic cells in vitro by the stimulation of growth factors and small molecules [38,40]. DFAT cells differentiate into osteoblasts much earlier than bone marrowderived mesenchymal stem cells or ADSC [41,42].…”