3, 3′, 5‐triiodo‐L‐thyronine Increases In Vitro Chondrogenesis of Mesenchymal Stem Cells From Human Umbilical Cord Stroma Through SRC2

Abstract

Our group focuses on the study of mesenchymal stem cells (MSCs) from human umbilical cord stroma or Warthońs jelly and their directed differentiation toward chondrocyte-like cells capable of regenerating damaged cartilage when transplanted into an injured joint. This study aimed to determine whether lactogenic hormone prolactin (PRL) or 3, 3', 5-triiodo-L-thyronine (T3), the active thyroid hormone, modulates chondrogenesis in our in vitro model of directed chondrogenic differentiation, and whether Wnt signalling is involved in this modulation. MSCs from human umbilical cord stroma underwent directed differentiation toward chondrocyte-like cells by spheroid formation. The addition of T3 to the chondrogenic medium increased the expression of genes linked to chondrogenesis like collagen type 2, integrin alpha 10 beta 1, and Sox9 measured by quantitative real time polymerase chain reaction (qRT-PCR) analysis. Levels of collagen type 2 and aggrecane analyzed by immunohistochemistry, and staining by Safranin O were increased after 14 days in spheroid culture with T3 compared to those without T3 or only with PRL. B-catenin, Frizzled, and GSK-3β gene expressions were significantly higher in spheroids cultured with chondrogenic medium (CM) plus T3 compared to CM alone after 14 days in culture. The increase of chondrogenic differentiation was inhibited when the cells were treated with T3 plus ML151, an inhibitor of the T3 steroid receptor. This work demonstrates, for first time, that T3 promotes differentiation towards chondrocytes-like cells in our in vitro model, that this differentiation is mediated by steroid receptor co-activator 2 (SRC2) and does not induce hypertrophy. J. Cell. Biochem. 117: 2097-2108, 2016. © 2016 Wiley Periodicals, Inc.

Dose-dependent effect of triiodothyronine on the chondrogenic differentiation of mesenchymal stem cells from the bone marrow of female rats

“…However, the expression of Col II did not change over time in any of the tested groups, demonstrating that the effect of T3 on Col II expression was only dose‐dependent. Another study also demonstrated the beneficial effect of T3 on the chondrogenic differentiation of human umbilical cord SCs, reporting that T3 at the dose of 100 ng/ml increased the expression of Col II at 14 days of differentiation . There are some differences in the study by Fernández‐Pernas et al ., in relation to the present research; the effect of T3 on chondrogenic differentiation was tested in human umbilical cord SCs at a dose much higher than that tested in the present study.…”

“…Another study also demonstrated the beneficial effect of T3 on the chondrogenic differentiation of human umbilical cord SCs, reporting that T3 at the dose of 100 ng/ml increased the expression of Col II at 14 days of differentiation . There are some differences in the study by Fernández‐Pernas et al ., in relation to the present research; the effect of T3 on chondrogenic differentiation was tested in human umbilical cord SCs at a dose much higher than that tested in the present study. Thus, we can suggest that the donor site, as well as the donor species and T3 dose, may influence the effect of thyroid hormones on the chondrogenic differentiation of MSCs.…”

“…Among the studied doses of T3, the dose of 1000 n m was the only one that influenced the expression of Sox9, and this effect occurred only at 21 days of differentiation. T3 (100 ng/ml) in human umbilical cord MSC culture increased Sox9 expression at 14 days of chondrogenic differentiation . However, when studying the effect of T3 doses on Sox9 expression over the time of differentiation, it was found that the dose of 0.01 n m T3 promoted peak expression at 7 days, and the dose of 1000 n m T3 gradually increased the expression of this gene during the period of differentiation, with its peak expression at 21 days.…”

“…T3 (100 ng/ml) in human umbilical cord MSC culture increased Sox9 expression at 14 days of chondrogenic differentiation. [59] However, when studying the effect of T3 doses on Sox9 expression over the time of differentiation, it was found that the dose of 0.01 nM T3 promoted peak expression at 7 days, and the dose of 1000 nM T3 gradually increased the expression of this gene during the period of differentiation, with its peak expression at 21 days. Among the considerable literature consulted, no study verified how the expression of Sox9 behaved throughout the period of chondrogenic differentiation of female rat BMMSCs.…”

Cytotoxic Properties of HT-2 Toxin in Human Chondrocytes: Could T3 Inhibit Toxicity of HT-2?

“…It has also been exploited to enhance cartilage formation and improve the functional properties of tissue-engineered neocartilage [3]. Furthermore, T 3 enhances chondrogenesis of mesenchymal stem cells of the umbilical cord [4]. In addition, T 3 regulates the transition between proliferation and terminal differentiation of chondrocytes in the growth plate via the Wnt/β-catenin signaling pathway [5,6].…”