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EP143 Regeneration potency of tendon derived stem cell in tedinopathy can be suppressed by pain mediators: in vitro study
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  1. Yong-Taek Lee1 and
  2. Min-Jeong Kim2
  1. 1Physical Medicine and Rehabilitation, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, Korea
  2. 2Medical Research Institue, Kangbuk Samsung Hostpital, Sungkyunkwan University School of Medicine, Seoul, Korea

Abstract

Background and Aims Tendon-derived stem cells (TDSCs) in tendons are responsible for tenogenesis and tendon regeneration. Aberrant nontenogenic differentiation of TDSCs, such as chondrogenic metaplasia, have been suggested as a pathogenesis of tendinopathy. Additionally, pain mediators, such as substance P, calcitonin gene-related peptide (CGRP) and macrophage migration inhibitory factor (MIF), have been increasingly discussed as an important factor in the pathogenesis of tendinopathy. The purpose was to evaluate whether the pain mediator affects differentiation of TDSC.

Methods TDSC was isolated and cultured from the Achilles tendon of SD rats. TDSC were treated with recombinant MIF, recombinant substance P, or recombinant CGRP. For gene knockdown, TDSC were transfected with MIF small interfering RNA (siRNA), substance P siRNA, or CGRP siRNA. The TDSC culture mediums were prepared for RT-PCR. Expression of tenogenic genes (SCX, Egr1, Tnmd, Col type 1) and chondrogenic genes (BMP2, aggrecan, Sox9) of TDSC were compared with control group.

Results Treatment of recombinant pain mediators (MIF, Substance P or CGRP) in TDSC showed down-regulated tenogenic genes expression (figure 1A, 2A, 3A) and up-regulated chondrogenic genes expression (Fig 1B, 2B, 3B) compared with control (p<.05). Knockdown of pain mediator genes (MIF, Substance P or CGRP) in TDSC showed down-regulated chondrogenic gene expression (figure 1B, 2B, 3B) while expression was up-regulated in a few tenogenic gene (Col type 1 with MIF and SCX with Substance P).

Abstract EP143 Figure 1

A. The tenogenic Mrna expression levels of TDSC in recombinant MIF and knockdown of MIF. (*P <0.05 vs. control) B. The chondrogenic mRNA expression levels of TDSC in recombinant MIF and knockdown of MIF. (*P <0.05 vs. control)

Abstract EP143 Figure 2

A. The tenogenic mRNA expression levels of TDSC in recombinant Substance P and knockdown of Substance P. (*P <0.05 vs. control) B. The chondrogenic mRNA expression levels of TDSC in recombinant CGRP and knockdown of CGRP. (*P <0.05 vs. control)

Abstract EP143 Figure 3

A. The tenogenic mRNA expression levels of TDSC in recombinant CGRP and knockdown of CGRP. (*P <0.05 vs. control) B. The chondrogenic mRNA expression levels of TDSC in recombinant CGRP and knockdown of CGRP. (*P <0.05 vs. control)

Conclusions Pain mediators, such as Substance P, CGRP and MIF, appear to be associated with pathogenesis of tendinopathy via enhance the aberrant chondrogenic differentiation and suppression of tenogenic differentiation of TDSC.

[18-100-D3-N] Protocol Approval of Animal Study Plan

  • Tendon Derived Stem Cell
  • Pain Mediator
  • Tendinopathy

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