Effect of platelet mediator concentrate (PMC) on Achilles tenocytes: an in vitro study

BMC Musculoskelet Disord. 2016 Jul 22:17:307. doi: 10.1186/s12891-016-1160-2.

Abstract

Background: Although there are many studies discussing the etiological and pathological factors leading to both, acute and chronic tendon injuries, the pathophysiology of tendon injuries is still not clearly understood. Although most lesions are uncomplicated, treatment is long and unsatisfactory due to the poor vascularity of tendon tissue. Platelet mediator concentrate (PMC) contains many growth factors derived from platelets, which can promote wound healing. In this study we investigate the effects of PMC on tenocyte proliferation and differentiation in order to provide an experimental basis for tissue regeneration strategies and to develop new treatment concepts.

Methods: Using enzyme linked immunosorbent assay (ELISA) we were able to quantify the several growth factors and cytokines found in PMC. Tenocytes were isolated both from human and from mouse Achilles tendons and stimulated with PMC. CyQuant® and Cell Titer Blue® assays were carried out to analyze tendon growth and viability at different concentrations of PMC. Real time RT-PCR was used to analyze tenocyte gene expression with or without PMC treatment. Immunohistochemistry was carried out to detect the tenocyte-specific antibody tenomodulin (TNMD) and scleraxis (SCX).

Results: We were able to detect numerous mediators such as platelet derived growth factor BB (PDGF-BB), interleukin 6 (IL-6), vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF-α), transforming growth factor beta 1 (TGF-ß1), and bone morphogenetic proteins 2, 4 and 7 (BMP-4, BMP-2, BMP-7) in PMC. It was possible to show a positive effect of PMC on human tendon cell growth and viability in a dose-dependent manner. Furthermore, PMC treatment led to induction of gene expression of scleraxis (SCX), type I collagen A 1 (Col1A1) and TNMD by tenocytes.

Conclusions: We suggest that the use of autologous PMC may be a suitable addition to conventional tendon therapy that is capable of increasing and optimizing tendon healing and reducing the risk of recurrence.

Keywords: Platelet; Platelet mediator concentrate; Scleraxis; Tenocyte; Tenomodulin.

MeSH terms

  • Achilles Tendon / cytology
  • Adolescent
  • Adult
  • Angiogenesis Inducing Agents / therapeutic use*
  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Becaplermin
  • Blood Platelets / metabolism*
  • Bone Morphogenetic Proteins
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Collagen Type I / metabolism
  • Collagen Type I, alpha 1 Chain
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Gene Expression Profiling
  • Healthy Volunteers
  • Humans
  • Immunohistochemistry
  • Male
  • Membrane Proteins / metabolism
  • Mice
  • Middle Aged
  • Proto-Oncogene Proteins c-sis / therapeutic use
  • Regeneration / drug effects
  • Tendon Injuries / drug therapy*
  • Tenocytes / drug effects*
  • Tenocytes / metabolism
  • Tenocytes / physiology
  • Vascular Endothelial Growth Factor A
  • Wound Healing / drug effects*
  • Young Adult

Substances

  • Angiogenesis Inducing Agents
  • Basic Helix-Loop-Helix Transcription Factors
  • Bone Morphogenetic Proteins
  • Collagen Type I
  • Collagen Type I, alpha 1 Chain
  • Membrane Proteins
  • Proto-Oncogene Proteins c-sis
  • SCX protein, human
  • TNMD protein, human
  • Vascular Endothelial Growth Factor A
  • Becaplermin