Study design: in vitro and in vivo assessment of bone morphogenic protein 2 combined with platelet-rich plasma on treatment of disc degeneration

Int Orthop. 2016 Jun;40(6):1143-55. doi: 10.1007/s00264-015-2840-5. Epub 2015 Jul 14.

Abstract

Objective: Our aim was to investigate the biological effects of bone morphogenic protein 2 (BMP2) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocyte-like cells in platelet-rich plasma (PRP) gel in vitro. In addition, the effectiveness of BMP2-transduced BMSCs in combined with PRP gel to repair the degenerated intervertebral disc in a rabbit model was also evaluated. Previous studies have shown that tissue engineering provides many promising advantages to treating disc degeneration and may reverse the pathological process of disc degeneration.

Methods: The expressions of types I, II and X collagen, aggrecan and Sox9 of the BMP2-transduced BMSCs in monolayer or PRP gel were examined by reverse transcriptase polymerase chain reaction (RT-PCR). Sixty New Zealand white rabbits were divided into five groups: 12 normal controls; 12 puncture operated with only disc degeneration being induced; 12 PRP transplantation animals; 12 BMSC and PRP-transplantation animals; 12 BMP2-transduced BMSCs and PRP-transplantation animals. The effect of BMP2-transduced BMSCs on degenerated discs were evaluated by magnetic resonance image (MRI) scan, histology, immunohistochemistry and Western blot analysis.

Results: BMP2 could facilitate chondrogenic differentiation of BMSCs in monolayer or PRP gel. The discs treated with BMP2-transduced BMSCs exhibited relatively well-preserved nucleus pulposus (NP) structure. Significantly higher T2-weighted signal intensity and a greater amount of extracellular matrix were observed in the BMP2-transduced BMSC group compared with other groups. In addition, the presences of BMP2-transduced BMSCs were identified at week 12 postoperatively in vivo.

Conclusions: BMP2-transduced BMSCs can maintain the chondrocyte-like phenotype in PRP gel in vitro, and the combined use of these two agents can significantly promote repair of the degenerated discs in vivo.

Keywords: BMP2; BMSCs; Cell transplantation; Disc degeneration; Nucleus pulposus; PRP.

MeSH terms

  • Aggrecans / metabolism
  • Animals
  • Blotting, Western
  • Bone Morphogenetic Protein 2 / pharmacology*
  • Cell Differentiation
  • Chondrocytes / metabolism
  • Collagen / metabolism
  • Immunohistochemistry
  • Intervertebral Disc / drug effects
  • Intervertebral Disc / pathology
  • Intervertebral Disc Degeneration / therapy*
  • Magnetic Resonance Imaging
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects*
  • Mesenchymal Stem Cells / metabolism
  • Platelet-Rich Plasma / drug effects*
  • Platelet-Rich Plasma / metabolism
  • Rabbits
  • Reverse Transcriptase Polymerase Chain Reaction
  • SOX9 Transcription Factor / metabolism

Substances

  • Aggrecans
  • BMP2 protein, human
  • Bone Morphogenetic Protein 2
  • SOX9 Transcription Factor
  • SOX9 protein, human
  • Collagen