Intervertebral disc response to stem cell treatment is conditioned by disc state and cell carrier: An ex vivo study

Abstract

SummaryIn vitro and in vivo studies evidenced that mesenchymal stem cells (MSCs) contribute to intervertebral disc (IVD) regeneration by differentiation towards the disc phenotype and matrix synthesis and/or by paracrine signalling to endogenous cells, thereby promoting a healthier disc phenotype in degenerative discs. The aim of this study was to investigate IVD response to human MSC (hMSC) treatment based on the disc degenerative state and hMSC carrier. Bovine caudal IVDs with endplates were cultured in a bioreactor under simulated physiological (0.1 Hz load and sufficient glucose) or degenerative (10 Hz load and limited glucose) conditions for 7 days. Discs were partially nucleotomised, restored with hMSCs in either fibrin gel or saline solution and cultured under physiological conditions for 7 days. Controls included fibrin and saline without hMSCs. Cell viability, histology, disc height, and gene expression analyses were performed to evaluate regeneration. hMSCs in fibrin were viable and homogenously distributed following 7 days of culture under dynamic loading in partially nucleotomised discs. IVD response to hMSCs was conditioned by both disc degenerative state and hMSC carrier. The effect of the regenerative treatment was stronger on simulated-degenerative discs than on simulated-physiological discs. hMSCs in fibrin induced a superior anabolic response in degenerative IVDs compared with fibrin alone, thus suggesting an added value of the cellular therapy compared with an acellular solution. When comparing fibrin and saline as a hMSC carrier, a significantly higher anabolic response was observed in IVDs treated with hMSCs in fibrin. Moreover, it was found that the degenerative state of the disc influenced hMSC differentiation. Indeed, a significantly higher expression of specific discogenic markers (ACAN and CA12) was observed in hMSCs implanted into physiological discs than in those implanted into degenerative discs. In conclusion, host disc cells and donor hMSC response depend on the degenerative state of the host disc and carrier used for hMSC delivery, and these two aspects need to be considered for a successful translation of hMSC therapies for the treatment of IVD degeneration.

An intervertebral disc whole organ culture system to investigate proinflammatory and degenerative disc disease condition

“…Approximately 150 mg of each AF and NP tissue was used for RNA extraction; 50 mg of each AF and NP tissue was digested with 0.5 mg/ml proteinase K at 56 °C overnight for glycosaminoglycan (GAG), collagen, and DNA content measurement. For RNA extraction, tissue samples were digested with 2 mg/ml pronase for 1 hr at 37 °C, flash frozen, pulverized in liquid nitrogen, and homogenized using a TissueLyser (Qiagen, Venlo, Netherlands; Lee et al, ; Peroglio et al, ). Total RNA was extracted with TRI Reagent (Molecular Research Center), and reverse transcription was performed with SuperScript VILO cDNA Synthesis Kit (Life Technologies, Carlsbad, CA).…”

Adipose Tissue Stem Cells for Therapy: An Update on the Progress of Isolation, Culture, Storage, and Clinical Application

“…For five years recently, most ASCs therapy has been studied in animal [100]. In human, ASCs showed the ability to promote cell proliferation and cell viability of nucleus pulposus cells in the intervertebral disc [101,102]. SVF, non-insolated ASCs from adipose tissue, can also improve the outcome in degenerative disc disease with no adverse effects [103].…”

Intervertebral Disc Nucleus Repair: Hype or Hope?

“…Concomitantly, Abbott et al proposed NP cells do possess regeneration potential even in severe status of degeneration [87]. Several cell-based strategies have been investigated to retard NP degeneration, using different IVD model system [88,89,90,91], with NP and/or AF cells, articular chondrocytes, or mesenchymal stem/stromal cells (MSCs). Feng et al summarized GDF-5 a suitable growth factor for inducing NP-like cells based on its positive effect on the differentiation of MSCs towards an NP-like phenotype [92].…”