Abstract
Introduction
Augmentation of spinal fusion using bone grafts is largely mediated by the osteoinductive potential of mesenchymal stem cells (MSC) that reside in cancellous bone. Iliac crest (IC) is a common autograft, but its use presents an increased risk for donor-site pain, morbidity and infection. Degenerative facet joints (FJ) harvested during facetectomy might servce as alternative local grafts. In this study, we conducted an intra-individual comparison of the osteogenic potential of MSC from both sources.
Methods
IC and degenerative FJ were harvested from 8 consecutive patients undergoing transforaminal lumbar interbody fusion surgery for spinal stenosis. MSC were isolated by collagenase digestion, selected by plastic adherence and minimally expanded for downstream assays. Clonogenic and osteogenic potential was evaluated by colony formation assays in control and osteogenic culture medium. Osteogenic properties, including alkaline phosphatase (ALP) induction, matrix mineralization and type I collagen mRNA and protein expression were characterized using quantitative histochemical staining and reverse transcription PCR. Spontaneous adipogenesis was analysed by adipocyte enumeration and gene expression analysis of adipogenic markers.
Results
Average colony-forming efficiency in osteogenic medium was equal between IC (38±12%) and FJ (36±11%). Osteogenic potential at the clonal level was 55±26 and 68±17% for IC and FJ MSC, respectively. Clonogenic and osteogenic potential were significantly negatively associated with donor age. Osteogenic differentiation led to significant induction of ALP activity in IC (6-fold) and FJ (8-fold) MSC. Matrix mineralization quantified by Alizarin red staining was increased by osteogenic differentiation, yet similar between both MSC sources. Protein expression of type I collagen was enhanced during osteogenesis and significantly greater in IC MSC. Correspondingly, COL1A2 mRNA expression was higher in osteogenically differentiated MSC from IC. Adipocyte numbers showed significant differences between IC (63±60) and FJ (18±15) MSC under osteogenic conditions. Negative (GREM1) and positive (FABP4) adipogenic markers were not differentially expressed between sources.
Conclusion
MSC from IC and degenerative FJ largely display similar clonogenic and osteogenic properties in vitro. Differences at the molecular level are not likely to impair the osteoinductive capacity of FJ MSC. Facetectomy samples are viable bone autografts for intervertebral spinal fusion.
