The Journal of Bone and Joint Surgery (American). 2010;92:675-685.
doi:10.2106/JBJS.H.01672
© 2010 The Journal of Bone and Joint Surgery, Inc.
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Multipotential Differentiation of Human Anulus Fibrosus Cells

An in Vitro Study

Gang Feng, MD, PhD1, Xinlin Yang, PhD2, Hulan Shang, BS2, Ian W. Marks, BS, PA2, Francis H. Shen, MD2, Adam Katz, MD2, Vincent Arlet, MD2, Cato T. Laurencin, MD, PhD3 and Xudong Li, MD, PhD2

1 Department of Orthopaedic Surgery, Nanchong Central Hospital, North Sichuan Medical College, Nanchong 637000, P.R. China
2 Departments of Orthopaedic Surgery (X.Y., I.W.M., F.H.S., V.A., and X.L.) and Plastic Surgery (H.S. and A.K.), University of Virginia School of Medicine, P.O. Box 800374, Charlottesville, VA 22908. E-mail address for X. Li: XL2N{at}virginia.edu
3 Departments of Orthopaedic Surgery, and Chemical, Materials and Biomolecular Engineering, The University of Connecticut, Farmington, CT 06032

Investigation performed at the Orthopaedic Research Laboratories, University of Virginia School of Medicine, Charlottesville, Virginia

Disclosure: The authors did not receive any outside funding or grants in support of their research for or preparation of this work. Neither they nor a member of their immediate families received payments or other benefits or a commitment or agreement to provide such benefits from a commercial entity.

Background The existence of fibrocartilage, bone-like tissues, nerves, and blood vessels in the anulus fibrosus during intervertebral disc degeneration has been well documented. Migration of differentiated cells from outside the intervertebral disc has been hypothesized as a possible mechanism for the formation of these tissues. We hypothesized that the normal anulus fibrosus tissue contains multipotent progenitor cells, which are able to differentiate into cartilage and/or fibrocartilage cells, osteoblasts, neurons, and blood vessel cells.

Methods We isolated anulus fibrosus cells from the nondegenerative intervertebral discs of adolescent (thirteen to sixteen-year-old) patients with idiopathic scoliosis and cultured the cells in vitro in induction media containing different stimuli. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Expression of markers of adipogenesis, osteogenesis, chondrogenesis, neurogenesis, and differentiation into endothelial lineages was determined with use of immunostaining, cytohistological staining, and reverse transcription-polymerase chain reaction.

Results Anulus fibrosus cells expressed several of the cell surface antigens that are sometimes associated with mesenchymal stem cells, including CD29, CD49e, CD51, CD73, CD90, CD105, CD166, CD184, and Stro-1, and two neuronal stem cell markers, nestin and neuron-specific enolase. Furthermore, varying the stimulants added to the induction media determined whether anulus fibrosus cells differentiated into adipocytes, osteoblasts, chondrocytes, neurons, or endothelial cells.

Conclusions Anulus fibrosus cells isolated from nondegenerative intervertebral discs can differentiate into adipocytes, osteoblasts, chondrocytes, neurons, and endothelial cells in vitro.

Clinical Relevance Our results, by offering new insights into the biology of anulus fibrosus cells, may assist in future strategies to treat intervertebral disc diseases.


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