Effect of perfluorotributylamine-enriched alginate on nucleus pulposus cell: Implications for intervertebral disc regeneration.

Various scaffolds have been attempted for intervertebral disc regeneration, but their effectiveness was limited by loss of nutrients within the scaffolds. It has been suggested that the disc is not severely hypoxic and limited availability of oxygen results in disc degeneration. Therefore, a certain oxygen level might be beneficial for disc regeneration, which has not been given enough attention in previous studies. Here, we used perfluorotributylamine (PFTBA) for the first time as an oxygen regulator in alginate scaffold for disc regeneration in vitro and in vivo. We found that the characteristics of alginate were not affected by PFTBA and the oxygen level of the scaffold was regulated. Then, human nucleus pulposus (NP) cells were cultured in the PFTBA-enriched alginates. It was found that PFTBA could promote NP cell survival and proliferation. In addition, 2.5% PFTBA was capable of regulating extracellular matrix (ECM) to a disc-like tissue graft with little effect on the expression of NP cell markers. Finally, 2.5% PFTBA-enriched alginate was found to restore the disc height and the ECM in a mouse disc degeneration model, indicating its beneficial effect on alleviating disc degeneration. These findings highlight the promising application of PFTBA in further intervertebral disc regeneration.

[1]  Justin J Cooper-White,et al.  An injectable hydrogel incorporating mesenchymal precursor cells and pentosan polysulphate for intervertebral disc regeneration. , 2013, Biomaterials.

[2]  S. Sigrist,et al.  Perfluorocarbon Emulsions Prevent Hypoxia of Pancreatic β-Cells , 2012, Cell transplantation.

[3]  I. Shapiro,et al.  Hypoxic regulation of nucleus pulposus cell survival: from niche to notch. , 2010, The American journal of pathology.

[4]  S. M. Naqvi,et al.  Differential response of encapsulated nucleus pulposus and bone marrow stem cells in isolation and coculture in alginate and chitosan hydrogels. , 2015, Tissue engineering. Part A.

[5]  S. Richardson,et al.  Understanding the native nucleus pulposus cell phenotype has important implications for intervertebral disc regeneration strategies. , 2013, Regenerative medicine.

[6]  Fan Yang,et al.  Mesenchymal stem cells arrest intervertebral disc degeneration through chondrocytic differentiation and stimulation of endogenous cells. , 2009, Molecular therapy : the journal of the American Society of Gene Therapy.

[7]  Yongqiang Chen,et al.  Starvation-induced autophagy is regulated by mitochondrial reactive oxygen species leading to AMPK activation. , 2013, Cellular signalling.

[8]  W. Tawackoli,et al.  The use of a synthetic oxygen carrier-enriched hydrogel to enhance mesenchymal stem cell-based bone formation in vivo. , 2009, Biomaterials.

[9]  J. Kadlowec,et al.  Thermogelling bioadhesive scaffolds for intervertebral disk tissue engineering: preliminary in vitro comparison of aldehyde-based versus alginate microparticle-mediated adhesion. , 2015, Acta biomaterialia.

[10]  R. Fessler,et al.  Molecular Biology of Degenerative Disc Disease , 2000, Neurosurgery.

[11]  R. G. Richards,et al.  A combined biomaterial and cellular approach for annulus fibrosus rupture repair. , 2015, Biomaterials.

[12]  F. Qi,et al.  A synthetic oxygen carrier in fibrin matrices promotes sciatic nerve regeneration in rats. , 2013, Acta biomaterialia.

[13]  S. Roberts,et al.  The cartilage end‐plate and intervertebral disc in scoliosis: Calcification and other sequelae , 1993, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[14]  Yin-gang Zhang,et al.  Animal models of disc degeneration and major genetic strategies. , 2013, Pain physician.

[15]  M. Lamy,et al.  Effects of perfluorocarbon emulsions on cultured human endothelial cells. , 1997, Artificial cells, blood substitutes, and immobilization biotechnology.

[16]  J. Riess.  Perfluorocarbon-based Oxygen Delivery , 2006, Artificial cells, blood substitutes, and immobilization biotechnology.

[17]  Wen-Jer Chen,et al.  Effect of Hyperbaric Oxygenation on Intervertebral Disc Degeneration: An In Vivo Study With Sprague-Dawley Rats , 2013, Spine.

[18]  Xuanyi Ma,et al.  Photochemically crosslinked collagen annulus plug: a potential solution solving the leakage problem of cell-based therapies for disc degeneration. , 2013, Acta biomaterialia.

[19]  Xinyan Tang,et al.  Changes in the Molecular Phenotype of Nucleus Pulposus Cells with Intervertebral Disc Aging , 2012, PloS one.

[20]  Damien Lacroix,et al.  The Effect of Sustained Compression on Oxygen Metabolic Transport in the Intervertebral Disc Decreases with Degenerative Changes , 2011, PLoS Comput. Biol..

[21]  T. Steffen,et al.  Effect of Oxygen Levels on Proteoglycan Synthesis by Intervertebral Disc Cells , 2011, Spine.

[22]  Li Li,et al.  Hypoxia promotes nucleus pulposus phenotype in 3D scaffolds in vitro and in vivo: laboratory investigation. , 2014, Journal of neurosurgery. Spine.

[23]  Zhen Sun,et al.  Impact of direct cell co‐cultures on human adipose‐derived stromal cells and nucleus pulposus cells , 2013, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[24]  A. Penttilä,et al.  Lumbar Disc Degeneration and Atherosclerosis of the Abdominal Aorta , 1994, Spine.

[25]  George M. Wahba,et al.  Intervertebral Disc Cell Therapy for Regeneration: Mesenchymal Stem Cell Implantation in Rat Intervertebral Discs , 2004, Annals of Biomedical Engineering.

[26]  ZhouYue,et al.  Utilization of stem cells in alginate for nucleus pulposus tissue engineering. , 2014 .

[27]  E. Wachtel,et al.  Structure, function, aging and turnover of aggrecan in the intervertebral disc. , 2014, Biochimica et biophysica acta.

[28]  A. Turgut,et al.  Pineal gland calcification, lumbar intervertebral disc degeneration and abdominal aorta calcifying atherosclerosis correlate in low back pain subjects: A cross-sectional observational CT study. , 2008, Pathophysiology : the official journal of the International Society for Pathophysiology.

[29]  T. Ma,et al.  The effect of synthetic oxygen carrier-enriched fibrin hydrogel on Schwann cells under hypoxia condition in vitro. , 2013, Biomaterials.

[30]  H. Matsuzaki,et al.  Alteration of Gene Expression in Intervertebral Disc Degeneration of Passive Cigarette- Smoking Rats: Separate Quantitation in Separated Nucleus pulposus and Annulus fibrosus , 2005, Pathobiology.

[31]  L. Setton,et al.  Screening of hyaluronic acid-poly(ethylene glycol) composite hydrogels to support intervertebral disc cell biosynthesis using artificial neural network analysis. , 2014, Acta biomaterialia.

[32]  Benjamin Gantenbein,et al.  Activation of intervertebral disc cells by co-culture with notochordal cells, conditioned medium and hypoxia , 2014, BMC Musculoskeletal Disorders.

[33]  G. Waddell.  Low Back Pain: A Twentieth Century Health Care Enigma , 1996, Spine.

[34]  Vincenzo Denaro,et al.  Mesenchymal stem cells injection in degenerated intervertebral disc: cell leakage may induce osteophyte formation , 2012, Journal of tissue engineering and regenerative medicine.

[35]  L. Lescaudron,et al.  The lumbar intervertebral disc: from embryonic development to degeneration. , 2014, Joint, bone, spine : revue du rhumatisme.

[36]  T. Ma,et al.  A synthetic oxygen carrier-olfactory ensheathing cell composition system for the promotion of sciatic nerve regeneration. , 2014, Biomaterials.

[37]  Sally Roberts,et al.  Histology and pathology of the human intervertebral disc. , 2006, The Journal of bone and joint surgery. American volume.

[38]  K. Luk,et al.  Injury‐induced sequential transformation of notochordal nucleus pulposus to chondrogenic and fibrocartilaginous phenotype in the mouse , 2009, The Journal of pathology.

[39]  Lei-Sheng Jiang,et al.  Hypoxia facilitates the survival of nucleus pulposus cells in serum deprivation by down-regulating excessive autophagy through restricting ROS generation. , 2015, The international journal of biochemistry & cell biology.

[40]  A Shirazi-Adl,et al.  Computation of coupled diffusion of oxygen, glucose and lactic acid in an intervertebral disc. , 2007, Journal of biomechanics.

[41]  K. Lowe,et al.  Perfluorochemicals: their applications and benefits to cell culture. , 1998, Trends in biotechnology.

[42]  A. Nachemson,et al.  In vitro diffusion of dye through the end-plates and the annulus fibrosus of human lumbar inter-vertebral discs. , 1970, Acta orthopaedica Scandinavica.

[43]  S. Nicoll,et al.  Photo-crosslinked alginate hydrogels support enhanced matrix accumulation by nucleus pulposus cells in vivo. , 2009, Osteoarthritis and cartilage.

[44]  Hamid Bahramian Renani,et al.  Determination and comparison of specifics of nucleus pulposus cells of human intervertebral disc in alginate and chitosan–gelatin scaffolds , 2012, Advanced biomedical research.

[45]  J. Urban,et al.  Intervertebral disc regeneration: do nutrients lead the way? , 2014, Nature Reviews Rheumatology.

[46]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[47]  J. Urban,et al.  Effects of low oxygen concentrations and metabolic inhibitors on proteoglycan and protein synthesis rates in the intervertebral disc , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[48]  P. Ma,et al.  Hypoxia differentially regulates human nucleus pulposus and annulus fibrosus cell extracellular matrix production in 3D scaffolds. , 2013, Osteoarthritis and cartilage.

[49]  S Holm,et al.  Nutrition of the intervertebral disc: solute transport and metabolism. , 1981, Connective tissue research.

[50]  F. Lin,et al.  Differential phenotypic behaviors of human degenerative nucleus pulposus cells under normoxic and hypoxic conditions: influence of oxygen concentration during isolation, expansion, and cultivation. , 2013, The spine journal : official journal of the North American Spine Society.

[51]  C. Koch,et al.  In situ oxygen utilization in the rat intervertebral disc , 2007, Journal of anatomy.

[52]  C. Niu,et al.  Hyperbaric oxygen treatment suppresses MAPK signaling and mitochondrial apoptotic pathway in degenerated human intervertebral disc cells , 2013, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.