Fabrication and properties of acellular porcine anulus fibrosus for tissue engineering in spine surgery
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RESEARCH ARTICLE
Open Access
Fabrication and properties of acellular porcine anulus fibrosus for tissue engineering in spine surgery Lien-Chen Wu1,2, Chang-Jung Chiang2, Zen-Hao Liu1, Yang-Hwei Tsuang2, Jui-Sheng Sun3,4 and Yi-You Huang1*
Abstract Background: Over the last few years, new treatments for a damaged intervertebral disc (IVD) have included strategies to repair, replace, or regenerate the degenerative disc. However, these techniques are likely to have limited success, due to insufficiently effective means to address the damaged anulus fibrosus (AF). Here, we try to develop a bioprocess method for decellularization of the xenogeneic AF tissue, with a view to developing a scaffold as a potential candidate for clinical application in spinal surgery. Methods: Porcine AFs were decellularized using freeze-thaw cycles, followed by various combined treatments with 0.1% sodium dodecyl sulfate (SDS) and nucleases. Results: Hematoxylin and eosin (H & E) staining showed that decellularization was achieved through the decellularization protocols. Biochemical analyses revealed 86% reduction in DNA, but only 15.9% reduction in glycosaminoglycan (GAG) content, with no significant difference in the hydroxyproline content. There was no appreciable cytotoxicity of the acellular AF. Biomechanical testing of the acellular AF found no significant decline in stiffness or Young’s modulus. Conclusions: Porcine AF tissues were effectively decellularized with the preservation of biologic composition and mechanical properties. These results demonstrate that acellular AF scaffolds would be a potential candidate for clinical application in spinal surgery. Keywords: Anulus fibrosus, Decellularization, Acellular, Disc degeneration, Intervertebral disc, Tissue engineering
Introduction As the global population ages, degenerative disc disease (DDD) and lower back pain affect millions of people worldwide. Chronic back pain can manifest in relation to several clinical conditions, including disc herniation, radiculopathy, myelopathy, spinal stenosis, and instability. There is significant evidence of links between DDD and lower back pain although the pathogenesis of DDD has yet to be fully delineated. Although degeneration of the intervertebral disc (IVD) is associated with the majority of cases of lower back pain, current treatment options are palliative rather than curative. Although the prevalent surgical procedures can * Correspondence: [email protected] 1 Institute of Biomedical Engineering, College of Engineering, College of Medicine, National Taiwan University, No.1, Sec.1, Jen-Ai Road, Taipei, Taiwan Full list of author information is available at the end of the article
be successful in relieving back pain in the short term, but they do not repair the disc nor restore the normal biological and mechanical properties of the human spine. Such procedures limit mobility or may otherwise further alter the biomechanics of the spine, leading to further degeneration of adjacent segments. Discectomy is the most common surgical treatment o
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