Fabrication and properties of acellular porcine anulus fibrosus for tissue engineering in spine surgery

Lien Chen Wu, Chang Jung Chiang, Zen Hao Liu, Yang Hwei Tsuang, Jui Sheng Sun, Yi You Huang

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish
Number of pages1
JournalJournal of Orthopaedic Surgery and Research
Volume9
DOIs
Publication statusPublished - Dec 3 2014

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Tissue Engineering
Spine
Swine
Biological Preservation
Elastic Modulus
Intervertebral Disc
Hydroxyproline
Hematoxylin
Eosine Yellowish-(YS)
Glycosaminoglycans
Sodium Dodecyl Sulfate
Annulus Fibrosus
Staining and Labeling
DNA
Therapeutics

ASJC Scopus subject areas

  • Surgery
  • Orthopedics and Sports Medicine

Cite this

Fabrication and properties of acellular porcine anulus fibrosus for tissue engineering in spine surgery. / Wu, Lien Chen; Chiang, Chang Jung; Liu, Zen Hao; Tsuang, Yang Hwei; Sun, Jui Sheng; Huang, Yi You.

In: Journal of Orthopaedic Surgery and Research, Vol. 9, 03.12.2014.

Research output: Contribution to journalArticle

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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.",
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AU - Wu, Lien Chen

AU - Chiang, Chang Jung

AU - Liu, Zen Hao

AU - Tsuang, Yang Hwei

AU - Sun, Jui Sheng

AU - Huang, Yi You

PY - 2014/12/3

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N2 - 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.

AB - 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.

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