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

Wu LC, Chiang CJ, Liu ZH, Tsuang YH, Sun JS, Huang YY - J Orthop Surg Res (2014)

Bottom Line: Biochemical analyses revealed 86% reduction in DNA, but only 15.9% reduction in glycosaminoglycan (GAG) content, with no significant difference in the hydroxyproline content.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.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Engineering, College of Engineering, College of Medicine, National Taiwan University, No.1, Sec.1, Jen-Ai Road, Taipei, Taiwan. lewis826@gmail.com.

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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Cytotoxicity studies of the control and decellularized anulus fibrosus (AF). Data revealed no statistically significant difference of the decellularized AF in comparison with the negative control. Control negative control, DAF decellularized AF.
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Fig6: Cytotoxicity studies of the control and decellularized anulus fibrosus (AF). Data revealed no statistically significant difference of the decellularized AF in comparison with the negative control. Control negative control, DAF decellularized AF.

Mentions: The in vitro toxicity studies of the control and decellularized AF were conducted by MTS assay with NIH3T3 cells. There was no statistically significant difference in in vitro cytotoxicity of the decellularized AF in comparison with the negative control (p > 0.05) (Figure 6).Figure 6


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

Wu LC, Chiang CJ, Liu ZH, Tsuang YH, Sun JS, Huang YY - J Orthop Surg Res (2014)

Cytotoxicity studies of the control and decellularized anulus fibrosus (AF). Data revealed no statistically significant difference of the decellularized AF in comparison with the negative control. Control negative control, DAF decellularized AF.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4264257&req=5

Fig6: Cytotoxicity studies of the control and decellularized anulus fibrosus (AF). Data revealed no statistically significant difference of the decellularized AF in comparison with the negative control. Control negative control, DAF decellularized AF.
Mentions: The in vitro toxicity studies of the control and decellularized AF were conducted by MTS assay with NIH3T3 cells. There was no statistically significant difference in in vitro cytotoxicity of the decellularized AF in comparison with the negative control (p > 0.05) (Figure 6).Figure 6

Bottom Line: Biochemical analyses revealed 86% reduction in DNA, but only 15.9% reduction in glycosaminoglycan (GAG) content, with no significant difference in the hydroxyproline content.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.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Engineering, College of Engineering, College of Medicine, National Taiwan University, No.1, Sec.1, Jen-Ai Road, Taipei, Taiwan. lewis826@gmail.com.

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.

Show MeSH
Related in: MedlinePlus