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A minimally invasive rabbit model of progressive and reproducible disc degeneration confirmed by radiology, gene expression, and histology.

Kwon YJ - J Korean Neurosurg Soc (2013)

Bottom Line: Significant disc space narrowing compared to preoperative disc height was observed during the time period (p<0.001).The MRI grade, aggrecan, and matrix metalloprotease-13 mRNA expression and hematoxylin and eosin/safranin O/anti-collagen II staining were consistently indicative of degeneration, supporting the results of the X-ray data.This in vivo model can be used to study and evaluate the safety and efficacy of biologic treatments for degenerative disc disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Sungkyunkwan University School of Medicine, Kangbuk Samsung Hospital, Seoul, Korea.

ABSTRACT

Objective: To develop a simple, reproducible model of disc degeneration in rabbits through percutaneous annular puncture and to confirm the degree of degeneration over time.

Methods: Fifteen New Zealand white rabbits (4 to 5 months old and weighing approximately 3 to 3.5 kg each) underwent annular puncture of the L2-L3, L3-L4, and L4-L5 discs. Rabbits were sacrificed at 4, 8, or 20 weeks after puncture. For a longitudinal study to assess changes in disc height over time, serial X-rays were performed at 0, 2, 4, 8, and 20 weeks for rabbits in the 20-week group. Upon sacrifice, the whole spinal column and discs were extracted and analyzed with magnetic resonance imaging (MRI), real time reverse transcriptase-polymerase chain reaction, and histological staining.

Results: The X-rays showed a slow, progressive decrease in disc height over time. Significant disc space narrowing compared to preoperative disc height was observed during the time period (p<0.001). The MRI grade, aggrecan, and matrix metalloprotease-13 mRNA expression and hematoxylin and eosin/safranin O/anti-collagen II staining were consistently indicative of degeneration, supporting the results of the X-ray data.

Conclusion: Percutaneous annular puncture resulted in slow, reproducible disc degeneration that was confirmed by radiology, biochemistry, and histology. This in vivo model can be used to study and evaluate the safety and efficacy of biologic treatments for degenerative disc disease.

No MeSH data available.


Related in: MedlinePlus

Percutaneous needle puncture into the rabbit discs. A : An angiography needle is inserted 3-4 cm ventral from the midline into the disc space using a fluoroscope. B : Constant amount of nucleus material is checked during the puncture to create even disc degeneration.
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Figure 1: Percutaneous needle puncture into the rabbit discs. A : An angiography needle is inserted 3-4 cm ventral from the midline into the disc space using a fluoroscope. B : Constant amount of nucleus material is checked during the puncture to create even disc degeneration.

Mentions: Each rabbit was anesthetized with intramuscular injection of xylazine (5 mg/kg) and ketamine (35 mg/kg), and the fur was shaved from the mid back and right flank. After anesthesia, a lateral plain X-ray was obtained to establish the pre-injection baseline height of the IVDs. The rabbit was then placed in the lateral oblique prone position, and the injection field was sterilized with an alcohol sponge. Initially, the L5-L6 disc was identified through manual palpation of the interspinous space from the mid back and pelvic rim with fluoroscopy (VPX-200; Toshiba Corporation, Tokyo, Japan). After confirmation of the exact level, an 18-gauge angiography needle was inserted 3-4 cm ventral from the midline into the disc space under fluoroscopic control (Fig. 1A). After brief confirmation of the needle position in the center of the disc space by anteroposterior and lateral fluoroscopy, the needle was held in the disc space for 10 seconds. Before removal of the punctured needle, the needle was rotated 180 degrees. A constant amount of nucleus was leaked within the needle and identified by pushing the needle with an empty syringe after removal of the stylet (Fig. 1B). In each rabbit, each of three discs (L2-L3, L3-L4, and L4-L5) was punctured. The L1-L2 and L5-L6 levels were designated as the non-punctured, internal controls. For each level, all procedures for identification and puncture were performed within 20 seconds. Special care was taken to minimize contact with the periosteal tissues of the vertebrae because this could cause hypertrophy of the soft tissues and bony structures around the discs. No antibiotics and analgesic were used after puncture. No rabbits showed neurological symptoms, and all tolerated the procedure well. The rabbits were placed in their cages after observation for recovery. At the planned time, they were sacrificed with intramuscular injection of ketamine (25.0 mg/kg) followed by intravenous injection of sodium pentobarbital (1.2 g/kg).


A minimally invasive rabbit model of progressive and reproducible disc degeneration confirmed by radiology, gene expression, and histology.

Kwon YJ - J Korean Neurosurg Soc (2013)

Percutaneous needle puncture into the rabbit discs. A : An angiography needle is inserted 3-4 cm ventral from the midline into the disc space using a fluoroscope. B : Constant amount of nucleus material is checked during the puncture to create even disc degeneration.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3756123&req=5

Figure 1: Percutaneous needle puncture into the rabbit discs. A : An angiography needle is inserted 3-4 cm ventral from the midline into the disc space using a fluoroscope. B : Constant amount of nucleus material is checked during the puncture to create even disc degeneration.
Mentions: Each rabbit was anesthetized with intramuscular injection of xylazine (5 mg/kg) and ketamine (35 mg/kg), and the fur was shaved from the mid back and right flank. After anesthesia, a lateral plain X-ray was obtained to establish the pre-injection baseline height of the IVDs. The rabbit was then placed in the lateral oblique prone position, and the injection field was sterilized with an alcohol sponge. Initially, the L5-L6 disc was identified through manual palpation of the interspinous space from the mid back and pelvic rim with fluoroscopy (VPX-200; Toshiba Corporation, Tokyo, Japan). After confirmation of the exact level, an 18-gauge angiography needle was inserted 3-4 cm ventral from the midline into the disc space under fluoroscopic control (Fig. 1A). After brief confirmation of the needle position in the center of the disc space by anteroposterior and lateral fluoroscopy, the needle was held in the disc space for 10 seconds. Before removal of the punctured needle, the needle was rotated 180 degrees. A constant amount of nucleus was leaked within the needle and identified by pushing the needle with an empty syringe after removal of the stylet (Fig. 1B). In each rabbit, each of three discs (L2-L3, L3-L4, and L4-L5) was punctured. The L1-L2 and L5-L6 levels were designated as the non-punctured, internal controls. For each level, all procedures for identification and puncture were performed within 20 seconds. Special care was taken to minimize contact with the periosteal tissues of the vertebrae because this could cause hypertrophy of the soft tissues and bony structures around the discs. No antibiotics and analgesic were used after puncture. No rabbits showed neurological symptoms, and all tolerated the procedure well. The rabbits were placed in their cages after observation for recovery. At the planned time, they were sacrificed with intramuscular injection of ketamine (25.0 mg/kg) followed by intravenous injection of sodium pentobarbital (1.2 g/kg).

Bottom Line: Significant disc space narrowing compared to preoperative disc height was observed during the time period (p<0.001).The MRI grade, aggrecan, and matrix metalloprotease-13 mRNA expression and hematoxylin and eosin/safranin O/anti-collagen II staining were consistently indicative of degeneration, supporting the results of the X-ray data.This in vivo model can be used to study and evaluate the safety and efficacy of biologic treatments for degenerative disc disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Sungkyunkwan University School of Medicine, Kangbuk Samsung Hospital, Seoul, Korea.

ABSTRACT

Objective: To develop a simple, reproducible model of disc degeneration in rabbits through percutaneous annular puncture and to confirm the degree of degeneration over time.

Methods: Fifteen New Zealand white rabbits (4 to 5 months old and weighing approximately 3 to 3.5 kg each) underwent annular puncture of the L2-L3, L3-L4, and L4-L5 discs. Rabbits were sacrificed at 4, 8, or 20 weeks after puncture. For a longitudinal study to assess changes in disc height over time, serial X-rays were performed at 0, 2, 4, 8, and 20 weeks for rabbits in the 20-week group. Upon sacrifice, the whole spinal column and discs were extracted and analyzed with magnetic resonance imaging (MRI), real time reverse transcriptase-polymerase chain reaction, and histological staining.

Results: The X-rays showed a slow, progressive decrease in disc height over time. Significant disc space narrowing compared to preoperative disc height was observed during the time period (p<0.001). The MRI grade, aggrecan, and matrix metalloprotease-13 mRNA expression and hematoxylin and eosin/safranin O/anti-collagen II staining were consistently indicative of degeneration, supporting the results of the X-ray data.

Conclusion: Percutaneous annular puncture resulted in slow, reproducible disc degeneration that was confirmed by radiology, biochemistry, and histology. This in vivo model can be used to study and evaluate the safety and efficacy of biologic treatments for degenerative disc disease.

No MeSH data available.


Related in: MedlinePlus