Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Lim, Kai-Zheong; Daly, Christopher; Ghosh, Peter; Jenkin, Graham; Oehme, David; Cooper-White, Justin J.; Naidoo, Taryn; Goldschlager, Tony

doi:10.3791/55753

Cited by 14 publications

(13 citation statements)

References 19 publications

(29 reference statements)

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“…Only a significant increase in NGF for statically loaded repaired discs was found, which might be responsible for innervation and was often associated with painful discs [ 26 ]. Other approaches of IVD damage involving AF damage and disc repair models were lately tested in vivo ovine using defined drill bit injuries induced to the AF [ 27 ]. It would be interesting to see how a composite approach using the silk-membrane fleece and fibrin hydrogel for such a model would perform [ 3 ].…”

Section: Discussionmentioning

confidence: 99%

Genipin-Enhanced Fibrin Hydrogel and Novel Silk for Intervertebral Disc Repair in a Loaded Bovine Organ Culture Model

Frauchiger

May

Bakırcı

et al. 2018

JFB

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(1) Background: Intervertebral disc (IVD) repair represents a major challenge. Using functionalised biomaterials such as silk combined with enforced hydrogels might be a promising approach for disc repair. We aimed to test an IVD repair approach by combining a genipin-enhanced fibrin hydrogel with an engineered silk scaffold under complex load, after inducing an injury in a bovine whole organ IVD culture; (2) Methods: Bovine coccygeal IVDs were isolated from ~1-year-old animals within four hours post-mortem. Then, an injury in the annulus fibrosus was induced by a 2 mm biopsy punch. The repair approach consisted of genipin-enhanced fibrin hydrogel that was used to fill up the cavity. To seal the injury, a Good Manufacturing Practise (GMP)-compliant engineered silk fleece-membrane composite was applied and secured by the cross-linked hydrogel. Then, IVDs were exposed to one of three loading conditions: no load, static load and complex load in a two-degree-of-freedom bioreactor for 14 days. Followed by assessing DNA and matrix content, qPCR and histology, the injured discs were compared to an uninjured control IVD that underwent the same loading profiles. In addition, the genipin-enhanced fibrin hydrogel was further investigated with respect to cytotoxicity on human stem cells, annulus fibrosus, and nucleus pulposus cells; (3) Results: The repair was successful as no herniation could be detected for any of the three loading conditions. Disc height was not recovered by the repair DNA and matrix contents were comparable to a healthy, untreated control disc. Genipin resulted being cytotoxic in the in vitro test but did not show adverse effects when used for the organ culture model; (4) Conclusions: The current study indicated that the combination of the two biomaterials, i.e., genipin-enhanced fibrin hydrogel and an engineered silk scaffold, was a promising approach for IVD repair. Furthermore, genipin-enhanced fibrin hydrogel was not suitable for cell cultures; however, it was highly applicable as a filler material.

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Section: Discussionmentioning

confidence: 99%

Genipin-Enhanced Fibrin Hydrogel and Novel Silk for Intervertebral Disc Repair in a Loaded Bovine Organ Culture Model

Frauchiger

May

Bakırcı

et al. 2018

JFB

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“…Local anesthetic (bupivacaine 0.5%) (AstraZeneca Australia, Macquarie Park, New South Wales, Australia) was administered subcutaneously and the L2-3 and L3-4 lumbar intervertebral discs exposed via left lateral retroperitoneal approach, as previously described. 24,25 Intraoperative lateral radiographs (Radlink, Atomscope HF200A, Redondo Beach, CA, USA) were performed to confirm the correct levels. Six sheep underwent microdiscectomy APN injury, performed by the creation of a 3 mm × 5 mm annular window followed by disc resection using pituitary rongeurs.…”

Section: Methodsmentioning

confidence: 99%

“…DBI was performed on the L2/3 and L3/4 intervertebral discs of the remaining 6 sheep using a 3.5-mm Brad point drill bit (Carbatec, Melbourne, Victoria, Australia) with a drill bit stop applied at 12 mm drill bit length (Drill Warehouse, Amazon, Seattle, WA, USA) as described previously. 25 Following intervertebral disc injury, the wound was closed using a routine layered procedure performed using absorbable sutures (Vicryl, Ethicon, Somerville, NJ, USA). Animals received a fentanyl patch (Duragesic 75 mg/h, Jannsen LLC., North Ryde, New South Wales, Australia) and intravenous paracetamol (Pfizer Ltd, West Ryde, New South Wales, Australia) for postoperative analgesia.…”

Section: Surgical Proceduresmentioning

confidence: 99%

A Comparison of Two Ovine Lumbar Intervertebral Disc Injury Models for the Evaluation and Development of Novel Regenerative Therapies

Daly

Ghosh

Badal

et al. 2018

Global Spine Journal

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Study Design:Large animal research.Objective:Lumbar discectomy is the most commonly performed spinal surgical procedure. We investigated 2 large animal models of lumbar discectomy in order to study the regenerative capacity of mesenchymal stem cells following disc injury.Methods:Twelve adult ewes underwent baseline 3-T magnetic resonance imaging (MRI) followed by lumbar intervertebral disc injury by either drill bit (n = 6) or annulotomy and partial nucleotomy (APN) (n = 6). Necropsies were performed 6 months later. Lumbar spines underwent 3-T and 9.4-T MRI prior to histological, morphological and biochemical analysis.Results:Drill bit-injured (DBI) and APN-injured discs demonstrated increased Pfirrmann grades relative to uninjured controls (P < .005), with no difference between the 2 models. Disc height index loss was greater in the APN group compared with the DBI group (P < .005). Gross morphology injury scores were higher in APN than DBI discs (P < .05) and both were higher than controls (P < .005). Proteoglycan was reduced in the discs of both injury models relative to controls (P < .005), but lower in the APN group (P < .05). Total collagen of the APN group disc regions was higher than DBI and control discs (P < .05). Histology revealed more matrix degeneration, vascular infiltration, and granulation in the APN model.Conclusion:Although both models produced disc degeneration, the APN model better replicated the pathobiology of human discs postdiscectomy. We therefore concluded that the APN model was a more appropriate model for the investigation of the regenerative capacity of mesenchymal stem cells administered postdiscectomy.

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“…Intervertebral discs are the primary component of the human spine, which provides mechanical support and spinal motion for daily activities (1). Intervertebral disc degeneration (Idd) is an important contributor to neck, back and radicular pain, and a leading cause of disability worldwide (2). commonly, Idd is characterized by disc space narrowing, water content reduction in the nucleus pulposus (NP) tissues, the appearance of fibrosis that is reflected in an increase in thickness and spacing of collagen fibers, and in the secretion of proinflammatory cytokines (3,4).…”

Section: Introductionmentioning

confidence: 99%

Bone morphogenetic protein 2 alleviated intervertebral disc degeneration through mediating the degradation of ECM and apoptosis of nucleus pulposus cells via the PI3K/Akt pathway

et al. 2018

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The present study aimed to explore the underlying mechanisms of bone morphogenetic protein 2 (BMP2) in alleviating intervertebral disc degeneration (Idd). A rat puncture Idd model was constructed, and the rats were randomly divided into six groups: control; Idd (model); Idd+PBS [containing 10 10 adeno-associated virus serotype 2 (AAV)]; and Idd + AAV2-BMP2 (10 6 , 10 8 and 10 10 ). IL-1β was used to treat primary nucleus pulposus (NP) cells to mimic Idd in vitro. The effects of BMP2 in Idd were determined by magnetic resonance imaging (MRI), hematoxylin and eosin staining and Alcian Blue staining in vivo. The levels of collagen II, aggrecan, transcription factor SOX9 (SOX9) and matrix metalloproteinase 13 (MMP-13) were examined using western blot analysis and reverse transcription quantitative polymerase chain reaction (RT-qPcR) in NP tissues and cells. The expression of c-telopeptide of type II collagen (cTX-II) in the sera or cell supernatants was determined by ELISA. In addition, the levels of phosphorylation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt), and the levels of apoptosis-associated proteins and apoptosis ratio of NP cells were also determined by western blot analysis and flow cytometry, respectively. LY29400, an inhibitor of PI3K, was used to additionally confirm the signal pathway mechanism of BMP2 treatment in IDD. BMP2 significantly extended the interval between discs and alleviated the fibrous ring rupture and the decrease in the levels of glycoproteins in Idd rats, as determined by MRI and histological staining. Additionally, BMP2 treatment significantly upregulated the levels of collagen II, aggrecan and SOX9, but downregulated the levels of MMP-13 and cTX-II in Idd rats and NP cells in a dose-dependent manner. concurrently, recombinant human (rh)BMP2 pretreatment also significantly decreased the apoptosis ratio of interleukin (IL)-1β-treated NP cells via downregulating the level of cleaved caspase-3 and upregulating the level of uncleaved poly (adenosine 5'-diphosphate-ribose) polymerase. It was demonstrated that rhBMP2 also significantly decreased the inflammatory response in NP tissues and cells, based on levels of IL-6, TNF-α and IL-10. In addition, rhBMP2 inhibited cell apoptosis via upregulating the phosphorylation levels of the PI3K/Akt signaling pathway, and LY29400 pretreatment inhibited the effects of BMP2 in IL-1β treated NP cells. BMP2 alleviated Idd via the PI3K/Akt signaling pathway by inhibiting NP cell apoptosis and decreasing the levels of matrix proteins.

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Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Cited by 14 publications

References 19 publications

Genipin-Enhanced Fibrin Hydrogel and Novel Silk for Intervertebral Disc Repair in a Loaded Bovine Organ Culture Model

Genipin-Enhanced Fibrin Hydrogel and Novel Silk for Intervertebral Disc Repair in a Loaded Bovine Organ Culture Model

A Comparison of Two Ovine Lumbar Intervertebral Disc Injury Models for the Evaluation and Development of Novel Regenerative Therapies

Bone morphogenetic protein 2 alleviated intervertebral disc degeneration through mediating the degradation of ECM and apoptosis of nucleus pulposus cells via the PI3K/Akt pathway

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