Developing consistently reproducible intervertebral disc degeneration at rat caudal spine by using needle puncture

2009 ◽  
Vol 10 (6) ◽  
pp. 522-530 ◽  
Author(s):  
Huina Zhang ◽  
Frank La Marca ◽  
Scott J. Hollister ◽  
Steven A. Goldstein ◽  
Chia-Ying Lin
Keyword(s):  
Animal Model ◽  
Mr Imaging ◽  
Intervertebral Disc ◽  
Disc Degeneration ◽  
Collagen Type ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Anulus Fibrosus ◽  
Safranin O ◽  
Ivd Degeneration

Object The goal in this study was to develop a convenient, less-invasive animal model to monitor progression of intervertebral disc (IVD) degeneration for future testing of new treatments for disc degeneration. Methods Level 5/6 and 7/8 IVDs of rat caudal spine were stabbed laterally with 18- or 21-gauge hypodermic needles to a depth of 5 mm from the subcutaneous surface with the aid of fluoroscopy. In vivo MR imaging studies were performed at 4, 8, and 12 weeks postsurgery to monitor progression of IVD degeneration. Histological analysis including H & E and safranin O staining, and immunohistochemical studies of collagen type II and bone morphogenetic protein receptor type II (BMPRII) were assessed at 12 weeks postsurgery. Results The 18- and 21-gauge needle–stabbed discs illustrated decreases in both the T2 density and MR imaging index starting at 4 weeks, with no evidence of spontaneous recovery by 12 weeks. Histological staining demonstrated a decreased nucleus pulposus (NP) area, and the NP–anulus fibrosus border became unclear during the progression of disc degeneration. Similar patterns of degenerative signs were also shown in both safranin O– and collagen type II–stained sections. The BMPRII immunohistochemical analysis of stabbed discs demonstrated an increase in BMPRII expression in the remaining NP cells and became stronger in anulus fibrosus with the severity of disc degeneration. Conclusions After introducing an 18- or 21-gauge needle into the NP area of discs in the rat tail, the stabbed disc showed signs of degeneration in terms of MR imaging and histological outcome measurements. Changes in BMPRII expression in this animal model provide an insight for the effectiveness of delivering BMPs into the region responsible for chondrogenesis for disc repair. This convenient, less-invasive, reproducible, and cost-effective model may be a useful choice for testing novel treatments for disc degeneration.

126. LMP1 overexpression in the rabbit intervertebral disc upregulates proteoglycans and collagen type ii synthesis in vivo

2004 ◽  
Vol 4 (5) ◽  
pp. S64-S65
Author(s):  
S.Tim Yoon ◽  
Emad Attallah-Wasif ◽  
Jun Li ◽  
William Hutton ◽  
Scott Boden
Keyword(s):  
Intervertebral Disc ◽  
Collagen Type ◽  
Type Ii ◽  
Collagen Type Ii

scholarly journals Intervertebral Disc Degeneration Induced by Needle Puncture and Ovariectomy: A Rat Coccygeal Model

2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Tao Tian ◽  
Haidong Wang ◽  
Zhaohui Li ◽  
Sidong Yang ◽  
Wenyuan Ding
Keyword(s):  
Animal Model ◽  
Intervertebral Disc ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Corn Oil ◽  
Female Rats ◽  
Needle Puncture ◽  
Time Points ◽  
17Β Estradiol ◽  
Safranin O

Objectives. To establish a novel animal model of intervertebral disc degeneration (IVDD) in rats and to investigate the effect of 17β-estradiol (E2) intervention in this model. Methods. This study was divided into two parts: animal model (four groups: Sham, ovariectomy (OVX), Puncture, and OVX+Puncture; three-time points: 4, 8, and 12 weeks; three female rats/group/time point) and drug intervention (Sham, OVX+Puncture+corn oil, and OVX+Puncture+E2; three female rats/group). The rats were analyzed by micromagnetic resonance imaging (MRI), hematoxylin and eosin (HE) staining, and safranin-O staining. Results. MRI and histological scores significantly differed among the four groups at the three-time points (all P < 0.05 ). IVDD progressed with time in the OVX, Puncture, and OVX+Puncture groups (all P < 0.05 ). The changes were the most obvious in the OVX+Puncture group. In the E2 intervention part, the Veh group had the worst MRI signals and histological scores ( P < 0.05 ). The MRI scores in the E2 group were less obvious compared to the Sham group ( P > 0.05 ). Also, the histological scores were significantly different between the Sham and E2 groups ( P < 0.05 ). Conclusions. The combination of ovariectomy and needle puncture can synergically induce IVDD in rat coccygeal discs. Estrogen treatment can effectively ameliorate IVDD progression in the newly established IVDD models.

Stromal Cell Derived Factor-1-Mediated Migration of Mesenchymal Stem Cells Enhances Collagen Type II Expression in Intervertebral Disc

2018 ◽  
Vol 24 (23-24) ◽  
pp. 1818-1830 ◽  
Author(s):  
Catarina Leite Pereira ◽  
Graciosa Quelhas Teixeira ◽  
Joana Rita Ferreira ◽  
Matteo D'Este ◽  
David Eglin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Intervertebral Disc ◽  
Stromal Cell ◽  
Collagen Type ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Stromal Cell Derived Factor

The presence of collagen types II and X in medial coronoid processes of 21 dogs

2000 ◽  
Vol 13 (04) ◽  
pp. 178-184 ◽  
Author(s):  
D. T. Crouch ◽  
D. D. Lewis ◽  
J. M. Kreeger ◽  
J. L. Tomlinson ◽  
J. L. Cook
Keyword(s):  
Immunohistochemical Staining ◽  
Collagen Type ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Collagen Types ◽  
Definitive Answer ◽  
Naturally Occurring ◽  
Significant Difference ◽  
Safranin O

SummaryThe purpose of this study was to determine collagen type II and X immunohistochemical staining characteristics of naturally-occurring fragmented medial coronoid processes (FMCP) in order to help delineate potential pathophysiological events associated with FMCP.34 surgically excised FMCP from 21 client-owned dogs and 16 intact medial coronoid processes from 8 mongrel dogs were examined. The specimens were categorized by the dog's age: <12 months, 12–18 months, 18–24 months, >24 months.The excised FMCP and normal medial coronoid processes were sectioned and stained with haematoxylin and eosin, safranin-O fast green, and toluidine blue. Sections were subjectively evaluated for tissue morphology, cell and matrix content, and proteoglycan staining. Immunohistochemical staining was performed for collagen types II and X. Sections were then subjectively evaluated for the location and intensity of staining.FMCP tissue demonstrated a wide variety of histological and immunohistochemical characteristics compared to normal medial coronoid processes. Significantly (P = 0.016) more normal dogs stained positive for collagen type X than dogs with FMCP in the <12 months old group. No other significant differences between affected and normal dogs were noted for either collagen type in any age group. No significant difference in age was noted for the presence or absence of collagen type II among affected dogs, and no statistically significant correlation was observed between age of those with collagens type X and type II present. Although not statistically significant (P = 0.100), there was a trend for the presence of collagen type II when collagen type X was present. The results of this study have not provided a definitive answer regarding the role of collagen type X in the etiopathogenesis of FMCP, but suggest that it may be an important factor in some cases, warranting further investigation. Excised fragmented medial coronoid processes were examined to determine the collagen immunohistochemical staining characteristics when compared to intact processes.

scholarly journals Aberrant spinal mechanical loading stress triggers intervertebral disc degeneration by inducing pyroptosis and nerve ingrowth

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Fangda Fu ◽  
Ronghua Bao ◽  
Sai Yao ◽  
Chengcong Zhou ◽  
Huan Luo ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Mechanical Loading ◽  
Disc Degeneration ◽  
Sensory Nerves ◽  
Pcr Analysis ◽  
Mechanical Instability ◽  
The Matrix ◽  
Matrix Metabolism ◽  
Safranin O ◽  
Ivd Degeneration

AbstractAberrant mechanical factor is one of the etiologies of the intervertebral disc (IVD) degeneration (IVDD). However, the exact molecular mechanism of spinal mechanical loading stress-induced IVDD has yet to be elucidated due to a lack of an ideal and stable IVDD animal model. The present study aimed to establish a stable IVDD mouse model and evaluated the effect of aberrant spinal mechanical loading on the pathogenesis of IVDD. Eight-week-old male mice were treated with lumbar spine instability (LSI) surgery to induce IVDD. The progression of IVDD was evaluated by μCT and Safranin O/Fast green staining analysis. The metabolism of extracellular matrix, ingrowth of sensory nerves, pyroptosis in IVDs tissues were determined by immunohistological or real-time PCR analysis. The apoptosis of IVD cells was tested by TUNEL assay. IVDD modeling was successfully produced by LSI surgery, with substantial reductions in IVD height, BS/TV, Tb.N. and lower IVD score. LSI administration led to the histologic change of disc degeneration, disruption of the matrix metabolism, promotion of apoptosis of IVD cells and invasion of sensory nerves into annulus fibrosus, as well as induction of pyroptosis. Moreover, LSI surgery activated Wnt signaling in IVD tissues. Mechanical instability caused by LSI surgery accelerates the disc matrix degradation, nerve invasion, pyroptosis, and eventually lead to IVDD, which provided an alternative mouse IVDD model.

scholarly journals Fexofenadine Protects Against Intervertebral Disc Degeneration Through TNF Signaling

2021 ◽  
Vol 9 ◽  
Author(s):  
Kaiwen Liu ◽  
Jianlu Wei ◽  
Guohua Li ◽  
Ronghan Liu ◽  
Dawang Zhao ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Disc Degeneration ◽  
Pathological Process ◽  
Mechanism Study ◽  
Tnf Α ◽  
Safranin O ◽  
Factor Α ◽  
Ivd Degeneration

Objective: Fexofenadine (FFD) is an antihistamine drug with an anti-inflammatory effect. The intervertebral disc (IVD) degeneration process is involved in inflammation in which tumor necrosis factor-α (TNF-α) plays an important role. This study aims to investigate the role of FFD in the pathological process of IVD degeneration.Methods: Safranin O staining was used for the measurement of cartilageous tissue in the disc. Hematoxylin-Eosin (H&amp;E) staining was used to determine the disc construction. A rat needle puncture model was taken advantage of to examine the role of FFD in disc degeneration in vivo. Western Blotting assay, immunochemistry, and immunoflurence staining were used for the determination of inflammatory molecules. ELISA assay was performed to detect the release of inflammatory cytokines. A real-time PCR assay was analyzed to determine the transcriptional expressions of molecules.Results: Elevated TNF-α resulted in inflammatory disc degeneration, while FFD protected against TNF-α-induced IVD degeneration. Mechanism study found FFD exhibited a disc protective effect through at least two pathways. (a) FFD inhibited TNF-α-mediated extracellular matrix (ECM) degradation and (b) FFD rescued TNF-α induced inflammation in disc degeneration. Furthermore, the present study found that FFD suppressed TNF-α mediated disc degeneration via the cPLA2/NF-κB signaling pathway.Conclusions: FFD provided another alternative for treating disc degeneration through a novel mechanism. Additionally, FFD may also be a potential target for the treatment of other inflammatory-related diseases, including IVD degeneration.

scholarly journals Inhibition of HDAC4 by GSK3β leads to downregulation of KLF5 and ASK1 and prevents the progression of intravertebral disc degeneration

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Lin Xiao ◽  
Dongping Gong ◽  
Loufeng Liang ◽  
Anwei Liang ◽  
Huaxin Liang ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Disc Degeneration ◽  
Cell Apoptosis ◽  
Thermal Sensitivity ◽  
Downstream Signaling ◽  
Thermal Pain ◽  
Histone Deacetylase 4 ◽  
Staining Methods ◽  
Hematoxylin And Eosin ◽  
Safranin O

Abstract Background Intervertebral disc degeneration (IDD) is a major cause of lower back pain. This study aimed at exploring the effects of histone deacetylase 4 (HDAC4) and its upstream and downstream signaling molecules on IDD development. Methods A murine IDD model was established by inducing a needle puncture injury to the vertebrate, whereupon we isolated and transfected of nucleus pulposus (NP) cells. Disc height index (DHI) of the mice was determined by X-ray tomography, while the pain experienced by the IDD mice was evaluated by mechanical and thermal sensitivity tests. Next, the interaction between GSK3β and HDAC4 as well as that between HDAC4 and KLF5 acetylation was assessed by co-immunoprecipitation, while the promoter region binding was assessed identified by chromatin immunoprecipitation. By staining methods with TUNEL, Safranin O fast green, and hematoxylin and eosin, the NP cell apoptosis, degradation of extracellular matrix, and morphology of intervertebral disc tissues were measured. Furthermore, mRNA and protein expressions of GSK3β, HDAC4, KLF5, and ASK1, as well as the extent of HDAC4 phosphorylation, were determined by RT-qPCR and Western blotting. Results GSK3β was identified to be downregulated in the intervertebral disc tissues obtained from IDD mice, while HDAC4, KLF5, and ASK1 were upregulated. HDAC4 silencing alleviated IDD symptoms. It was also found that GSK3β promoted the phosphorylation of HDAC4 to increase its degradation, while HDAC4 promoted ASK1 expression through upregulating the expression of KLF5. In IDD mice, GSK3β overexpression resulted in increased DHI, inhibition of NP cell apoptosis, alleviation of disc degeneration, and promoted mechanical and thermal pain thresholds. However, HDAC4 overexpression reversed these effects by promoting ASK1 expression. Conclusion Based on the key findings of the current study, we conclude that GSK3β can promote degradation of HDAC4, which lead to an overall downregulation of the downstream KLF5/ASK1 axis, thereby alleviating the development of IDD.

Specificity and T cell receptor β chain usage of a human collagen type II-reactive T cell clone derived from a healthy individual

1992 ◽  
Vol 22 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Tan Yan ◽  
Harald Burkhardt ◽  
Thomas Ritter ◽  
Barbara Bröker ◽  
Karl Heinz Mann ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Collagen Type ◽  
Cell Clone ◽  
Cell Receptor ◽  
Type Ii ◽  
Collagen Type Ii ◽  
T Cell Clone ◽  
Human Collagen ◽  
Β Chain
Sign in / Sign up

Export Citation Format

Share Document