The effect of needle size inducing degeneration in the rat caudal disc: evaluation using radiograph, magnetic resonance imaging, histology, and immunohistochemistry.
BACKGROUND
The rat caudal disc has been increasingly used in studying of disc degeneration because of its simplicity, low cost, and efficiency. However, the reproducibility and standardization are essential to facilitate the investigations of biologic therapeutics at different stages of degeneration.


OBJECTIVE
To identify the effect of different needle gauges to the degenerative response in rat caudal discs and to examine its pathogenesis by looking at the cellular and matrix changes.


METHODS
In vivo study of injury-induced rat caudal disc degeneration using needle puncture.


METHODS
Thirty-six Lewis rats aged 12-14 weeks.


METHODS
The induced degenerative discs were analyzed by plain radiograph, magnetic resonance imaging (MRI) and histological examination. Proteoglycan content was assessed by alcian blue stain. Immunohistochemistry using aggrecan, collagen II, and Sox-9 was also evaluated to investigate cell differentiation and matrix changes.


METHODS
All rats were divided into three groups according to different needle gauges (18G, 20G, and 22G). Caudal discs were punctured percutaneously under image guidance. Radiographs and MRI were obtained at 2 weeks interval until 8 weeks. At each time point, three rats from each group were sacrificed for histological analysis and immunohistochemistry.


RESULTS
Larger needle gauges, especially 18G, produced more deterioration of the disc when compared with smaller sizes, particularly with time. Significant differences were identified in almost all parameters compared between 18G and 22G at the 8-week time point. For the effect of time in the same needle size, the differences occurred between 2- or 4-week and 8-week time point in the 18G and 20G groups. The proteoglycan and aggrecan stain gradually decreased over time. Chondrogenic differentiation was identified within the degenerative disc by detecting Sox-9 positive cells and collagen II accumulation increased as degeneration progressed.


CONCLUSIONS
The puncture-induced degenerative changes in rat caudal discs can imitate the human degenerative cascade as observed in plain radiograph, MRI, histology, and immunohistochemistry. We suggest that needle size affects the occurrence of progression of degeneration; thus, the large needle size was required to accelerate the deterioration. The size of needle and time point after injury should be considered when investigating the effect of therapeutic materials to retard degeneration or regenerate the intervertebral disc.
Keorochana
Gun
G
Department of Orthopaedics, Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand.
Johnson
Jared S
JS
Taghavi
Cyrus E
CE
Liao
Jen-Chung
JC
Lee
Kwang-Bok
KB
Yoo
Jeong Hyun
JH
Ngo
Stephanie S
SS
Wang
Jeffrey C
JC
D000818:Animals; D004195:Disease Models, Animal; D007150:Immunohistochemistry; D007403:Intervertebral Disc / Q000293:injuries; D055959:Intervertebral Disc Degeneration; D008279:Magnetic Resonance Imaging; D008297:Male; D009339:Needles; D051381:Rats; D011917:Rats, Inbred Lew