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CHAPTER 3 hypersensitivity while improving sensorimotor function, and that these improvements were preceded by an anti-inflammatory microglia/macrophage cell population in the injury zone. 3.3 Methods 3.3.1 Hemicontusion spinal cord injury All animal work was approved by the ANU Animal Experimentation Ethics Committee. Hemicontusion spinal cord injuries were performed on 7 week old Wistar rats under isoflurane (1.7-2.3% v/v) anaesthesia. Following hair removal, a laminectomy of T10 vertebral body and removal of dura and arachnoid was performed, followed by a spinal cord hemicontusion using a customized impactor system (Vijayaprakash and Sridharan, 2013) comprising of a cylindrical 10 g weight with a 1 mm diameter tip that was guided onto the right dorsal horn and dropped from 25-50 mm above the spinal cord. 3.3.2 Treatment and experimental groups Injured animals were divided into 670 nm treated (SCI+670) and sham-treated (SCI) groups. SCI+670 rats received 30 min of 670 nm irradiation commencing 2 hrs after surgery, then every 24 hrs after locomotor assessment for the remainder of the recovery period. A commercially available 670 nm LED array (WARP 75A, Quantum Devices, Barneveld, WI; 75 mm2 treatment area) was used for treatment. Spectral characteristics and power output (Figure 3.1) of the LED were measured using a spectrometer (CCS175, Thorlabs) and custom made power meter that was calibrated against a commercially available power meter (PM100D, ThorLabs). Treatment was delivered through a transparent treatment box which was used to confine the animal within its home cage. This resulted in a 7 mm distance between the dorsal surface of the animal and the LED array, and delivery of 35 mW/cm2 (fluence = 63 J/cm2) of 670 nm at the contact surface of the animal’s dorsum. SCI rats (n=29) were restrained in the identical way as the SCI+670 group (n=29), but without the LED device switched on to control for 30 mins restraint in the transparent treatment box. Three additional control groups were included: an intact uninjured group (control; n=7) was untreated and did not receive any sham operations or sham treatment; a sham-injured group (shamSCI; n=8) underwent the spinal surgery, but without the contusion, and were subjected to sham treatment; a sham-injured 670 nm-treated group (shamSCI+670; n=10) underwent spinal surgery without the contusion and received daily 30 min treatments. 3.3.3 Light penetration Uninjured, unshaven animals (n=6) were euthanized with sodium pentobarbital solution (325 mg/ml; Virbac; dosage, 100 mg/kg). The overlaying heart, great vessels and muscles were detached from the anchoring connective tissues and retracted to the side to expose the underlying vertebral column. The T10 vertebral body was eroded with a dental drill to expose the spinal cord from the ventral surface. The sacrificed rat was placed on its back in an inverted transparent 58PDF Image | Effects of Red Light Treatment on Spinal Cord Injury
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