Table 3. Beneficial effects of Rolipram after CNS injury.

Reference	Animal and CNS Injury Paradigm	Rolipram Administration Protocol	Reported Beneficial effects on Neuroprotection and/or Behavioral Outcomes
Nikulina et al., 2004 [54]	Animal:Long Evan Hooded rats, 30 d old, 180–200 gInjury Model:C3–C4, spinal cord hemisection created using iridectomy scissors	In vivo- Rolipram/vehicle or vehicle only (saline and 16% DMSO) by mini osmotic pumps s.c. combined with embryonic spinal tissue implants:- 0.40 µmol/kg/h, 10 µl/h for 10 d- 0.80 µmol/kg/h, 10 µl/h for 10 d- SAC at 6–8 wk PI	Neuroprotective:Although neuroprotection was not assessed, Rolipram (lower dose) reduced levels of host GFAP (reduced astrogliosis) and increased numbers of serotonergic fibers in the transplant compared to vehicle only controlsBehavioral:- Improved forelimb paw placement in Rolipram-treated, implanted animals compared to vehicle controls
Pearse et al., 2004 [19]	Animal:Female Fischer rats, 160–180 gInjury Model:Thoracic (T8) moderate contusion SCI, MASCIS Impactor	In vivo- Rolipram/vehicle or vehicle only (DMSO) by mini osmotic pumps s.c., with or without Schwann cell implants:- 0.5 µl/h, 0.5 mg/kg/d immediately PI, then for 2 wk PI	Neuroprotective:- Increased the number of spared central myelinated axons in the lateral spinal cord- Increased the number of retrogradely labeled neurons in the reticular formation and the raphe nuclei with axon projections below the level of SCI- Rolipram also prevented injury-induced reductions in cyclic AMP levels in the rostral spinal cord (for 2 wk), brainstem (for 1 wk), and sensorimotor cortex (for 1 d); reduced TNF-α at both 3 h and 6 h PI and enhanced Schwann cell myelinationBehavioral:- Rolipram alone improved BBB subscores, foot exo-rotation, base of support and hindlimb footfall errors after SCI
Wang et al., 2006 [127]	Animal:Female Sprague–Dawley rats, 250–270 gInjury Model:Thoracic (T7) moderate contusion SCI, MASCIS Impactor	- Rolipram/vehicle or vehicle only (16% dimethylsulfoxide/PBS), mini osmotic pumps s.c.:- 1.2 mg/kg/d for 28 d	Behavioral:- Rolipram showed a trend for improvement on the BBB score.
Atkins et al., 2007 [20]	Animal:Male Sprague-Dawley rats, 270–320 gInjury Model:Moderate FPI (2.0±0.2atm) TBI; 3.8 mm posterior to bregma/2.5 mm lateral to the midline	- Rolipram/vehicle or vehicle only (5% DMSO/95% Saline) i.p.:- 0.3 or 3.0 mg/kg at 30 min PrI, then every 24 h PI, then SAC 30 min or 3 d later	Neuroprotective:- Reduced cortical contusion volume and contusion area near epicenter of injury and in penumbra region, improved total cortical neuron and CA3 neuron survival on ipsilateral side and modestly reduced β-APP deposits.- Rolipram also restored cyclic AMP levels in the hippocampus as well as total CREB levels in the parietal cortex and phosphorylated CREB levels in the hippocampus after TBI; Rolipram significantly reduced injury-induced increases in IL-1β in hippocampus and thalamus, but not parietal cortex. Significantly reduced injury-induced increases in TNF-α levels in parietal cortex and hippocampus, but not the thalamus
Kajana and Goshgarian, 2008 [128]	Animal:Male Sprague–Dawley rats, 3–6 mos, 295–480 gInjury Model:Unilateral cervical (C2) spinal cord hemisection	- Rolipram/vehicle or vehicle only (10% DMSO in saline) i.p.:- 2.0 mg/kg, 2x/d for 2 d with functional assessments at 5 d and 10 d after last dose	Behavioral:- Facilitated ipsilateral phrenic nerve recovery as exemplified by a recovery in motor activity (5 and 10 d) and enhanced contralateral phrenic nerve output (5 and 10 d); Rolipram-treated animals had higher respiratory rates than controls at 5 but not 10 d
Hatinen et al., 2008 [129]	Animal:Male Wistar rats, 3–4 mos, 262–359 gInjury Model:Stroke; transient occlusion of the middle cerebral artery (MCAO)	- Rolipram/vehicle or vehicle only [Macrogol 30%/0.9% NaCl (70%)] i.p.:- starting at 2 d PO for 13 d- Low-dose: a.m. 0.1 mg/kg (30 min before behavior), then p.m. 0.2 mg/kg- High-dose: a.m. 1.0 mg/kg (30 min before behavior), then p.m. 2.0 mg/kg	Behavioral:- Hindlimb function in the high-dose Rolipram group was associated with fewer slips on the beam walking task and high-dose Rolipram-MCAO rats resembled sham rats at 21 d PI indicating a possible delayed benefit of Rolipram- Rolipram-treated rats, however, showed decreased locomotor activity and rearing, atypical head twitches and possible hyperalgesia immediately after treatment (considered as acute side effects)
Whitaker et al., 2008 [64]	Animal:Female Sprague–Dawley rats, 228–267 gInjury Model:Cervical (C5–6) contusion SCI (180±7 kDyn), IH Impactor	- Rolipram/vehicle or vehicle only (DMSO), mini osmotic pumps s.c.:- 0.5 mg/kg/d, 0.5 ul/h for 12 h, 24 h, or 72 h PI	Neuroprotective:- Significantly reduced oligodendrocyte death at 24 h continuing through 72 h PI (assessed by APC immunohistochemistry)
Beaumont et al., 2009 [58]	Animal:Female Sprague–Dawley rats, 228–267 gInjury Model:Cervical (C5–6) contusion SCI (175±5 kDyn), IH Impactor	- Rolipram/vehicle or vehicle only (DMSO), mini osmotic pumps s.c.:- 0.5 mg/kg/d, 0.5 µl/h each pump for 2 wk- SAC at 5 wk PI	Neuroprotective:- Increased numbers of oligodendrocytes in the VLF compared to controls, though spared white matter area was unchanged- Axon conductivity through the VLF was better after Rolipram treatment than controls Behavioral:- Rolipram treated animals exhibited less hindlimb footfall errors than controls with no change in total number of steps, percentage of forelimb steps without footfall errors and the BBB score
Kajana and Goshgarian, 2009 [62]	Animal:Male Sprague–Dawley rats, 3–6 mos, 295–480 gInjury Model:Unilateral cervical (C2) spinal cord hemisection	- Rolipram/vehicle or vehicle only (10% DMSO in saline) i.v.:- 2 mg/kg at 1 wk PI/20 min before assessment- SAC 2 h after Rolipram	Behavioral- Increased both contralateral and ipsilateral phrenic nerve activity PI; electrophysiological measures indicated higher burst areas and amplitudes- Rolipram also increased cyclic AMP levels
Bretzner et al., 2010 [67]	Animal:Male Sprague-Dawley rats, 300–400 gInjury Model:Cervical (C4–5) dorsolateral funiculus crush (20 s) with fine surgical forceps, depth 2 mm	- Rolipram/vehicle or vehicle only (20 mM PBS/16% DMSO), osmotic pump s.c.:- 0.4 µmol/kg/h for 2 wk.- SAC at 5 wk PI	Neuroprotective:- Although neuroprotection was not assessed, Rolipram enhanced GFP-OEC cell density and axon density in the lesion siteBehavioral:- Rolipram in combination with OEC transplants enhanced forelimb function on the cylinder test compared to vehicle only controls
Ianotti et al., 2011 [66]	Animal:Female Sprague Dawley rats, 220–250 gInjury Model:Thoracic (T8–9) moderate contusion SCI, MASCIS Impactor	- Rolipram/vehicle or vehicle only (DMSO), mini osmotic pump, s.c.:- 0.5 mg/kg/d, 0.5 µL/h, immediately PI for 2 wk	Neuroprotective:- Decreased lesion volume and increased spared white matter; also increased corticospinal and supraspinal axon sparing/sprouting at 5 wk PIBehavioral:- Rolipram improved BBB scores and subscores from 1 to 4 wk PI
Downing et al., 2012 [130]	Animal:Adult female athymic rats, 170–243 gInjury Model:Cervical (C4–6) hemisection injury using a fine scalpel blade	- Rolipram administered through a drug-eluting microfibrous patch at 2 doses:- Low dose (25 µg/ml)- High dose (500 µg/ml)	Neuroprotective:- Although neuroprotection was not assessed, Rolipram (low dose) increased axon presence in the lesion site, increased numbers of oligodendrocytes and reduced the degree of astrocyte reactivityBehavioral:- Improved Martinez forelimb open-field scores with Rolipram (low dose) from weeks 1 through 4, 6 and 8, when compared to all other animals

This table summarizes those studies that have shown beneficial effects of Rolipram on anatomical (primarily cell or tissue protection) or functional outcomes following spinal cord or brain injury. Table abbreviations: FPI: parasagittal fluid-percussion brain injury, i.p.: intraperitoneal, i.v.: intravenous, PI: post injury, PO: post operation, PrI: prior to injury, PrS: prior to sacrifice, SAC: sacrificed, TBI: traumatic brain injury, VLF, ventrolateral funiculus.