Spinal cord repair in adult paraplegic rats: Partial restoration of hind limb function

Science

Volumn 273, Issue 5274, 1996, Pages 510-513

  Cheng, Henrich ^a,b,c   Cao, Yihai ^a   Olson, Lars ^a  

^a KAROLINSKA INSTITUTE   (Sweden)

^b TAIPEI VETERANS GENERAL HOSPITAL   (Taiwan)

^c NATIONAL YANG MING UNIVERSITY   (Taiwan)

Author keywords

[No Author keywords available]

Indexed keywords

ACIDIC FIBROBLAST GROWTH FACTOR; FIBRIN GLUE;

ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; CONTROLLED STUDY; FUNCTIONAL ASSESSMENT; HINDLIMB; HISTOLOGY; INTERCOSTAL NERVE; NERVE GRAFT; NERVE REGENERATION; NONHUMAN; PARAPLEGIA; PRIORITY JOURNAL; PYRAMIDAL TRACT; RAT; SCORING SYSTEM; SPINAL CORD SURGERY; SPINAL CORD TRANSSECTION; SURGICAL TECHNIQUE;

EID: 0029840835     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5274.510     Document Type: Article

References (29)

1. E. Puchala and W. Windle, Exp. Neurol. 55, 1 (1977); L. Olson et al., Brain Res. Bull. 9, 519 (1982).
2. E. Puchala and W. Windle, Exp. Neurol. 55, 1 (1977); L. Olson et al., Brain Res. Bull. 9, 519 (1982).
3. Y. Iwashita, S. Kawaguchi, M. Murata, Nature 367, 167 (1994); L. Schnell et al, ibid., p. 170; B. Bregman et al., ibid. 378, 498 (1995).
4. Y. Iwashita, S. Kawaguchi, M. Murata, Nature 367, 167 (1994); L. Schnell et al, ibid., p. 170; B. Bregman et al., ibid. 378, 498 (1995).
5. Y. Iwashita, S. Kawaguchi, M. Murata, Nature 367, 167 (1994); L. Schnell et al, ibid., p. 170; B. Bregman et al., ibid. 378, 498 (1995).
6. 2. The T7 and T10 spinal processes were fixed in dorsiflexion with an S-shaped monofilament surgical steel (DS-20, Ethicon) loop and fastened to the spinal column with nonabsorbable circumspinal threads. One experimental group was subjected to unilateral redirection, and aFGF was included in the glue (URDaFGF); the other two groups were subjected to bilateral redirection and aFGF glue through use of autografts (50%) mixed with allografts (BRDaFGF 1) or through use of only autografts (BRDaFGF II). Control animals were subjected to transection at T8 (C1), to removal of a 5-mm cord segment (C2), or to grafting with the same methods as in the experimental groups except that HBSS replaced aFGF (BRD) or that grafting was done with only white matter-to-white matter connections (RBaFGF). Animals were caged on thick soft bedding, with heating from below during the first postoperative days. The rats' bladders were emptied manually twice daily as long as needed. Antibiotics (Borgal, Hoechst, 15 mg per kilogram of body weight, subcutaneously) were injected once daily for 7 days. Decubitus sores on hind limbs were treated with iodine-soaked dressings. Animals were killed if severe sepsis (urinary tract infection), infected decubitus sores, or other wounds occurred. For the major experiments described here, mortality was <10%. Experiments were approved by the animal research ethical committee of Stockholm. Animals were killed at different time points for histological analyses but no earlier than 1 month after surgery to ensure complete degeneration of the cut descending fibers in the distal stump (9).
7. S. David and A. Aguayo, Science 214, 931 (1981); M. Benfey and A. Aguayo, Nature 296, 150 (1982); R. Bunge, Curr. Opin. Neurobiol. 3, 805 (1993).
8. S. David and A. Aguayo, Science 214, 931 (1981); M. Benfey and A. Aguayo, Nature 296, 150 (1982); R. Bunge, Curr. Opin. Neurobiol. 3, 805 (1993).
9. S. David and A. Aguayo, Science 214, 931 (1981); M. Benfey and A. Aguayo, Nature 296, 150 (1982); R. Bunge, Curr. Opin. Neurobiol. 3, 805 (1993).
10. T. Savio and M. Schwab, Proc. Natl. Acad. Sci. U.S.A. 87, 4130 (1990); M. Schwab, Trends Neurosci. 13, 452 (1990).
11. T. Savio and M. Schwab, Proc. Natl. Acad. Sci. U.S.A. 87, 4130 (1990); M. Schwab, Trends Neurosci. 13, 452 (1990).
12. L. Nygren, K. Fuxe, G. Jonsson, L. Olson, Brain Res. 78, 377 (1974); R. Kuang, M. Merline, K. Kalil, Development 120, 1937 (1994).
13. L. Nygren, K. Fuxe, G. Jonsson, L. Olson, Brain Res. 78, 377 (1974); R. Kuang, M. Merline, K. Kalil, Development 120, 1937 (1994).
14. L. Brown, Exp. Brain Res. 13, 432 (1971).
15. H. Cheng, S. Almström, L. Olson, J. Neural Transplant. Plast. 5, 233 (1995).
16. H. Cheng and L. Olson, Exp. Neurol. 136, 149 (1995).
17. R. Elde et al., Neuron 7, 349 (1991); Y. Cao, thesis, Karolinska Institute (1993); M. Koshinaga, H. Sanon, S. Whittemore, Exp. Neurol. 120, 32 (1993).
18. R. Elde et al., Neuron 7, 349 (1991); Y. Cao, thesis, Karolinska Institute (1993); M. Koshinaga, H. Sanon, S. Whittemore, Exp. Neurol. 120, 32 (1993).
19. R. Elde et al., Neuron 7, 349 (1991); Y. Cao, thesis, Karolinska Institute (1993); M. Koshinaga, H. Sanon, S. Whittemore, Exp. Neurol. 120, 32 (1993).
20. P. McNeil, L. Muthukrishnan, E. Warder, P. D'Amore, J. Cell Biol. 109, 811 (1989).
21. M. Giacobini, B. Hoffer, G. Zerbe, L. Olson, Exp. Brain Res. 86, 73 (1991).
22. H. Cheng et al., ibid. 104, 199 (1995).
23. K. Gale, H. Kerasidis, J. Wrathall, Exp. Neurol. 88, 123 (1985). Percentage functional deficit as indicated by the CBS was used for behavioral evaluation. The CBS (except hot plate), which ranges from 100 points (complete paralysis) to 0 (normal), comprises scores of motor skills (0 to 45), toe spread (0 to 5), righting reflex (0 to 15), extension withdrawal (0 to 5), placing reflex (0 to 5), climbing on an inclined plane (0 to 15), and swimming (0 to 10).
24. M. Beattie and J. Bresnahan, in Criteria for the Assessment of Recovery of Function: Behavioral Methods, M. Brown et al., Eds. (American Paralysis Association, Springfield, NJ, 1989), pp. 16-25; D. Behrmann, J. Bresnahan, M. Beattie, B. Shah, J. Neurotrauma 9, 197 (1992). Open-field walking measured the animal's gross locomotor ability during a 5-min testing period. Two coded animals were observed simultaneously to stimulate locomotor activity and were scored according to a modified Tarlov scale, which ranged from 0 (flaccid paralysis) to 5 (normal walking). This scale was subdivided to more accurately reflect the pattern of recovery of locomotor function as follows: 0, no spontaneous movement; 1, movement that was not reflexive and movement in the hip or knee but not ankle; 2, movement of the limb in all three major joints; 3, active support and an uncoordinated gait or occasional short bouts of coordinated gait; 4, coordination of forelimbs and hind limbs in gait (trotting), including some walking on knuckles or the medial surface of the foot or a few toe drags; and 5, normal.
25. M. Beattie and J. Bresnahan, in Criteria for the Assessment of Recovery of Function: Behavioral Methods, M. Brown et al., Eds. (American Paralysis Association, Springfield, NJ, 1989), pp. 16-25; D. Behrmann, J. Bresnahan, M. Beattie, B. Shah, J. Neurotrauma 9, 197 (1992). Open-field walking measured the animal's gross locomotor ability during a 5-min testing period. Two coded animals were observed simultaneously to stimulate locomotor activity and were scored according to a modified Tarlov scale, which ranged from 0 (flaccid paralysis) to 5 (normal walking). This scale was subdivided to more accurately reflect the pattern of recovery of locomotor function as follows: 0, no spontaneous movement; 1, movement that was not reflexive and movement in the hip or knee but not ankle; 2, movement of the limb in all three major joints; 3, active support and an uncoordinated gait or occasional short bouts of coordinated gait; 4, coordination of forelimbs and hind limbs in gait (trotting), including some walking on knuckles or the medial surface of the foot or a few toe drags; and 5, normal.
26. L. Heimer, in Neuroanatomical Tract-Tracing Methods, L. Heimer, M. Robards, L. Zaborszky, Eds. (Plenum, New York, 1989). For retrograde HRP tracing, six 0.1-μl injections of 5% WGA-HRP (Sigma) were made into the lumbar enlargement (L4 to L5, 25 mm distal to the distal graft-host interface) and combined with an L6 transection to enhance HRP uptake. Positive and negative controls included injection of WGA-HRP with and without colchicin blockage or an acute cut at T8 (Table 1). For anterograde HRP tracing, a total of 2 μl of 5% WGA-HRP was injected into the sensorimotor cortices as five 0.2-μl injections per side. Positive and negative controls are also given in Table 1. Animals were perfused 48 hours later (Ringer's solution, 2% paraformaldehyde, and 10% sucrose). For immunohistochemistry of grafted areas, tissues were further treated with 4% paraformaldehyde for 2 hours and then with polyvinylpyrrolidone (Sigma) for 6 weeks to allow sectioning of soft and bony tissue together. Cryostat 14-μm sections for immunohistochemistry and 60- (brain and brainstem) or 90-μm (spinal cord) free-floating sections for tetramethylbenzidine histochemistry were collected.
27. M. Abercrombie, Anat Rec. 94, 239 (1946). HRP-labeled neuron counts required the presence of the nucleus. The total number of neurons (N) equals nt/(t + d), where n is the number of nuclei of HRP-labeled neurons counted times the section period, t is the section thickness, and d is the average diameter of counted nuclei.
28. S. Grillner, in Control of Posture and Locomotion, R. Stein, K. Person, R. Smith, J. Redford, Eds. (Plenum, New York, 1973), pp. 515-535.
29. We dedicate this paper to Arne Nylander. Supported by the American Paralysis Association, the Swedish Medical Research Council, USPHS grants, the Taiwan Chin-Lin Fund, and the National Science Council. We thank the late J. Arvidsson for advice and S. Almström, M. Casserlöv, K. Erlandsson, and I. Engqvist for assistance. aFGF was provided by Y. Cao and R. Pettersson, Ludwig Institute for Cancer Research, Stockholm.