Lasers in Surgery and Medicine 7:441-443 (1987)

Response of Peripheral Nerve to He-Ne

Laser: Experimental Studies
S. Rochkind, MD, M. Nissan, PhD, L. Barr-Nea, PhD, N. Razon, MD,
M. Schwartz, PhD, and A. Bartal, MD
Neurosurgery Department, Tel-Aviv Medical Center, lchilov Hospital, Tel Aviv (S. R., N. R.,
A. 6.1, Center for Technological Education, Holon (M.N.), Department of Histology and Cell
Biology, Tel-Aviv University, Tel Aviv (L.6.-N.) and Department of Neurobiology, The
Weizmann Institute of Science, Rehovot (M.S.), Israel

Low-energy He-Ne laser irradiation (LELI) was found to affect the electric
activity and morphology in both intact and severely injured peripheral nerves in
rats. Action potential (AP) in the healthy nerve increased by 33% following a
single transcutaneous irradiation. Similar irradiation in crushed nerves caused AP
to increase significantly over the AP of nonirradiated crushed nerve. Morpholog-
ical observations revealed that a laser-irradiated injured nerve had diminished scar
tissue as compared to an injured but not an irradiated nerve.

Key words: low-energy laser irradiation, crush injury, action potential, scar tissue

INTRODUCTION
Neuronal response to injury is one of the major
problems in neurobiology and medicine. Broadly speak-
ing, peripheral nerve injuries can be categorized into
three groups: 1) transection, a complete block of all
axonal flow; 2) tight ligation, a partial block of axonal
flow; and 3) crush injury, some axons, Schwann cells,
and basal lamina remain intact. The lack of adequate
treatment in traumatic injuries to the peripheral nerve on
the one hand and the hints of the potential that lies in
low-energy laser irradiation (LELI) on the other hand led
us to the present work.
Research on low-energy lasers revealed the poten-
tial applications of laser radiation in biology and medi-
cine when operated as a photobiological stimulator rather
than as a means to deliver high-energy to a small area
over a short time. To mention just a few examples:
affecting brain cell excitability [Olson et al, 19811, stim-
ulating wound healing [Mester et al, 19711, epithelizing
tympanic membrane [Ribari, 19811, and healing of Portio
surface [Kovacs, 19811. We choose crush injury as our
traumatic model, this type of injury being the most fre-
quently encountered in surgical wards.
MATERIALS AND METHODS
Research was conducted on 64 healthy adult Spra-
gue-Dawley rats, weighing -250-300 g. A11 rats were
Address reprint requests to Dr. S. Rochkind, Department of Neuro-
anesthetised with 2 mg nernbutaU100 g BW to allow surgery, Ichilov Hospital, Tel-Aviv Medical Center, Tel-Aviv, Israel
surgery, irradiation, and recording. Irradiation was per- 69978.
formed by delivering 5 J ( - 10 J/cm2) of energy trans-
cutaneously along the nerve projection via a shaven skin.
The laser used was a contineous wave (CW) He-Ne
(632.8 nm, 16mW; Aerotech Co.).
The standard crush injury was done under a Zeiss
surgical microscope; the nerve was exposed, and a he-
mostat was closed completely on the nerve for 30 sec
after which the slun was closed and sutured. Action
potentials (AP) were recorded using a Beckman dyno-
graph and a Grass stimulator (model 54B), delivering the
activating stimulus (1 V, 1 msec) and collecting the AP
via bipolar needle electrodes. The AP amplitude was
used for the present work. Rats were treated according
to the schedule given in Table I. The rats that were used
for morphological studies were sacrificed at the end of
the planned periods and the sciatic nerves were fixed in
Bouin’s solution. The tissue was embedded in paraplast,
cut at 6 pm, and stained with hematoxylin and eosin.
RESULTS AND DISCUSSION
The results presented in Figure 1 show that LELI
has an effect on noninjured sciatic nerve. AP were mea-
Accepted for publication May 19, 1987.
This work is dedicated to Dr. Rochkind’s teacher, Y.P. Melman,
Professor of Anatomy, Ivano-Frankovsk Medical School, Ukraine,
U.S.S.R. He was an example of intelligence and integrity we would
like to follow.

0 1987 Alan R. Liss, Inc.

442 Rochkind et a1
TABLE 1. Treatment Schedule
Period of Recording
No. of Crush irradiation period
Group rats injury (days) (days)
la 7 -. 0 1
Ib 13 -. 1 1
2a 9 + 0 360
2b 11 +- 20 360
3a 12 +- 0 -

3b 12 + 20 - 21

sured in seven untreated rats (group la). The measure-

ments were repeated 20 min later, and AP were compared
for each rat. The change was minimal. We repeated the
same experiment with a second group of 13 healthy rats
(group lb), with the sole difference of applying LELI
transcutaneously between the two measurements. The
results in this group indicate an increase in AP in 13 rats
by an average of 43 % .
A large effect of the laser irradiation was observed
in crushed nerves (Fig. 2). A follow-up study for 1 year
provided evidence that, with the laser irradiation, the AP
of the injured nerve was reversed to normal values or
even higher, whereas the injured, nonirradiated nerve
showed the expected pattern of response to injury (AP of
0.4-0.11 of the original value during the first day, 0.2-
0.06 3-5 days after injury, remaining low for 3-4 months
and then gradually increasing to 0.7-0.24 after 1 year).
Our previous studies provide evidence that the recorded
NON IRRADIATED, l a I IRRADIATED, l b
Fig. 1. Action potential (AP) in the rat sciatic nerve. AP is presented
as percentage of the primary AP recorded in the same rat.
AP reflects the activity of the nerve itself, since, in a
totally transected sciatic nerve (six rats), no AP could be
detected with or without irradiation [Rochkind et al,
19861.
The electrophysiological effects of the laser irradia-
tion were accompanied by clear morphological manifes-
tations in all sciatic nerves examined-12 from control
rats (group 3a) and 12 from irradiated rats (group 3b).A
typical picture of the crush area is shown in Figure 3a
(controls) and 3b (irradiated): A prominent scar tissue

3.2

2.8

2.6 LI T

2.41 I

I
.- 0.6 1

3 7 10 14 21 30 45 60 90 150 240 360

days

Fig. 2. AP in the rat crushed sciatic nerve as compare to the AP in

the same nerve before injury. The upper line represents irradiated
group (2b), the lower line the control group (2a)
 Response of Peripheral Nerve to He-Ne Laser
Fig. 3 . Histological section through the crush area of the sciatic
nerve. Hematoxylin and eosin staining. X 150. p, Proximal; d, distal.
a: Nonirradiated animal. Note the ring of constricting scar of fibrous
characterized the controls; this was minimal in the irra-
diated nerves, in which the fibers seemed to maintain
their morphological continuity.
CONCLUSIONS
A number of conclusions can be deducted from our
work so far. 1) LELI causes an increase in the electrical
activity of noninjured sciatic nerve and to a larger extent
of injured sciatic nerve, manifested by the size of the AP.
2) The LELI effect is instantaneous. 3) The LELI effect
persists for at least 1 year following a short series of
irradiations starting at the time of the injury. 4) Scar
tissue in injured sciatic nerve is minimized after LELI.
LELI was found to enhance the rate of regeneration in
crush-injured nerves in rats. Further studies are now
being conducted to assess the clinical application of LELI
in neuromuscular disorders.
ACKNOWLEDGMENTS
The authors acknowledge the technical assistance
of Mr. G . Sheiner, Mr. N. Brusovanic, and Mr. M.
Lerer. This work was supported by a grant from Ramot,
443
tissue and the disorganization of the nerve. b: Irradiated animal. No
visible scar. The proximal part of the nerve is a continuation of the
distal part.
University Authority for Applied Research and Industrial
Development, Ltd., Israel.
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