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Experimental Study on Facial Nerve Regeneration with Or Without Geniculate Ganglionectomy

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Experimental Study on Facial Nerve Regeneration with Or Without Geniculate Ganglionectomy ORIGINAL ARTICLE Experimental Study on Facial Nerve Regeneration With or Without Geniculate Ganglionectomy Zhengmin Wang, MD; Chunfu Dai, MD, PhD; Yuhai Zhang, MD Objective: To investigate regeneration of the distal fa- postoperatively for histological examination by elec- cial nerve following nerve grafting within the tympanic tron microscopy. segment with geniculate ganglion preservation or dis- section. Results: The number of myelinated axons in normal fa- cial nerves was 1819.6±535.6. In group 1, the number Design: Randomized controlled trial. of myelinated axons was 123.6±31.1, and, compared with normal facial nerves, the diameter of the regenerative Subjects: Twenty-three adult New Zealand albino rab- axons was decreased and the sheath thickness in the re- bits were used in this study. generative fiber was diminished. In group 2, the num- ber of myelinated axons was 515.1±103.1, while the my- Interventions: A 2-mm tympanic segment of the elin sheath thickness was proportionate to axon diameter. facial nerve was removed, and the greater auricular (Data are given as mean±SD.) nerve was harvested for grafting in all animals. In group 1 (10 rabbits), the geniculate ganglion was pre- Conclusion: Geniculate ganglionectomy may improve served. In group 2 (13 rabbits), the geniculate gan- motor axon regeneration. glion was dissected. Mastoidal and extratemporal segments of the facial nerve were harvested 3 months Arch Otolaryngol Head Neck Surg. 2001;127:422-425 EVERAL ATTEMPTS have been that focus on axon regeneration and func- made at morphological and tional recovery following facial nerve in- quantitative analysis of the jury. Nerve regeneration involves a com- geniculate ganglion.1-3 Previ- plex interaction of neurons, Schwann cells, ous studies2 indicated that the elements of the extracellular matrix, and Stotal number of ganglion cells in a single a host of neurotrophic substances. With temporal bone ranged from 589 to 4183 respect to surgical repair, suturing the sev- (mean, 2162 cells). In 88% of patients, ered nerve ends and nerve grafting have most of these cells were found in the ge- remained the procedures of choice.4 The niculate ganglion.2 However, in 8% of pa- immediate neural environment plays an tients, most of these cells were in the in- important role. The success of neural re- ternal acoustic meatus; in 4%, the meatus generation depends on the cellular ma- and geniculate ganglion contained an equal trix components (lamina, type IV colla- number of cells.2 There was no correla- gen, neural adhesive molecular, and tion between total ganglion cell number others) and on neurotrophic factors, such and age or sex of the patient.1 The gan- as nerve growth factor, that are produced glion cell bodies were aggregated at the by the denervated target nerve.5-7 A recent apex of the genu, close to the origin of the study8 showed that electromagnetic stimu- greater superficial petrosal nerve.2 These lation enhances early regeneration and fa- findings suggested a possible therapeutic cial movement. Most investigations have benefit from geniculate ganglionectomy in focused on motor fiber regeneration and patients with facial paralysis. facial movement. From the Department of Relative to other cranial nerves, the It is poorly understood how the se- Otolaryngology, Eye, Ear, facial nerve is particularly prone to in- cretomotor fibers and gustatory fibers Nose, and Throat Hospital, jury because of the long distance it regenerate after facial nerve injury and Shanghai Medical University, traverses intratemporally and extratem- whether their regeneration affects motor People’s Republic of China. porally. Studies4-8 have been conducted fiber regeneration. The purpose of the pres- (REPRINTED) ARCH OTOLARYNGOL HEAD NECK SURG/ VOL 127, APR 2001 WWW.ARCHOTO.COM 422 ©2001 American Medical Association. All rights reserved. Downloaded From: https://jamanetwork.com/ on 09/23/2021 MATERIALS AND METHODS superficial petrosal nerve were excised, and the main trunk of the facial nerve was preserved. To prevent regeneration of the intermediate nerve to the distal segment of the fa- ANIMALS cial nerve, bone wax was replaced in the geniculate gan- glion. Antibiotics were administered to minimize the Twenty-three adult New Zealand albino rabbits of both sexes possibility of infectious complications. Three months post- weighing 2.5 to 3.2 kg were randomly assigned to 2 groups. operatively, all animals were placed under deep general an- All animals underwent removal of a 2-mm tympanic seg- esthesia, the otic vesicle was opened, the parotid gland was ment of the facial nerve, and the greater auricular nerve was retracted forward slightly, the stylomastoid foramen was harvested for grafting. In group 1 (10 rabbits), the genic- dissected, and the mastoidal and extratemporal segments ulate ganglion was preserved. In group 2 (13 rabbits), the of the facial nerve were harvested for electron micro- geniculate ganglion was dissected. Mastoidal and extra- scopic evaluation. The methods and protocol of the study temporal segments of the facial nerve were harvested 3 were reviewed and approved by the Institutional Animal months postoperatively for histological examination. Care and Use Committee of Shanghai Medical University, People’s Republic of China. SURGICAL PROCEDURE HISTOLOGICAL EXAMINATION All operations were performed on the left side with an operating microscope (Carl Zeiss, Inc, Oberkochen, For electron microscopic evaluation, tissues were im- Germany). The right side served as a control. All surgical mersed in 3% glutaraldehyde for 2 hours and then in 1% procedures were performed under aseptic conditions. The osmium tetroxide for 1 hour. Dehydration in a series of animals were anesthetized with inhalation through intu- graded ethanols was followed by gradual infiltration with bation of 0.5% enflurane with equal parts of nitrous oxide epoxy (Embed-812; Electron Microscope Sciences, To- and oxygen. The otic vesicle was exposed postauricularly. kyo, Japan). Semithin sections were made and stained with Through the surgical fenestration, the incus was removed toluidine blue O for light microscopic examination, and ul- and the tympanic fallopian canal was identified above the trathin sections were stained with uranyl acetate and lead footplate of the stapes. The fallopian canal was opened citrate for electron microscopy (JEM-2000CX; JEOL Co, with a diamond burr, and 2.0 mm of the facial nerve was Tokyo, Japan). Axon count and size and distribution of removed. The nerve graft was carefully inserted in the unmyelinated and myelinated fibers were determined by opened canal between the cut ends of the facial nerve. electron microscopic examination. The donor nerve for grafting was the greater auricular Analyses to determine the statistical significance of the nerve. difference in the axon count of the facial nerve between In addition to the above procedures, the animals in group 1 and group 2 were performed using t tests with com- group 2 had the malleus head removed to expose the ge- mercially available software (STAT-VIEW,version 5.0; Aba- niculate ganglion. The geniculate ganglion and the greater cus Concepts Inc, Berkeley, Calif). ent study was to observe regeneration of the distal facial elin sheath thickness was proportionate to the diameter nerve following nerve grafting within the tympanic seg- of the axons (Figure 3). No unmyelinated axons were ment with geniculate ganglion preservation or dissec- seen. In the extratemporal segments, myelinated axons tion. were evenly distributed. The number of myelinated axons was 515.1±103.1, which is about one quarter to one third RESULTS of that in normal facial nerve. Proliferation of connec- tive tissue was noted among axons. However, compar- In normal facial nerves, myelinated axons were evenly ing group 1 with group 2, the number of regenerative my- distributed. Sheath thickness was proportionate to axon elinated axons was increased significantly (P,.001) in diameter (Figure 1), and the number of axons was group 2. 1819.6±535.6. Data are given as mean±SD. In group 1, regenerative myelinated fibers and un- myelinated fibers were identified in the mastoidal seg- COMMENT ments. However, the number of myelinated axons was 123.6±31.1, which was much fewer than were found in It is well-known that there are somatic motor, gusta- normal facial nerves. In addition, the diameter of regen- tory, and secretomotor fibers within the facial nerve. Early erative axons was decreased, and the sheath thickness in regenerative gustatory fibers in chorda tympani and se- regenerative fibers was diminished (Figure 2). Extra- cretomotor fibers are unmyelinated. Mature regenera- temporal segments were almost completely composed of tive gustatory fibers and somatic motor fibers are my- connective tissue, with fewer myelinated axons than were elinated. An anatomical study9 has indicated that the total found in the mastoidal segments. Myelinated axons were number of myelinated nerve fibers in the facial nerve var- diffusely distributed throughout the extratemporal seg- ies from 7500 to 9370, depending on the anatomical level ments. It is estimated that only one eighteenth to one of the nerve segment. The greatest number of nerve axons twelfth of myelinated axons are used to innervate muscle. was found at the level of the middle of the mastoidal por- In group 2, a large number of regenerative myelin- tion. The peak diameter of the facial nerve axon was ated axons was found in the mastoidal segments. The my- between 4 and 6 µm. The number of facial nerve fibers (REPRINTED) ARCH OTOLARYNGOL HEAD NECK SURG/ VOL 127, APR 2001 WWW.ARCHOTO.COM 423 ©2001 American Medical Association. All rights reserved. Downloaded From: https://jamanetwork.com/ on 09/23/2021 matolytic technique in cats to trace the distribution of the afferent fibers through the facial nerve branches, found that 20% of fibers traversed the greater superficial pe- trosal nerve. The remainder was distributed in the chorda tympani (45%), posterior auricular rami (21%), branches to mimetic muscle (8%), deep cervical branch (5%), and nerve to the stapedius (1%).
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