Int J Stem Cells.  2015 Nov;8(2):146-154. 10.15283/ijsc.2015.8.2.146.

Role of Endogenous Bone Marrow Stem Cells Mobilization in Repair of Damaged Inner Ear in Rats

• Elbana AM
• Abdel-Salam S
• Morad GM
• Omran AA

• KMID: 2380809
• DOI: http://doi.org/10.15283/ijsc.2015.8.2.146

Abstract

BACKGROUND AND OBJECTIVES
The utilization of the stem cells is widely used in the last few years in different fields of medicine, either by external transplantation or endogenous mobilization, most of these studies still experimental on animals; few were tried on human as in the spinal cord injury or myocardial infarction. As regard its use in the inner ear, stem cell transplantation was examined in many previous studies, while the mobilization idea is a new method to be experimented in inner ear hair cell regeneration. The present work assessed the possibility of mobilizing endogenous bone marrow derived stem cells (SCs) in rats using granulocyte colony stimulating factor (G-CSF) to induce regeneration and repair to experimentally damaged inner ear hair cells by Amikacin injection.
METHODS
The study included thirty adult Sprague Dawley male rats. Experimental induction of inner ear damage was done by repeated intratympanic injection of amikacin sulfate. Mobilization of bone marrow SCs was provoked by subcutaneous injection of GCSF. Cochlear integrity, induction of hearing loss and functional recovery of sensory hearing loss were assessed using Distortion Product Otoacoustic Emission (DPOAEs). The morphological alteration and recovery of the organ of Corti was assessed histologically using the light and scanning electron microscopes.
RESULTS
After six month duration, there was improvement in 50% of the sensorineural DPOAE results. Functional recovery coincided with the repair of structural components of organ of Corti.
CONCLUSIONS
SCs mobilization by G-CSF is a promising alternative method for replacement therapy in sensorineural hearing loss.

Keyword

Stem cells; Mobilization; Inner ear; Rats; Hearing; Amikacin

MeSH Terms

Adult
Amikacin
Animals
Bone Marrow*
Colony-Stimulating Factors
Ear, Inner*
Granulocyte Colony-Stimulating Factor
Granulocytes
Hair
Hearing
Hearing Loss
Hearing Loss, Sensorineural
Humans
Injections, Subcutaneous
Male
Myocardial Infarction
Organ of Corti
Rats*
Regeneration
Spinal Cord Injuries
Stem Cell Transplantation
Stem Cells*
Amikacin
Colony-Stimulating Factors
Granulocyte Colony-Stimulating Factor

Figure

• 

  Fig. 1 Light photomicrographs of control rat inner ear cavity demonstrating the cochlear cavity divided by Reissener’s membrane (RM) into scala media (SM) and scala vestibule(SV). The organ of Corti (OC) intervenes between the scala media and the scala tympani (ST). It is bounded laterally by the stria vascularis (St V) and medially by the spiral ganglion (SG). The semicircular duct (SD) shows the macula utriculi (U) and the crista ampullaris (CA) (H&E stain. Mic. Mag. ×40).

• 

  Fig. 2 Higher magnification of the cellular components of the control organ of Corti demonstrating: A single row of inner hair cells (IHC) supported by the underlying inner phalangeal cells (IPhC), and several rows of outer hair cells (OHC) supported by the outer phalangeal cells (OPhC). The inner tunnel (IT) is bounded between the inner (IP) and outer pillar (OP) cells (H&E stain. Mic. Mag. ×1000).

• 

  Fig. 3 (A) Light photomicrographs of control rat Organ of Corti showing: The main structural components including the spiral limbus (SL), tectorial membrane (TM), inner supporting cells (IS), inner hair cells (IHC), inner tunnel (IT), outer hair cells (OHC), outer supporting cells (OS), basilar membrane (BM) and the related cochlear nerve (CN). (B) An OAE negative rat (deaf), 6 months after administration of amikacin revealing destruction of the organ of Corti (OC), disappearance of the inner and outer supporting cells and infiltration of the cochlear cavity by red blood corpuscles (arrow). Note the intact basilar membrane (BM) (H&E stain. Mic. Mag. ×400).

• 

  Fig. 4 (A) The crista ampullaris shows an intact superficial layer of hair cells (HC) supported by an underlying layer of supporting cells (SC) and connective tissue stroma (ct). (B) The macula utriculi show an intact superficial layer of hair cells (HC) and the overlying otoconia (O). It is supported by a deeper layer of supporting cells (SC) and connective tissue stroma (ct). (C, D) Light photomicrograph of OAE negative rat, 6 months after injection with amikacin: The crista ampullaris reveals vacuolation and widening of interstitial spaces between hair cells and supporting cells (thick arrow). Note that many hair cells (HC) are still intact. The macula utriculi show intact layers of hair and supporting cells (arrow) (H&E stain. Mic. Mag. ×400).

• 

  Fig. 5 (A) The stria vascularis of rat recovering after GCSF therapeutic regimen reveals a complete sheet of intact superficial epithelium (arrow) with residual interstitial vacuolation in the supporting connective tissue stroma (*). (B) The macula utriculi of a rat recovering after GCSF therapeutic regimen show intact structural components: (HC) hair cells, (SC) supporting cells, (O) otoconia. (C) The crista ampullaris of rat recovering after GCSF therapeutic regimen shows intact layers of hair cells (HC) and supporting cells (SC). (D) The spiral ganglion of rat recovering after GCSF therapeutic regimen (arrow) is composed of intact bipolar cells with apparently average cell density (H&E stain. Mic Mag. ×100).

• 

  Fig. 6 (A) Light photomicrographs of organ of Corti of OAE positive rats recovering within variable durations from receiving GCSF therapeutic regimen: After 4 weeks, the organ of Corti shows few outer hair cells (arrow). No other cellular components of the organ of Corti are identifiable. (B) After 8 weeks, the scala tympani reveals group of cells (arrow) that are apparently organized into a primitive Organ of Corti. SL=spiral limbus, BM=basilar membrane. (C) After 12 weeks, the organ of Corti is formed of well structured rows of inner supporting cells (IS), inner hair cells (IH), outer hair cells (OH) and the intervening tunnels (T). SL=spiral limbus, TM=tectorial membrane. (D) Light photomicrographs of the inner ear from OAE positive rats recovering after 24 weeks from receiving GCSF therapeutic regimen: showing a normal, fully structured organ of Corti composed of intact rows of outer hair cells (OH), outer pillar cells (OP) and an intact row of inner hair cells (IH) supported by inner pillar cells (IP). The outer (OPh) and inner (IPh) phalangeal cells border the inner tunnel (IT). (IS) inner supporting cells (H&E stain. Mic Mag. ×400).

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  Fig. 7 (A) Scanning electron photomicrograph of control rat organ of Corti demonstrating: Normal hair cells (HC) with intact stereocilia (*) projecting from their apices. (B) Surface view of the reticular lamina (RL) showing three parallel rows of intact stereocilia of the outer hair cells. (Ph C) bodies of supporting outer phalangeal cells. (C) Scanning electron photomicrographs of the cochlear cavity in an OAE negative (deaf) rat, 6 months after injecting amikacin: The cochlear cavity is infiltrated by many red blood cells (arrow). No details of hair cells neither of its stereocilia can be identified. (D) Surface view of the reticular lamina (RL) of the organ of Corti showing parallel rows of empty indentations (arrow) marking the lost outer hair cells (Mic. Mag. ×5000).

Reference

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