J Korean Med Sci.  2011 Apr;26(4):492-498. 10.3346/jkms.2011.26.4.492.

Transplantation of Neural Differentiated Human Mesenchymal Stem Cells into the Cochlea of an Auditory-neuropathy Guinea Pig Model

• Cho YB
• Cho HH
• Jang S
• Jeong HS
• Park JS

• KMID: 1777878
• DOI: http://doi.org/10.3346/jkms.2011.26.4.492

Abstract

The aim of this study was to determine the effects of transplanted neural differentiated human mesenchymal stem cells (hMSCs) in a guinea pig model of auditory neuropathy. In this study, hMSCs were pretreated with a neural-induction protocol and transplanted into the scala tympani of the guinea pig cochlea 7 days after ouabain injury. A control model was made by injection of Hanks balanced salt solution alone into the scala tympani of the guinea pig cochlea 7 days after ouabain injury. We established the auditory neuropathy guinea pig model using 1 mM ouabain application to the round window niche. After application of ouabain to the round window niche, degeneration of most spiral ganglion neurons (SGNs) without the loss of hair cells within the organ of Corti and increasing the auditory brain responses (ABR) threshold were found. After transplantation of neural differentiated hMSCs, the number of SGNs was increased, and some of the SGNs expressed immunoreactivity with human nuclear antibody under confocal laser scanning microscopy. ABR results showed mild hearing recovery after transplantation. Based on an auditory neuropathy animal model, these findings suggest that it may be possible to replace degenerated SGNs by grafting stem cells into the scala tympani.

Keyword

Mesenchymal Stem Cells; Auditory neuropathy; Hearing Loss; Spiral ganglion neuron

MeSH Terms

Animals
Cardiotonic Agents/toxicity
Cochlea/drug effects/pathology
Disease Models, Animal
Female
Guinea Pigs
Hearing Loss, Central/chemically induced/pathology/*therapy
Humans
*Mesenchymal Stem Cell Transplantation
Mesenchymal Stem Cells/*cytology
Neurogenesis
Ouabain/toxicity
Spiral Ganglion/pathology
Transplantation, Heterologous

Figure

• 

  Fig. 1 Photomicrographs of stem cell transplantation method. (A) Visualization of the round window after opening the bulla. (B, C) Note the tip of a needle of a Hamilton syringe is inserted into the most basal turn of the scala tympani, where neural-induced human mesenchymal stem cells (nhMSCs) or Hanks balanced salt solution (HBSS) were injected (C, H&E staining, × 40).

• 

  Fig. 2 Ouabain-induced spiral ganglion neuropathy. (A) There is a severe loss of spiral ganglion neurons (SGNs) without a loss of hair cells after 7 days of ouabain application (H&E staining, original magnification × 40 and × 200). (B) Also, an increase of the hearing threshold is noted (dB, decibel). V, wave v.

• 

  Fig. 3 Histologic findings of cochlea after stem cell transplantation. (A) Severe loss of SGNs and sensory hair cells from the basal to the apical turn of the cochlea is noted 6 weeks after HBSS injection (control group). (B) Numerous SGNs and sensory hair cells are observed in all turns of the cochlea 6 weeks after stem cell grafting (stem cell group). (H&E staining, × 40 and × 200).

• 

  Fig. 4 Confocal microscope image after immu- nostaining for anti-human nuclei and NF-H. (A) Numerous neurons are observed in the spiral ganglion (black arrow) after 6 weeks of stem cell transplantation (H&E staining, × 40). (B) the expression of anti-human nuclei (red) and NF-H (green) co-localizes with that of DAPI (blue)-positive cells in the spiral ganglion. Grafted stem cells (white arrow) are located in the spiral ganglion (× 200).

• 

  Fig. 5 Average neuron counts in each turn of the spiral ganglion 6 weeks after transplantation. Average number of neurons in each turn of the spiral ganglion is significantly greater in the stem cell group than in the control group (*P < 0.05).

• 

  Fig. 6 Auditory brainstem response (ABR) results after nhMSC transplantation. Click-evoked ABR waves are recorded up to -10 ± 10 dB in normal guinea pigs. Seven days after application of 1 mM ouabain to the round window, an increase of ABR threshold is noted. After receiving a nhMSC transplantation, the auditory neuropathy guinea pigs show a significant improvement in hearing threshold compared to the control group (*P < 0.05).

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