Mesenchymal stem cell transplantation to the mouse cochlea as a treatment for childhood sensorineural hearing loss

https://doi.org/10.1016/j.ijporl.2013.03.011Get rights and content

Abstract

Objective

There is no treatment established for congenital sensorineural hearing loss because the majority of the cases are hereditary. Although congenital sensorineural hearing loss is thought to be hereditary, this hearing loss occur postnatally. We hypothesized that the transplantation of MSCs (mesenchymal stem cells) to the cochlea would be an effective therapy for stopping or delaying the progression of sensorineural hearing loss in childhood.

Methods

Cultured mouse MSCs were labeled with EGFP (enhanced green fluorescence protein) using retroviruses. EGFP-MSCs were transplanted into the posterior semicircular canal of mice at 2–3 weeks (young group) and 24–26 weeks (adult group) of age by a novel perilymphatic perfusion technique. Engraftment of MSCs was evaluated immunohistologically at 1 week and 2 weeks after transplantation.

Results

In young mice, migrated MSCs were detected in the cochlea tissue by immunofluorescence for EGFP and by immunohistochemistry for fibronectin. The differentiation of migrated MSCs into fibrocyte-like cells was demonstrated by immunofluorescence for connexin 26. There were no adverse effects on auditory function by MSC transplantation, and the auditory brain stem responses threshold did not significantly shift after surgery. In contrast, neither MSC migration nor differentiation was detected in the adult mice canal after MSC transplantation.

Conclusion

The bone marrow derived MSCs were successfully transplanted into the cochlea of young mice by the perilymphatic perfusion technique and were further differentiated into fibrocyte-like cells without any adverse effects on auditory function.

Introduction

There is no treatment established for congenital sensorineural hearing loss because the majority of the cases are hereditary [1]. However, recent clinical studies suggested that such sensorineural hearing loss may develop postnatally, at least in some cases. For example, apoptosis can be activated in the auditory system by loud noise or therapeutic drugs and cause hearing loss at any age, including infants [2]. We hypothesized that for those patients, hearing loss can be stopped or delayed during childhood by transplantation of mesenchymal stem cells (MSCs) to the cochlea; these cells may then differentiate into other cells, including fibrocytes.

Although MSC transplantation has not been tried for hearing loss, MSCs have been implanted to treat many other organ disorders. Those clinical trials have examined hematological disorders, cardiovascular diseases, osteogenesis imperfecta, neurological pathologies, and cancer. In comparison with classical treatment for acute myocardial infarction, MSC transplantation increased left-ventricular ejection fraction and systolic wall motion [3], [4]. MSC transplantation has been used to treat multiple sclerosis and amyotrophic lateral sclerosis [4], [5], [6], [7]. The previous studies demonstrated that treatment with MSCs is feasible and safe and may contribute to the regeneration of myocardial tissue. Furthermore, MSC treatment for osteogenesis imperfecta increased bone formation, bone mineralization and total body mineral content [4], [8]. In contrast, the efficacy of MSC transplantation for inherited diseases has not been shown.

Differentiation of implanted MSCs to fibrocytes is critical for the efficacy of the transplantation treatment. Fibrocytes, particularly in the mesenchymal non-sensory regions, play an important role in cochlear physiology. Fibrocytes constitute a significant proportion of the cells in the cochlea, and five types of fibrocytes are recognized based on morphological and cytochemical characteristics [9]. Fibrocytes in the spiral ligament (SL) of the cochlea are connected with gap junctions [10] and play key roles in potassium uptake from the perilymph and recycling of those ions back to endolymph. In aging human with hearing difficulty, fibrocytes are progressively lost from the SL of the cochlea, suggesting the importance of fibrocytes for auditory capacity [11].

In the present study, bone marrow-derived MSCs were successfully transplanted into the cochlea of young (2–3 week old) mice and differentiated into fibrocyte-like cells without any adverse effects on auditory function.

Section snippets

Mice

The study protocols were approved by the Institutional Animal Care and Use Committee at Juntendo University and all the procedures on animals were performed in compliance with relevant laws and regulations. C57BL/6J mice (Sankyo Labo Service Corporation, Inc., Tokyo, Japan) were maintained in the animal research facility of Juntendo University School of Medicine and utilized for the present study. The mice were maintained in a limited-access barrier and housed in a humidity- (55 ± 10%) and

MSCs engraftment to the cochlea

The engraftment of MSCs was detected in the cochlea by immunofluorescence for EGFP in the young group 1 week (Fig. 1A-1), and 2 weeks (Fig. 1A-2) after transplantation. In contrast, MSC migration was hardly detected in the adult group 1 week after MSC perfusion (Fig. 1B-1). For the adult group, there were no changes in engraftment between 1 and 2 weeks after MSC transplantation (Fig. 1B-2).

MSC migration to the cochlea was confirmed by immunohistochemistry for fibronectin (FN). FN is a major

Discussion

The changes in the auditory system with age have been well studied in C57BL/6J mice. The hearing ability, as well as auditory conduction pathway, is fully matured at 4 weeks of age in C57BL/6J mice. While ABR thresholds in the high-frequency gradually increase from 8 weeks of age, ABR thresholds in the low-frequency do not change until 28 weeks of age [21]. In the cochlea, histological aging changes occur after 24 weeks of age. Conventional hematoxylin and eosin (H&E) staining showed

Grants

None.

Disclosures/conflicts of interest

None.

Acknowledgement

The authors thank the Department of Reproductive Biology, National Institute for Child Health and Development, Tokyo, Japan for providing the MSCs.

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