[Show abstract][Hide abstract] ABSTRACT: Tympanic membrane (TM) perforation, in particular chronic otitis media, is one of the most common clinical problems in the world and can present with sensorineural healing loss. Here, we explored an approach for TM regeneration where the latent progenitor or stem cells within TM epithelial layers may play an important regulatory role. We showed that potential TM stem cells present highly positive staining for epithelial stem cell markers in all areas of normal TM tissue. Additionally, they are present at high levels in perforated TMs, especially in proximity to the holes, regardless of acute or chronic status, suggesting that TM stem cells may be a potential factor for TM regeneration. Our study suggests that latent TM stem cells could be potential regulators of regeneration, which provides a new insight into this clinically important process and a potential target for new therapies for chronic otitis media and other eardrum injuries.
[Show abstract][Hide abstract] ABSTRACT: Since Korean red ginseng (KRG) has been proven to protect against gentamicin-induced vestibular and hearing dysfunction, the effects of KRG on age-related inner ear disorder in C57BL/6 mice were investigated. While age-related hearing loss was detected at the age of 6months (32kHz) and 9months (16kHz) in the control group, it was significantly delayed (p<0.05) in the 150mg/kg KRG-treated group. Vestibular dysfunction was observed in the tail-hanging and swimming tests, with significantly different severity scores and swimming times detected between the control and 150mg/kg KRG-treated group at the age of 12months (p<0.05). Mice treated with 500mg/kg KRG exhibited irritability and aggravated inner ear dysfunction. Histological observation supported the findings of hearing and vestibular function defects. In conclusion, C57BL/6 mice showed early-onset hearing loss and progressive vestibular dysfunction with aging, which were delayed by treatment with 150mg/kg KRG. However, 500mg/kg KRG treatment may induce aggressive behavior.
[Show abstract][Hide abstract] ABSTRACT: Cis-diamminedichloroplatinum (cisplatin) is an effective chemotherapeutic drug for cancer therapy. However, most patients treated with cisplatin are at a high risk of ototoxicity, which causes severe hearing loss. Inspired by the "Good Samaritan effect" or "bystander effect" from gap junction coupling, we investigated the role of gap junctions in cisplatin-induced ototoxicity as a potential therapeutic method. We showed that connexin 43 (Cx43) was highly expressed in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells, mediating cell-cell communication. The viability of HEI-OC1 cells was greatly decreased by cisplatin treatment, and cisplatin-treated HEI-OC1 cells showed lower Cx43 expression compared to that of untreated HEI-OC1 cells. In particular, high accumulation of Cx43 was observed around the nucleus of cisplatin-treated cells, whereas scattered punctuate expression of Cx43 was observed in the cytoplasm and membrane in normal cells, suggesting that cisplatin may interrupt the normal gap junction communication by inhibiting the trafficking of Cx43 to cell membranes in HEI-OC1 cells. Interestingly, we found that the inhibition of gap junction activity reduced cisplatin-induced apoptosis of auditory hair cells. Cx43 siRNA- or 18α-GA-treated HEI-OC1 cells showed higher cell viability compared to control HEI-OC1 cells during cisplatin treatment; this was also supported by fluorescence recovery after photobleaching studies. Inhibition of gap junction activity reduced recovery of calcein acetoxymethyl ester fluorescence compared to control cells. Additionally, analysis of the mechanisms involved demonstrated that highly activate extracellular signal-regulated kinase and protein kinase B, combined with inhibition of gap junctions may promote cell viability during cisplatin treatment.
Cellular and Molecular Life Sciences CMLS 03/2014; 71(19). DOI:10.1007/s00018-014-1594-3 · 5.86 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Abstract Conclusion: Gankyrin seems to be a better biomarker for cholesteatoma compared with Ki-67. Objective: Gankyrin is an oncoprotein, and occurs in cancers but not in benign diseases. The goal of this study was to compare expression of gankyrin, p53, and a proliferation marker (Ki-67) in cholesteatoma and retroauricular skin (RAS), and to evaluate their significance as clinical parameters. Methods: The levels of expression of gankyrin, Ki-67, and p53 in 10 cholesteatoma and 10 paired samples of normal RAS were evaluated by immunohistochemical staining and Western blot. The results were compared with clinical profiles to investigate a correlation. Results: The expression of gankyrin, Ki-67, and p53 proteins was observed in both basal and suprabasal layers of cholesteatoma. The intensity of gankyrin expression was 'positive' in two cases (20%) and 'strongly positive' in eight cases (80%); p53 expression in the suprabasal layer was 'positive' in 70% of cases; and the Ki-67 staining was 'focal' in 80% of cases. In RAS, these proteins were expressed dominantly in the basal layer. Western blot analysis showed that the gankyrin band was more intense in cholesteatoma than in RAS for three of four cases (p < 0.05). However, there was no significant difference in the expression of gankyrin, Ki-67, and p53 according to clinical variables.
[Show abstract][Hide abstract] ABSTRACT: Gap junctional intercellular communication (GJIC) may play an important role in the hearing process. Cisplatin is an anticancer drug that causes hearing loss and Gingko biloba extracts (EGb 761) have been used as an antioxidant and enhancer for GJIC. The purpose of this study was to examine the efficiency of EGb 761 in protecting against cisplatin-induced apoptosis and disturbance of GJIC. HEI-OC1 auditory cells were cultured and treated with cisplatin (50μM) and EGb (300μg/ml) for 24 h, and then analyzed by immunocytochemistry (Annexin V/ propidium iodide) and Western blots. GJIC was evaluated by scrape-loading dye transfer (SLDT). Basal turn organ of Corti (oC) explants from neonatal (p3) rats were exposed to cisplatin (1-10μM) and EGb (50-400μg/ml). The number of intact hair cells was counted by co-labeling with phalloidin and MyoVIIa. EGb prevented cisplatin-induced apoptosis in immunostaining and decreased caspase 3 and poly-ADP- ribose polymerase (PARP) bands, which were increased in cisplatin-treated cells in Western blots. EGb prevented abnormal intracellular locations of Cx 26, 30, 31, and 43 in cells treated with cisplatin and increased quantities of Cx bands. EGb also prevented cisplatin-induced disturbance of GJIC in SLDT. In oC explants, EGb significantly prevented hair cell damage induced by cisplatin. In animal studies, EGb significantly prevented cisplatin-induced hearing loss across 16 kHz and 32 kHz. These results show that cisplatin induces ototoxicity including hearing loss as well as down-regulation of GJIC and inhibition of Cxs in auditory cells. EGb prevents hearing loss in cisplatin-treated rats by inhibiting down-regulation of Cx expression and GJIC. The disturbance of GJIC or Cx expression may be one of the important mechanisms of cisplatin-induced ototoxicity.
[Show abstract][Hide abstract] ABSTRACT: 3-Nitropropionic acid (3-NP), a mitochondrial toxin, has been reported to induce an acute cochlear damage. Korean red ginseng (KRG) is known to have protective effects from some types of hearing loss. This study aimed to observe the protective effect of KRG in an ototoxic animal model using 3-NP intratympanic injection. BALB/c mice were classified into 5 groups (n=15) and dose-dependent toxic effects after intratympanic injection with 3-NP (300 to 5000mM) on the left ear were investigated to determine the appropriate toxicity level of 3-NP. For observation of the protective effects of KRG, 23 mice were grouped into 3-NP (500mM, n=12) and KRG+3-NP groups (300mg/kg KRG for 7 days before 500mM 3-NP administration, n=11). Auditory brain response (ABR) and cochlear morphological evaluations were performed before and after drug administration. The ABR thresholds in the 800 to 5000mM groups exceeded the maximum recording limit at 16 and 32kHz 1 day after 3-NP administration. The ABR threshold in the 500mM 3-NP+KRG group was significantly lower than that in the 500mM 3-NP group from post 1 week to 1 month. The mean type II fibrocyte counts significantly differed between the control and 3-NP groups and between the 3-NP and 3-NP+KRG groups. Spiral ganglion cell degeneration in the 3-NP group was more severe than that in the 3-NP+KRG group. This animal model exhibited a dose-dependent hearing loss with histological changes. KRG administration ameliorated the deterioration of hearing by 3-NP.
[Show abstract][Hide abstract] ABSTRACT: To evaluate the preventive effects of Korean red ginseng (KRG) on gentamicin (GM)-induced ototoxicity and to identify the effective components of KRG.
In vivo and in vitro studies.
Thirty-five Sprague-Dawley rats were divided into four groups. The GM group (n = 15) received intraperitoneal injections (IPI) of GM (160 mg/kg) for 5 days. The KRG + GM group (n = 12) was treated with intragastric feeding of KRG (500 mg/kg) for 12 days with 5 days of IPIs of GM. KRG (n = 4) and control (n = 4) groups were treated with KRG and saline, respectively. Auditory brainstem response (16 or 32 kHz) and Rotarod treadmill tests were done before and after treatments. Cochleas were evaluated by a scanning electron microscope (SEM) and phalloidin staining. Ginsenosides Rb1, Rb2, Rg1, and Re were evaluated as the water-soluble terazolium salt assay, annexin V/propidium iodide assay, and Western blots in HEI-OC1 cells.
Posttreatment hearing thresholds in GM, KRG + GM, KRG, and control groups were: 27.7 ± 7.2 dB, 23.1 ± 4.1 dB, 16.9 ± 2.6 dB, and 21.3 ± 3.5 dB, respectively, for 16 kHz, 30.5 ± 6.6 dB, 25.2 ± 4.3 dB, 22.5 ± 2.7 dB, and 22.5 ± 3.8 dB, respectively for 32 kHz. The KRG + GM group had significantly better hearing than the GM group (P < .05). On SEM and phalloidin staining, the GM group showed severe loss of stereocilia in the basal outer hair cells and a few losses in the middle turns, whereas the KRG + GM group showed relatively intact hair cells. Balance impairment in treadmill tests was not definite in any group. Rb1 and Rb2 showed more effective protection than other components.
KRG protects against GM-induced hearing loss and hair cell death in rats. Laryngoscope, 2011.
The Laryngoscope 06/2011; 121(6):1294-302. DOI:10.1002/lary.21756 · 2.03 Impact Factor