The biocompatibility and mineralization potential of mineral trioxide aggregate containing calcium fluoride-An in vitro study.

BACKGROUND/PURPOSE: MTA is used to induce hard tissue regeneration in various procedures. This study evaluated the biocompatibility and mineralization potential of mineral trioxide aggregate (MTA) containing calcium fluoride (CaF2). To verify if the change of components affected physical properties, the setting time, solubility, and surface roughness were measured. MATERIALS AND METHODS: Human dental pulp cells (HDPCs) were treated with powder and set MTA containing CaF2 (0, 1, 5, and 10 wt %). The proliferation of HDPCs was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The mineralization potential of HDPCs was investigated with the relative gene expression of alkaline phosphatase (ALP), collagen type I (ColI), osteocalcin (OCN), and runt-related transcription factor 2 (Runx2) using real-time reverse transcription polymerase chain reaction (RT-PCR). For investigating the physical properties, setting time and solubility were tested. Surface profiles of material were analyzed by a non-contact surface profiler and a scanning electron microscope (SEM). RESULTS: MTA-5% CaF2 mixtures increased the proliferation and the mineralization-related gene expression of HDPCs to a greater degree than pure MTA. The addition of CaF2 to MTA delayed the setting, but the difference was only significant in the MTA-10% CaF2. Solubility and surface roughness was not altered. CONCLUSION: The addition of more than 5% CaF2 can be considered to increase the regeneration potential of pulp cells without adverse effects on physical property.

Calcium silicate-based root canal sealers: a literature review.

Epoxy resin-based sealers are currently widely used, and several studies have considered AH Plus to be the gold-standard sealer. However, it still has limitations, including possible mutagenicity, cytotoxicity, inflammatory response, and hydrophobicity. Drawing upon the advantages of mineral trioxide aggregate, calcium silicate-based sealers were introduced with high levels of biocompatibility and hydrophilicity. Because of the hydrophilic environment in root canals, water resorption and solubility of root canal sealers are important factors contributing to their stability. Sealers displaying lower microleakage and stronger push-out bond strength are also needed to endure the dynamic tooth environment. Although the physical properties of calcium silicate-based sealers meet International Organization for Standardization recommendations, and they have consistently reported to be biocompatible, they have not overcome conventional resin-based sealers in actual practice. Therefore, further studies aiming to improve the physical properties of calcium silicate-based sealers are needed.

Pulp-dentin regeneration: current approaches and challenges.

Regenerative endodontic procedures for immature permanent teeth with apical periodontitis confer biological advantages such as tooth homeostasis, enhanced immune defense system, and a functional pulp-dentin complex, in addition to clinical advantages such as the facilitation of root development. Currently, this procedure is recognized as a paradigm shift from restoration using materials to regenerate pulp-dentin tissues. Many studies have been conducted with regard to stem/progenitor cells, scaffolds, and biomolecules, associated with pulp tissue engineering. However, preclinical and clinical studies have evidently revealed several drawbacks in the current clinical approach to revascularization that may lead to unfavorable outcomes. Therefore, our review examines the challenges encountered under clinical conditions and summarizes current research findings in an attempt to provide direction for transition from basic research to clinical practice.

Reformulated mineral trioxide aggregate components and the assessments for use as future dental regenerative cements.

Mineral trioxide aggregate, which comprises three major inorganic components, namely, tricalcium silicate (C3S), dicalcium silicate (C2S), and tricalcium aluminate (C3A), is promising regenerative cement for dentistry. While mineral trioxide aggregate has been successfully applied in retrograde filling, the exact role of each component in the mineral trioxide aggregate system is largely unexplored. In this study, we individually synthesized the three components, namely, C3S, C2A, and C3A, and then mixed them to achieve various compositions (a total of 14 compositions including those similar to mineral trioxide aggregate). All powders were fabricated to obtain high purity. The setting reaction of all cement compositions was within 40 min, which is shorter than for commercial mineral trioxide aggregate (~150 min). Over time, the pH of the composed cements initially showed an abrupt increase and then plateaued (pH 10-12), which is a typical behavior of mineral trioxide aggregate. The compression and tensile strength of the composed cements increased (2-4 times the initial values) with time for up to 21 days in an aqueous medium, the degree to which largely depended on the composition. The cell viability test with rat mesenchymal stem cells revealed no toxicity for any composition except C3A, which contained aluminum. To confirm the in vivo biological response, cement was retro-filled into an extracted rat tooth and the complex was re-implanted. Four weeks post-operation, histological assessments revealed that C3A caused significant tissue toxicity, while good tissue compatibility was observed with the other compositions. Taken together, these results reveal that of the three major constituents of mineral trioxide aggregate, C3A generated significant toxicity in vitro and in vivo, although it accelerated setting time. This study highlights the need for careful consideration with regard to the composition of mineral trioxide aggregate, and if possible (when other properties are satisfactory), the C3A component should be avoided, which can be achieved by the mixture of individual components.

Transplantation of human adipose tissue-derived stem cells for repair of injured spiral ganglion neurons in deaf guinea pigs.

Excessive noise, ototoxic drugs, infections, autoimmune diseases, and aging can cause loss of spiral ganglion neurons, leading to permanent sensorineural hearing loss in mammals. Stem cells have been confirmed to be able to differentiate into spiral ganglion neurons. Little has been reported on adipose tissue-derived stem cells (ADSCs) for repair of injured spiral ganglion neurons. In this study, we hypothesized that transplantation of neural induced-human ADSCs (NI-hADSCs) can repair the injured spiral ganglion neurons in guinea pigs with neomycin-induced sensorineural hearing loss. NI-hADSCs were induced with culture medium containing basic fibroblast growth factor and forskolin and then injected to the injured cochleae. Guinea pigs that received injection of Hanks' balanced salt solution into the cochleae were used as controls. Hematoxylin-eosin staining showed that at 8 weeks after cell transplantation, the number of surviving spiral ganglion neurons in the cell transplantation group was significantly increased than that in the control group. Also at 8 weeks after cell transplantation, immunohistochemical staining showed that a greater number of NI-hADSCs in the spiral ganglions were detected in the cell transplantation group than in the control group, and these NI-hADSCs expressed neuronal markers neurofilament protein and microtubule-associated protein 2. Within 8 weeks after cell transplantation, the guinea pigs in the cell transplantation group had a gradually decreased auditory brainstem response threshold, while those in the control group had almost no response to 80 dB of clicks or pure tone burst. These findings suggest that a large amount of NI-hADSCs migrated to the spiral ganglions, survived for a period of time, repaired the injured spiral ganglion cells, and thereby contributed to the recovery of sensorineural hearing loss in guinea pigs.

Mastoid obliteration with silicone blocks after canal wall down mastoidectomy.

OBJECTIVES: To evaluate the usefulness of silicone blocks as graft material for mastoid cavity obliteration in the prevention of problematic mastoid cavities after canal wall down mastoidectomies. METHODS: Retrospective evaluation of 20 patients who underwent mastoid obliteration with silicone blocks between 2002 and 2009 at the Chonnam National University Hospital. The cases consisted of 17 patients with chronic otitis media with cholesteatoma and 3 patients with adhesive otitis media. The postoperative follow-up period was an average 49 months (range, 6 to 90 months). The surgical technique used at our institution composed four major steps: First, the canal wall down mastoidectomy was performed and the middle ear procedure was completed. The silicone blocks were used to fill up the mastoidectomized cavity. Then, a cortical bone pate was used to cover the surface of the silicone blocks. Finally, temporalis fascia and a split musculoperiosteal flap were used to surround the bone pate for reinforcement of the reconstructed canal wall. We examined postoperative success rate and hearing outcomes. RESULTS: In 19 cases (95%), the reconstructed canal wall maintained a cylindrical shape and the ear drum healed without perforation. In only 1 case (5%), the reconstructed canal wall was destroyed with ear drum perforation. The mean improvement in air-bone gap was about 12 dB (P<0.05), and the mean improvement in air-conduction was about 16 dB (P<0.05). CONCLUSION: We suggest that silicone blocks could be valuable resources as graft materials for mastoid obliteration after canal wall down mastoidectomies.

Matrix metalloproteinase inhibitor attenuates cochlear lateral wall damage induced by intratympanic instillation of endotoxin.

OBJECTIVE: Oxytetracycline and ilomastat are inhibitors of matrix metalloproteinases (MMPs). Their efficacy in protecting against cochlear damage induced by the intratympanic instillation of lipopolysaccharide (LPS), as a means of inducing labyrinthitis, was investigated. MATERIALS AND METHODS: Experiments were performed in 21 young male guinea pigs. Intratympanic instillation of LPS was done in the control group (n=7). Intratympanic instillation of oxytetracycline or ilomastat was done after LPS instillation in the experimental group. Measurements of auditory brainstem response (ABR) and cochlear blood flow (CBF) were performed. The organ of Corti was evaluated by field emission scanning electron microscopy (FE-SEM). The blood-labyrinth barrier (BLB) integrity was evaluated with Evans blue uptake. Gelatin zymography was used to assess the expression of active MMP-2 and MMP-9. RESULTS: Ears treated with MMP inhibitors were significantly protected from hearing loss compared to the LPS group. In LPS group, there was a significant decrease of CBF. However, experimental group displayed a statistically significant recovery of CBF. FE-SEM revealed hair cell damage in the LPS-treated group, but hair cells presented a normal appearance in MMP inhibitors. The LPS group showed a marked increase of Evans blue extravasation in the cochlea. However, MMP inhibitors significantly reduced the BLB opening. Active MMP-9 was expressed in the LPS group. Treatment with MMP inhibitors attenuated active MMP-9 expression. CONCLUSION: The MMP inhibitors oxytetracycline and ilomastat protect from cochlear lateral wall damage caused by LPS-induced labyrinthitis.

A comparison of the shaping ability of reciprocating NiTi instruments in simulated curved canals.

OBJECTIVES: The study was to compare the shaping ability of Reciproc (VDW) and WaveOne (Dentsply Maillefer) instruments compared with ProTaper, Profile and hand instrument during the preparation of simulated root canals. MATERIALS AND METHODS: Five groups (n = 5) were established. Reciproc, WaveOne, ProTaper, Profile and K file (K-flexo file) were used to prepare the resin simulated canals. A series of preoperative and postoperative images were taken by a microscope and superimposed in 2 different layers. The amount of resin removed from both the inner and the outer sides of the canal was measured to the level of 10 mm from the apical tip, with a 1 mm increment. RESULTS: The mean of resin removal from the inner canal wall was not different from the outer canal wall for Reciproc and WaveOne groups at apical third (1 - 3 mm level). There was no difference in the change of working length and maintenance of canal curvature. NiTi instruments are superior to stainless-steel K file in their shaping ability. CONCLUSIONS: Within the limitation of this present study, Reciproc and WaveOne instruments maintained the original canal curvature in curved canals better than ProTaper and Profile, which tend to transport towards the outer canal wall of the curve in the apical part of the canal.

Cavernous hemangioma of the tympanic membrane.

Cavernous hemangioma seems to most frequently arise in the posterior portion of the external auditory canal. However, they rarely occur in the tympanic membrane. A 49-year-old male patient was referred for evaluation of right-sided pulsatile tinnitus that he'd experienced for the previous 2 years. Temporal bone computerized tomography showed an isolated soft tissue mass just lateral to the tympanic membrane. There was no evidence of bony erosion or middle ear invasion. The patient underwent excision of the mass using a postauricular approach. The mass was removed en bloc and the defect of the tympanic membrane was repaired by tympanoplasty type I. There was no recurrence after 1 year of follow-up.

Dentinogenic potential of human adult dental pulp cells during the extended primary culture.

Despite the frequent use of primary dental pulp cells in dental regenerative research, few systematic studies of stemness for osteogenic and dentinogenic differentiation of human adult pulp cells have been reported. To investigate the stemness of human adult dental pulp cells, pulp tissues were obtained from extracted third molars and used as a source of pulp cells. In FACS analysis and immunophenotyping, the general mesenchymal stem cell markers CD44, CD90, and CD146 were highly expressed in early passages of the pulp cell culture. The stem cell population was dramatically decreased in an expansion culture of human dental pulp cells. When pulp cells were treated with additives such as ß-glycerophosphate, ascorbic acid, and dexamethasone, nodule formation was facilitated and mineralization occurred within 2 weeks. Expression of osteogenic markers such as alkaline phosphatase, osteocalcin, and osteonectin was relatively low in undifferentiated cells, but increased significantly under differentiation conditions in whole passages. Dentinogenic markers such as dentin sialophosphoprotein and dentin matrix protein-1 appeared to decrease in their expression with increasing passage number; however, peak levels of expression occurred at around passage 5. These data suggested that stem cells with differentiation potential might exist in the dental pulp primary culture, and that their phenotypes were changed during expansion culture over 8-9 passages. Under these conditions, a dentinogenic population of pulp cells occurred in limited early passages, whereas osteogenic cells occurred throughout the whole passage range.

Effect of Ginkgo biloba extract on endotoxin-induced labyrinthitis.

OBJECTIVE: There are no reports on the therapeutic effect of Ginkgo biloba extract (GBE) on otitis media-induced labyrinthitis. The present study examined whether GBE can protect against cochlear damage induced by intratympanic instillation of lipopolysaccharide (LPS)-induced labyrinthitis. MATERIALS AND METHODS: Experiments were performed in 20 healthy young male guinea pigs. The control group (n=10) received an intratympanic instillation of LPS (20 µl, 3mg/ml). The experimental group (n=10) received intratympanic instillation of LPS immediately after instillation of GBE (10mg/kg) and then experimental groups received GBE (100mg/kg) by intraperitoneal injection every day for 3 days. Instillation of LPS or LPS immediately after GBE was done in the right ear; the untreated left ear was considered normal. Physiological and morphological changes were evaluated. RESULTS: Statistical analysis of treatment of GBE revealed significantly less hearing loss than LPS group (p<0.05). The ratio of the value of cochlear blood flow (CBF) compared to untreated left side was significantly higher in the GBE treated group than in the LPS-treated group (p<0.05). This result indicated the recovery of CBF in GBE treated group compared to LPS treated group. In the LPS group, scanning electron microscopy revealed hair cell damage with edema. Missing stereocilia in the third layer of the outer hair cell was revealed. However, both the inner hair cells and the outer hair cells had normal appearance in the GBE group. LPS group showed that cochlear Evans blue extravasation was increased strongly in the stria vascularis, spiral limbus, and in the spiral ligament compared with the GBE treated group. CONCLUSION: GBE significantly minimizes cochlear damage against LPS-induced otitis media with labyrinthitis in a guinea pig model. GBE has potential as an adjunctive therapy to antibiotics in the treatment of acute otitis media with complicated labyrinthitis.

Transplantation of neural differentiated human mesenchymal stem cells into the cochlea of an auditory-neuropathy guinea pig model.

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.

Effect of piperacillin-tazobactam coated ß-tricalcium phosphate for mastoid obliteration in otitis media.

BACKGROUND AND OBJECTIVE: ß-Tricalcium phosphate (TCP) has good biodegradability and osteoconductivity as a scaffold material for bone tissue engineering. Both block and granular forms are available; however, it has been associated with risk of infection and exposure. To this end, the study evaluated the effect of piperacillin-tazobactam coated ß-TCPs for mastoid obliteration in otitis media. MATERIALS AND METHODS: Ten guinea pigs were divided into the experimental (piperacillin-tazobactam coated ß-TCP granules, n=5) and control groups (uncoated ß-TCP granules, n=5). After mastoid obliteration, transtympanic injection with a saline suspension of lipopolysaccharide established inflammation. The animals were sacrificed 5 weeks later. Tissue sections were stained with hematoxylin and eosin and examined. RESULTS: Encapsulation and formation of fibrous capsule by foreign material in the bulla were not evident. The histological evaluation did not reveal inflammatory cells and fibrosis in the piperacillin-tazobactam coated ß-TCP group. In contrast, the control group showed numerous inflammatory cells around the implanted uncoated ß-TCP granules and incomplete new bone formation. CONCLUSION: ß-TCP is an effective carrier material for piperacillin-tazobactam. The use of piperacillin-tazobactam coated ß-TCP may be optimal for mastoid obliteration.

Effect of nebulized bovine surfactant for experimental otitis media with effusion.

OBJECTIVES: In this study, we evaluated the efficacy of nebulized bovine pulmonary surfactant on experimentally induced otitis media with effusion (OME) in guinea pigs. METHODS: Twenty guinea pigs were divided into three groups. Four untreated animals served as normal controls. Experimental OME was established in both ears of the remaining 16 animals by a transbullar injection of 10 microL of Pseudomonas aeruginosa lipopolysaccharide in saline. Thereafter, the guinea pigs received nebulized phosphate buffered saline (n=8) or nebulized bovine pulmonary surfactant (n=8). Nebulization was given daily for 7 days. On day 8, all the animals' passive opening pressure (POP) of the Eustachian tube was measured and histopathological observations of the bulla were made by light microscopy. RESULTS: Nebulized bovine pulmonary surfactant significantly reduced the POP compared to that of saline nebulization. The bovine pulmonary surfactant improved the tubal patency and produced less histopathologcally-evident edematous bullar mucosa. CONCLUSION: Nebulization of bovine pulmonary surfactant plays an important role in treating otitis media with effusion in guinea pigs. Our results suggest that the chosen nebulized bovine pulmonary surfactant can be of good clinical benefit for treating OME in the future.

Functional neural differentiation of human adipose tissue-derived stem cells using bFGF and forskolin.

BACKGROUND: Adult mesenchymal stem cells (MSCs) derived from adipose tissue have the capacity to differentiate into mesenchymal as well as endodermal and ectodermal cell lineage in vitro. We characterized the multipotent ability of human adipose tissue-derived stem cells (hADSCs) as MSCs and investigated the neural differentiation potential of these cells. RESULTS: Human ADSCs from earlobe fat maintained self-renewing capacity and differentiated into adipocytes, osteoblasts, or chondrocytes under specific culture conditions. Following neural induction with bFGF and forskolin, hADSCs were differentiated into various types of neural cells including neurons and glia in vitro. In neural differentiated-hADSCs (NI-hADSCs), the immunoreactivities for neural stem cell marker (nestin), neuronal markers (Tuj1, MAP2, NFL, NFM, NFH, NSE, and NeuN), astrocyte marker (GFAP), and oligodendrocyte marker (CNPase) were significantly increased than in the primary hADSCs. RT-PCR analysis demonstrated that the mRNA levels encoding for ABCG2, nestin, Tuj1, MAP2, NFL, NFM, NSE, GAP43, SNAP25, GFAP, and CNPase were also highly increased in NI-hADSCs. Moreover, NI-hADSCs acquired neuron-like functions characterized by the display of voltage-dependent tetrodotoxin (TTX)-sensitive sodium currents, outward potassium currents, and prominent negative resting membrane potentials under whole-cell patch clamp recordings. Further examination by RT-PCR showed that NI-hADSCs expressed high level of ionic channel genes for sodium (SCN5A), potassium (MaxiK, Kv4.2, and EAG2), and calcium channels (CACNA1C and CACNA1G), which were expressed constitutively in the primary hADSCs. In addition, we demonstrated that Kv4.3 and Eag1, potassium channel genes, and NE-Na, a TTX-sensitive sodium channel gene, were highly induced following neural differentiation. CONCLUSIONS: These combined results indicate that hADSCs have the same self-renewing capacity and multipotency as stem cells, and can be differentiated into functional neurons using bFGF and forskolin.

Effect of neural-induced mesenchymal stem cells and platelet-rich plasma on facial nerve regeneration in an acute nerve injury model.

OBJECTIVES/HYPOTHESIS: The purpose of this study was to investigate the effects of platelet-rich plasma (PRP) and neural-induced human mesenchymal stem cells (nMSCs) on axonal regeneration from a facial nerve axotomy injury in a guinea pig model. STUDY DESIGN: Prospective, controlled animal study. METHODS: Experiments involved the transection and repair of the facial nerve in 24 albino guinea pigs. Four groups were created based on the method of repair: suture only (group I, control group); PRP with suture (group II); nMSCs with suture (group III); and PRP and nMSCs with suture (group IV). Each method of repair was applied immediately after nerve transection. The outcomes measured were: 1) functional outcome measurement (vibrissae and eyelid closure movements); 2) electrophysiologic evaluation; 3) neurotrophic factors assay; and 4) histologic evaluation. RESULTS: With respect to the functional outcome measurement, the functional outcomes improved after transection and reanastomosis in all groups. The control group was the slowest to demonstrate recovery of movement after transection and reanastomosis. The other three groups (groups II, III, and IV) had significant improvement in function compared to the control group 4 weeks after surgery (P < .05). On the electrophysiologic evaluation, there was significantly better performances in groups II, III, and IV when compared to group I with respect to the amplitude and excitation area of the compound motor action potentials (MAPs) 4 and 6 weeks after surgery (P < .05); group IV had the best performance. A Western blot assay showed that group II had marked expression of several neurotrophic factors. Groups II, III, and IV demonstrated better results in axon counts and myelin thickness when compared with group I. Based on quantitative histology analysis, group IV had the greatest myelinated axon fibers compared to the other groups (P < .05). CONCLUSIONS: The use of PRP and/or nMSCs promotes facial nerve regeneration in an animal model of facial nerve axotomy. The use of nMSCs showed no benefit over the use of PRP in facial nerve regeneration, but the combined use of PRP and nMSCs showed a greater beneficial effect than use of either alone. This study provides evidence for the potential clinical application of PRP and nMSCs in peripheral nerve regeneration of an acute nerve injury. Laryngoscope, 2010.

Fate of sudden deafness occurring in the only hearing ear: outcomes and timing to consider cochlear implantation.

The present study was undertaken to learn the outcome of patients with idiopathic sudden sensorineural hearing loss (ISSNHL) in their only hearing ear. Timing to conduct a cochlear implantation was also determined in those who did not recover the hearing. The study group comprised 25 patients who confronted ISSNHL in their only hearing ear. A total of 192 patients, who had ISSNHL in one ear and had normal contralateral ear, served as the control. Demographically there were no significant differences between the groups. The recovery rate was similar between the groups: 64.0% in the experimental and 62.5% in the control group. The duration until the recovery of ISSNHL in the only hearing ear was 5-90 days (average 17.6 days). In the experimental group, 8 patients did not recover from ISSNHL, and underwent cochlear implantation in 6 with satisfactory results. These results suggest that the same treatment is applicable for patients with ISSNHL regardless of whether their contralateral ear is deaf or normal. For those who do not recover from ISSNHL in their only hearing ear, culminating in bilateral deafness, we may consider further definitive treatment including cochlear implantation as early as 3 months after initiating the treatment of ISSNHL.

Efficacy of transnasal nebulized surfactant on experimental otitis media with effusion in guinea pig.

OBJECTIVE: Recently experimental trials have demonstrated that nebulized natural surfactant is effective in the treatment of experimental otitis media with effusion (OME). Two methodologically incomplete studies using animal models of induced OME have reported that nebulized surfactant can improve Eustachian tube function. There have been no reports that investigated the efficacy of the nebulized surfactant for OME using analysis of tympanic membrane vibration. In this study, we evaluated the therapeutic effect of nebulized natural surfactant by investigation of tympanic membrane vibration. MATERIALS AND METHODS: The guinea pigs (n=20) were divided into three groups. Four normal guinea pigs without any treatment were used as normal controls. Sixteen pigs were given experimental OME in both ear and then divided into two groups. The OME model of guinea pig was created by transbullar injection 10 microl of lipopolysaccharide in saline. Experimental group 1 (n=8) was nebulized phosphate buffered saline (PBS). Experimental group 2 (n=8) was nebulized natural bovine surfactant. The transnasal nebulization was repeated for 7 days in both experimental groups. On the 8th day, both experimental groups and normal control group were measured the tympanic membrane vibration by laser Doppler vibrometer (LDV), and additionally histopathologic observation was performed by scanning electron microscope (SEM). RESULTS: The TM vibration velocity in nebulized PBS group showed significantly reduced mainly at low frequencies. However, nebulized natural surfactant group recovered the reduction of tympanic membrane vibration. In nebulized PBS group, SEM showed severe thickened subepithelial layer and hyperplasia of gland structure. However, in nebulized natural surfactant group, SEM showed the reduced the thickness of subepithelial layer and the loss of glandular hyperplasia. CONCLUSION: From our results, the nebulized natural surfactant is postulated to be effective in the treatment of intractable OME in animal model. The measurement of tympanic membrane vibration for analysis of middle ear mechanics was significantly corresponding to the middle ear mucosal changes. LDV is a useful tool for investigate the therapeutic effect of nebulized surfactant in experimental OME. With supporting clinical studies, nebulization of natural surfactant may become noninvasive treatment of OME in future.

Assessment and treatment of dynamic obstruction in anomalous right coronary artery using dynamic diastolic pressure gradient change during dobutamine challenge with rapid atrial pacing.

In anomalous right coronary artery (RCA), extravascular coronary compression results in dynamic obstruction which can cause effort angina as well as syncope. Measurement of dynamic intracoronary pressure gradient (DeltaP) change across the lesion using dobutamine challenge with rapid atrial pacing (RAP) can be used for assessment of dynamic obstruction in anomalous coronary artery. We report a case of anomalous origin of RCA which dynamic diastolic pressure gradient (DeltaP) was measured using this method. During dobutamine challenge with RAP, diastolic DeltaP significantly increased from 6 mm Hg at baseline to 13 mm Hg, and when diastolic DeltaP reached to 13 mm Hg, T wave inversion was observed, the chest pain was developed. After intracoronary stenting, diastolic DeltaP decreased to 2 mm Hg, chest pain was relieved, and T wave inversion was disappeared. The patient was asymptomatic at two months' follow-up.

Cholesterol granuloma of the tympanic membrane presenting as a blue eardrum.

Intramembranous tympanic membrane cholesterol granuloma (CG) occurs infrequently. Here, the authors report a case of CG in the tympanic membrane presenting as a blue eardrum in the right ear. In addition, a pinhole perforation noted in the anterosuperior area revealed a brown discharge. High-resolution temporal bone CT showed a bulging mass shadow in the middle ear and a soft tissue dense lesion that filled both the epitympanum and mastoid cavity. Tympanomastoidectomy was performed under general anesthesia. New bone formation was confirmed in the mastoid antrum and epitympanum, and the epitympanum was blocked by new bone. The tympanic membrane revealed a round, brownish mass with a glistening surface and a severely thickened pars tensa. We herein report this case and review pertinent medical literature.