Bone Regeneration Capability of 3D Printed Ceramic Scaffolds.

In this study, we evaluated the bone regenerative capability of a customizable hydroxyapatite (HA) and tricalcium phosphate (TCP) scaffold using a digital light processing (DLP)-type 3D printing system. Twelve healthy adult male beagle dogs were the study subjects. A total of 48 defects were created, with two defects on each side of the mandible in all the dogs. The defect sites in the negative control group (sixteen defects) were left untreated (the NS group), whereas those in the positive control group (sixteen defects) were filled with a particle-type substitute (the PS group). The defect sites in the experimental groups (sixteen defects) were filled with a 3D printed substitute (the 3DS group). Six dogs each were exterminated after healing periods of 4 and 8 weeks. Radiological and histomorphometrical evaluations were then performed. None of the groups showed any specific problems. In radiological evaluation, there was a significant difference in the amount of new bone formation after 4 weeks (p < 0.05) between the PS and 3DS groups. For both of the evaluations, the difference in the total amount of bone after 8 weeks was statistically significant (p < 0.05). There was no statistically significant difference in new bone between the PS and 3DS groups in both evaluations after 8 weeks (p > 0.05). The proposed HA/TCP scaffold without polymers, obtained using the DLP-type 3D printing system, can be applied for bone regeneration. The 3D printing of a HA/TCP scaffold without polymers can be used for fabricating customized bone grafting substitutes.

3D-Printed Ceramic Bone Scaffolds with Variable Pore Architectures.

This study evaluated the mechanical properties and bone regeneration ability of 3D-printed pure hydroxyapatite (HA)/tricalcium phosphate (TCP) pure ceramic scaffolds with variable pore architectures. A digital light processing (DLP) 3D printer was used to construct block-type scaffolds containing only HA and TCP after the polymer binder was completely removed by heat treatment. The compressive strength and porosity of the blocks with various structures were measured; scaffolds with different pore sizes were implanted in rabbit calvarial models. The animals were observed for eight weeks, and six animals were euthanized in the fourth and eighth weeks. Then, the specimens were evaluated using radiological and histological analyses. Larger scaffold pore sizes resulted in enhanced bone formation after four weeks (p < 0.05). However, in the eighth week, a correlation between pore size and bone formation was not observed (p > 0.05). The findings showed that various pore architectures of HA/TCP scaffolds can be achieved using DLP 3D printing, which can be a valuable tool for optimizing bone-scaffold properties for specific clinical treatments. As the pore size only influenced bone regeneration in the initial stage, further studies are required for pore-size optimization to balance the initial bone regeneration and mechanical strength of the scaffold.

Feasibility of injectable thermoreversible gels for use in intramuscular injection of parathyroid autotransplantation.

Surgical transplantation of parathyroid tissue into the forearm muscle is one of the most commonly used surgical techniques. While simple, the procedure suffers from drawbacks. This study evaluated the feasibility of thermoreversible gel as an injectable carrier for parathyroid autotransplantation. Polyethyleneglycol-polyalanine-co-phenylalanine (PEG-PAF) thermoreversible gel (sol form at 4 °C, gel form at 37 °C) were manufactured. Thirty-eight Sprague-Dawley rats were divided into two groups (19 control, C group; 19 experimental, P group). The parathyroid glands of rats were excised. Parathyroid tissues were transplanted into the muscle pocket in sternocleidomastoid muscle in the C group. In the P group, the tissues were injected into the same muscle mixed with 0.3 ml thermoreversible gel. The serum levels of parathyroid hormone (PTH), ionized calcium, and phosphorous were measured before surgical procedure, on 7, 21, 56, and 70 days after surgery. Histology and immunohistochemistry were performed. Preoperative median PTH level of the C and the P group were 60.80 and 43.85 pg/ml, respectively (p = 0.641). Seventy days after surgery, median PTH level was 32.8 and 25.61 pg/ml, respectively. On day 70, the PTH level was restored by 54 % in the C group and 56 % in the P group compared to the preoperative value (p = 0.620). There were no significant intergroup differences in the ionized calcium/phosphorous level. Histology and immunohistochemistry revealed the successful transplantation of parathyroid tissues into the muscles in both groups. In conclusion, the PEG-PAF-based thermoreversible gel is a good candidate carrier material for intramuscular parathyroid autotransplantation.

Development of hypoparathyroidism animal model and the feasibility of small intestinal submucosa application on the parathyroid autotransplantation.

The purpose of this study is to evaluate the feasibility of small intestinal submucosa (SIS) application on the parathyroid autotransplantation in a rat model of hypoparathyroidism. The rats were divided into four groups: NC (no procedure, n = 5), PTX (total parathyroidectomy, n = 6), PT (total parathyroidectomy and parathyroid autotransplantation, n = 10) and PT + SIS group (total parathyroidectomy and parathyroid autotransplantation with SIS, n = 10). The levels of parathyroid hormone (PTH), calcium, and phosphorous were measured on 0, 3, 7, 21, 56 and 84 days after surgery. PTH level was expressed as median (interquartile range) and histological and immunohistochemical examinations were performed. PTH levels were significantly decreased to "not detectable level" from day 3 in PTX group. PTH was not detected in both PT and PT + SIS groups on the 21st day. On the 56th day, PTH levels were increased in both groups: 3 out of 8 rats (37.5%) in the PT group, 6 out of 9 rats (66.7%) in the PT + SIS group. The PTH level was fully recovered to its preoperative range on the day 84 as 6 of 8 rats (75%) of the PT group and 7 of 9 rats (77.8%) of the PT + SIS group were recovered; the PTH levels were 117.84 and 178.36 pg/ml, respectively. The neo-vascularization was well observed around the parathyroid tissue, and the number of new vessels formed was higher in the PT + SIS group (15 vessels/high power field) as compared to the PT group (10 vessels/high power field). This study showed the feasibility and the treatment effect of SIS as the success rate of autotransplantation of parathyroid tissue was significantly increased without severe inflammatory response in hypothyroidism animal model.

Tonsil-derived mesenchymal stromal cells: evaluation of biologic, immunologic and genetic factors for successful banking.

BACKGROUND AIMS: Although mesenchymal stromal cells (MSC) from human palatine tonsils (tonsillar MSC, T-MSC) have been isolated, whether T-MSC isolated from multiple donors are feasible for cell banking has not been studied. METHODS: T-MSC before and after a standard protocol of cryopreservation and thawing were assessed regarding several basic characteristics, including colony-forming unit-fibroblast features, MSC-specific surface antigen profiles, and inhibition of alloreactive T-cell proliferation. In vitro mesodermal differentiation potentials to adipocytes, osteocytes and chondrocytes were detected by staining with either cell-specific dyes or antibody after incubation with each appropriate differentiation medium. Expression of mesoderm-specific genes was also quantified by real-time polymerase chain reaction (PCR) assay. Expression profiles of endoderm-specific genes were identified by reverse transcription PCR assay. The feasibility of T-MSC in future engraftment was tested by short tandem repeat (STR) analysis using genomic DNA isolated randomly from three independent subjects. RESULTS: Both fresh and cryopreserved-thawed T-MSC showed a similar high proliferation capacity and expressed primitive cell-surface markers. Hematopoietic cell markers, HLA-DR, co-stimulatory molecules and follicular dendritic cell markers were not detected. In addition to mesodermal differentiation, fresh and cryopreserved-thawed cells also underwent endodermal differentiation, as evidenced by the expression of endoderm-specific genes including forkhead box A2 (FoxA2), SIX homeobox 1 (Six1) and chemokine (C-C motif) ligand 21 (CCL21). Both cells significantly decreased phorbol 12- myristate 13-acetate (PMA)-induced T-cell proliferation. T-MSC from three independent donors formed chimerism in STR analysis. CONCLUSIONS: Our results demonstrate for the first time that T-MSC are a potentially good source for MSC banking.

Three-dimensional CT with virtual bronchoscopy: a useful modality for bronchial foreign bodies in pediatric patients.

Bronchial foreign bodies (FB) in children are difficult to diagnose when there is no witness to the aspiration event. The experience with three-dimensional CT and virtual bronchoscopic reconstruction (3D-CTVB) was reviewed for the diagnosis of bronchial FB in pediatric patients in this study. A retrospective review of ten pediatric patients (M:F = 5:5) who underwent rigid bronchoscopy by one otolaryngologist for the impression of a bronchial FB, at a tertiary training hospital, from February 2006 to September 2010 is reported. All patients had a plain chest X-ray and 3D-CTVB. The physical, radiological and bronchoscopic findings of the patients were analyzed. The mean duration from the aspiration event to hospital evaluation was 7 days (0-21 days). Four patients (44%) had witnesses to the aspiration event. Hyperaeration of the affected lung was noted in nine patients (90%) on the plain chest X-ray. In all of the patients, the foreign body location, suspected on the preoperative 3D-CTVB, was consistent with the intra-operative bronchoscopic findings. All FBs were successfully removed by rigid bronchoscopy without any complications. In conclusion, the 3D-CTVB was a very safe and useful diagnostic modality for bronchial FBs. The findings of this study suggest that the 3D-CTVB should be considered for the diagnosis of all patients with suspected bronchial FBs.

Apatite nanosheets inhibit initial smooth muscle cell proliferation by damaging cell membrane.

Understanding how nanostructured coatings interact with cells is related to how they manipulate cell behaviors and is therefore critical for designing better biomaterials. The apatite nanosheets were deposited on metallic substrates via biomimetic precipitation. Cell viability of apatite nanosheets towards to smooth muscle cells (SMCs) were investigated, and the underlying mechanism was proposed. Apatite nanosheets presented inhibitory activity on SMC growth, and caused rupture of cell membranes. On the basis of measuring changes in intracellular calcium ([Ca2+]i), observing cell contraction and apatite nanosheets - SMC interaction, it was found that calcium ions released from apatite led to rises in [Ca2+]i, which induced vigorous SMC contraction on apatite nanosheets. Consequently, the cell membrane of individual SMCs was cut/penetrated by the sharp edges of apatite nanosheets, resulting in cell inactivation. This damage of cell membranes suggests a novel mechanism to manipulate cell viability, and may offer insights for the better design of calcium-based nanostructured coatings or other biomedical applications.

Endoscope assisted submandibular sialadenectomy: the face-lift approach.

This study evaluated the benefits of a new approach, endoscopic resection of a submandibular gland through the face-lift approach. The study comprised the clinical trial of new procedure. Six submandibular sialadenectomies were carried out (4 women, 2 men; mean age 28 years) via the face-lift approach by endoscopy. All six submandibular gland resections were successfully performed endoscopically, and no conversion to conventional open resection was necessary. Five patients had a mixed tumor and one patient had sialadenitis. The procedures lasted 55-115 min (mean 74 min). No complications associated with the operation occurred, and the scar was almost invisible. Endoscope assisted submandibular sialadenectomy via the face-lift approach was safe and effective treatment of benign lesions. The main advantage of this procedure is that the operative scar is concealed by the hairline.

Bone regeneration in ceramic scaffolds with variable concentrations of PDRN and rhBMP-2.

This study evaluated the bone regeneration capacity and mechanical properties of block-type hydroxyapatite (HA)/tricalcium phosphate (TCP) scaffolds in response to different concentrations of polydeoxyribonucleotide (PDRN) and recombinant human bone morphogenic protein 2 (rhBMP-2). Thirty-two male white rabbits were used as a model of calvarial bone defect and classified into eight groups according to type and concentration of growth factor administered, viz., control group (only HA/TCP scaffold), scaffold + PDRN (0.1, 1, 5, and 10 mg/mL each) and scaffold + rhBMP-2 (0.01, 0.05, and 0.1 mg/mL each). The specimens were evaluated using histomorphometric and radiological analyses. Histomorphometric analyses indicated that the administration of PDRN did not increase bone formation. However, significant increases in bone formation were observed with the administration of rhBMP-2 at 0.05 and 0.10 mg/mL on week 8 compared to the control (p < 0.05). Radiological analyses revealed a significant increase in bone formation at week 8 with the administration of PDRN at 5 mg/mL and 10 mg/mL, and rhBMP-2 at 0.05 or 0.10 mg/mL compared to the control (p < 0.05). Our findings show that block-type HA/TCP scaffolds possess sufficient mechanical strength and bone regeneration capacity when used with optimal concentrations of growth factors.

Trends in the Incidence and Treatment of Benign Vocal Fold Lesions in Korea, 2006-2015: A Nationwide Population-based Study.

OBJECTIVE: The aim of this study was to determine the prevalence and demographics of benign vocal fold lesions (BVFL) and trends in its treatment in Korea based on data collected from the National Health Insurance Service database. MATERIAL AND METHODS: Data for patients diagnosed with BVFL (ICM-10 codes J381, J382, J384) from 2006 to 2015 were selected for analysis. Patient characteristics, including sex, age, income, area of residence, and comorbidity, were analyzed. Treatment was divided into surgical management and conservative management using operation codes. RESULTS: The prevalence and incidence of BVFL increased from 7.07% and 5.29%, respectively, in 2006 to 12.47% and 7.98% in 2015. Compared with the non-BVFL population, patients with BVFL were more likely to be female, reside in an urban area, and have gastroesophageal reflux disease. There was no significant change in the incidence of surgical treatment during the study period (around 6000 per year); however, the surgical treatment rate decreased from 19.29% to 8.38%. The probability of undergoing surgical treatment for BVFL was higher in men, those aged 50-59 years, and those in the lowest quartile for income, except for the medical aid group. CONCLUSION: In Korea, there was an increase in the number of patients diagnosed with BVFL and a decrease in the operation rate for this condition between 2006 and 2015. Diagnosis of BVFL varied significantly based on income and sex; however, the only variable affecting the operation rate was patient age.

Lateral Window Design for Maxillary Sinus Graft Based on the Implant Position.

The purpose of this study was to devise a classification and lateral window design method based on implants and to evaluate whether these classifications and methods are applicable to clinical practice. When applying the maxillary sinus elevation technique using the lateral window, possible situations were classified into four: (A) two or more sites for implants are required for maxillary sinus augmentation, (B) a single implant is required when there are no adjacent teeth, (C) a single implant is required when one adjacent tooth is present at the mesial or distal area, and (D) a single implant is required when both mesial and distal adjacent teeth are present. In order to verify whether this classification can be used in all situations, 76 patients who underwent maxillary sinus elevation with a lateral window were selected and investigated. Of them, 47 (62%) were included in Group A, 9 (12%) in Group B, 8 (11%) in Group C, and 12 (15%) in Group D. Lateral window designing in the lateral approach of sinus augmentation can be classified into four clinical situations. There were no unclassified cases. This classification and window positioning method can be applied to most cases.

Development of guideline for rating the physical impairment of otolaryngologic field.

We develop a guideline for rating the physical impairment of otolaryngologic fields. Assessment of hearing disturbance and tinnitus required physical examination, pure tone audiometry, speech audiometry, impedance audiometry, brainstem evoked response audiometry, Bekesy audiometry, otoacoustic emission test, and imaging examination. History taking, physical examination, and radiological examination for the vestibular organ and brain, righting reflex test, electronystagmography, and caloric test are taken for evaluation of balance disorder. Olfactory function tests include University of Pennsylvania Smell Identification test, Connecticut Chemosensory Clinical Research Center test, T and T olfactometry and Korean Version of Sniffin's Sticks test. Medical history and physical examination is mandatory to evaluatezseverity of respiration difficulty. Examinations include flexible fiberoptic nasopharyngoscope, bronchoscopy, simple soft-tissue radiography films of upper airway and high resolution computed tomography. Evaluation of mastication and swallowing are history taking, physical examination, examination for upper jaw, lower jaw, and temporomandibular joint, dental examination and radiological studies. Endoscopy and esophagography are also needed. Voice disorder is evaluated based on physical examination, oral pharynx and larynx endoscopy, larynx stroboscopy, hearing assessment, laryngeal electromyography, sound analysis test, aerodynamic test, electroglottography, and radiologic examination. Articulation disorder is assessed by picture consonant articulation test. These are position articulation test, Lee-Kim Korean articulation picture and speech intelligibility assessment.

Cystic lymphangioma: trauma may be a significant cause?

Cystic lymphangioma is a rare congenital anomaly of the lymphatic system. Its embryopathogenesis is still controversial. Most lesions present in infancy or early childhood with a swelling found in the head and neck region. We report the case of a woman who presented with a cystic swelling in the left supraclavicular region that appeared after left side thyroidectomy. Surgical excision achieved complete removal. Histology showed the lesion to be a cystic lymphangioma. It can be hypothesized that an embryogenetic origin of the adult lymphangioma is associated with some precipitating trauma such as surgery.

Silk fibroin membrane used for guided bone tissue regeneration.

With the aim to develop a novel membrane with an appropriate mechanical property and degradation rate for guided bone tissue regeneration, lyophilized and densified silk fibroin membrane was fabricated and its mechanical behavior as well as biodegradation property were investigated. The osteoconductive potency of the silk fibroin membranes were evaluated in a defect rabbit calvarial model. Silk fibroin membrane showed the modulated biodegradable and mechanical properties via ethanol treatment with different concentration. The membrane could prevent soft tissue invasion from normal tissue healing, and the amounts of new bone and defect closure with silk fibroin membrane were similar to those of commercially available collagen membrane.

Biomimetic apatite formed on cobalt-chromium alloy: A polymer-free carrier for drug eluting stent.

In this study, sirolimus (SRL) was loaded within biomimetic apatite formed on cobalt-chromium (Co-Cr) alloy, which has been reported for the first time, to inhibit the in-stent restenosis. Two different groups of loading SRL within biomimetic apatite were prepared: Group A (mono-layer of apatite/SRL) and Group B (bi-layer of apatite/SRL). Group A and Group B showed the biphasic pattern of SRL release up to 40 and 90days, respectively. The attachment of human artery smooth muscle cell (HASMC) for both Group A and Group B was significantly inhibited, and proliferation dramatically decreased with the release of SRL. Noteworthily, biomimetic apatite alone also suppressed the SMC proliferation. The porous biomimetic apatite uniformly covered Co-Cr stent without crack or webbings. After balloon expansion, the integrity of biomimetic apatite was sufficient to resist delamination or destruction. Thus, this study demonstrated that biomimetic apatite is a promising drug carrier for potential use in stents.

Laryngeal myofascial pain syndrome as a new diagnostic entity of dysphonia.

OBJECTIVE: To consider the feasibility of diagnosing intrinsic laryngeal muscle myofascial pain syndrome (MPS) in dysphonic patients who demonstrated immediate symptom and stroboscopic finding improvement after laryngeal electromyography (LEMG) without further treatment. METHODS: A chart review of patients who showed subtle vocal fold movement abnormalities on a stroboscopic examination and underwent ultrasonography (US)-guided LEMG was performed. Patients with vocal fold paralysis, mucosal lesions, spasmodic dysphonia, and vocal tremor on stroboscopic examination were excluded. Among them, patients with normal EMG findings were included in this study. The patients who reported voice symptom improvement after LEMG without further treatment were placed in laryngeal MPS (LMPS) group and the other patients were placed in non-laryngeal MPS (non-MPS) group. Predisposing factors, voice symptom, symptom-duration, and stroboscopic findings of these patients were reviewed. RESULTS: Among the 16 patients, LEMG findings were normal, five (31%) were included in the LMPS group and the other 11 patients (69%) were included in the non-MPS group. All LMPS group patients had a history of voice abuse and reported odynophonia. The Korean Voice Handicap Index-10 score decreased significantly after US-guided LEMG without additional treatment in the LMPS group. The stroboscopic findings revealed that vocal fold hypomobility was the most common finding in the LMPS group, and two patients showed a muscle tension dysphonia pattern. The LMPS groups showed improvement of vocal fold mobility on 1-week stroboscopic evaluation. CONCLUSION: LMPS is a potential diagnosis for patients with vocal fold hypomobility finding on stroboscopic findings but with normal EMG results. Diagnosis of LMPS could be considered in patients who showed symptom and vocal fold movement improvement after LEMG.

Standardization of A Physiologic Hypoparathyroidism Animal Model.

Ideal hypoparathyroidism animal models are a prerequisite to developing new treatment modalities for this disorder. The purpose of this study was to evaluate the feasibility of a model whereby rats were parathyroidectomized (PTX) using a fluorescent-identification method and the ideal calcium content of the diet was determined. Thirty male rats were divided into surgical sham (SHAM, n = 5) and PTX plus 0, 0.5, and 2% calcium diet groups (PTX-FC (n = 5), PTX-NC (n = 10), and PTX-HC (n = 10), respectively). Serum parathyroid hormone levels decreased to non-detectable levels in all PTX groups. All animals in the PTX-FC group died within 4 days after the operation. All animals survived when supplied calcium in the diet. However, serum calcium levels were higher in the PTX-HC than the SHAM group. The PTX-NC group demonstrated the most representative modeling of primary hypothyroidism. Serum calcium levels decreased and phosphorus levels increased, and bone volume was increased. All animals survived without further treatment and did not show nephrotoxicity including calcium deposits. These findings demonstrate that PTX animal models produced by using the fluorescent-identification method, and fed a 0.5% calcium diet, are appropriate for hypoparathyroidism treatment studies.

3D printed polyurethane prosthesis for partial tracheal reconstruction: a pilot animal study.

A ready-made, acellular patch-type prosthesis is desirable in repairing partial tracheal defects in the clinical setting. However, many of these prostheses may not show proper biological integration and biomechanical function when they are transplanted. In this study, we developed a novel 3D printed polyurethane (PU) tracheal scaffold with micro-scale architecture to allow host tissue infiltration and adequate biomechanical properties to withstand physiological tracheal condition. A half-pipe shaped PU scaffold (1.8 cm of height, 0.18 cm thickness, and 2 cm of diameter) was fabricated by 3D printing of PU 200 µm PU beam. The 3D printed tracheal scaffolds consisted of a porous inner microstructure with 200 × 200 × 200 µm3 sized pores and a non-porous outer layer. The mechanical properties of the scaffolds were 3.21 ± 1.02 MPa of ultimate tensile strength, 2.81 ± 0.58 MPa of Young's modulus, and 725% ± 41% of elongation at break. To examine the function of the 3D printed tracheal scaffolds in vivo, the scaffolds were implanted into 1.0 × 0.7 cm2 sized anterior tracheal defect of rabbits. After implantation, bronchoscopic examinations revealed that the implanted tracheal scaffolds were patent for a 16 week-period. Histologic findings showed that re-epithelialization after 4 weeks of implantation and ciliated respiratory epithelium with ciliary beating after 8 weeks of implantation were observed at the lumen of the implanted tracheal scaffolds. The ingrowth of the connective tissue into the scaffolds was observed at 4 weeks after implantation. The biomechanical properties of the implanted tracheal scaffolds were continually maintained for 16 week-period. The results demonstrated that 3D printed tracheal scaffold could provide an alternative solution as a therapeutic treatment for partial tracheal defects.

Evaluation of Different Combinations of Biphasic Calcium Phosphate and Growth Factors for Bone Formation in Calvarial Defects in a Rabbit Model.

The aim of the present preclinical study was to investigate the capability of a new formulation of biphasic calcium phosphate (BCP) in achieving new bone formation either by itself or in combination with different concentrations of growth factors. Twenty-four 3-month-old male New Zealand white rabbits (weight range, 2.5 to 3.0 kg) that had been bred exclusively for biomedical research purposes and obtained from a licensed vendor were used. Four calvarial defects were created in each animal, for a total of 96 defects. Each defect received alloplastic BCP (Osteon III, Genoss) that was composed of 60% hydroxyapatite and 40% ß-tricalcium phosphate) (porosity, ~80%; macropore size, 200 to 400 µm; crystallinity, 95%) combined with different concentrations of recombinant human platelet-derived growth factor BB (rhPDGF-BB), human recombinant basic fibroblast growth factor-2 (rhFGF-2), or recombinant human bone morphogenetic protein-2 (rhBMP-2). A custom-made polycarbonate tube was fixed to each defect site by applying slight pressure, and a mixture of bone graft and growth factor was implanted into the tubes. Data were collected 2, 4, and 8 weeks after creation of the defects to assess early and late healing. Various amounts of newly formed bone and remnant BCP particles formed inside of the tube throughout the study period. The BCP + 0.5 mg/mL rhBMP-2 group exhibited the most bone formation. At 8 weeks, more new bone formation was noted in the Osteon III + rhBMP-2 combined group than in other groups. The present study results indicate that BCP can be combined with different concentrations of rhBMP-2, rhFGF-2, and rhPDGF-BB to produce new bone formation within a polycarbonate tube in calvarial defects in a rabbit model.

Clinical Usefulness of Ultrasonography-Guided Laryngeal Electromyography.

OBJECTIVES/HYPOTHESIS: To investigate the proper approach and technical method of ultrasonography-guided laryngeal electromyography (US-guided LEMG). STUDY DESIGN: This is a prospective study. METHODS: Twenty patients who underwent US-guided LEMG were enrolled. US-guided LEMG was cooperatively performed by one otolaryngologist, one neurologist, and one radiologist. The location of the needle electrode was confirmed with US after electrode insertion into the laryngeal intrinsic muscle. The US transducer was applied on the neck by a transverse/midline and transverse/oblique approach to identify the cricothyroid (CT), thyroarytenoid (TA) muscles, and the location of the needle electrode. RESULTS: CT muscles were easily identified on US in all 20 patients. TA muscles were identified in 17 patients (85%). The transverse/oblique approach was helpful to detect TA muscle in case of calcified thyroid cartilage or anatomic variation. CONCLUSIONS: US-guided LEMG, which enables the exact insertion of the needle electrode, improves the reliability of examination and is helpful in early detection and to prevent complications.