Multimodal imaging using optical coherence tomography and endolaryngeal ultrasonography in a new rabbit VX2 laryngeal cancer model.

BACKGROUND AND OBJECTIVE: Optical coherence tomography (OCT) provides ultrahigh-resolution imaging of tissues within a depth of a few millimeters, whereas ultrasonography provides good imaging further below the surface. We aimed to develop a minimally invasive rabbit model of VX2 laryngeal cancer, suitable for these two imaging modalities through a transoral approach. We also sought to study the utility of combined OCT and endolaryngeal ultrasonography (EUS) for evaluation of early and advanced laryngeal cancer, using this model. MATERIALS AND METHODS: VX2 tumor suspension was inoculated into the vocal folds of ten rabbits by injection through the trans-thyrohyoid membrane. The tumor model was characterized by rigid laryngoscopy and the tumor generation rate was 80% (8/10). Correlation between frequency-domain OCT and high-frequency EUS were used to visualize laryngeal tumors in the area of protruding mass formation in four rabbits, one week after injection (group A) and the remaining four rabbits two weeks after injection (group B). RESULTS: A small submucosal tumor was observed with rigid laryngoscopy in group A, and pathologic evaluation showed that the tumor was close to the basement membrane of the vocal fold mucosa, but had not invaded. OCT confirmed that the lining of the mucosa and basement membrane of the vocal fold was not broken, but the mucosa had thinned at the most elevated ridge. However, these lesions were not detected by EUS, and the overall shape of the tumor could not be clearly identified by EUS. A large tumor filling the laryngeal lumen was observed with rigid laryngoscopy in group B, and nearly the entire vocal fold, including the paraglottic space, was found to be involved on pathologic analysis. Distinguishing between normal structures and tumor was difficult using OCT; however, EUS confirmed the overall shape, size, and extent of the tumor, and the paraglottic space and thyroid cartilage were shown to be intact. CONCLUSIONS: This study is the first experimental trial, assessing the value of multimodal imaging using OCT and EUS in a rabbit VX2 laryngeal tumor model. Combining OCT and EUS helped to identify changes in laryngeal mucous membranes, and could potentially be used to identify laryngeal tumors and predict how tumors progress. This combined modality could help in determining tumor extent, assisting in diagnosis, and establishing a treatment plan for laryngeal cancer.

In vivo real-time imaging of airway dynamics during bronchial challenge test.

BACKGROUND AND OBJECTIVE: Asthmatic patients exhibit airway hyper-responsiveness, which induces bronchoconstriction and results in a ventilation defect. The bronchial challenge test using methacholine is a useful way to measure airway hyper-responsiveness with airway constriction. Anatomical optical coherence tomography has been used to image airway hyper-responsiveness of medium sized bronchus with the aid of an endoscopic probe. Recently, a thoracic window was reported that allows direct visualization of terminal airway such as alveolus. A multi-scale integrated airway dynamics was assessed in this study. We imaged in vivo changes in the right intermedius bronchus and alveolar structure during the bronchial challenge test using two optical coherence tomography systems and correlated the changes with airway resistance. MATERIALS AND METHODS: Rabbits intubated with a non-cuffed endotracheal tube on a ventilator sequentially inhaled normal saline and methacholine (2 or 5 µg/ml). The airway resistance was measured by mechanical ventilation and airway structures were monitored by a commercial endoscopic optical coherence tomography system (1,310 nm) and a house-made table-top spectral-domain optical coherence tomography system (850 nm). RESULTS: We demonstrated an early decrease in the size of the right intermedius bronchus and alveoli in accordance with increased airway resistance after methacholine inhalation. OCT image after inhalation of 2 µg/ml methacholine showed some segmental narrowing of the right intermedius bronchus and the image after inhalation of 5 µg/ml methacholine showed even greater segmental narrowing. The cross-sectional areas were 7.2 ± 3.3 mm2 (normal saline), 3.7 ± 2.1 mm2 (2 µg/ml methacholine), and 2.4 ± 1.1 mm2 (5 µg/ml methacholine), respectively (P = 0.04). Most of the alveolar space was collapsed under elevated airway resistance with methacholine inhalation. The averaged areas per alveolus at the end of inspiration were 0.0244 ±0.0142 mm2 (normal saline), 0.0046 ±0.0026 mm2 (2 µg/ml methacholine), and 0.0048 ±0.0028 mm2 (5 µg/ml methacholine), respectively (P = 0.03). Methacholine induced a dose-dependent increase in airway resistance (1.1 ± 0.3 cm H2O sec/ml for 2 µg/ml methacholine, 1.5 ± 0.5 cm H2O sec/ml for 5 µg/ml methacholine) (P = 0.03). These results were obtained from normal rabbits during the bronchial challenge test with a non-cuffed endotracheal tube on a ventilator. With this setup increased airway resistance possibly resulted in larger leakage around the endotracheal tube, decreased inhaled volumes, and, in turn, alveolar collapse. CONCLUSION: We performed a feasibility study of in vivo visualization of real-time airway dynamics. To our best knowledge, this is the first report of real-time integrated airway dynamics including the right intermedius bronchus and alveoli during a bronchial challenge test. OCT showed bronchial constriction and alveolar collapse with a higher methacholine dose. OCT images correlated with the measured airway resistance. Therefore, OCT could be a potential diagnostic device for airway hyper-responsiveness and airway remodeling.

New model of subconjunctival tumor development in rabbits.

Conjunctival squamous cell carcinoma (SCC) is an uncommon disease. However, SCC has recently become an important clinical problem due the identification of a significantly high incidence of SCC among a group of black African patients with AIDS. However, basic research concerning SCC, including both intraepithelial and invasive squamous neoplasia, is limited due to the lack of an ocular tumor animal model. Specifically, current ocular imaging and treatment modalities are insufficient for investigating currently available small animal models, because the conjunctival space is not comparable to that of humans. We describe the development of a reproducible model of subconjunctival squamous carcinoma in moderate-sized immunocompetent rabbits. Under optical coherence tomography guidance, 1×107 VX2 carcinoma cells are inoculated into the subconjunctival space of 3 to 4-kg New Zealand white rabbits. Malignant tumor involvement developed on the subconjunctival space after an average of 1 to 2 weeks. This subconjunctival tumor model induction method will likely facilitate a broad range of investigation of subconjunctival cancer diagnostics and therapeutics.

Optical imaging of subacute airway remodeling and adipose stem cell engraftment after airway injury.

Acquired airway injury is frequently caused by endotracheal intubations, long-term tracheostomies, trauma, airway burns, and some systemic diseases. An effective and less invasive technique for both the early assessment and the early interventional treatment of acquired airway stenosis is therefore needed. Optical coherence tomography (OCT) has been proposed to have unique potential for early monitoring from the proliferative epithelium to the cartilage in acute airway injury. Additionally, stem cell therapy using adipose stem cells is being investigated as an option for early interventional treatment in airway and lung injury. Over the past decade, it has become possible to monitor the level of injury using OCT and to track the engraftment of stem cells using stem cell imaging in regenerative tissue. The purpose of this study was to assess the engraftment of exogenous adipose stem cells in injured tracheal epithelium with fluorescent microscopy and to detect and monitor the degree of airway injury in the same tracheal epithelium with OCT. OCT detected thickening of both the epithelium and basement membrane after tracheal scraping. The engraftment of adipose stem cells was successfully detected by fluorescent staining in the regenerative epithelium of injured tracheas. OCT has the potential to be a high-resolution imaging modality capable of detecting airway injury in combination with stem cell imaging in the same tracheal mucosa.