Alternative non-oral nutrition in a rat model: a novel modified gastrostomy technique.

The gastrostomy technique is essential for esophageal reconstruction using a scaffold. To date, there are no established methods to supply nutrients through a gastrostomy tube in rats. The purpose of this study was to analyze the feasibility of a newly modified gastrostomy technique for non-oral nutrition in an adult rat model. We modified the gastrostomy technique for adult rats in a few different ways. (1) The external opening for food injection was made at the midpoint between the ears to prevent damage due to self-harm behaviour. (2) An imbedded subcutaneous tunnel was created between the internal and external openings of the gastrostomy. We compared the efficacy and safety between groups with a T-tube for biliary drainage (TT group, n=14) and a conventional silicone Foley catheter (FC group, n=7) as optimal gastrostomy tubes for in a rat model. We also evaluated the feasibility of the heparin cap connector at the end of gastrostomy tube to control food supply in the TT group (with a cap, n=7; without a cap, n=7). No mortality was observed in the TT group with a cap, whereas most rats in the FC group died within 2 weeks after the procedure. Weight loss decreased significantly in the TT group with a cap compared with all the other groups. The appearance and attitude scores were significantly better in the TT group with a cap. In addition, histologic analysis showed that the TT group a cap showed a marked decrease over time in tissue fibrosis and macrophages compared with the other experimental groups. Therefore, gastrostomy using a silicone T-tube plugged with a cap proved to be a stable and effective option for non-oral feeding in an adult rat model.

Tissue-Engineered Esophagus via Bioreactor Cultivation for Circumferential Esophageal Reconstruction.

The use of biomaterials for circumferential esophageal repair is technically challenging in a rat model, and an optimal scaffold implantation technique with nutritional support is essential. The purpose of this study was to investigate the effects of three-dimensional printed esophageal grafts and bioreactor cultivation on muscle regeneration and reepithelialization from circumferential esophageal defects in a rat model. Here, we designed an artificial esophagus that can enhance the regeneration of esophageal mucosa and muscle through the optimal combination of a two-layered tubular scaffold and mesenchymal stem cell-based bioreactor system. The graft was verified by the performance comparison with an omentum-cultured esophageal scaffold. We also applied a new surgical anastomosis technique and a thyroid gland flap over the implanted scaffold to improve graft survival. Although no regenerated mucosal layer was observed around the implants of the control group, histological examination of the regenerative esophagi along the scaffold revealed that the bioreactor system and omentum-cultured groups showed more than 80% of the mucosal regeneration without a fistula. The regenerated tissues showed that the integration of the esophageal scaffold and its native esophageal tissue was intact and were covered with layers of stratified squamous epithelium with several newly developed blood vessels. Therefore, this study describes a novel approach for circumferential esophageal reconstruction. Impact Statement In vivo functional esophageal reconstruction remains challenging due to anastomosis site leakage and necrosis of the implanted scaffold in a circumferential esophageal defect. Therefore, it is necessary to develop a tissue-engineered esophagus that enables regeneration of esophageal mucosa and muscle without leakage of the esophageal anastomosis. In this study, we proposed an intriguing strategy that combines a mesenchymal stem cell-seeded tubular scaffold with a bioreactor system for esophageal reconstruction and introduced a new surgical anastomosis technique with the thyroid gland flap over the implanted scaffold to improve graft survival. We believe that this system should be a powerful platform for circumferential replacement of the esophagus in a rat model.

Healing of the nasal septal mucosa in an experimental rabbit model of mucosal injury.

OBJECTIVE: The aim of this study was to investigate the regeneration process of the nasal mucosa after a surgically created mucosal defect in the rabbit nasal septum, and to evaluate the effects of different interventions. METHODS: A 7 mm-diameter circular mucosal defect was made in the septum of forty New Zealand white rabbits. The rabbits were divided into four groups (ten rabbits in each group) according to the type of intervention; no treatment (control), silastic sheet (SS), hyaluronic acid (HA), and silastic sheet and hyaluronic acid (SS + HA) group. The diameter of the defect, mucosal thickness, epithelial thickness, and ciliated cell count were evaluated every week for five weeks. RESULTS: The average diameter of the defect in the control group were 5.1, 3.65, 1.2, 0.75, and 0.05 mm at postoperative 1, 2, 3, 4, and 5 weeks. In the SS group, the diameter decreased to 4.35, 2.1, 0.35, 0.15, and 0 mm at postoperative 1, 2, 3, 4, and 5 weeks, respectively, in which the mean diameter of the postoperative week 2 was significantly smaller compared to control (3.65 mm vs. 2.1 mm, P = 0.039). For the HA group and SS + HA group, the diameter of the defect did not show a significant difference from the control group during the five weeks. The mucosal thickness, epithelial thickness, and ciliated cell count of the regenerated mucosa were not significantly different among the groups. CONCLUSION: The regeneration process of the nasal septal mucosa was identified using a novel rabbit model. Mucosal regeneration can be accelerated by applying silastic sheets.

MicroRNAs 218a-5p, 219a-5p, and 221-3p regulate vestibular compensation.

Unilateral vestibular deafferentation (UVD) interrupts afferent signals from one side, resulting in an imbalance of the resting activity between bilateral vestibular nuclei. Vestibular compensation is the process of balancing the resting activity to reestablish homeostasis. Here, we investigated microRNAs (miRNAs) that regulate vestibular compensation using the Sprague-Dawley rat. After determining the progression of vestibular compensation following UVD, microarray analysis was performed and nine miRNAs were selected as candidates. Following validation by quantitative reverse transcription-PCR, three miRNAs remained. We assessed the effect of these miRNAs on vestibular compensation using miRNA oligomers. We compared the results of the rotarod test and 5-bromo-2'-deoxyuridine immunohistochemistry following UVD between the control group and the groups in which the candidate miRNA oligomers were administered. Administration of miR-218a-5p, 219a-5p, and 221-3p oligomers significantly affected vestibular compensation. Target pathway analysis of these miRNAs supported our results. Our findings suggest that the miRNAs 218a-5p, 219a-5p, and 221-3p regulate vestibular compensation.

Disappearance of contralateral dominant neural activity of auditory cortex after single-sided deafness in adult rats.

Hearing loss in mature ears can cause functional reorganization of the auditory cortex. The functional reorganization is speculated to negatively affect the outcome of hearing rehabilitation. Therefore, once hearing loss occurs, it is important to provide auditory input before extensive reorganization in the auditory pathways. We investigated the neural plasticity in auditory cortex after single-sided deafness (SSD) in an adult rat model. The animals were divided into two groups: a normal hearing (NH) and the SSD group. The neural recordings of the SSD group were conducted at different time points (2, 4, 6 and 8 weeks) after cochlear ablation. The multi-unit activity was discriminated on the sum of spikes, peak amplitude, onset latency, peak latency, and responsive area based on the peak amplitude. The auditory cortical reorganization was observed after SSD. The contralateral dominance of peak amplitude and latency that normally occur in NH group were not present in the SSD group, replaced by higher amplitude and faster response in ipsilateral cortex. According to serial recordings at different time points after SSD, different phases in the response of the auditory cortex were speculated. Compared with normal hearing, alteration of contralateral dominance was observed because of the functional reorganization of the auditory cortex after SSD.

The Effect of Systemic Steroid on Hearing Preservation After Cochlear Implantation via Round Window Approach: A Guinea Pig Model.

HYPOTHESIS: When administered perioperatively, systemic dexamethasone will reduce the hearing loss associated with cochlear implantation (CI) performed via the round window approach. BACKGROUND: The benefits of electroacoustic stimulation have led to interest in pharmacological interventions to preserve hearing after CI. METHODS: Thirty guinea pigs were randomly divided into three experimental groups: a control group; a 3-day infusion group; and a 7-day infusion group. Dexamethasone was delivered via a mini-osmotic pump for either 3 or 7 days after CI via the round window. Pure tone-evoked auditory brainstem response (ABR) thresholds were monitored for a period of 12 weeks after CI. The cochleae were then collected for histology. RESULTS: At 4 and 12 weeks after CI, ABR threshold shifts were significantly reduced in both 7-day and 3-day infusion groups compared with the control group. Furthermore, the 7-day infusion group has significantly reduced ABR threshold shifts compared with the 3-day infusion group. The total tissue response, including fibrosis and ossification, was significantly reduced in the 7-day infusion group compared with the control group. On multiple regression the extent of fibrosis predicted hearing loss across most frequencies, while hair cell counts predicted ABR thresholds at 32 kHz. CONCLUSION: Hearing protection after systemic administration of steroids is more effective when continued for at least a week after CI. Similarly, this treatment approach was more effective in reducing the fibrosis that encapsulates the CI electrode. Reduced fibrosis seemed to be the most likely explanation for the hearing protection.

Celecoxib enhances the inhibitory effect of 5-FU on human squamous cell carcinoma proliferation by ROS production.

OBJECTIVES: The role of celecoxib in preventing and treating tumors has attracted broad attention in recent years because of its selective and specific inhibition of COX-2 activity. We investigated the inhibitory effects and mechanisms of celecoxib combined with 5-fluorouracil (5-FU) on proliferation of squamous cell carcinoma cells in vivo and in vitro. STUDY DESIGN: Animal study and basic research. METHODS: SNU-1041 and SNU-1076 squamous cell lines and an orthotopic tongue cancer mouse model were used to study growth inhibition with 5-FU enhanced by celecoxib. Sensitivity of cells to drug treatment was analyzed by the MTT assay, and generation of reactive oxygen species (ROS) was measured using dichlorofluorescein diacetate. Phosphorylation of AKT was detected by Western blotting. Survival analysis in the mouse model was assessed according to combination treatment with 5-FU and celecoxib. RESULTS: Reactive oxygen species production in vitro was highest when celecoxib was administered 48 hours after 5-FU treatment. 5-FU-induced inhibition of cell proliferation was enhanced when combined with celecoxib, which was positively correlated with ROS production. Antioxidant treatment reversed 5-FU-inhibited cell proliferation by up to 60%. Cotreatment with celecoxib and 5-FU partially blocked AKT phosphorylation, although no significant changes in total AKT protein levels were detected. An increased survival time was observed in an orthotopic mouse model treated with a combination of celecoxib and 5-FU compared to treatment with either agent alone. CONCLUSION: Celecoxib may have an enhanced anticancer effect in combination with 5-FU. Reactive oxygen species production may be a key mechanism in this combination therapy by inhibiting the AKT pathway. LEVEL OF EVIDENCE: N/A. Laryngoscope, 127:E117-E123, 2017.

Continuous topical drug delivery using osmotic pump in animal cochlear implant model: Continuous steroid delivery is effective for hearing preservation.

CONCLUSIONS: Continuous topical drug delivery using an osmotic pump is an effective supplementary technique for hearing preservation after cochlear implantation, as demonstrated in a guinea pig model. OBJECTIVE: To evaluate the effect of continuous topical steroid delivery via an osmotic pump in an animal cochlear implant model. METHODS: Twenty-three guinea pigs were used for the study. The animals were divided into three groups: control group (n = 8), simple topical dexamethasone delivery group (sDEXA group, n = 7) and continuous topical dexamethasone delivery group (cDEXA, n = 8). The hearing thresholds of all animals were measured by pre-operative auditory brain stem responses (ABRs) at 2, 8, 16, 24, and 32 kHz. ABRs were re-evaluated after cochlear implantation, and the animals were sacrificed for hematoxylin and eosin staining. RESULTS: The ABR threshold at 1 week post-operatively was significantly lower in the cDEXA group than in the control and sDEXA groups at most frequencies. Threshold shifts from baseline were statistically smaller in the cDEXA group than in the control and sDEXA groups at all frequencies. Histological analysis revealed decreased numbers of multi-nucleated giant cells and thinner histiocyte layers.

Diverse patterns of perilymphatic space enhancement in the rat inner ear after intratympanic injection of two different types of gadolinium: a 9.4-tesla magnetic resonance study.

OBJECTIVE: To compare the quality of perilymphatic enhancement in the rat inner ear after intratympanic injection of two types of gadolinium with a 9.4-tesla micro-MRI. MATERIALS AND METHODS: Gadolinium was injected into the middle ear in 6 Sprague-Dawley rats via the transtympanic route. The left ear was injected with Gd-DO3A-butrol first, and then the right ear was injected with Gd-DOTA. MR images of the inner ear were acquired 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, and 4 h after intratympanic (IT) injection using an Agilent MRI system 9.4T/160/AS. The normalized signal intensity was quantitatively analyzed at the scala vestibuli (SV), scala media, and scala tympani (ST) using a Marosis M-view system. Then the normalized signal intensities (SIs) were compared between the two contrast agents. RESULTS: For Gd-DO3A-butrol, the SI was as low as 1.0-1.5 throughout 1-4 h at the SV and ST of the basal turn. The maximum SI was 1.5 ± 0.5 at the SV (2 h) and 1.3 ± 0.5 at the ST (2 h). For Gd-DOTA, the 1-hour postinjection SI at the basal turn was 2.5 ± 0.5 at the SV, 1.6 ± 0.3 at the ST, and 1.2 ± 0.3 at the scala media. In the apical turn, the maximum SI was reached after 2.5 h. The maximum SI in the apical turn was 1.8 ± 0.4 at the SV (3.5 h), 1.8 ± 0.4 at the ST (4 h), and 1.4 ± 0.3 at the scala media (4 h). CONCLUSIONS: We were able to clearly visualize and separate the ST and SV using IT Gd and 9.4-tesla micro-MRI. We recommend using Gd-DO3A-butrol over Gd-DOTA and to perform the MRI 2.5 h after using IT Gd in the rat inner ear.