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Objectives/hypothesis Phonation threshold pressure (Pth ) is the minimum subglottic pressure required to reach phonation onset and is considered a marker for vocal efficiency and health. We investigated the effects of intrinsic laryngeal muscle (ILM) activation on Pth . Study design In vivo animal study. Methods In an in vivo canine phonation model, laryngeal adductor muscles were activated together by stimulation of the recurrent laryngeal nerves (RLNs) and individually via stimulation of respective terminal nerve branches. Cricothyroid (CT) muscles were activated via stimulation of the superior laryngeal nerves. ILMs were activated in a graded manner at various combinations as transglottal airflow was gradually increased. Aerodynamic and glottal posture parameters were measured at phonation onset. Results Graded RLN stimulation decreased glottal distance and increased Pth . Thyroarytenoid (TA) muscle activation alone increased Pth . Lateral cricoarytenoid (LCA) muscle activation alone had minimal effects. However, graded TA activation as a function of LCA activation level revealed a synergistic relationship between the two muscles in increasing Pth . Effects of CT activation were dependent on adductor stimulation level: CT activation increased Pth at low RLN stimulation levels and decreased Pth at high RLN levels. Conclusions The effects of ILM activation on Pth were consistent with their expected effects on vocal fold stiffness and tension. TA was the primary adductor controlling Pth . While LCA alone had minimal effects on Pth , it enhanced the role of TA in controlling Pth . TA and CT have antagonistic roles in controlling Pth . These relationships should be considered in clinical efforts to improve ease of phonation and vocal efficiency. Level of evidence N/A, basic science Laryngoscope, 2021.

PURPOSE To optimize and apply deep neural network based CEST (deepCEST) and apparent exchange dependent-relaxation (deepAREX) for imaging the mouse brain with Alzheimer's disease (AD) at 3T MRI. METHODS CEST and T1 data of central and anterior brain slices of 10 AD mice and 10 age-matched wild type (WT) mice were acquired at a 3T animal MRI scanner. The networks of deepCEST/deepAREX were optimized and trained on the WT data. The CEST/AREX contrasts of AD and WT mice predicted by the networks were analyzed and further validated by immunohistochemistry. RESULTS After optimization and training on CEST data of WT mice, deepCEST/deepAREX could rapidly (~1 s) generate precise CEST and AREX results for unseen CEST data of AD mice, indicating the accuracy and generalization of the networks. Significant lower amide weighted (3.5 ppm) signal related to amyloid β-peptide (Aβ) plaque depositions, which was validated by immunohistochemistry results, was detected in both central and anterior brain slices of AD mice compared to WT mice. Decreased magnetization transfer (MT) signal was also found in AD mice especially in the anterior slice. CONCLUSION DeepCEST/deepAREX could rapidly generate accurate CEST/AREX contrasts in animal study. The well-optimized deepCEST/deepAREX have potential for AD differentiation at 3T MRI.

Simvastatin (SMV) could increase tooth anchorage during orthodontic tooth movement (OTM). However, previous studies on its bone‐specific anabolic and anti‐inflammation properties were based on static in vitro and in vivo conditions. AMPK is a stress‐activated kinase that protects tissue against serious damage from overloading inflammation. Rat periodontal ligament cells (PDLCs) were subjected to a serial of SMV concentrations to investigate the optimization that promoted osteogenic differentiation. The PDLCs in static and/or tensile culturing conditions then received the proper concentration SMV. Related factors expression was measured by the protein array, real‐time PCR and Western blot. The 0.05UM SMV triggered osteogenic differentiation of PDLCs. The inhibition of AMPK activation through a pharmacological approach (Compound C) caused dramatic decrease in osteogenic/angiogenic gene expression and significant increase in inflammatory NF‐κB phosphorylation. In contrast, pharmacological activation of AMPK by AICAR significantly inhibited inflammatory factors expression and activated ERK1/2, P38 MAPK phosphorylation. Moreover, AMPK activation induced by SMV delivery significantly attenuated the osteoclastogenesis and decreased the expression of pro‐inflammatory TNF‐α and NF‐κB in a rodent model of OTM. The current studies suggested that SMV could intrigue intrinsic activation of AMPK in PDLCs that promote attenuate the inflammation which occurred under tensile irritation through AMPK/MAPK/NF‐kB Inhibition.

The response of the dentin-pulp complex in rat teeth was investigated after direct capping with biodentine with or without bone marrow-derived stem cells (BMDSCs). Following mechanical exposure, pulps were randomly capped with one of the followings materials: calcium hydroxide, biodentine or 1 × 105 BMDSCs mL-1 + biodentine. Histological examination was performed by light microscopy after 1, 3 and 5 weeks. Inflammatory reaction, necrotic tissue formation and calcific bridge formation were scored. Analysis showed that compared with the effects of calcium hydroxide or biodentine, BMDSCs + biodentine substantially reduced inflammatory reaction and necrotic tissue while promoting calcified tissue formation. Therefore, the combination of biodentine and BMDSCs could potentially stimulate pulp tissue regeneration after direct pulp capping.

Small molecules including sodium butyrate (SB) and dimethyloxalylglycine (DMOG) can promote bone regeneration via inhibitive effects eliciting cellular responses through signaling cascades. The purpose of this study was to determine the synergistic effects of SB and DMOG loaded on calcium sulfate (CaS) on bone regeneration in the challenging vertical augmentation model in the rabbit calvarium. Four plastic cylinders screwed on the calvarium of each of 10 rabbits were randomly grafted with CaS, CaS/SB, CaS/DMOG, or CaS/DMOG/SB. All specimens were assessed by radiographic, histologic, and histomorphometric analyses. In the radiographic analysis, three different layers (new bone, degraded CaS, and pristine CaS layers) could be distinguished within the cylinder in all groups at 2 weeks. Newly formed bone grew up from basal bone, and CaS in contact with newly formed bone was degraded into small particles to form a different layer. At 8 weeks, most of the pristine CaS had been absorbed and hardly seen within the cylinder. In the histomorphometric analysis, all groups showed comparable new bone areas and heights at 2 and 8 weeks. The DMOG group showed a significant increase in new bone area at 8 weeks compared with 2 weeks, but there was no significant difference among the groups at 8 weeks. The DMOG group showed significantly lower values for the residual material area than the control group at 2 weeks. Within the limitations of this study, SB and DMOG seem to exert smaller synergistic effects on bone regeneration compared to CaS alone in vertical bone augmentation.

The aim of this study was to investigate the outcomes of direct pulp capping performed with two types of tricalcium silicate-based materials (mineral trioxide aggregate/MTA and Biodentine/BD); nano-hydroxyapatite (nHAP) crystals or calcium hydroxide (CH) in dogs. Following mechanical exposure, the pulps were randomly capped with one of the four materials. Histological analyses were performed to examine the outcomes after 7 days or 3 months. At 7 days, BD and nHAP showed significantly less inflammatory cell response than MTA and CH. At 3 months, the inflammatory cell response and tissue necrosis were significantly higher in the CH group. There was no significant difference between the tested materials in the calcific bridge formation after 7 days; however, a significant difference was noticed at the 3-month period. Tricalcium silicate-based cements and nHAP are potential alternatives to CH in vital pulp therapy following accidental pulp exposure.

Introduction Despite technical progress, ventricular tachycardia (VT) recurrence after unipolar ablation remains relatively high (12%-47%). Bipolar ablation has been proposed as an appealing solution that may overcome limitations associated with unipolar ablation settings. We designed an animal study to compare bipolar (BPA) vs sequential unipolar ablation (UPA) using contact force-sensing technology on both ablation catheters. Methods Twenty large white female pigs (6-months-old, 50-60 kg) underwent multiple RF ablations (30 W, 60 seconds, 30 mL/min irrigation) on the ventricular myocardium from the epicardial and endocardial sides. The hearts were fixed and scanned with high-resolution cardiac magnetic resonance imaging. Thermal lesions were located and characterized in volume, depth, width, and transmurality. Results Lesion volume was calculated as the sum of epicardial or endocardial conjoined/isolated lesions at one location. Linear dimensions (width and depth) were measured twice for each location, on the endocardial and epicardial side. We evaluated 35 lesions across the intraventricular septum (UPA, N = 17 vs BPA, N = 18). No difference in volume, linear dimensions or impedance drop was observed in this area between UPA and BPA. However, BPA required half RF time and showed an increased transmurality trend. We then analyzed 73 lesions from the endocardial side (UPA, N = 35 vs BPA, N = 38) and 50 from the epicardial side (UPA, N = 11 vs BPA N = 39) of the ventricular free walls. Lesion transmurality was markedly improved by BPA (P = .030, odds ratio, 23.73 [4.71,31.96]). Ventricular BPA lesions were significantly deeper on the epicardial side (P < .0001) and endocardial side (P = .015). Conclusion Bipolar ablation is more likely to create transmural and epicardial lesions in the ventricle wall. Half the time is needed for the creation of comparably deep and large lesions.

Micropigmentation, also termed medical tattooing, can be a useful alternative treatment for patients with vitiligo who are resistant to conventional treatments. To assess the benefits and risks of micropigmentation in the treatment of refractory vitiligo, 25 lesions of 14 patients with vitiligo (Fitzpatrick skin types III and IV) were subjected to micropigmentation using an electric tattooing machine between December 2018 and March 2019. The procedure was repeated until satisfactory results were obtained. Treatment response was assessed by color matching of the treated lesion and surrounding skin using a 4-point scale (poor, fair, good and excellent). Excellent color matching was achieved in 80% (20/25) of cases after a median of three (range, 1-5) treatment sessions. Procedure-associated pain was considerable, but no anesthetic injection was needed. Immediate erythema and swelling were noticed after each procedure, but resolved within a few days. Overall, the treatment was tolerable. This study was limited by a small sample, no control group and a short follow-up period. This study revealed that micropigmentation was beneficial for patients with refractory vitiligo who had light to moderately colored skin. Pigment selection, implantation depth and selection of body parts amenable to treatment were critical.

Coptisine is a natural small‐molecular compound extracted from Coptis chinensis (CC) with a history of using for thousands of years. This work aimed at summarizing coptisine's activity and providing advice for its clinical use. We analysed the online papers in the database of SciFinder, Web of Science, PubMed, Google scholar and CNKI by setting keywords as ‘coptisine’ in combination of ‘each pivotal pathway target’. Based on the existing literatures, we find (a) coptisine exerted potential to be an anti‐cancer, anti‐inflammatory, CAD ameliorating or anti‐bacterial drug through regulating the signalling transduction of pathways such as NF‐κB, MAPK, PI3K/Akt, NLRP3 inflammasome, RANKL/RANK and Beclin 1/Sirt1. However, we also (b) observe that the plasma concentration of coptisine demonstrates obvious non‐liner relationship with dosage, and even the highest dosage used in animal study actually cannot reach the minimum concentration level used in cell experiments owing to the poor absorption and low availability of coptisine. We conclude (a) further investigations can focus on coptisine's effect on caspase‐1‐involved inflammasome assembling and pyroptosis activation, as well as autophagy. (b) Under circumstance of promoting coptisine availability by pursuing nano‐ or microrods strategies or applying salt‐forming process to coptisine, can it be introduced to clinical trial.

PURPOSE The three-dimensional (3D) principal strain represents the major direction and magnitude of the deformation by its definition and can be measured using 3D echocardiography. We aimed to validate 3D echocardiographic left ventricular (LV) global principal strains as an assessment of LV contractility by comparison with gold standard invasive measurements. METHODS In 14 beagles, the LV pressure-volume loop was recorded to invasively measure the end-systolic pressure-volume relationship (ESPVR) and dP/dt as reference indicators representing LV contractility. The echocardiographic image was obtained simultaneously, and endocardial motions and volume changes were extracted in the form of speckle-tracking point grids to calculate strains. High or low inotropic states were induced pharmacologically by using an intravenous infusion of dobutamine and esmolol, respectively. RESULTS The direction of 3D endocardial global principal strain (GP1S) appeared to be circumferential. The dP/dt showed the highest Pearson's correlation coefficients with GP1S (r = -0.845, P < 0.001), whereas ESPVR showed the best correlation with global secondary strain (GP2S; r = -0.819, P < 0.001). In comparison with GP1S and GP2S, global circumferential (GCS) and longitudinal strains (GLS) tended to correlate less with invasive measurements, respectively. LV ejection fraction showed excellent correlations with GP1S or GCS, but the correlation with GLS was relatively weak. The correlations between invasive measurements and GP2S or GLS were strengthened when strains were corrected by the LV residual volume ratio, whereas those of GP1S or GCS were weakened after correction. CONCLUSIONS The principal direction of the LV endocardial contraction presents circumferential behavior reflecting LV volume change. The 3D principal strains derived from 3D echocardiography are reliable indicators for LV contractility and seem better than conventional strains.

In septorhinoplasty, septal, auricular, and costal cartilage are often used as autologous graft. Autologous grafts are preferred in nasal reconstruction. The aim of this study was to histopathologically examine the tissue compatibility and the effect on the stability and cartilage vitality of poly-p-dioxanone (PDS) plates.Ten adult New Zealand rabbits were used.Ten New Zealand rabbits were used. Septal and auricular cartilage sections were removed; one of the two cartilage grafts was left plain, and the other was sutured to a PDS plate. Grafts were placed into the back of the rabbits. After 12 weeks, the graft material was examined microscopically.The specimens did not cause any significant foreign body reaction. Within 3 months, a significant degree of color, stability, and stiffness was lost. Microscopically, inflammation, necrosis, and cartilage cell degeneration scores were statistically significantly lower in the grafts using PDS (P.05), and the vascularization, collagen, and cartilage proliferation scores were found to be statistically significantly higher (P.05). No statistically significant difference was determined in respect of the bone proliferation scores (P.05).The use of PDS plates together with cartilage provided mechanical support to the graft. Therefore, changes that disrupt the integrity of the graft, such as inflammation, necrosis, and cartilage cell degeneration, were reduced, and changes that provide greater stability, such as vascularization, collagen, and cartilage proliferation, were increased.NA Laryngoscope, 129:E129-E134, 2019.

Traumatic brain injury (TBI) is a major health challenge, is very difficult to diagnose, and currently has no accepted treatment options. The difficulty in diagnosis is due to the complex pathophysiology of TBI, where the damage mechanisms span multiple spatial and temporal scales and ultimately manifest as diverse cognitive impairments in brain function. The speed and dynamic nature with which the mechanical insults occur in TBI means that computational models are the best candidate to mimic the in vivo event and to investigate the injury progression in detail. A current weakness in TBI computational models is the poor understanding of how material properties within different regions of the brain change after damage. Here, we developed an animal experiment pipeline where a controlled and reproducible mechanical insult can be applied to a sheep brain in vivo followed by ex vivo compression mechanical testing. For each brain, the compression mechanical testing was conducted across the length of the brain to assess baseline regional variations in material properties and how these properties changed after mechanical impact. Using this pipeline, we have characterized locational variations in the brain material properties both pre and post‐TBI. Our preliminary results confirm that there exist locational variations in the brain material properties across all regions of cerebrum. The analysis of post‐TBI tissues showed that compressive strength was reduced in the area nearest the direct impact.

Background Polydioxanone (PDO) sutures have been widely used to tighten and lift the face. However, why the complexion brightens and skin elasticity is maintained with a smaller facial outline after a PDO monofilament thread treatment remains unclear. Aims We aimed to determine what significant changes occur in the tissue over time when a PDO suture is inserted. Methods We selected four White Yucatan variety pygmy pigs with skin that most closely resembles the structure of human skin. 4-0 PDO thread was inserted into the subcutaneous fat. Tissue samples were obtained at 4, 12, 24, and 48 weeks. For the histologic analysis, H&E staining, Masson trichrome staining, and anti-smooth muscle actin immunohistochemical staining techniques were used. Results Nine histological findings appeared over time, and these findings are summarized as five tissue changes. Conclusions PDO sutures cause specific changes to the surrounding tissues that result in neo-collagenesis, a fibrous merging effect, fat reduction, tissue contracture, and an improved vascular environment. The results of this study explain the positive changes described in previous clinical research.

Objectives To evaluate the effect of real-time three-dimensional (3D) ultrasonography (US) in guiding percutaneous nephrostomy (PCN). Materials and methods A hydronephrosis model was devised in which the ureters of 16 beagles were obstructed. The beagles were divided equally into groups 1 and 2. In group 1, the PCN was performed using real-time 3D US guidance, while in group 2 the PCN was guided using two-dimensional (2D) US. Visualization of the needle tract, length of puncture time and number of puncture times were recorded for the two groups. Results In group 1, score for visualization of the needle tract, length of puncture time and number of puncture times were 3, 7.3 ± 3.1 s and one time, respectively. In group 2, the respective results were 1.4 ± 0.5, 21.4 ± 5.8 s and 2.1 ± 0.6 times. The visualization of needle tract in group 1 was superior to that in group 2, and length of puncture time and number of puncture times were both lower in group 1 than in group 2. Conclusions Real-time 3D US-guided PCN is superior to 2D US-guided PCN in terms of visualization of needle tract and the targeted pelvicalyceal system, leading to quick puncture. Real-time 3D US-guided puncture of the kidney holds great promise for clinical implementation in PCN.

OBJECTIVE In this study, we aimed to functionally and morphologically demonstrate the effectiveness of platelet-rich plasma (PRP) on anosmia in a mouse model of anosmia. STUDY DESIGN Animal study. METHODS A total of 16 male mice were included. When selecting the mice, the food-finding test (FFT) was used to make sure that the animals could smell, and anosmia was induced by administration of intraperitoneal 3-methylindole. The mice were randomly divided into two groups of eight (groups A and B). After 1 week, topical PRP was administered to the mice in group A and topical saline was administered to the mice in group B. The FFT was again administered at 7, 14, and 21 days. The mice were sacrificed on day 21, the olfactory neuroepithelium was histopathologically examined, and the epithelial damage scores and epithelial thickness were measured. RESULTS After topical administration of PRP and saline, the difference in the average FFT values of the groups was statistically significant at 7, 14, and 21 days (P < 0,005). During the histopathological examination, the epithelial damage score was statistically significantly lower in the PRP group (P = 0.001) than in the saline group, and epithelial thickness was statistically significantly greater in the PRP group compared to the saline group (P = 0.003). CONCLUSION We showed that PRP administration has a curative effect on olfactory functions in an anosmia-induced mice model. However, there is a need for further research before PRP can be considered for use in patients with anosmia in clinical practice. LEVEL OF EVIDENCE NA. Laryngoscope, 128:E157-E162, 2018.

Purpose To propose a simple method to correct vascular input function (VIF) due to inflow effects and to test whether the proposed method can provide more accurate VIFs for improved pharmacokinetic modeling. Methods A spoiled gradient echo sequence-based inflow quantification and contrast agent concentration correction method was proposed. Simulations were conducted to illustrate improvement in the accuracy of VIF estimation and pharmacokinetic fitting. Animal studies with dynamic contrast-enhanced MR scans were conducted before, 1 week after, and 2 weeks after portal vein embolization (PVE) was performed in the left portal circulation of pigs. The proposed method was applied to correct the VIFs for model fitting. Pharmacokinetic parameters fitted using corrected and uncorrected VIFs were compared between different lobes and visits. Results Simulation results demonstrated that the proposed method can improve accuracy of VIF estimation and pharmacokinetic fitting. In animal study results, pharmacokinetic fitting using corrected VIFs demonstrated changes in perfusion consistent with changes expected after PVE, whereas the perfusion estimates derived by uncorrected VIFs showed no significant changes. Conclusion The proposed correction method improves accuracy of VIFs and therefore provides more precise pharmacokinetic fitting. This method may be promising in improving the reliability of perfusion quantification. Magn Reson Med 79:3093-3102, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

Objectives/hypothesis Evaluate the effect of in vitro exposure of mice laryngeal mucosa to solutions that simulated human gastric juice and to assess the topical protective effect of cashew gum on mice laryngeal mucosal integrity in vitro. Study design Animal study. Methods Murine (Swiss) laryngeal samples were mounted in Ussing chambers. The luminal side of biopsies was exposed to solutions of different acidity with or without pepsin and/or taurodeoxycholic acid (TDC). Transepithelial electrical resistance (TER) was continuously recorded. The topical protective effect of cashew gum solution was evaluated by precoating the biopsies before the exposure with a solution at pH 5 containing 5 mM TDC. Changes in TER and mucosal permeability to fluorescein were measured. Results Exposure of laryngeal mucosa to acidic solutions containing pepsin and TDC provoked a pH-dependent drop in TER with the maximal effect at pH 1, but still present at pH 5 (weakly acidic). The exposure of the laryngeal mucosa to a solution of pH 5 with TDC, but not with pepsin, produced a dose-dependent decrease in TER. Precoating the mucosa with cashew gum prevented the reduction of TER and increased transepithelial permeability by exposure to a solution at pH5 containing TDC. Conclusions Weakly acidic solutions containing bile acids can produce impairment of laryngeal epithelial barrier, which may be protected by topical treatment with cashew gum. Level of evidence NA. Laryngoscope, 128:1157-1162, 2018.

Opening a chest in an experimental echocardiographic animal study eliminates ultrasound signal attenuation by the chest wall. We developed a scanning technique that involves the use of an attenuative pad created from a mixture of urethane and titanium dioxide. The pad was interposed within transmission gel between the transducer face and cardiac surface in open-chest scans in a porcine model. Comparative measurements of left ventricular echogenicity without and with the pad demonstrate that the pad reproducibly causes ultrasound signal attenuation that closely mimics chest attenuation in clinical transthoracic echocardiographic studies.

Background Osteotomies have been performed for centuries yet there remains a remarkable lack of consensus on optimal methods for cutting bone. There is universal agreement, however, that preserving cell viability is critical. Purpose To identify mechanobiological parameters influencing bone formation after osteotomy site preparation. Materials and methods A murine maxillary osteotomy model was used to evaluate healing. Computational modeling characterized stress and strain distributions in the osteotomy, as well as the magnitude and distribution of heat generated by drilling. The impact of osteocyte death and bone composition were assessed using molecular and cellular assays. Results The phases of osteotomy healing in mice align closely with results in large animals; in addition, molecular analyses extended our understanding of osteoprogenitor cell proliferation, differentiation, and mineralization. Computational analyses provided insights into temperature changes caused by drilling and the mechanobiological state in the healing osteotomies, while concomitant cellular assays correlate drill speed with osteocyte apoptosis and bone resorption. Even when drilling was controlled, trabeculated, spongy (Type III) bone healed faster than densely lamellar (Type I) bone because of the abundance of Wnt responsive osteoprogenitor cells in the former. Conclusions These data provide a mechanobiological framework for evaluating tools and technologies designed to improve osteotomy site preparation.

OBJECTIVE: To evaluate the healing after elevation of the sinus mucosa when a collagen membrane was placed between the sinus mucosa and a xenograft used as filler. MATERIALS AND METHODS: Eighteen rabbits were used. Sinus mucosa elevation was performed bilaterally, and a collagen membrane was applied subjacent to the sinus mucosa only at a randomly selected test site. At both sites, a collagenated corticocancellous porcine bone was placed within the elevated space and the access window was covered with a collagen membrane. The animals were sacrificed after 2, 4, and 8 weeks of healing, six animals for group. Ground sections were prepared. RESULTS: At the histomorphometric evaluation, the elevated area after 2 and 8 weeks was 11.8 and 8.8 mm(2) at the test, and 10.0 and 5.3 mm(2) at the control sites, respectively. The available area was obtained subtracting the remaining area occupied by the membrane from the elevated area and, after 8 weeks, was 6.7 +/- 0.9 mm(2) . After 8 weeks of healing, the mineralized new bone within the elevated space was 18.2 +/- 5.5% at the test and 26.7 +/- 7.7% at the control sites. Within the available space at the test site, the percentage was 27.3 +/- 7.0% after 8 weeks of healing. At 2 and 8 weeks of healing, within the elevated space, the xenograft proportion was 30.9 +/- 4.4% and 6.9 +/- 2.8% at the test, and 35.2 +/- 7.3% and 9.6 +/- 4.9% at the control sites, respectively. CONCLUSIONS: The placement of a collagen membrane subjacent the sinus mucosa did not reveal any major morphometric difference. The collagen membrane was not completely resorbed after 8 weeks.

Objective Human adipose-derived mesenchymal stem cells (ADSCs) were used to rehabilitate bone damaged by osteoradionecrosis (ORN) in an established animal model. Study Design Prospective animal study. Setting Academic department laboratory. Subjects and Methods After institutional review board and Institutional Animal Care and Use Committee approval, 24 athymic nude rats were divided into 5 groups: 4 groups irradiated (20 Gy) by brachytherapy catheter placed at the left hemimandible and 1 mock irradiation control (n = 4). For all groups, ORN was initiated by extraction of the central molar 1 week later. After 28 days, animals (n = 5/group) received injection at the extraction site with saline (SAL), ADSCs, platelet-rich plasma and collagen (PRP/COL), or ADSCs + PRP/COL. Rats were sacrificed 28 days later and their mandibles harvested for histopathology analysis (osteoblasts, osteoclasts, and fibrosis) and bone volume measurement using 3-dimensional micro-computed tomography. Results All but 1 rat survived the experiment period (23/24). Radiographic and histological analysis revealed 60% bone loss in the SAL group compared with the nonirradiated control. Injection of ADSCs increased jaw region bone volume by up to 36% ( P.01). All experimental groups (ADSC, PRP/COL, and ADSC + PRP/COL) showed dramatically decreased osteoclast counts ( P.001) while injection of PRP/COL with or without ADSCs increased osteoblasts. Increased fibrosis was observed after ADSC injection ( P.05). Conclusion The application of human ADSCs to an induced mandibular osteoradionecrosis model in athymic rats results in increased deposition or preservation of bone, demonstrated both histologically and radiographically. This offers an encouraging possible treatment option for translational research in this difficult disease.

The reproducibility of laboratory experiments is fundamental to the scientific process. There have been increasing reports regarding challenges in reproducing and translating preclinical experiments in animal models. In Alzheimer's disease and related dementias, there have been similar reports and growing interest from funding organizations, researchers, and the broader scientific community to set parameters around experimental design, statistical power, and reporting requirements. A number of efforts in recent years have attempted to develop standard guidelines; however, these have not yet been widely implemented by researchers or by funding agencies. A workgroup of the International Alzheimer's disease Research Funder Consortium, a group of over 30 research funding agencies from around the world, worked to compile the best practices identified in these prior efforts for preclinical biomedical research. This article represents a consensus of this work group's review and includes recommendations for researchers and funding agencies on designing, performing, reviewing, and funding preclinical research studies.

Implants with deep thread depth have been developed for the purpose of increasing total implant surface area. However, effects of implant thread depth remain controversial. The aim of this study is to examine effects of thread depth on peri-implant tissues in terms of bone-implant contact (BIC), bone-implant volume (BIV), and hard and soft tissue dimensions using comprehensive analyses, including microcomputed tomography (micro-CT).Five beagle dogs received experimental intramandibular implants 3 months after removal of their premolars and first molars (PThere were no significant differences in resonance frequency test results between the two groups. Although radiographic analysis showed no group differences, micro-CT (P = 0.01) and histomorphometry (P = 0.003) revealed the DT group had significantly lower BIC values than the ST group at 4 weeks. However, by 8 weeks, BIC values of the two groups did not differ significantly. No significant differences in BIV or soft tissue height were observed between the two groups at either time point.DT implants showed no benefits over ST implants when inserted in dog mandibles.

SA37 inhibits protein tyrosine phosphatase 1B (PTP1B) with a medium potency, and inhibits IκB kinase β (IKKβ) at a submicromolar half-maximal inhibitory concentration. In animal study, SA37 reduced diet-induced weight gain in mice, without significantly ameliorating glucose tolerance and lipid parameters. These observations coincide partly with PTP1B knockout but are inconsistent with IKKβ depletion in mice. The effect of SA37 on high-fat diet (HFD)-induced obese mice was compared with previous results obtained by the authors’ group on a series of SA compounds that exhibited different inhibitory potencies toward IKKβ and PTP1B.

Orthodontic implants may fracture at the cortical bone level upon rotational torque. The impacted fragment can be detached by a range of methods, which are all more or less time‐consuming and injurious to the cortical bone. The aim of this study was to compare three different methods for detaching an orthodontic implant impacted in cortical bone. Health Sciences University of Hokkaido animal ethics committee approved the study protocol. Orthodontic titanium‐alloy (Ti‐6Al‐4 V) implants were placed bilaterally on the buccal side of the mandible of beagle dogs. Subsequently, the implants were detached using either a low‐speed handpiece with a round bur, alternatively by use of a low‐power or a high‐power ultrasonic instrument. In the first experiment, 56 orthodontic implants were placed into the dissected mandible from 7 animals. The methods for detachment were compared with respect to time interval, as well as associated undesirable bone loss as appraised by use of cone‐beam computed tomography. In experiment two, 2x2 implants were placed bilaterally in the mandible of 8 animals and subsequently detached by manual rotational torque, and the described three methods for detachment. The implant socket was investigated histologically as a function of removal method immediately after removal, and after 1, 3 and 8 weeks and contrasted with the healing of the socket of the implant that was detached by manual rotational torque. Statistical significance was appraised by the use of non‐parametric Kruskal‐Wallis one‐way analysis of variance. The method using the low‐power ultrasonic required significantly longer removal time versus the two other methods, i.e. high‐power ultrasonic and low‐speed handpiece with a round bur (p

Background : Bone healing processes following osteotomy may vary according to the type of selected surgical procedures. Frictional heat caused by traditional bur osteotomies may result in thermal osteonecrosis if the temperature arises above the critical temperature of 47º C. Therefore, less invasive and precise tools for bone tissue surgery with reduced collateral tissue thermal trauma, force and friction are developed: laser- assisted bone surgery and piezoelectric surgery. Aim/Hypothesis : The purpose of the present in vivo experimental study was to determine the thermal changes of the bone tissue following osteotomies performed by piezoelectric surgery, surgical drill and Er:YAG laser- assisted ablation used in contact and NO- contact modes using infrared thermographic camera. Materials and Methods : A total number of 24 Wistar rats were randomly divided into 4 groups, each consisting of 6 rats (I- sacrificed immediately after surgery ; II- 7 days after surgery ; III- 14 days after surgery ; IV- 21 days after surgery). Osteotomies on both rat ' s tibiae were performed under general anesthesia and always in the same sequence: digitally controlled NO- contact Er:YAG laser (7.5 W, 750 mJ, 10 Hz, QSP mode), piezosurgery (round diamond tip, 1.2 mm diameter, average pressure 15 N), low- speed surgical drill (1200 rpm, round steel surgical bur 2 mm diameter) and contact Er:YAG laser (H- 14N handpiece, fiber of core diameter of 940 μm, 7.5 W, 375 mJ, 20 Hz, MSP mode). The osteotomies were 5 mm away from each other and ~2 mm deep, with a diameter ranging between 1.5–2.0 mm. Temperature measurements during each osteotomy were done using an infrared thermographic camera with a detection range of - 20º to + 650º C, a thermal sensitivity of < 50 mK, and IR resolution of 320x240 pixels. Results : For each osteotomy a thermal camera movie was recorded and movie frames were extracted as thermal camera images, and the position of the osteotomy was determined by visual inspection. Temperature readout was performed for this point in all recorded movie frames, providing the information on the temperature at the position of osteotomy while osteotomy was performed. Time evolution of the bone temperature at the point of osteotomy was measured. For each measurement, baseline temperature (Tbase), and maximum temperature measured during the osteotomy (Tmax) were determined. ΔT=Tmax- Tbase was then calculated for each measurement. Mean baseline temperature for each group of osteotomies was 29.96°C for non- contact laser, 27.98°C for contact laser, 28.34°C for surgical drill and 29.39°C for piezosurgery. Mean Tmax for contact laser was 29.92 °C (ΔT=1.94 °C), for non- contact laser was 79.11°C (ΔT=49.15 °C), for surgical drill was 27.36°C and 29.15 °C (ΔT=- 0.24 °C) for piezosurgery. Conclusions and Clinical Implications : The results of the present study showed beneficial effects of the piezosurgery and Er:YAG laser used in contact mode of working on heat generation of bone tissue during osteotomy, reducing potential overheating of the bone as registered by means of infrared thermography. Digitally controlled non- contact Er:YAG laser results in the production of temperature that is higher than the allowed border for bone tissue during osteotomy.

Objectives/hypothesis Dehydrated vocal folds are inefficient sound generators. Although systemic dehydration of the body is believed to induce vocal fold dehydration, this causative relationship has not been demonstrated in vivo. Here we investigate the feasibility of using in vivo proton density (PD)-weighted magnetic resonance imaging (MRI) to demonstrate hydration changes in vocal fold tissue following systemic dehydration in rats. Study design Animal study. Methods Sprague-Dawley rats (n = 10) were imaged at baseline and following a 10% reduction in body weight secondary to withholding water. In vivo, high-field (7 T), PD-weighted MRI was used to successfully resolve vocal fold and salivary gland tissue structures. Results Normalized signal intensities within the vocal fold decreased postdehydration by an average of 11.38% ± 3.95% (mean ± standard error of the mean [SEM], P = .0098) as compared to predehydration levels. The salivary glands experienced a similar decrease in normalized signal intensity by an average of 10.74% ± 4.14% (mean ± SEM, P = .0195) following dehydration. The correlation coefficient (percent change from dehydration) between vocal folds and salivary glands was 0.7145 (P = .0202). Conclusions Ten percent systemic dehydration induced vocal fold dehydration as assessed by PD-weighted MRI. Changes in the hydration state of vocal fold tissue were highly correlated with that of the salivary glands in dehydrated rats in vivo. These preliminary findings demonstrate the feasibility of using PD-weighted MRI to quantify hydration states of the vocal folds and lay the foundation for further studies that explore more routine and realistic magnitudes of systemic dehydration and rehydration. Level of evidence NA. Laryngoscope, 128:E222-E227, 2018.

Objectives Healing characteristics as well as level of tissue integration and degradation of two different nanostructured hydroxyapatite bone substitute materials (BSM) in comparison with a deproteinized hydroxyapatite bovine BSM were evaluated in an in vivo animal experiment. Material and methods In the posterior mandible of 18 minipigs, bilateral mono cortical critical size bone defects were created. Randomized augmentation procedures with NanoBone(®) (NHA1), Ostim(®) (NHA2) or Bio-Oss(®) (DBBM) were conducted (each material n = 12). Samples were analyzed after five (each material n = 6) and 8 months (each material n = 6). Defect healing, formation of soft tissue and bone as well as the amount of remaining respective BSM were quantified both macro- and microscopically. Results For NHA2, the residual bone defect after 5 weeks was significantly less compared to NHA1 or DBBM. There was no difference in residual BSM between NHA1 and DBBM, but the amount in NHA2 was significantly lower. NHA2 also showed the least amount of soft tissue and the highest amount of new bone after 5 weeks. Eight months after implantation, no significant differences in the amount of residual bone defects, in soft tissue or in bone formation were detected between the groups. Again, NHA2 showed significant less residual material than NHA1 and DBBM. Discussion We observed non-significant differences in the biological hard tissue response of NHA1 and DBBM. The water-soluble NHA2 initially induced an increased amount of new bone but was highly compressed which may have a negative effect in less stable augmentations of the jaw.

Microtubules are an ideal target for anticancer drugs because of their essential role in mitosis. Colchicine is a microtubule destabilizer. Whether the clinically acceptable colchicine concentrations had anticancer effects on human cholangiocarcinoma cells was investigated. Two human cholangiocarcinoma cell lines (C14/KMUH, C51/KMUH) were investigated using clinically acceptable plasma colchicine concentrations (2 ng/mL and 6 ng/mL for the in vitro experiment, 0.07 mg colchicine/kg/d × 14 days for the nude mouse experiment). Our results showed that colchicine caused significantly dose-dependent antiproliferative effects on both cell lines (all p

OBJECTIVES Creation of an atraumatic, hearing-preservation cochleostomy is integral to the future of minimally invasive inner ear surgery. The goal of this study was to develop and characterize a novel chemical approach to cochleostomy. STUDY DESIGN Prospective animal study. SETTING Laboratory. METHODS Experimental animal study in which phosphoric acid gel (PAG) was used to decalcify the otic capsule in 25 Hartley guinea pigs. Five animals in each of 5 surgical groups were studied: (1) mechanically opening the auditory bulla alone, (2) PAG thinning of the basal turn otic capsule, leaving endosteum covered by a layer of bone, (3) micro-pick manual cochleostomy, (4) PAG chemical cochleostomy, exposing the endosteum, and (5) combined PAG/micro-pick cochleostomy, with initial chemical thinning and subsequent manual removal of the last osseous layer. Preoperative and postoperative auditory brainstem responses and otoacoustic emissions were obtained at 2, 6, 10, and 16 kHz. Hematoxylin and eosin-stained paraffin sections were compared. RESULTS Surgical and histologic findings confirmed that application of PAG provided reproducible local bone removal, and cochlear access was enabled. Statistically significant auditory threshold shifts were observed at 10 kHz (P = .048) and 16 kHz (P = .0013) following cochleostomy using PAG alone (group 4) and at 16 kHz using manual cochleostomy (group 3) (P = .028). No statistically significant, postoperative auditory threshold shifts were observed in the other groups, including PAG thinning with manual completion cochleostomy (group 5). CONCLUSION Hearing preservation cochleostomy can be performed in an animal model using a novel technique of thinning cochlear bone with PAG and manually completing cochleostomy.

Summary Aim Anticoagulants are commonly used to treat ischemic stroke. Its impact on cerebellar infarction has not been fully understood. Methods In the clinical study, we reviewed a consecutive series of patients with large-sized cerebellar infarction (diameter > 3 cm, n = 30) treated with or without anticoagulation. In animal study, cerebellar infarction operation was performed in 12 Cynomolgus monkeys. Then the animals were administrated with low molecular weight heparin (LMWH) or vehicle for 14 days. Results Six patients died during the following treatment. All the subjects that died received anticoagulation therapy, while nobody in the survival group received such a therapy. Compared with sham-operated animals, all monkeys with cerebellar infarction have obvious neurological deficits. The number and size of the Purkinje cells in the cerebellar area were also reduced. Two animals in the LMWH group (33%) died, while all animals in the vehicle control group survived. Compared with the vehicle group, the neurological score in the LMWH group was significantly increased (P

Objective The aim of this study was to assess the bone regeneration of critical size defects in rabbit calvarias filled with β-TCP doped with silicon. Materials and methods Twenty-one New Zealand rabbits were used in this study. Two critical size defects were created in the parietal bones. Three experimental groups were evaluated: Test A (HA/β-TCP granules alone), Test B (HA/β-TCP granules plus 3% silicon), Control (empty defect). The animals were sacrificed at 8 and 12 weeks. Evaluation was performed by μCT analysis and histomorphometry. Results μCT evaluation showed higher volume reduction in Test A group compared with Test B (P

Objective: The purpose of this study was to determine the ability of oral submucosal tissue to serve as a temporary storage medium for the maintenance of periodontal ligament (PDL) cell viability of avulsed teeth. Materials and methods: Thirty cats were divided into five groups. After extraction of three teeth in each cat, one tooth was put in the depth of cat's oral submucosal tissue and the other two teeth were put in Hanks' balanced salt solution (HBSS) and tap water. The teeth were removed after 8, 24, 48, 72, and 168 h from their mediums and sent for laboratory processing and counting of vital periodontal cells. Results: Statistical analysis demonstrated that submucosal tissue kept PDL cells viable as good as HBSS. Conclusion: This animal study showed that the efficacy of oral submucosal tissue in maintaining the viability of human PDL cells is similar to that of HBSS. © 2014 John Wiley & Sons A/S.

Endometriosis (EM) impacts the healthcare and the quality of life for women of reproductive age. However, there is no reliable noninvasive diagnosis method for either animal study or clinical use. In this work, a novel imaging method, photoacoustic microscopy (PAM) was employed to study the EM on the mouse model. Our results demonstrated the PAM noninvasively provided the high contrast and 3D imaging of subcutaneously implanted EM tissue in the nude mouse in vivo. The statistical study also indicated PAM had high sensitivity and specificity in the diagnosis of EM in this animal study. In addition, we also discussed the potential clinical application for PAM in the diagnosis of EM.

Summary Background UV-blocking contact lenses were evaluated to determine if they could provide adequate ocular protection during narrowband UVB phototherapy treatment. Theoretical safe exposure durations for the crystalline lens, cornea and conjunctiva were determined. Methods A Cary 500 spectrophometer generated transmittance data for six UV-blocking and two non-UV-blocking contact lenses. An IL-1700 radiometer measured the lenses' radiation transmittance within the NB-UVB phototherapy unit. The lenses were exposed to a 1500-mJ/cm2 dose of radiation from a 308-nm excimer laser to determine if the radiation would alter their protective properties. Theoretical safe exposure durations for eye structures were calculated using previous human and animal study data. Results All UV-blocking contact lenses showed less than 1E-7 W/cm2 of radiation transmittance within the narrowband phototherapy unit. The excimer laser did not significantly alter the lenses' UV-blocking capabilities. The safe exposure durations for the cornea and crystalline lens were greater than 11 min with UV-blocking lenses, and that for the unprotected conjunctiva was approximately 11 s. Conclusion Some UV-blocking contact lenses potentially provide sufficient ocular protection during narrowband UVB phototherapy treatment, as the crystalline lens and cornea are adequately protected should a patient open his or her eyes for a short time.

Background and Aim To evaluate the effectiveness and outcomes of endoscopic closure of a gastric fundus perforation using over-the-scope clips (OTSCs) system in a surviving canine model. Methods Gastric fundus perforations (20-mm diameter) were created by an endoscopic needle-knife in six dogs. The perforations then were closed by the OTSC system. Gastroscopy was performed to evaluate the postoperative perforation healing every week. The animals were sacrificed 4 weeks later to examine the possible intraperitoneal complications, and the healing of the perforation was examined histopathologically. Results The gastric fundus perforations could primarily be closed using one OTSC in each experimental dog, and the mean time of the procedure was 17.3 ± 7.6 min (9–26 min). All animals survived without postoperative complications. The OTSC retention was observed in one dog at the end of 4 weeks, and the apparent foreign-body reaction was examined pathologically. Conclusions Our surviving animal study demonstrated that the OTSC clip system could reliably close gastric fundus perforations without complications.

Purpose The aim of this in vivo animal study was to investigate changes in the surface roughness of soft liners over time. Materials and Methods Forty adult Wistar rats (Rattus norvergicus albinus) were fitted with acrylic custom-made palatal plates relined by dynamic impressions and tested with the following soft liners: Dentuflex (DF), Trusoft (TS), Dentusoft (DS), and Ufi Gel P (UG). Half of the animals for each tested material had the plates fitted during the material reline procedure. Their surface roughness was read immediately (IRa group, n = 5). The other half used the palatal plates for 14 days before roughness readings were performed (FRa group, n = 5). The surface roughness (Ra) of the inner surface from the relined dentures was recorded using a Surftest SJ-401 with eight readings per specimen, and mean values were obtained. Data (μm) were analyzed by two-way ANOVA and Tukey's test (α = 0.05). Results IRa means (2.92 ± 0.87 μm) and FRa means (3.35 ± 0.65 μm) were significantly different (p = 0.016). UG showed a lower (p = 0.01) Ra mean (2.1 ± 0.52 μm) than DF (3.94 ± 0.81 μm), TS (4.12 ± 0.64 μm), and DS (3.27 ± 0.64 μm). Conclusions Ufi Gel P showed the smoothest surface among the materials evaluated. The period of use resulted in changes in the surface roughness of the materials tested.

BACKGROUND: High irrigation rates during radiofrequency (RF) ablation may cause fluid overload and limit lesion size. This in vivo animal study assessed the safety and efficacy of RF ablation at low irrigation rates using a novel 12-hole gold catheter. METHODS: A total of 103 lesions, created on the thigh of five mongrel dogs, were analyzed. Lesions were created using a 12-hole irrigated gold-tip (Au) and a six-hole irrigated platinum-iridium (PtIr) catheter (both 7F/3.5-mm electrode; BIOTRONIK SE & CO, KG, Berlin, Germany) in parallel and perpendicular orientation. RF current was delivered for 60 seconds at 30 W using 8 mL/min and 15 mL/min irrigation. Electrode temperature, steam pops, lesion dimensions, and coagulum formation were recorded. RESULTS: Electrode temperatures were lower for Au compared to PtIr in parallel (8 mL/min: 38.1 +/- 1.7 degrees C vs 48.0 +/- 4.8 degrees C, P < 0.0001; 15 mL/min: 36.0 +/- 1.5 degrees C vs 46.9 +/- 5.4 degrees C, P < 0.0001) and perpendicular position (15 mL/min: 35.5 +/- 1.2 degrees C vs 38.4 +/- 2.5 degrees C, P = 0.003). The number of steam pops between Au and PtIr was comparable for parallel (8 mL/min: 14% vs 27%, P = 0.65; 15 mL/min: 14% vs 43%, P = 0.21) and perpendicular orientation (8 mL/min: 25% vs 17%, P = 1.00; 15 mL/min: 18% vs 0%, P = 0.48). Au created larger volumes than PtIr at 8 mL/min irrigation (861 +/- 251 mm3 vs 504 +/- 212 mm3 , P = 0.004); however, for 15 mL/min, volumes were comparable (624 +/- 269 mm3 vs 768 +/- 466 mm3 , P = 0.46). No coagulum formation was observed for any of the catheters on the surface and catheter tip. CONCLUSION: RF ablation at low flow rate using a novel 12-hole irrigation Au catheter is safe and results in larger lesions than with a PtIr electrode.

Summary What is already known about this subject Red cell distribution width correlates with outcomes in numerous clinical populations. RDW is elevated in inflammatory states. Obesity is associated with inflammation. What this study adds RDW is elevated in obese adolescents. RDW represents a potential marker for the presence of metabolic syndrome. Background Obesity and the metabolic syndrome are dramatically increasing problems. Red blood cell distribution width (RDW), the variability in size of circulating red blood cells, has been demonstrated to be altered in different clinical settings. This analysis aimed to investigate the relationship between RDW and obesity in adolescents and in an animal model of diet-induced obesity (DIO). Methods Seventy-nine male adolescents (aged 13–17 years) were studied. Thirty-seven of them were overweight (body mass index ≥ 90th percentile). RDW, markers of inflammation and stem cell factor (SCF) were determined. In an animal study, mice were fed with different diets for 15 weeks. RDW was determined using an animal blood count machine. Results RDW differed significantly between normal-weight adolescents (13.07 ± 0.09) and overweight adolescents (13.39 ± 0.10, P = 0.015), whereas erythrocyte counts and haematocrit did not differ. RDW correlated to markers of inflammation and inversely to SCF. In the mice animal model, nutritional changes increased RDW, whereas overweight per se did not change RDW. Conclusions RDW is elevated in overweight and reflects the inflammatory state. RDW potentially represents an additional and cost-effective tool to indicate inflammation. Future studies are needed to understand the differential influences of nutrition and overweight on RDW.

11th International Congress of the European-Society-of-Pediatric-Otorhinolaryngology (ESPO) -- MAY 20-23, 2012 -- Amsterdam, NETHERLANDS Atmaca, Enes/0000-0002-8978-3755; Aksoy, Abdurrahman/0000-0001-9486-312X; /0000-0002-0636-4214 WOS: 000318361700022 PubMed: 23232827 Objectives. The aim of this study is to investigate the effect of early intranasal steroid administration on wound healing after sinus surgery. Study Design. Randomized, placebo-controlled, blinded animal study. Setting. Animal laboratory. Subjects and Methods. Forty-two male New Zealand rabbits underwent bilateral 3-mm punch resection of the concha nasalis ventralis. The animals were divided into 3 groups: saline, late steroid, and early steroid. The saline group received saline drops, the late steroid group received saline drops for 7 days followed by intranasal mometasone furoate 50 mu g/nostril/d, and the early steroid group received intranasal mometasone furoate (same dose) starting on postoperative day 1. The animals were sacrificed on postoperative days 10 and 21. Left nasal specimens were examined histopathologically with hematoxylin-eosin and matrix metalloproteinase-9 (MMP-9) stains. Right nasal specimens were examined with high-performance liquid chromatography, and hydroxyproline levels were measured as mg/g in wet tissue. Results. Late steroid and early steroid groups were similar with regard to MMP-9 staining on days 10 and 21. On day 10, the early steroid group revealed significantly intense MMP-9 staining when compared with the saline group, and the late steroid and early steroid groups revealed significant fibrosis when compared with the saline group. Hydroxyproline levels were similar in all groups on day 10. The early steroid group revealed significantly higher hydroxyproline levels when compared with the late steroid group on day 21. Conclusion. Our findings showed that early intranasal steroid administration after sinus surgery in an animal model has no detrimental effects with regard to wound-healing parameters. European Soc Pediat Otorhinolaryngol (ESPO)

Tear glucose sensing for diabetes management has long been sought as an alternative to more invasive self-monitoring of blood glucose (SMBG). However, tear glucose sensors were known to have limitations, including correlation issues with blood glucose due to low sample volume, low concentration of glucose in the tear fluid, and evaporation of the tear sample. An engineering design approach to solve these problems led to the development of an integrated device capable of collecting the tear sample from the ocular surface with little to no stress on the eye, with an extremely low limit of detection, broad dynamic range, and rapid detection and analysis of sample. Here we present the development of a prototypical self-monitoring of tear glucose (SMTG) sensor, summarizing bench studies on the enzymes and their specificity, the development of the fluid capture device and its manufacture and performance and results of system testing in an animal study where safety, lag time and tear glucose to blood glucose correlation were assessed.