Sialendoscopy for Sublingual Gland Sialolithiasis.

OBJECTIVES: To present a unique series of sublingual gland sialolithiasis treated with sialendoscopy. METHODS: A retrospective chart review of patients at a single tertiary care academic center undergoing sialendoscopy. RESULTS: We present a series of 4 patients who presented with 1 episode each of sialadenitis without preceding postprandial symptoms. In each case, sialendoscopy identified the calculi embedded within the sublingual duct of Bartholin at its emptying point into Wharton's duct. The large size of the stones did not permit endoscopic removal, and transoral excision was performed via a floor of mouth incision. All patients recovered well with resolution of preoperative symptoms. CONCLUSIONS: Sublingual gland sialolithiasis has a distinct presentation that is not associated with obstructive symptoms and may be successfully treated with a combined endoscopic and transoral approach. Sialendoscopy allows identification of the location of the sialolith to guide removal and helps identify stone fragments and additional stones remaining after sialolithotomy. Transoral resection of the sublingual gland in this scenario facilitates stone removal and may minimize long-term chance of ranula formation.

Nonvisualization of Sialoliths during Sialendoscopy.

OBJECTIVES: Analyze the characteristics of patients undergoing interventional sialendoscopy for sialolithiasis whose stones were not visualized intraoperatively. STUDY DESIGN: Case series with chart review. SETTING: Tertiary care hospital. SUBJECTS AND METHODS: Patients (n = 276) undergoing sialendoscopy between June 2008 and December 2014 were reviewed for patient characteristics, imaging characteristics, and outcomes. Nonvisualization was defined as a sialolith that was documented on imaging preoperatively but not visualized intraoperatively during sialendoscopy, despite successful ductal cannulation and evaluation. RESULTS: A total of 337 sialendoscopy procedures were preformed. Preoperative imaging documented a sialolith in 203 (60%) cases. Nonvisualization occurred in 31 (15%) cases with sialolith. The parotid gland was involved in 58% (18 of 31) of nonvisualization cases, as opposed to 21% (43 of 203) of all sialolith cases. The submandibular gland was involved in 42% (13 of 31) of nonvisualization cases, as compared with 79% (160 of 203) of all sialolith cases. Nonvisualization occurred in 42% (18 of 43) of parotid cases versus 8% (13 of 160) of submandibular cases, a statistically significant difference (P < .001). Parotid stones located posterior to the plane of insertion of the posterolateral edge of the masseter were significantly more likely to experience nonvisualization (73%) than those along and anterior to the masseter (25% and 0%, respectively; P = .009). CONCLUSIONS: Intraoperative nonvisualization of a sialolith is more likely to occur in the parotid gland. Proximal stone location may predict nonvisualization. These factors should be considered during treatment planning and counseling for patients with sialolithiasis.

Increased yield of endothelial cells from peripheral blood for cell therapies and tissue engineering.

AIM: Peripheral blood-derived endothelial cells (pBD-ECs) are an attractive tool for cell therapies and tissue engineering, but have been limited by their low isolation yield. We increase pBD-EC yield via administration of the chemokine receptor type 4 antagonist AMD3100, as well as via a diluted whole blood incubation (DWBI). MATERIALS & METHODS: Porcine pBD-ECs were isolated using AMD3100 and DWBI and tested for EC markers, acetylated LDL uptake, growth kinetics, metabolic activity, flow-mediated nitric oxide production and seeded onto titanium tubes implanted into vessels of pigs. RESULTS: DWBI increased the yield of porcine pBD-ECs 6.6-fold, and AMD3100 increased the yield 4.5-fold. AMD3100-mobilized ECs were phenotypically indistinguishable from nonmobilized ECs. In porcine implants, the cells expressed endothelial nitric oxide synthase, reduced thrombin-antithrombin complex systemically and prevented thrombosis. CONCLUSION: Administration of AMD3100 and the DWBI method both increase pBD-EC yield.

Learning Progression in the Use of Sialendoscopy for Sialolithiasis: Effect on Gland Preservation.

OBJECTIVE: Evaluate how learning progression affects outcomes for the use of sialendoscopy for sialolithiasis. STUDY DESIGN: Case series with chart review. SETTING: Academic tertiary care center. SUBJECTS AND METHODS: A retrospective chart review was conducted on 81 patients presenting with sialolithiasis between 2008 and 2012 who underwent surgical intervention on 85 salivary glands. Outcomes compared between the first 43 and subsequent 42 consecutive glands included successful removal, surgical method, and need for further intervention. RESULTS: For the first 43 patients (group A), mean age was 47.3 years (range, 15-77), and 44.2% were male. For the second 38 patients (group B), mean age was 49.9 (range, 23-76), and 50% were male. Mean stone size was similar in both groups; 8.3 versus 7.6 mm, respectively. In cases of stone visualization and removal, complete removal without gland excision was accomplished in 78.4% of cases in group A versus 94.3% in group B (P = .04). Endoscopic removal occurred in 24.3% of patients in group A, versus 45.8% in group B. A combined approach was used in 54.1% of patients in group A versus 48.6% in group B. In group A, 18.9% underwent gland removal as part of initial intervention versus 0% in group B (P = .007). Overall gland preservation was 81.4% in group A versus 97.6% in group B (P = .015). CONCLUSION: This study documents the effect of learning curve on sialendoscopy for the management of sialolithiasis. With experience, the success of sialendoscopy increases with a significant decrease in the number of gland excisions.

Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress.

The overall goal of this method is to describe a technique to subject adherent cells to laminar flow conditions and evaluate their response to well quantifiable fluid shear stresses. Our flow chamber design and flow circuit (Fig. 1) contains a transparent viewing region that enables testing of cell adhesion and imaging of cell morphology immediately before flow (Fig. 11A, B), at various time points during flow (Fig. 11C), and after flow (Fig. 11D). These experiments are illustrated with human umbilical cord blood-derived endothelial progenitor cells (EPCs) and porcine EPCs. This method is also applicable to other adherent cell types, e.g. smooth muscle cells (SMCs) or fibroblasts. The chamber and all parts of the circuit are easily sterilized with steam autoclaving. In contrast to other chambers, e.g. microfluidic chambers, large numbers of cells (> 1 million depending on cell size) can be recovered after the flow experiment under sterile conditions for cell culture or other experiments, e.g. DNA or RNA extraction, or immunohistochemistry (Fig. 11E), or scanning electron microscopy. The shear stress can be adjusted by varying the flow rate of the perfusate, the fluid viscosity, or the channel height and width. The latter can reduce fluid volume or cell needs while ensuring that one-dimensional flow is maintained. It is not necessary to measure chamber height between experiments, since the chamber height does not depend on the use of gaskets, which greatly increases the ease of multiple experiments. Furthermore, the circuit design easily enables the collection of perfusate samples for analysis and/or quantification of metabolites secreted by cells under fluid shear stress exposure, e.g. nitric oxide (Fig. 12).

Autologous endothelial progenitor cell-seeding technology and biocompatibility testing for cardiovascular devices in large animal model.

Implantable cardiovascular devices are manufactured from artificial materials (e.g. titanium (Ti), expanded polytetrafluoroethylene), which pose the risk of thromboemboli formation. We have developed a method to line the inside surface of Ti tubes with autologous blood-derived human or porcine endothelial progenitor cells (EPCs). By implanting Ti tubes containing a confluent layer of porcine EPCs in the inferior vena cava (IVC) of pigs, we tested the improved biocompatibility of the cell-seeded surface in the prothrombotic environment of a large animal model and compared it to unmodified bare metal surfaces (Figure 1). This method can be used to endothelialize devices within minutes of implantation and test their antithrombotic function in vivo. Peripheral blood was obtained from 50 kg Yorkshire swine and its mononuclear cell fraction cultured to isolate EPCs. Ti tubes (9.4 mm ID) were pre-cut into three 4.5 cm longitudinal sections and reassembled with heat-shrink tubing. A seeding device was built, which allows for slow rotation of the Ti tubes. We performed a laparotomy on the pigs and externalized the intestine and urinary bladder. Sharp and blunt dissection was used to skeletonize the IVC from its bifurcation distal to the right renal artery proximal. The Ti tubes were then filled with fluorescently-labeled autologous EPC suspension and rotated at 10 RPH x 30 min to achieve uniform cell-coating. After administration of 100 USP/kg heparin, both ends of the IVC and a lumbar vein were clamped. A 4 cm veinotomy was performed and the device inserted and filled with phosphate-buffered saline. As the veinotomy was closed with a 4-0 Prolene running suture, one clamp was removed to de-air the IVC. At the end of the procedure, the fascia was approximated with 0-PDS (polydioxanone suture), the subcutaneous space closed with 2-0 Vicryl and the skin stapled closed. After 3 - 21 days, pigs were euthanized, the device explanted en-block and fixed. The Ti tubes were disassembled and the inner surfaces imaged with a fluorescent microscope. We found that the bare metal Ti tubes fully occluded whereas the EPC-seeded tubes remained patent. Further, we were able to demonstrate a confluent layer of EPCs on the inside blood-contacting surface. Concluding, our technology can be used to endothelialize Ti tubes within minutes of implantation with autologous EPCs to prevent thrombosis of the device. Our surgical method allows for testing the improved biocompatibility of such modified devices with minimal blood loss and EPC-seeded surface disruption.

Use of autologous blood-derived endothelial progenitor cells at point-of-care to protect against implant thrombosis in a large animal model.

Titanium (Ti) is commonly utilized in many cardiovascular devices, e.g. as a component of Nitinol stents, intra- and extracorporeal mechanical circulatory assist devices, but is associated with the risk of thromboemboli formation. We propose to solve this problem by lining the Ti blood-contacting surfaces with autologous peripheral blood-derived late outgrowth endothelial progenitor cells (EPCs) after having previously demonstrated that these EPCs adhere to and grow on Ti under physiological shear stresses and functionally adapt to their environment under flow conditions ex vivo. Autologous fluorescently-labeled porcine EPCs were seeded at the point-of-care in the operating room onto Ti tubes for 30 min and implanted into the pro-thrombotic environment of the inferior vena cava of swine (n = 8). After 3 days, Ti tubes were explanted, disassembled, and the blood-contacting surface was imaged. A blinded analysis found all 4 cell-seeded implants to be free of clot, whereas 4 controls without EPCs were either entirely occluded or partially thrombosed. Pre-labeled EPCs had spread and were present on all 4 cell-seeded implants while no endothelial cells were observed on control implants. These results suggest that late outgrowth autologous EPCs represent a promising source of lining Ti implants to reduce thrombosis in vivo.

The biocompatibility of titanium cardiovascular devices seeded with autologous blood-derived endothelial progenitor cells: EPC-seeded antithrombotic Ti implants.

Implantable and extracorporeal cardiovascular devices are commonly made from titanium (Ti) (e.g. Ti-coated Nitinol stents and mechanical circulatory assist devices). Endothelializing the blood-contacting Ti surfaces of these devices would provide them with an antithrombogenic coating that mimics the native lining of blood vessels and the heart. We evaluated the viability and adherence of peripheral blood-derived porcine endothelial progenitor cells (EPCs), seeded onto thin Ti layers on glass slides under static conditions and after exposure to fluid shear stresses. EPCs attached and grew to confluence on Ti in serum-free medium, without preadsorption of proteins. After attachment to Ti for 15 min, less than 5% of the cells detached at a shear stress of 100 dyne / cm(2). Confluent monolayers of EPCs on smooth Ti surfaces (Rq of 10 nm), exposed to 15 or 100 dyne/cm(2) for 48 h, aligned and elongated in the direction of flow and produced nitric oxide dependent on the level of shear stress. EPC-coated Ti surfaces had dramatically reduced platelet adhesion when compared to uncoated Ti surfaces. These results indicate that peripheral blood-derived EPCs adhere and function normally on Ti surfaces. Therefore EPCs may be used to seed cardiovascular devices prior to implantation to ameliorate platelet activation and thrombus formation.