Cadaveric Tissue Research Articles

Simulator Availability in Meeting the Requirements of United States Army Ophthalmology Individual Critical Task Lists.

IntroductionIndividual Critical Task Lists (ICTLs) are a list of requirements set forth by the United States Army which each soldier must fulfill to maintain competency in a specialty. By providing senior leadership objective criteria with which to evaluate the competency of each service member, ICTLs support commanders in ensuring that soldiers are mission ready and deployable. Board-certified ophthalmologists can meet ICTL requirements by demonstrating skills on an actual patient, a simulator, and/or cadaveric or live tissue. We sought to determine the availability of simulators that can be used to meet Army ophthalmology ICTL requirements.MethodsWe reviewed the current Army ICTLs for ophthalmologists. We performed an online search, as well as an extensive review of Pubmed, AccessMedicine, Academic Search Elite, Thieme, and ScienceDirect, to identify available simulators for each ICTL. We did not use any date or language restrictions in the electronic search for trials. We last searched the electronic databases on April 27, 2019.ResultsArmy Ophthalmologists are required to maintain current status in 19 areas based on ICTLs established by the Critical Task Site and Selection Board. Eight of these requirements are not amenable to a simulation of any kind. Of the 11 remaining ICTLs, approximately 82% can be satisfied with a simulator alone based on current simulator availability. The remaining 18% of applicable ICTLs can be satisfied using cadaveric or live tissue training.ConclusionsArmy ophthalmologists can keep current with their ICTLs, and thus maintain mission readiness, by using either simulators or cadaveric or live tissues. This is particularly important for ophthalmologists who are either located in remote or austere locations without resources or areas with low surgical volumes. Several tasks are applicable to other medical specialties which can benefit from the same simulators.

Promotion of Cadaver-Based Military Trauma Education: Integration of Civilian and Military Trauma Systems.

For the past few decades, there has been an emphasis on encouraging the partnership of civilian and military trauma care systems which would allow military personnel to maintain competency in life-saving skills and gain experience prior to or in between deployments. Currently, there are only five primary military-civilian training centers in the United States. Thus, the majority of service members do not get the opportunity to train at these facilities prior to scheduled deployments. To bridge this gap, a joint military-civilian hands-on supplemental training program was established to allow deploying National Guard Combat Medics to practice life-saving techniques on human cadaver tissue. The purpose of this report is to provide the feedback survey from this pilot training session in hopes to expand and improve the curriculum and encourage partnerships between military and civilian trauma centers. Survey respondents include the 13 National Guard Combat Medic Specialists who attended the 6-hour voluntary training session. The survey asked qualitative and quantitative (using a 5-point Likert rating scale; 1 = insufficient/unacceptable/disagree; 5 = outstanding/exceeded expectations/strongly agree) questions regarding the structure of the session, likes and improvements about the classroom and cadaver lab portions, implementation into training, confidence in techniques before and after the session and overall rating of the session. This report has been reviewed and approved by the Institutional Review Board (IRB) at Rutgers University. Most attendees (12 of 13) agreed that the 6-hour total time length and division of classroom versus lab time was appropriate. Most attendees (12 of 13) agreed that this type of supplemental hands-on cadaver training should be a mandatory part of their medical training. The attendees' rated confidence in covered techniques was significantly increased from 3.77 ± 0.73 to 4.85 ± 0.38 (p < 0.01) before and after the session, respectively. The overall satisfaction rating of the session was 4.92 ± 0.28. This collaboration between military and civilian trauma care system was successful in giving National Guard combat medic specialists the opportunity to practice life-saving techniques on human tissue prior to deployment. This type of partnership can assist in maintaining readiness for trauma care and increasing the confidence of military pre-hospital providers in performing life-saving techniques.

Ultrathin, Strong, and Cell-Adhesive Agarose-Based Membranes Engineered as Substrates for Corneal Endothelial Cells.

We aimed to bioengineer a scaffold that can facilitate the transplantation of corneal endothelial cells (CEC), given the global shortage of cadaveric donor tissues. Although agarose (A) has outstanding biocompatibility and mechanical properties, it natively does not permit cell adhesion. In this study, agarose was modified with different attachment signals: GRGD (giving AR as product), lysine (AK), poly lysine (AP), and fish-derived gelatin (AG). Samples with varying conjugation ratios were prepared. All products formed bulk hydrogels, which were then collapsed into ultrathin membranes in a controlled environment. Membranes were evaluated for their ability to support attachment of various cell types. Cells, however, preferred the AG series of membrane. Notably, primary rabbit CEC remained attached and viable for ⩾4 weeks. The cells also stained positive for CD166, ZO-1 and Na+/K+ ATPase, indicative of function. The hydrated AG membranes allowed >96% transmittance of visible light. The membranes were typically ∼15 μm thick and did not swell significantly after immersion in PBS. Tensile strength was 49-60 MPa, while young's modulus was 525-596 MPa. This membrane thus offers great promise as a scaffold for CEC during endothelial keratoplasty.

Computer-Based Video Instruction for Training Medical Students on Skin Biopsies.

The development of procedural skills is necessary for medical students. Computer-based video instruction (CBVI) increases knowledge and procedural skills. This pilot study's aim was to investigate the usefulness of CBVI in dermatologic procedure training for medical students and secondarily assess students' overall perception of the field of dermatology. Twenty-nine first- and second-year medical students were randomly assigned to the CVBI group or control group, in addition to in-person instructor demonstration of shave and punch biopsies using fresh cadaver tissue. Blinded evaluators graded student performances using a five-point Likert scale immediately after demonstration, and 1 week later to assess knowledge retention. In overall performance, the CBVI group demonstrated higher scores both in shave (3.54 vs 2.59, p = .01) and punch biopsies (3.63 vs 2.88, p = .01) at immediate recall and knowledge retention (3.68 vs 2.67, p = .01; 4.00 vs 2.99, p < .001, respectively). Approximately 33.3% of the students stated that the experience increased their interest in the field of dermatology. Incorporation of CBVI into the dermatology curriculum augments medical students' procedural skills. The CBVI group performed significantly better in all 7 grading categories for shave biopsy and in 5 of 7 categories for punch biopsy. Integration of procedural laboratory tests raises students' interest in dermatology.

HP-03-004 Establishing the safety profile in sexual partners of a new topical glyceryl trinitrate gel for the treatment of erectile dysfunction

MED2005 is a fast-acting, topical, glyceryl trinitrate (GTN) gel for the treatment of erectile dysfunction (ED), applied directly to the glans penis by the male or his partner.1 As there is potential for transfer to the female partner, the safety of MED2005 in sexual partners was investigated. (A) In a 232 ED patient Phase IIA study, adverse events (AEs) were recorded in female partners following the application of 0.2% MED2005 (n=229) to the penis before intercourse.1 (B) A pharmacokinetic study measured GTN absorption by tandem mass spectrometry on penile swabs taken 5 minutes after administration of 0.8% MED2005 to the glans penis (n=10). (C) Disruption to the epidermal membrane 2 hours after administration of MED2005 was measured by impedance testing on cadaveric human epidermal tissue and significance compared to commercial lubricants using the Wilcoxon signed rank test. (A) In over 1003 intercourse events, only 4 mild AEs in females were related to their partners’ MED2005 treatment; two cases of headache and one case each of vulvovaginal pruritis and nasopharyngitis. (B) Five minutes after application, an average of 27% of MED2005 remained on the penis, indicating absorption of 73% of the gel. Linear input rates predicted that virtually no MED2005 would remain on the penis 10 minutes after application. (C) Impedance testing showed no significant difference between MED2005 and commercial lubricants on integrity of human epidermal membranes, p=0.58 and 0.30 compared to KY warming and Astroglide, respectively.

Prevalence of Anatomical Variants of the Circle of Willis

The Circle of Willis is an anastomotic structure that connects the anterior and posterior arterial networks that supply blood to the brain. It functions to maintain adequate blood supply to the brain in the event of obstruction due to injury or disease. Although it is well‐known that variants in the “textbook” structure of the Circle of Willis are common, previous studies characterizing the CoW have been limited by small sample sizes and, in some cases, reliance on cadaveric tissue in which preservation of delicate CoW structures is challenging. These shortfalls have made investigating the relationships between variants and neurological pathology difficult. Therefore, the purpose of the present study was to characterize the CoW in a large sample of living subjects to determine the prevalence of CoW variants and to identify relationships between variants and pathology. After approval for this study was obtained from the AdventHealth IRB, computed tomography angiography (CTA) scans of 1,494 cases were assessed for CoW variants and all CoW vessels measured. Inclusion criteria included patients aged 18–89 that received CTA imaging through AdventHealth Radiology between 11/15/12 and 11/14/17. One examiner assessed all CTA images via imaging software (Impax) for variants. Six other investigators measured each of the CoW vessels on the selected cases. Patient demographic information was collected via AdventHealth's medical record system. Our results indicate that CoW variants were present in 47.99% of the subjects. Of the cases with variants, 8.93% possessed symmetrical bilateral variations and 39.05% possessed multiple variants. The most commonly occurring variant was a non‐dominant vertebral artery at 17.27%. Of the non‐diminutive variants, meaning there were additions to or subtractions from the standard CoW architecture, the most commonly occurring variant was fetal origin posterior cerebral artery at 12.38%. Our results suggest that anatomical variants found in the Circle of Willis occur in just over under half of the population. Future analyses will characterize vessel diameters for the cases under study and relationships of each of the variants to neurological pathology. This study demonstrates the need for further investigation into the nature of altered CoW circulation patterns, how CoW variants may relate to brain function and pathology, and whether diminutive variants can be altered by non‐invasive interventions.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Sex, but not skin tone affects penetration of red-light (660 nm) through sites susceptible to sports injury in lean live and cadaveric tissues.

Red-light treatment is emerging as a novel therapy for promoting tissue recovery but data on red-light penetration through human tissues are lacking. We aimed to: (1) determine the effect of light irradiance, tissue thickness, skin tone, sex and bone/muscle content on 660 nm light penetration through common sites of sports injuries, and (2) establish if cadaver tissues serve as a useful model for predicting red-light penetration in live tissues. Live and cadaver human tissues were exposed to 660 nm light at locations across the skull, spinal cord and upper and lower limbs. Red-light was produced by a light emitting diode array of various irradiances (15-500 mW/cm2 ) and measured by a light-probe positioned on the tissue surface opposite to the light emitting diodes. 100 mW/cm2 successfully penetrated tissue <50 mm thick; a disproportionate irradiance increase was required to achieve deeper penetration. Penetration was unaffected by skin tone, increased with irradiance and relative bone/muscle composition, and decreased with greater tissue thickness and in males. Live and cadaveric tissue penetration did not differ statistically for tissues <50 mm but cadavers required more red-light to penetrate >50 mm. These results assist clinicians and researchers in determining red-light treatment intensities for penetrating human tissues.

Minimally Invasive Muscle Embedding (MIME), Facilitates the Development of Functional Muscle Fibers of Human Cadaveric Origin, in Host Mice

BACKGROUNDMinimally Invasive Muscle Embedding (MIME), is a technique developed in our laboratory, to facilitate the development of donor‐cell‐derived muscle fibers in a host muscle. MIME involves passing a sterile needle through a host muscle, and then implanting a segment of donor muscle tissue along with its myogenic satellite cells, in the needle track within the host muscle.METHODSWe validated MIME, by implanting a single extensor digitorum longus (EDL) muscle, from donor mice (N=2), which expressed a red fluorescent protein (RFP), into the left tibialis anterior muscle (TA), of immunodeficient host mice (N=4), which expressed a green fluorescent protein (GFP). In the same host mice, we performed a SHAM procedure on the right TA. In another set of host mice, we implanted by MIME, segments of human cadaveric TA (3 days post‐mortem), or performed a SHAM procedure (N=4 per group). We triggered concerted muscle degeneration and regeneration by injecting 1.2% barium chloride (BACL) into the TAs after MIME or SHAM. We collected TAs at 14 days after implanting RFP+ mouse donor muscle, and at 12 weeks after implanting human donor muscle. We performed histological and physiological assays to compare MIME‐ and SHAM‐treated host TA muscles. We studied histological images with Image J software to obtain quantitative data. We performed statistical analyses (ANOVAs) with SigmaStat or SPSS software.RESULTSIn host TAs implanted with RFP+ mouse EDL, 22±4% and 78±4% muscle fibers were RFP+ and GFP+, respectively. There were no RFP+ fibers in SHAM‐treated muscles. All RFP+ fibers were positive for desmin and dystrophin, and 65±4 % fibers were centrally nucleated. Donor‐derived fibers had multiple sarcomeres and nuclear domains in series. In host TAs implanted with human muscle, ~30% of the host muscle was of donor origin (GFP−), and the majority of GFP− fibers were positive for human‐specific markers. The contractile torque produced by MIME and SHAM TAs was not statistically different at 12 weeks post‐MIME, and there was no statistical difference compared to torque produced before MIME or SHAM procedures. Data are reported as Mean±SEM. P&lt;0.05 considered significant.CONCLUSIONThrough MIME, we confirm that, human cadaveric muscle, implanted up to 3 days post‐mortem, can generate donor‐derived muscle fibers that are morphologically and functionally healthy. Our studies are translationally‐relevant; since MIME can be adapted to implant myogenic tissue of human cadaveric origin, to generate donor‐derived muscle fibers in human recipients (example: patients with muscle loss from muscle disease, trauma, or aging), with suitable immunosuppressive and rehabilitative therapies.Support or Funding InformationFunded by a Pilot Grant from the Alliance for Regenerative Rehabilitation Research and Training (AR3T), NIH 1R03HD091648‐01 and 5R03HD091648‐02 from NICHD, and a Faculty Startup Package and FRAP award from Wayne State University, to JAR. De‐identified, cadaveric human muscle tissue, was made available through the Body Bequest Program at Wayne State University. MIME facilitates donorderived myogenesisimageMIME facilitates donorderived myogenesisThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Use of Magnetic Induction to Activate a “Touchless” Shape Memory Alloy Implantable Penile Prosthesis

IntroductionWe describe a novel physiologic penile prosthesis that uses shape memory alloy properties to mimic the transition between a flaccid and erect penis using magnetic induction instead of hydraulic pressure. AimTo evaluate the parameters of magnetic induction to activate a shape memory alloy implantable penile prosthesis. MethodsWe prototyped an implantable penile prosthesis cylinder using temperature-tuned nickel titanium alloy tubes laser cut to specifications.The device was then tested implanted in an animal tissue model and in cadaveric tissue. Testing consisted of placing the device deactivated in its more malleable and compressed state, then activating it using an external inducer wand while measuring temperature changes that occur on the surface of the device, within the tissue, and at the skin surface. Main Outcome MeasuresOur main outcome measures were the efficiency of activation and thermal safety of this approach. ResultsUsing a handheld magnetic inductor, we were able to successfully activate the SMA penile prosthesis with no direct contact under 45 seconds. This handheld wand produced a magnetic field that penetrated tissue and caused the appropriate phase change within the prosthesis. Tissue temperature (middle and surface probes) in the animal tissue model increased only a few degrees Celsius during the activation process, and never exceeded 28 degrees Celsius from a baseline at room temperature ∼ 25 degrees Celsius. We encountered similar results without a notable change in tissue temperature in the cadaveric tests. The fully activated device resisted buckling forces of 2.66 kgf ± 0.045. Clinical ImplicationsThis non-hydraulic shape memory prosthesis obviates the needs for reservoirs and pumps, and the wand-based interaction with the device may be easier to use. Strength & LimitationsThis technology represents a fundamental departure from the hydraulic-based penile prosthesis and has comparable mechanical properties as current-marked devices. It appears to show thermal safety in controlled environments, however real-world use would need further studies. Further optimization of prototypes needs to be done prior to human clinical trials. ConclusionA shape memory penile prosthesis is a promising alternative to hydraulic-based penile prostheses and can be activated safely and efficiently using magnetic induction in our models of the human penis.Le BV, McVary KT, McKenna K, et al. Use of Magnetic Induction to Activate a “Touchless” Shape Memory Alloy Implantable Penile Prosthesis. J Sex Med 2019;16:596–601.

Performance and microbiological safety testing after multiple use cycles and hydrogen peroxide sterilization of a 5-mm vessel-sealing device.

To evaluate the functional and microbiological safety of a single-use laparoscopic vessel-sealing device in a multiuse setting. Ex vivo study. Twelve 5-mm LigaSure Maryland jaw devices. Handsets underwent repeated test cycles until failure. The handset packaging was opened, and handsets were agitated in phosphate-buffered saline (PBS). The PBS was removed, centrifuged, and submitted for culture. Canine ovariectomy was simulated on cadaveric tissue, after which vascular seal quality was evaluated by pressure testing of sealed porcine carotid arteries. The handsets were cleaned and sterilized with hydrogen peroxide gas and repackaged. Mean ± SD cycles to failure was 7.7 ± 2.8, with a minimum of 4 and a maximum of 12 use cycles achieved. Eleven of the 12 handsets failed by failure of handset activation after depression of the activation trigger. Only 1 handset failed to hold an adequate vascular seal under 300 mm Hg of pressure. No handset exhibited positive bacterial culture at any cycle. The sterilization method used in this study resulted in an excellent microbiological safety profile. Most of the handsets failed by activation button failure. Under the conditions of this study, hydrogen peroxide sterilization achieved microbiological safety. Handsets can successfully be reused until activation button failure without negative effects on the vascular seal or increased risk of infection to the patient.

Developing a dynamic simulator for endoscopic intraventricular surgeries

A novel dynamic simulator brain model with hydrocephalus has been developed for endoscopic intraventricular procedures. Detachable components allow enhancement of the walls of the ventricle by choroid plexus, ependymal veins and the membranous floor of the third ventricle which are derived from cadaveric lab animal tissues to give a lifelike appearance. These can be changed for every exercise. Ventricles are filled with injection of saline to give appropriate transparent medium and connected to a device transmitting pulsations creating conditions similar to live surgeries. Thirty-five participants have used this model over the last 1year and found it to be useful for conducting third ventriculostomy. Further development of the model for septostomy, aqueductoplasty and tumour biopsy has also been recently tested successfully by 12 participants. It is hoped that this simulator model for intraventricular endoscopy is comprehensive as a learning tool in carrying out most of thethe surgical procedures currently practised.

Forensic toxicological study on adipocere formation in submerged cadavers of female albino rats intoxicated with cadmium

There is a dearth of information on the mutual interaction between metal intoxication and adipocere formation. Herein, 40 adult female albino rats were distributed into two equal groups, one used as control while the other orally administered single dose of cadmium chloride (CdCl2) 225 mg/kg·bw (LDmin). Control group was killed by cervical dislocation. Half of dead rats of both groups were subjected for determination of iodine value and estimation of cadmium (Cd) residues while the other half of both groups were submerged in opened glass container previously filled with 4 L dechlorinated tap water kept in closed room with an open air access (one cadaver/container). Gross morphological changes of submerged cadavers were recorded weekly along the experiment. At the end of the experiment, after 3 months, samples were collected again for iodine value determination and estimation of Cd residues. The obtained results revealed the depressant effect of Cd toxicity on development of adipocere. Cd residues were found in different tissues of cadavers at time of death with the highest amount in the intestines followed by the liver and kidneys, then lungs, adipose tissue, muscles, and finally the bones. After 3 months of water submersion, tissues exhibited significant decrease in the amount of Cd residues but to a limit that was still detected. This study concluded the possibility of detection of Cd residues even after adipocere formation. Additionally, it shed light on the possibility of the interference of environmental pollution with the natural rate of decomposition especially adipocere formation.

An implanted pH sensor read using radiography.

A biomedical sensor was developed to measure local pH near orthopedic implants to detect and study implant-associated infection. The sensor is read using plain radiography, a technique which is noninvasive, inexpensive, ubiquitously available in medical facilities, and routinely used in diagnosis and follow-up. The sensor comprises a radiopaque tungsten indicator pin embedded within a chemically responsive hydrogel that exhibits a pH-dependent swelling. A stainless steel well holds this hydrogel and attaches to an orthopedic plate. The local pH may be determined from the extent of hydrogel swelling by radiographically measuring the indicator position relative to the well. We calibrated the sensor in a series of standard pH buffers and tested it during bacterial growth in culture. The sensor was robust: its response was negligibly affected by changes in temperature, ionic strength within the normal physiological range, or long-term incubation with reactive oxygen species generated from hydrogen peroxide and copper. Pooled data from several sensors fabricated at different times and tested in different conditions had a root-mean-square deviation from a pH electrode reading of 0.24 pH units. Radiographic measurements were also performed in cadaveric tissue with the sensor attached to an orthopedic plate fixed to a tibia. Pin position readings varied by 100 μm between observers surveying the same radiographs, corresponding to 0.065 pH units precision in the range pH 4-8. The sensor was designed to augment standard radiographs of tissue, bony anatomy, and hardware by also indicating local chemical concentrations.

Effects of dynamic loading on solute transport through the human cartilage endplate

Nutrient and metabolite transport through the cartilage endplate (CEP) is important for maintaining proper disc nutrition, but the mechanisms of solute transport remain unclear. One unresolved issue is the role of dynamic loading. In comparison to static loading, dynamic loading is thought to enhance transport by increasing convection. However, the CEP has a high resistance to fluid flow, which could limit solute convection. Here we measure solute transport through site-matched cadaveric human lumbar CEP tissues under static vs. dynamic loading, and we determine how the degree of transport enhancement from dynamic loading depends on CEP porosity and solute size. We found that dynamic loading significantly increased small and large solute transport through the CEP: on average, dynamic loading increased the transport of sodium fluorescein (376 Da) by a factor of 1.85 ± 0.64 and the transport of a large dextran (4000 Da) by a factor of 4.97 ± 3.05. Importantly, CEP porosity (0.65 ± 0.07; range: 0.47–0.76) strongly influenced the degree of transport enhancement. Specifically, for both solutes, transport enhancement was greater for CEPs with low porosity than for CEPs with high porosity. This is because the CEPs with low porosity were susceptible to larger improvements in fluid flow under dynamic loading. The CEP becomes less porous and less hydrated with aging and as disc degeneration progresses. Together, these findings suggest that as those changes occur, dynamic loading has a greater effect on solute transport through the CEP compared to static loading, and thus may play a larger role in disc nutrition.

Diets of animal origin and their influence on the development of the immatures of Chrysomya albiceps (Diptera: Calliphoridae): implications for forensic entomology

AbstractIn forensic entomology, the minimum and maximum post‐mortem interval (PMI) is estimated on the basis of the developmental stages of necrophagous Diptera larvae on a corpse. The nutritional composition of the decomposing tissue, which can influence larval development rates, may impact the precision of this estimate. We compared the post‐embryonic development of Chrysomya albiceps reared on bovine muscle and swine lung. Forty newly hatched fly larvae, per treatment, were placed on 60 g of the following tissues: bovine muscle (control) and swine lung (T1). Each treatment had three replicates. The experiments were maintained under controlled conditions (28°C day/26°C night, 70 ± 10% RH and 12 h photophase). Flies reared on bovine muscle took significantly longer to complete their larval phase (8.1 days) than did larvae reared on swine lung (6.8 days). The total duration of development in bovine muscle was significantly slower (12.3 days) than in swine lung. Larvae that were reared on swine lungs were significantly heavier than larvae reared on bovine muscle. Total larval viability was high on pig lung (over 49%). The sex ratio was closer to the 1:1 ratio in bovine muscle (rs = 0.55), with a slightly greater proportion of female larvae. The percentage of abnormality remained within the predicted range. The results show that the different tissues influenced larval development. This underlines the importance of considering the type of cadaveric tissue when larval development is used to calculate the minimum PMI in forensic investigations.

Evolution of a Novel Tissue Preservation Protocol to Optimize Osteochondral Transplantation Outcomes.

Osteochondral allograft transplantation is a procedure to treat focal osteochondral lesions (OCLs), but is limited by tissue availability, the quality of transplanted tissue, and inconsistent storage protocols. The objective of this study was to assess the clinical outcomes of a novel tissue procurement, storage, and quality control protocol in treating OCLs. Prospective case series. Donor cadaveric tissue was processed, stored, and the tissue quality analyzed using the unique tissue preservation protocol developed at our institution. Advanced cross-sectional imaging was used to size match donor tissue with recipient patients. Osteochondral allografts were transplanted using the Arthrex Allograft OATS. Patients were evaluated with the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Knee injury and Osteoarthritis Outcome Score (KOOS), visual analog scale (VAS), and 36-Item Short Form Survey (SF-36) preoperatively and at 1 year and 2 years postoperatively. Twenty patients (17 knees, 3 shoulders) were included in the study. There was a significant improvement in the following scores: overall WOMAC score, WOMAC function and pain subcategories; KOOS pain, knee-related symptoms, activities of daily living, sports and recreation, and quality of life; SF-36 physical functioning, physical role, pain, and social functioning subcategories; and VAS at all time points postoperatively. There was a significant improvement in WOMAC stiffness at 2 years postoperatively. There were 2 failures, defined by graft subsidence and persistent pain requiring reoperation. The protocol developed at our institution for OAT resulted in significant clinical improvement in patients with OCLs and is an improvement on existing tissue storage techniques.

Implementation and Evaluation of Training for Ultrasound-Guided Vascular Access to Small Vessels Using a Low-Cost Cadaver Model.

Critically ill neonatal and pediatric patients often require central vascular access. Real-time ultrasound guidance for central venous catheterization is beneficial. Because the diameter of central veins is much smaller in neonates than in adults, extensive training is needed to master the visualization and catheterization of central veins in neonates. This study assessed the learning effect of a standardized simulation-based teaching program on ultrasound-guided cannulation in a low-cost cadaver tissue model. This simulation-based prospective study assessed physician competence in the ultrasound-guided central venous catheterization procedure. Analyses were conducted before and after the teaching course. Pediatric simulation center at a tertiary care center. Staff physicians from the Neonatal ICU and PICUs at the Medical University of Vienna. Two latex tubes, with internal diameters of 2 and 4 mm, were inserted in parallel into cadaver tissue to mimic vessels and create a model for central venous catheterization. Under ultrasound guidance, each participant attempted to puncture and insert a guide-wire into each of the latex tubes using in-plane and out-of-plane techniques, both before and after the teaching course. The training program was assessed using a questionnaire and a performance checklist. Thirty-nine physicians participated in this study. The rates of failure of guide-wire insertion into 2-mm tubes were significantly lower after than before the teaching course, using both in-plane (p = 0.001) and out-of-plane (p = 0.004) techniques. Teaching, however, did not significantly reduce the insertion failure rate into 4-mm tubes, either in-plane (p = 0.148) or out-of-plane (p = 0.069). The numbers of successful cannulations on the first attempt increased after the teaching in all methods (p = 0.001). Implementation of a skills training program for ultrasound-guided central venous catheterization in a cadaver tissue model was feasible and cost- and time-effective. The number of attempts until successful cannulation of small vessels (2-mm tube) was significantly lower after than before the standardized teaching program.

Agili-C implant promotes the regenerative capacity of articular cartilage defects in an ex vivo model.

Osteochondral implants are currently adopted for the treatment of symptomatic full-thickness chondral and osteochondral defects. Agili-C™ is a cell-free aragonite-based scaffold which aims to reproduce the original structure and function of the articular joint while directing the growth and regeneration of both cartilage and its underlying subchondral bone. The goal of the present study was to investigate the ex vivo mechanisms of action (MOA) of the Agili-C™ implant in the repair of full-thickness cartilage defects. In particular, we tested whether Agili-C™ implant has the potential to stimulate cartilage ingrowth through chondrocytes migration into the 3D interconnected porous structure of the scaffold, along with maintaining theirviability and phenotype and the deposition of hyaline cartilage matrix. Articular cartilage samples were collected through the Gift of Hope Organ and Tissue Donor Network (Itasca, IL) within 24h from death. For this study, cartilage from a total of 14 donors was used. To model a chondral defect, donut-shaped cartilage explants were prepared from each tissue specimen. The chondral phase of the Agili-C™ implant was placed inside the tissue in full contact and press fit manner. Cartilage explants with the Agili-C™ implant inside were cultured for 60 days. As a control, the same donut-shaped cartilage explants were cultured without Agili-C™, under the same culture conditions. Using fresh human cadaveric articular cartilage tissue in a 60-day culture, it was demonstrated that chondrocytes were able to migrate into the Agili-C™ scaffold and contribute to the deposition of theextracellular matrix (ECM) rich in collagen type II and aggrecan, and lacking collagen type I. Additionally, we were able to show the formation of a layer populated by progenitor-like cells on the articular surface of the implant. The analysis of samples taken from knee and ankle joints of human donors with a wide age range and both genders supports the potential of Agili-C™ scaffold to stimulate cartilage regeneration and repair. Based on these results, the present scaffold can be used in the clinical practice as a one-step procedure to treat full-thickness chondral defects.

Decreased expression of folate transport proteins in oral cancer

The purpose of this study was to assess the expression of the 3 major folate transporters-folate receptors (FRs), reduced folate carrier (RFC), and proton-coupled folate transporter (PCFT)-in oral squamous cell carcinoma (OSCC). We hypothesized that patterns of expression of folate transporters would be different in OSCC compared with normal oral epithelium. We used immunohistochemistry to examine the expression of FR, RFC, and PCFT in 15 primary specimens collected from patients with OSCC, 2 human cadaveric samples of OSCC, and 12 normal human cadaveric oral tissues from a medical gross anatomy laboratory. Possible correlations between the expression of each folate transporter and patients' clinical data were determined. All 3 folate transporters were highly expressed in normal oral epithelium. In contrast, OSCC samples generally demonstrated low expression of FR, RFC, and PCFT, with wide distribution in the invading cancer cells. There were no differences in folate transporter expression between OSCC samples collected from patients and from human cadavers. The lowest expression of FR and PCFT characterized less-differentiated tumors, and the lowest expression of RFC correlated with higher lymph node involvement. Human oral cancer samples expressed decreased amounts of all 3 major folate transport proteins compared with controls from normal cadaveric oral tissues.

Chondrogenic differentiation of synovial fluid mesenchymal stem cells on human meniscus-derived decellularized matrix requires exogenous growth factors

The objective of this study was to investigate whether meniscus-derived decellularized matrix (DCM) has the capacity to induce differentiation of synovial fluid-derived mesenchymal stem cells (SF-MSCs) towards a meniscus fibrochondrocyte (MFC) phenotype. The potential roles of transforming growth factor beta-3 (TGF-β3) and insulin-like growth factor 1 (IGF-1) in the differentiation of SF-MSCs towards an MFC phenotype were also investigated.SF-MSCs were isolated via plastic adherence cell culture from the synovial fluid of five donors (5 male, average age 34 years). Porous DCM was generated by homogenizing and freeze-drying fresh normal human cadaveric meniscus tissue. SF-MSCs were seeded and cultured on the DCM scaffold in a defined serum-free media (SFM) supplemented with or without the combination of TGF-β3 and IGF-1. Cell pellets of SF-MSCs were cultured in SFM with either TGF-β3 or IGF-1 or their combination as controls. The duration of culture was 3 weeks for both experimental configurations. We assessed newly-formed tissues by biochemical assays, scanning electron microscopy (SEM), immunofluorescence and quantitative real-time PCR (qPCR).The combination of TGF-β3 and IGF-1 induced production of the cartilaginous matrix in DCM and upregulated the expression of aggrecan, collagens I and II. Moreover, the SF-MSCs exhibited a round morphology in the DCM scaffolds in the presence of the growth factors. In pellets, combined TGF-β3 and IGF-1 synergistically enhanced cartilaginous matrix production. In contrast to bone marrow mesenchymal stem cells (BM-MSCs), the differentiated SF-MSCs showed little evidence of the expression of the hypertrophic differentiation marker, collagen X.In conclusion, meniscus-derived DCM appears to require exogenous growth factor supplementation to direct differentiation of SF-MSCs. Statement of significanceMeniscus tears are the most common injury of the knee joint. These tears pose a major risk factor for the early development of knee osteoarthritis. Unfortunately, the majority of these tears occur in the inner region of the meniscus and lacks blood supply with no reparative or regenerative capacity.The goal of this study was to determine if the native extracellular matrix (ECM) of human meniscus has the capacity to differentiate human knee synovial fluid resident mesenchymal stem cells (SF-MSCs) towards a meniscus phenotype as a potential strategy to repair avascular meniscal tears.Our findings show that the human meniscus-derived ECM without supplementation with growth factors (TGF-β3 and IGF-1) cannot differentiate SF-MSCs towards a meniscus phenotype.The use of meniscus-derived scaffolds as a material to stimulate endogenous repair of meniscus tears via differentiation of SF-MSCs may require supplementation with TGF-β3 and IGF-1.