Yelena Akelina

The Rat Model in Microsurgery Education: Classical Exercises and New Horizons

Microsurgery is a precise surgical skill that requires an extensive training period and the supervision of expert instructors. The classical training schemes in microsurgery have started with multiday experimental courses on the rat model. These courses have offered a low threat supervised high fidelity laboratory setting in which students can steadily and rapidly progress. This simulated environment allows students to make and recognise mistakes in microsurgery techniques and thus shifts any related risks of the early training period from the operating room to the lab. To achieve a high level of skill acquisition before beginning clinical practice, students are trained on a comprehensive set of exercises the rat model can uniquely provide, with progressive complexity as competency improves. This paper presents the utility of the classical rat model in three of the earliest microsurgery training centres and the new prospects that this versatile and expansive training model offers.

Histologic and functional outcomes of nerve defects treated with acellular allograft versus cabled autograft in a rat model.

Acellular nerve allograft is a new option for bridging nerve defects that allows appropriate diameter matching. The aim of the study was to compare the histologic and functional recovery of nerve defects treated with acellular nerve allograft versus cabled sural nerve autograft.

Collagen nerve protector in rat sciatic nerve repair: A morphometric and histological analysis

Peripheral nerve repair is often complicated by fibroblastic scar formation, nerve dysfunction, and traumatic neuroma formation. Use of bio‐absorbable protective wraps may improve outcomes of these repairs. This study histologically compared the incidence of neuroma formation, connective tissue proliferation, and axonal regrowth in transected rat sciatic nerves repaired with and without tubular collagen nerve sleeves. Twenty Sprague‐Dawley rats underwent unilateral sharp sciatic nerve transection and microscopic nerve repair with four epineural sutures and were randomly treated with or without an encircling collagen nerve sleeve. Normal nerves from the contralateral sciatic nerve were also examined. At sacrifice three months later, the nerves were evaluated for traumatic neuroma formation, perineural scar formation, and morphometric analysis. Histological examination of normal and repaired nerves by a neuropathologist demonstrated healing, minimal Wallerian degeneration and no traumatic neuroma formation. Distal section analysis (nine nonwrapped, 10 wrapped), revealed no significant differences in total fascicular area, myelinated fibers per nerve, fiber density, myelin area per nerve, myelinated fiber diameter, axon diameter, myelin thickness, or G‐ratio. Significantly greater (P = 0.005) inner epineural connective tissue formation was observed in nonwrapped nerves (0.62 mm2 ± 0.2) versus wrapped nerves (0.35 mm2 ± 0.16). The ratio of connective tissue to fascicular area was larger in nonwrapped (1.08 ± 0.26) versus wrapped nerves (0.63 ± 0.22) (P < 0.001). This study demonstrated decreased inner epineural connective tissue formation with use of a collagen nerve wrap during primary repair of peripheral nerve transection in a rat sciatic nerve model.

Effect of white adipose tissue flap and insulin‐like growth factor‐1 on nerve regeneration in rats

Adipose tissue‐derived stem cells and insulin‐like growth factor‐1 (IGF‐1) have shown potential to enhance peripheral nerve regeneration. The purpose of this study was to investigate the effect of an in vivo biologic scaffold, consisting of white adipose tissue flap (WATF) and/or IGF‐1 on nerve regeneration in a crush injury model. Forty rats all underwent a sciatic nerve crush injury and then received: a pedicled WATF, a controlled local release of IGF‐1, both treatments, or no treatment at the injury site. Outcomes were the normalized maximum isometric tetanic force (ITF) of the tibialis anterior muscle and histomorphometric measurements. At 4 weeks, groups with WATF had a statistically significant improvement in maximum ITF recovery, as compared to those without (P < 0.05), and there was an increase in myelin thickness and total axon count in the WATF‐only group versus control (P < 0.01). Functional and histomorphometric data suggest that IGF‐1 suppressed the effect of the WATF. Use of a pedicled WATF improved the functional and histomorphometrical results after axonotmesis in a rat model. IGF‐1 does not appear to enhance nerve regeneration in this model. Utilizing the WATF may have a beneficial therapeutic role in peripheral nerve injuries.

Effect of Microvascular Anastomosis Technique on End Product Outcome in Simulated Training: A Prospective Blinded Randomized Controlled Trial.

Background The aim of this article is to evaluate the difference in skills acquisition of two end-to-end microvascular anastomosis techniques-the triangulation and biangulation-in early microsurgery training. Method In this study, 32 candidates ranging from medical students to higher surgical trainees underwent a 5-day basic microsurgery course. On days 3 and 5 of the course, candidates performed two end-to-end anastomoses on cryopreserved rat aortas. One anastomosis was performed using the biangulation technique and the other using the triangulation technique. Candidates were randomized to the order of technique performed. Structural patency, errors performed, and suture distribution were evaluated randomly by a blinded reviewer using the anastomosis lapse index score and ImageJ (U.S. National Institutes of Health, Bethesda, MD) Software. Results A total of 128 anastomoses were evaluated during the study period. A total of six anastomoses performed with the biangulation technique, and four anastomoses with the triangulation technique, were physically occluded on day 3 of the course. On day 5, two biangulation technique anastomoses and one triangulation technique produced a nonpatent outcome. There was a statistically significant difference of patency rate between the 2 days of evaluation confirming evidence of skill acquisition but no statistically significant difference between the two techniques in relation to anastomotic patency, errors performed, or suture placement quality. Conclusion The biangulation and triangulation techniques of microvascular anastomosis produce similar outcomes in relation to vessel structural patency and quality of anastomosis when taught in early stages of microsurgery training. Our results suggest that both techniques are equally suitable in training novices, basic microsurgical skills.

Topical cilostazol inhibits neointimal hyperplasia in a rat interposition vein graft model.

Background: Neointimal hyperplasia is a common cause of vein graft failure resulting from luminal narrowing and occlusion. Cilostazol is a U.S. Food and Drug Administration–approved phosphodiesterase III and platelet aggregation inhibitor commonly used in peripheral vascular disease. The authors studied whether topical cilostazol treatment at the time of vein grafting helps limit the development of neointimal hyperplasia in a rat model. Methods: Six experimental rats and six control rats underwent interposition vein grafting into the femoral artery, followed by a single topical dose of cilostazol applied around the vein graft in the experimental animals. After 4 weeks, grafts were harvested and underwent histologic staining of axial sections to visualize intima thickness and elastin/myocyte content. Quantification was performed to assess total intima area. The intima-to-media and the intima-to–sum of intima and media ratios were determined to control for discrepancies in overall graft size. Results: Cilostazol-treated grafts had a thinner intima layer with less myocyte content compared with control grafts, amounting to an 82 percent decrease in total intima area compared with controls. A similar trend was seen even after controlling for overall graft size, with 85 and 76 percent reductions seen in intima-to-media and intima-to–sum of intima and media ratios, respectively. Conclusions: A single intraoperative dose of cilostazol applied locally significantly reduced intima size and smooth muscle content in rat interposition vein grafts examined 4 weeks postoperatively. A topical dose of cilostazol at surgery may therefore be helpful in controlling neointimal hyperplasia and reducing the need for systemic medications to prolong vein graft patency.

Endogenous adipose tissue as a hemostatic: Use in microsurgery

Bleeding is a frequent complication of microsurgical repair of small blood vessels and time is spent while hemostasis is accomplished. We studied the hemostatic effect of endogenous adipose tissue on bleeding from rat femoral arterial anastomoses. We measured bleeding time (time from removal of clamps to cessation of active bleeding) and mean arterial blood velocity (using a micro‐Doppler system), the latter immediately after anastomosis, and again 7 days post‐anastomosis. Bleeding time for vessels with fat applied to the artery was 50% less than when no fat was applied. Blood velocity by day 7 post‐anastomosis returned to values equivalent to those for intact arteries. Histological evaluation of the anastomotic site demonstrated no significant differences in inflammatory response between fat‐treated and untreated arteries. These data suggest that endogenous adipose tissue may be a useful hemostatic agent devoid of significant effects on small artery blood velocity or histology.

Ipsilateral, cabled sural nerve for a sciatic nerve defect: An experimental model in the rat

The 10 mm rat sciatic nerve defect model is commonly used to investigate new strategies to improve functional recovery with segmental nerve defects. However, a lack of standardization makes comparisons between studies difficult. The present study aims to evaluate a standardized experimental model that can minimize the number of animals required for obtaining valid results and simulates a current treatment for human peripheral nerve injury defects. Eighteen cadaveric Sprague-Dawley rats were utilized in the anatomic arm of the study and 18 living Sprague-Dawley rats were used in the experimental arm. The results from the cadaveric study allowed us to create an ipsilateral, three-cable autologous sural nerve graft technique in the rat. This repair construct was evaluated with functional and histomorphometric analysis of nerve regeneration. The results support functional recovery of the sciatic nerve in all grafted animals. The use of an ipsilateral cabled sural nerve graft technique in the rat sciatic nerve defect model is a viable control group that utilizes a single incision, incurs minimal morbidity, and maintains muscle attachments. We conclude that this rat model can be used in various experimental trials in the field of peripheral nerve regeneration.

Predicting Thrombosis Formation in 1-mm-Diameter Arterial Anastomoses with Transit-Time Ultrasound Technology

Background: Thrombosis is a common source of failure in anastomoses, flaps, and vascular grafts. Ensuring vessel patency is critical to the success of microvascular procedures. Any tool that can accurately predict the patency of an anastomosis intraoperatively would enable the surgeon to detect and correct flow restrictions while the patient is still in the operating room. Methods: The authors used transit-time ultrasound technology in this pilot study to investigate whether a minimal cutoff value for quantitative postoperative blood flow (in milliliters per minute) could be established that would reliably predict sustained vessel patency at 24 hours postoperatively. Surgical end-to-end anastomoses were performed on 56 Sprague-Dawley rat femoral arteries with diameters ranging from 0.6 to 1.2 mm. Postoperative blood volume flow measurements were taken at 20-minute intervals up to 1 hour, and then again at 24 hours, to assess patency. Results: Forty-seven anastomoses (83.9 percent ) were patent 24 hours after surgery. Nine anastomoses (16.1 percent ) thrombosed within 24 hours. Based on a receiver operating characteristic curve analysis, the optimal cutoff value for immediate postoperative flow for predicting thrombosis within 24 hours of microvascular anastomosis is 0.21 ml/minute. Conclusions: At 20 minutes postoperatively, blood flows greater than 0.30 ml/minute are highly suggestive of patency, and flows less than 0.21 ml/minute are highly suggestive of failure. The authors therefore recommend a minimal cutoff flow value of 0.30 ml/minute for vessels ranging from 0.6 to 1.2 mm in diameter to predict long-term postoperative vascular patency.

Defining Standards in Experimental Microsurgical Training: Recommendations of the European Society for Surgical Research (ESSR) and the International Society for Experimental Microsurgery (ISEM)

Background: Expectations towards surgeons in modern surgical practice are extremely high with minimal complication rates and maximal patient safety as paramount objectives. Both of these aims are highly dependent on individual technical skills that require sustained, focused, and efficient training outside the clinical environment. At the same time, there is an increasing moral and ethical pressure to reduce the use of animals in research and training, which has fundamentally changed the practice of microsurgical training and research. Various animal models were introduced and widely used during the mid-20th century, the pioneering era of experimental microsurgery. Since then, high numbers of ex vivo training concepts and quality control measures have been proposed, all aiming to reduce the number of animals without compromising quality and outcome of training. Summary: Numerous microsurgical training courses are available worldwide, but there is no general agreement concerning the standardization of microsurgical training. The major aim of this literature review and recommendation is to give an overview of various aspects of microsurgical training. We introduce here the findings of a previous survey-based analysis of microsurgical courses within our network. Basic principles behind microsurgical training (3Rs, good laboratory practice, 3Cs), considerations around various microsurgical training models, as well as several skill assessment tools are discussed. Recommendations are formulated following intense discussions within the European Society for Surgical Research (ESSR) and the International Society for Experimental Microsurgery (ISEM), based on scientific literature as well as on several decades of experience in the field of experimental (micro)surgery and preclinical research, represented by the contributing authors. Key Messages: Although ex vivo models are crucial for the replacement and reduction of live animal use, living animals are still indispensable at every level of training which aims at more than just a basic introduction to microsurgical techniques. Modern, competency-based microsurgical training is multi-level, implementing different objective assessment tools as outcome measures. A clear consensus on fundamental principles of microsurgical training and more active international collaboration for the sake of standardization are urgently needed.