Electrical stimulation shifts healing/scarring towards regeneration in a rat limb amputation model.

Different species respond differently to severe injury, such as limb loss. In species that regenerate, limb loss is met with complete restoration of the limbs' form and function, whereas in mammals the amputated limb's stump heals and scars. In in vitro studies, electrical stimulation (EStim) has been shown to promote cell migration, and osteo- and chondrogenesis. In in vivo studies, after limb amputation, EStim causes significant new bone, cartilage and vessel growth. Here, in a rat model, the stumps of amputated rat limbs were exposed to EStim, and we measured extracellular matrix (ECM) deposition, macrophage distribution, cell proliferation and gene expression changes at early (3 and 7 days) and later stages (28 days). We found that EStim caused differences in ECM deposition, with less condensed collagen fibrils, and modified macrophage response by changing M1 to M2 macrophage ratio. The number of proliferating cells was increased in EStim treated stumps 7 days after amputation, and transcriptome data strongly supported our histological findings, with activated gene pathways known to play key roles in embryonic development and regeneration. In conclusion, our findings support the hypothesis that EStim shifts injury response from healing/scarring towards regeneration. A better understanding of if and how EStim controls these changes, could lead to strategies that replace scarring with regeneration.

Zebrafish fin regeneration involves transient serotonin synthesis.

The zebrafish is a vertebrate organism capable of regenerating many of its organs. Notably, it can undergo epimorphic regeneration of its fins after amputation. This process occurs through the formation of a wound epithelium and the dedifferentiation of mesenchymal and bone-forming cells, which form a proliferative blastema. Here, we report that the entry into the regenerative process involves the local synthesis of serotonin (5-hydroxytryptamine, 5-HT) in the injury-associated tissue. One day after wounding, intracellular accumulation of serotonin was induced in the stump below the amputation plane. During blastema formation, serotonin was detected in the mesenchyme at the vicinity of the amputation plane and in the apical wound epithelium. During the advanced outgrowth phase, this monoamine was no longer present in the blastema, suggesting a temporal involvement of serotonin in the postinjury area. We show the expression of two serotonin synthesizing enzymes, tryptophan hydroxylase 1a and 1b in the blastema, suggesting the local production of this monoamine. Neither depletion of serotonin by chemical inhibition of tryptophan hydroxylase, nor ectopic administration of this monoamine affected fin regeneration, indicating it does not play a role during this process. Finally, we found that the presence of serotonin during regeneration depends on fibroblast growth factor and retinoic acid signaling. Overall, our study demonstrates that the initiation of fin regeneration is associated with a transient synthesis of serotonin in the regrowing tissue.

Osteomyelitis in limb amputated by amniotic band sequence.

A preterm (30+2 week) neonate with below-knee amputation (right lower limb), constriction rings and syndactyly, subsequent to amniotic band sequence, developed pus discharge from the right tibial stump. The neonate did not have clinical features of systemic sepsis. Blood culture was sterile. The pus culture, however, grew methicillin-resistant coagulase-negative Staphylococcus and bone scan was suggestive of osteomyelitis of right proximal tibial stump. Osteomyelitis was likely caused by the contiguous spread of infection from the exposed stump. Neonate was treated with intravenous antibiotics for 4 weeks and discharged on oral feeds.

[Five-year survival and outcomes of chronic hip stump ischemia treatment]. / Piatiletniaia vyzhivaemost' i rezul'taty lecheniia khronicheskoi ishemii kul'ti bedra.

AIM: To investigate five-year survival, short- and long-term outcomes of hip stump ischemia treatment. MATERIAL AND METHODS: There were 383 patients with hip stump in 1997-2013. Critical hip stump ischemia in 3 months after amputation occurred in 45 patients. RESULTS: Two-fold decrease of 5-year survival was found. Five-year survival was higher 3 times in patients after arterial reconstruction compared with those without revascularization. The same tendency was confirmed in analysis of groups standardized by gender, age and type of arterial disease.

[Ischaemia of a femoral stump in short- and long-term periods after limb amputation]. / Ishemiia kul'ti bedra v rannem i pozdnem periodakh posle amputatsii.

The authors retrospectively analysed medical case histories of 287 patients subjected to femoral amputations over the period from January 1, 1998 to December 31, 2013. The studied parameters were as follows: the frequency of and risk factors for femoral stump ischaemia, as well as the effect on patients' survival after femoral amputation. Amongst 156 patients having endured transfemoral truncation of the limb performed as the first amputation, early femoral stump ischaemia (EFSI) within 3 postoperative months was found to have occurred in 43 (27.6%) patients, whereas amongst 127 patients first subjected to amputation of the crus and then to femoral truncation it occurred in 15 (13.2%) cases; p<0.05. The incidence rate of late femoral stump ischaemia (LFSI) was virtually similar in both groups, amounting to 5.8% (9 of 156) and 5.5% (7 of 127), respectively; p>0.05. The survival rate for patients without stump ischaemia at 12 months after amputation amounted to 79.4%, for those with EFSI to 50.0% (p=0.00928), and for those with LFSI to 71.4% (p=0.22576), whereas by the end of a 5-year follow up period these values appeared to equal 49.2%, 32.1% (p=0.13225) and 7.1% (p=0.01385), respectively. The obtained findings demonstrated that the risk factors for EFSI were as follows: the presence of a femoral stump on the contralateral side, grade III ischaemia, and proximal localization of the lesion of the arterial bed (odds ratio 3.3, 2.7 and 3.8, respectively); a risk factor for LFSI was the presence of a femoral stump on the contralateral side (odds ratio 6.0).

[Critical ischemia of hip stump. Incidence, clinical significance and diagnosis]. / Kriticheskaia ishemiia kul'ti bedra: chastota, klinicheskoe znachenie i diagnostika.

AIM: To study the incidence of hip stump ischemia, risk factors for 5-year survival after amputation, clinical features and diagnosis of this complication. MATERIAL AND METHODS: 1287 patients with peripheral artery disease followed by hip amputation were examined within 1997-2015. 50 of them had stump ischemia. RESULTS: It was found that hip stump ischemia occurs in 5.2% of cases and significantly aggravates 5-year survival. Transcutaneous oxygen tension less than 20 mm Hg is reliable hemodynamic criterion of ischemia.

Amputation stump perfusion is predictive of post-operative necrotic eschar formation.

BACKGROUND: A large proportion of patients develop poor amputation stump healing. We hypothesize that Laser-Assisted Fluorescent Angiography (LAFA) can predict inadequate tissue perfusion and healing. METHODS: Over an 8-month period we reviewed all patients who underwent lower extremity amputation and LAFA. We evaluated intra-operative LAFA global and segmental stump perfusion, and post-operative modified Bates-Jensen (mBJS) wound healing scores. RESULTS: In 15 patients, amputation stumps with lower global perfusion demonstrated higher mBJS (P = 0.01). Lower suture-line perfusion also correlated with more eschar formation (P < 0.001). Diabetic patients had higher mBJS (P = 0.009), lower stump perfusion (P = 0.02), and increased eschar volume (P < 0.001). CONCLUSION: LAFA is a useful adjunct for intra-operative stump perfusion assessment and can predict areas of poor stump healing and eschar formation. Diabetic patients seem to be at higher risk of stump eschar formation.

Femoral Artery Bypass Graft Pseudoaneurysm Rupture in a Transfemoral Amputee.

The case of a patient with an actively bleeding pseudoaneurysm associated with remnants of a polytetrafluoroethylene femoral bypass graft in his transfemoral residual limb is described. Initial graft placement was due to peripheral arterial disease. During subsequent transfemoral amputation, remnants of the nonpatent graft were retained in the residuum. After 4 yrs of lower limb prosthesis use, a proximal anastomosis pseudoaneurysm developed (with avulsion of graft remnants). The patient presented to clinic with a 5-day history of increased left groin fullness and largely nonradiating pain (rated 10/10). He was diagnosed with a pseudoaneurysm (1.9 cm) originating from the left common femoral artery and an associated hematoma (8 cm) on computed tomography; this required emergent reoperation. This case highlights the importance surrounding the decision to leave or explant neovascularization materials, which may carry significant risk for infection or physical disruption complications in residual limbs.

Intra-arterial fibrinolysis for the management of acute ischemia on a below-knee amputation stump. Case report.

Preservation of the knee joint has enormous advantages in terms of mobility and rehabilitation of an amputee. Any cause of breakdown requiring revision to an above-knee amputation is a major setback because it reduces the patient's rehabilitative potential. We report a case of intra-arterial thrombolysis use to save a below-knee amputation stump with acute ischemia. A 56-year-old man who sought the emergency department with 1-day history of acute pain on his right below-knee stump. The angiography confirmed popliteal artery occlusion. Pharmacomechanical thrombectomy, with Aspirex (rotational catheter to restore blood flow in occluded vessel, by removing occlusion material from the vessel) and recombinant tissue plasminogen activator, was performed. After 9 years of follow-up the patient remained asymptomatic, capable of independent ambulation with prosthetic limb. Intra-arterial fibrinolysis seems to be a safe and effective treatment for cases of acutely ischemic amputation stump.

Intra-arterial fibrinolysis for the management of acute ischemia on a below-knee amputation stump. Case report / Fibrinólise intra-arterial para tratamento da isquemia aguda em coto de amputação infrapatelar. Relato de caso

ABSTRACT Preservation of the knee joint has enormous advantages in terms of mobility and rehabilitation of an amputee. Any cause of breakdown requiring revision to an above-knee amputation is a major setback because it reduces the patient's rehabilitative potential. We report a case of intra-arterial thrombolysis use to save a below-knee amputation stump with acute ischemia. A 56-year-old man who sought the emergency department with 1-day history of acute pain on his right below-knee stump. The angiography confirmed popliteal artery occlusion. Pharmacomechanical thrombectomy, with Aspirex (rotational catheter to restore blood flow in occluded vessel, by removing occlusion material from the vessel) and recombinant tissue plasminogen activator, was performed. After 9 years of follow-up the patient remained asymptomatic, capable of independent ambulation with prosthetic limb. Intra-arterial fibrinolysis seems to be a safe and effective treatment for cases of acutely ischemic amputation stump.

RESUMO A preservação da articulação do joelho tem grandes vantagens para a mobilidade e a reabilitação de um amputado. Qualquer causa que exija revisão para uma amputação acima do joelho é um grande revés, porque reduz o potencial de reabilitação do paciente. O objetivo aqui foi descrever o uso de trombólise intra-arterial para salvar um coto de amputação abaixo do joelho com isquemia aguda. Homem, 56 anos, procurou pronto atendimento de nosso hospital com histórico de 1 dia de dor aguda em seu coto de amputação infrapatelar direito. A angiografia confirmou oclusão da artéria poplítea. Foi realizada trombectomia farmacomecânica com Aspirex (cateter rotativo para restabelecer o fluxo sanguíneo em vasos ocluídos, removendo material de oclusão do vaso) e ativador do plaminogênio tecidual recombinante. Após 9 anos de seguimento, o paciente permanecia assintomático, capaz de deambulação independente com membro protético. A fibrinólise intra-arterial parece ser um tratamento seguro e eficaz para casos selecionados de coto de amputação com isquêmica aguda.

Effect of C1-INH on ischemia/reperfusion injury in a porcine limb ex vivo perfusion model.

Revascularization of an amputated limb within 4-6h is essential to avoid extensive ischemia/reperfusion (I/R) injury leading to vascular leakage, edema and tissue necrosis. I/R injury is a pathological inflammatory condition that occurs during reperfusion of an organ or tissue after prolonged ischemia. It is characterized by a complex crosstalk between endothelial cell activation and the activation of plasma cascades. Vasculoprotective pharmacological intervention to prevent I/R injury might be an option to prolong the time window between limb amputation and successful replantation. We used C1-easterase inhibitor (C1-INH) in this study because of its known inhibitory effects on the activation of the complement, coagulation and kinin cascades. Forelimbs of 8 large white pigs were amputated, subjected to ischemia, and then reperfused with autologous whole blood. All limbs were exposed to 9h of cold ischemia at 4°C. After 2h of cold ischemia the limbs were either perfused with of C1-INH (1U/ml in hydroxyethyl starch, n=8) or hydroxyethyl starch alone (n=7). After completion of the 9-h ischemia period, all limbs were ex vivo perfused with heparinized autologous whole blood for 12h using a pediatric heart lung machine to simulate in vivo revascularization. Our results show that I/R injury in the control group led to a significant elevation of tissue deposition of IgG and IgM, complement C3b/c, C5b-9 and MBL. Also, activation of the kinin system was significantly increased, namely bradykinin in plasma, and expression of bradykinin receptors 1 and 2 in tissue. In addition, markers for endothelial integrity like expression of CD31, VE-cadherin and heparan sulfate proteoglycans were decreased in reperfused tissue. Limb I/R injury also led to activation of the coagulation cascade with a significant elevation of fibrin and thrombin deposition and increased fibrinogen-like protein-2 expression. C1-INH treated limbs showed much less activation of plasma cascades and better protection of endothelial integrity compared to the reperfused control limbs. In conclusion, the use of the cytoprotective drug C1-INH significantly reduced I/R injury by protecting the vascular endothelium as well as the muscle tissue from deposition of immunoglobulins, complement and fibrin.

Microcirculatory Assessment of Arterial Below-Knee Stumps: Near-Infrared Spectroscopy Versus Transcutaneous Oxygen Tension-A Preliminary Study in Prosthesis Users.

OBJECTIVE: To examine metrologic properties of near-infrared spectroscopy (NIRS) versus transcutaneous oxygen tension (TcPO2) for microcirculatory assessment of vascular transtibial stumps at the stabilized period of prosthesis fitting, as a preliminary step before exploring its ability to predict stump healing, considering the previously identified limits of TcPO2 (borderline area between 15 and 35mmHg). DESIGN: Prospective single-center observational study. SETTING: University-based rehabilitation center. PARTICIPANTS: Individuals with unilateral transtibial amputation for peripheral artery disease, at the definitive stage of prosthesis fitting, able to perform a 2-minute walk test (N=30). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Test-retest, with the stump being evaluated in supine and inclined positions, first by NIRS (tissue saturation index [TSI], oxyhemoglobin, deoxyhemoglobin, and total hemoglobin) and second by TcPO2. Subjects carried out a 2-minute walk test and visual analog scales (wound healing and pain). RESULTS: Feasibility and tolerance of NIRS were satisfactory. The reliability of NIRS and TcPO2 values was good (intraclass correlation coefficient >0.7; P<.05). No significant relation was found between NIRS and TcPO2. No responsiveness (inclined vs supine) was reported (P>.05). A significant relation between TSI and the 2-minute walk test (r>.49, P<.05) was found. CONCLUSIONS: NIRS is painless, complication-free, and feasible, with good reliability. NIRS evaluates others domain than TcPO2 that are more linked to metabolic adaptation. Its capacity to predict stump healing and tolerance to early prosthesis fitting is therefore interesting to estimate in future studies.

Amputation Stump Revision Using a Free Sural Neurocutaneous Perforator Flap.

BACKGROUND: The development of microsurgery and the concept of the surgical flap have enabled orthopedic surgeons to deal with various soft tissue defects and compositions during limb salvage procedures. However, severely damaged bony structures with accompanying soft tissue deficiencies often result in amputation. When performing amputations, surgeons must preserve the stump length to maximize functional ambulation, even during the posttraumatic phase and chronic period when a fillet flap is not an option. PATIENTS AND METHODS: Eleven patients with previous below-knee amputation were enrolled in our study from January 2006 to December 2011. Each patient underwent stump revision using a free sural neurocutaneous perforator flap. Preoperative and postoperative data were reviewed to evaluate the functional and aesthetic results. RESULTS: The 11 patients comprised 7 men and 4 women with an average age of 35.6 years (range, 26.0-49.0 years). The mean follow-up period was 15.3 months (range, 10.0 months-2.0 years). All free flap reconstructions of the amputation stump ultimately survived. All patients achieved both superficial and deep sensory recovery and are comfortable with their prostheses, and none have required further surgical intervention. CONCLUSIONS: The sural neurocutaneous perforator flap provided stable coverage and improved patients' quality of life by aiding in early rehabilitation and prosthetic fitting, function, and comfort.

[Iliac artery occlusion balloons for suspected placenta accreta during cesarean section]. / Inserción de balones intraarteriales ilíacos previa a la realización de cesárea ante la sospecha de acretismo placentario.

Massive obstetric hemorrhage still remains a major cause of maternal mortality and morbidity. The risk factors associated with this pathology must be identified in order to schedule the appropriate delivery with the necessary resources. A case is presented of an iliac artery occlusion with intravascular balloons for suspected placenta accreta during cesarean section. The perioperative treatment, as well as an analysis of the treatment options is described, along with their advantages and disadvantages, from the use of postpartum hemorrhage protocols, blood transfusion and procoagulant factors, and other maneuvers to control bleeding, until the hysterectomy.

[Stump problems after a lower limb amputation]. / Stompklachten na een beenamputatie.

Problems in the amputation stump occur frequently in lower limb amputees. These problems can range from skin defects to serious vascular insufficiency and have a major impact on the amputee's quality of life. We illustrate this with 2 patients aged 20 and 65. The first patient developed a Candida skin infection of the stump skin as a result of silicone liner use. She was successfully treated with miconazole/hydrocortisone and hygiene measures. The second patient developed progressive lower limb ischemia that resulted in a stump wound and claudication. He was treated with vascular surgery. Patients with stump problems should be referred to a rehabilitation physician for thorough examination of the stump in determining the cause of the symptoms and subsequent instalment of adequate treatment.

Acroangiodermatitis in a lower leg amputee related to suction socket use.

Acroangiodermatitis is a rare condition with numerous causes typically presenting as purple macules on the lower extremities. Although benign, it can mimic the presentation of more serious underlying conditions such as Kaposi's sarcoma. We present a case of acroangiodermatitis in the stump of an amputee related to suction socket use in order to raise awareness of an unusual setting for this vascular proliferation.

Latissimus dorsi flap coverage of soft tissue defect following below-knee amputation: emphasis on flap design and recipient vessels.

High-energy trauma to the lower extremity often results in amputation of the limb. For maximal preservation of limb length during amputation, free tissue transfer is often necessary. In this study, we report our experience of stump coverage using latissimus dorsi musculocutaneous flaps with an emphasis on flap design and recipient vessels. Between January 2005 and September 2010, twelve patients with severe traumatic injuries to the lower leg underwent below-knee amputations with stump coverage using latissimus dorsi free flaps. The primary and secondary cases were approached differently regarding the flap design and recipient vessels. All flaps survived completely. There were 8 primary cases and 4 secondary cases. In the primary cases, the anterior tibial artery was used as the recipient vessel in 6 cases, and in 2 cases, the descending geniculate artery was used. In the secondary cases, the descending geniculate artery was used in all cases. There were two cases of ulceration on the grafted non-weight-bearing site, but after the usage of collagen-elastin artificial dermis, no ulcerations were seen. The latissimus dorsi musculocutaneous flap is the most feasible option for coverage of amputation stumps. In flap design, the width of the skin paddle must match the anteroposterior diameter of the defect at the stump. The latissimus dorsi muscle must sufficiently wrap the bony stump for padding. We recommend using the anterior tibial artery as a recipient vessel in primary cases, and the descending geniculate artery in secondary cases.