Prolongation of the Time Window From Traumatic Limb Amputation to Replantation From 6 to 33 Hours Using Ex Vivo Limb Perfusion.

INTRODUCTION: Continuous extracorporeal perfusion (ECP), or machine perfusion, holds promise for prolonged skeletal muscle preservation in limb ischemia-reperfusion injury. This study aimed to extend the amputation-to-replantation time window from currently 6 hours to 33 hours using a 24-hour ECP approach. MATERIALS AND METHODS: Six large white pigs underwent surgical forelimb amputation under general anesthesia. After amputation, limbs were kept for 9 hours at room temperature and then perfused by 24-hour ECP with a modified histidine-tryptophan-ketoglutarate (HTK) solution. After ECP, limbs were orthotopically replanted and perfused in vivo for 12 hours. Clinical data, blood, and tissue samples were collected and analyzed. RESULTS: All 6 forelimbs could be successfully replanted and in vivo reperfused for 12 hours after 9 hours of room temperature ischemia followed by 24 hours ECP. Adequate limb perfusion was observed after replantation as shown by thermography and laser Doppler imaging. All pigs survived without severe organ failure, and no significant increase in inflammatory cytokines was found. Macroscopy and histology showed marked interstitial muscular edema of the limbs, whereas myofiber necrosis was not evident, implying the preservation of muscular integrity. CONCLUSIONS: The use of a 24-hour ECP has successfully extended limb preservation to 33 hours. The modified histidine-tryptophan-ketoglutarate perfusate demonstrated its ability for muscle protection. This innovative approach not only facilitates limb replantation after combat injuries, surmounting geographical barriers, but also broadens the prospects for well-matched limb allotransplants across countries and continents.

Carpal instability after partial trapeziectomy, total trapeziectomy and the resection of the distal scaphoid pole: a cadaveric study.

INTRODUCTION: Non-dissociative carpal instability (CIND) may lead to severe functional impairment. Destabilisation of the scapho-trapezial-trapezoidal (STT) ligament complex seems to result in a CIND. MATERIALS AND METHODS: In one group with eight cadaver arms, distal scaphoid pole was resected with the adjacent ligaments. In the other eight cadavers, hemitrapeziectomy was performed followed by total trapeziectomy. CT scans were performed in different wrist positions, and the changed positions of the scaphoid, lunate and capitate were measured in comparison to non-operated wrists. RESULTS: Mainly in clenched fist position, dissociation between proximal and distal row can be determined after total trapeziectomy and resection of distal scaphoid pole. Capitate rotates dorsally up to 24°, the scaphoid up to 17° and the lunate up to 7° compared to the non-operated wrists. Resection of the distal scaphoid pole results in dorsal rotation of capitate and scaphoid of about 14° and the lunate 8°. Relative scapholunate and capitolunate angle increased significantly after total trapeziectomy, especially in clenched fist position. After scaphoid pole resection, significant SL and CL angles changes could be seen in almost every wrist position. CONCLUSION: Destabilisation of the STT ligament complex by total trapeziectomy or distal scaphoid pole resection results in dissociation of the proximal and distal carpal row without instability within the proximal or distal row, corresponding to a CIND. LEVEL OF EVIDENCE: III.

Radio-luno-triquetral bone-ligament transfer as an additional stabilizer in scapholunate-instability.

INTRODUCTION: Reconstruction of the scapho-lunate (SL) ligament is still challenging. Many different techniques, such as capsulodesis, tendon graft and bone-ligament-bone graft have been described to stabilize reducible SL dissociation. If primary ligament repair alone is not possible, an additional stabilizer is needed to achieve scapho-lunate stability. A new local bone-ligament transfer using half of the radio-luno-triquetral ligament is performed. The direction of traction of the transposed ligament is very similar to the original ligament. Ideal tension can be attained by fixation of the bone block at the dorsal ridge of the scaphoid. The biomechanical stability of this bone-ligament transfer shall be examined biomechanically. MATERIAL AND METHODS: Computed tomography imaging was performed using eight cadaveric forearms with a defined position of the wrist. Axial load was accomplished with tension springs attached to the extensor and flexor tendons. Three series ([a] native, [b] divided SL ligament and [c]) after reconstruction with bone-ligament transfer] were reconstructed three-dimensionally to determine the angles between radius, scaphoid and lunate. The radial distal part including a bone fragment of the radio-luno-triquetral ligament was transferred from its insertion at the distal edge of the radius to be attached to the dorsal ridge of the scaphoid. RESULTS: SL gap was widened after its transection. Average SL distance was 6.6 ± 1.6 mm. After ligament reconstruction, the gap could be narrowed significantly to 4.2 mm (± 0.7 mm). The movement of the scaphoid and lunate showed significant changes, especially in wrist flexion, fist closure and radial deviation. These deviations could be corrected by the bone ligament transfer. CONCLUSION: Reconstruction of a transected SL ligament with a bone-ligament transfer from the radio-luno-triquetral ligament reduces SL dissociation under axial load. The described surgical technique causes low donor-side morbidity and can be considered in addition to improve stability if SL ligament suture alone does not appear sufficient. LEVEL OF EVIDENCE: Level II, therapeutic investigating experimental study.

Local Injections of Tacrolimus-loaded Hydrogel Reduce Systemic Immunosuppression-related Toxicity in Vascularized Composite Allotransplantation.

BACKGROUND: Routine application of vascularized composite allotransplantation is hampered by immunosuppression-related health comorbidities. To mitigate these, we developed an inflammation-responsive hydrogel for local immunosuppression. Here, we report on its long-term effect on graft survival, immunological, and toxicological impact. METHODS: Brown Norway-to-Lewis rat hindlimb transplantations were treated either systemically with daily injections of 1 mg/kg tacrolimus (TAC) or with subcutaneous intragraft injections of hydrogel containing 7 mg TAC, every 70 days. Animals were monitored for rejection or other pathology for 280 days. Systemic and graft TAC levels, regulatory T cells, and donor cell chimerism were measured periodically. At endpoint, markers for kidney, liver, and metabolic state were compared to naive age-matched rats. RESULTS: Both daily systemic TAC and subcutaneous intragraft TAC hydrogel at 70-day intervals were able to sustain graft survival longer than 280 days in 5 of 6 recipients. In the hydrogel group, 1 graft progressed to grade 3 rejection at postoperative day 149. In systemic TAC group, 1 animal was euthanized due to lymphoma on postoperative day 275. Hydrogel treatment provided stable graft and reduced systemic TAC levels, and a 4 times smaller total TAC dose compared with systemic immunosuppression. Hydrogel-treated animals showed preserved kidney function, absence of malignancies or opportunistic infections and increased hematopoietic chimerism compared with systemic immunosuppression. CONCLUSIONS: Our findings demonstrate that localized immunosuppression with TAC hydrogel is a long-term safe and reliable treatment. It may reduce the burden of systemic immunosuppression in vascularized composite allotransplantation, potentially boosting the clinical application of this surgical intervention.

Laser doppler imaging as additional monitoring after digital replanting: A prospective study.

BACKGROUND: Despite various exisiting monitoring methods, there is still a need for new technologies to improve the quality of post-operative evaluation of digital replantation. The purpose of the study is using a laser Doppler imaging device (Easy-LDI) as an additional tool to assess perfusion. In this method, the changes in the frequency of the laser ligth provide information regarding perfusion of the monitored tissue. PATIENTS AND METHODS: This study included seven patients (10 fingers; age of patients: 21-57 years) who suffered from a total (n = 6) or subtotal amputation (n = 4) due to accidents. In addition to hourly standard monitoring with clinical evaluation and skin thermometry, revascularized fingers were hourly monitored with Easy LDI for 48 h. RESULTS: LDI measurement values ranged between 0.8 and 223 (mean 90.62 ± 21.42) arbitrary perfusion units (APU). The mean LDI values before and after revascularization were 7.1 ± 2.85 and 65.30 ± 30.83 APU, respectively. For the successful revascularized fingers (8 of 10 fingers) values from 19 to 223 APU (mean 98.52 ± 15.48) were demonstrated. All of the replants survived, but due to venous occlusion two digits required revision 12 and 35 h after revascularization, respectively. In the two cases, Easy-LDI also showed a constant and slow decline of the perfusion values. Furthermore, Pearson normalized correlation coefficient showed a positive significant correlation between temperatures of the replants and LDI-values (P < .001, r = +0.392) and a negative significant correlation between Δtemperature and LDI-values (P < .001, r = -0.474). CONCLUSION: The LDI-device might be a promising additional monitoring technique in detection of perfusion disturbance in monitoring digital replantations.

Release of pig leukocytes and reduced human NK cell recruitment during ex vivo perfusion of HLA-E/human CD46 double-transgenic pig limbs with human blood.

BACKGROUND: In pig-to-human xenotransplantation, interactions between human natural killer (NK) cells and porcine endothelial cells (pEC) are characterized by recruitment and cytotoxicity. Protection from xenogeneic NK cytotoxicity can be achieved in vitro by the expression of the non-classical human leukocyte antigen-E (HLA-E) on pEC. Thus, the aim of this study was to analyze NK cell responses to vascularized xenografts using an ex vivo perfusion system of pig limbs with human blood. METHODS: Six pig forelimbs per group, respectively, stemming from either wild-type (wt) or HLA-E/hCD46 double-transgenic (tg) animals, were perfused ex vivo with heparinized human blood for 12 hours. Blood samples were collected at defined time intervals, cell numbers counted, and peripheral blood mononuclear cells analyzed for phenotype by flow cytometry. Muscle biopsies were analyzed for NK cell infiltration. In vitro NK cytotoxicity assays were performed using pEC derived from wt and tg animals as target cells. RESULTS: Ex vivo, a strong reduction in circulating human CD45 leukocytes was observed after 60 minutes of xenoperfusion in both wt and tg limb groups. NK cell numbers dropped significantly. Within the first 10 minutes, the decrease in NK cells was more significant in the wt limb perfusions as compared to tg limbs. Immunohistology of biopsies taken after 12 hours showed less NK cell tissue infiltration in the tg limbs. In vitro, NK cytotoxicity against hCD46 single tg pEC and wt pEC was similar, while lysis of double tg HLA-E/hCD46 pEC was significantly reduced. Finally, circulating cells of pig origin were observed during the ex vivo xenoperfusions. These cells expressed phenotypes mainly of monocytes, B and T lymphocytes, NK cells, as well as some activated endothelial cells. CONCLUSIONS: Ex vivo perfusion of pig forelimbs using whole human blood represents a powerful tool to study humoral and early cell-mediated rejection mechanisms of vascularized pig-to-human xenotransplantation, although there are several limitations of the model. Here, we show that (i) transgenic expression of HLA-E/hCD46 in pig limbs provides partial protection from human NK cell-mediated xeno responses and (ii) the emergence of a pig cell population during xenoperfusions with implications for the immunogenicity of xenografts.

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.

[Thumb Lengthening Following Amputation at the Proximal Phalanx using a Semicircular Distraction Ring Fixator: Results of 5 Cases]. / Daumenverlängerung nach traumatischer Amputation auf Grundphalanxhöhe mit semizirkulärem Distraktions-Fixateur: Ergebnisse von 5 Fällen.

Traumatic amputation of the thumb at the proximal phalanx or the metacarpophalangeal joint can be treated by distraction lengthening depending on the location of the amputation, the patient's age, occupation and functional demands. We report the results of proximal phalanx lengthening in 5 patients using a semicircular ring-type external fixator device. This prototype was developed at our clinic in collaboration with a specialised small company (Rotomed AG, Allmenstrasse 4, CH-4512 Bellach, www.rotomed.ch) based on the principles of Ilizarov's external fixator. In all patients, subjective and objective results were satisfactory (26.2 mm distraction length and an average Michigan Hand Outcome Score of 82.2%) without any rotational/axial deformities or complications requiring revision surgery.

Transgenic Expression of Human CD46 on Porcine Endothelium: Effect on Coagulation and Fibrinolytic Cascades During Ex Vivo Human-to-Pig Limb Xenoperfusions.

BACKGROUND: Dysregulation of the coagulation system due to inflammatory responses and cross-species molecular incompatibilities represents a major obstacle to successful xenotransplantation. We hypothesized that complement inhibition mediated by transgenic expression of human CD46 in pigs might also regulate the coagulation and fibrinolysis cascades and tested this in ex vivo human-to-pig xenoperfusions. METHODS: Forelimbs of wild-type and hCD46/HLA-E double transgenic pigs were ex vivo xenoperfused for 12 hours with whole heparinized human blood. Muscle biopsies were stained for galactose-α1,3-galactose, immunoglobulin M, immunoglobulin G, complement, fibrin, tissue factor, fibrinogen-like protein 2, tissue plasminogen activator (tPA), and plasminogen activator inhibitor (PAI)-1. The PAI-1/tPA complexes, D-dimers, and prothrombin fragment F1 + 2 were measured in plasma samples after ex vivo xenoperfusion. RESULTS: No differences of galactose expression or deposition of immunoglobulin M and immunoglobulin G were found in xenoperfused tissues of wild type and transgenic limbs. In contrast, significantly lower deposition of C5b-9 (P < 0.0001), fibrin (P = 0.009), and diminished expression of tissue factor (P = 0.005) and fibrinogen-like protein 2 (P = 0.028) were found in xenoperfused tissues of transgenic limbs. Levels of prothrombin fragment F1 + 2 (P = 0.031) and D-dimers (P = 0.044) were significantly lower in plasma samples obtained from transgenic as compared to wild-type pig limb perfusions. The expression of the fibrinolytic marker tPA was significantly higher (P = 0.009), whereas PAI-1 expression (P = 0.022) and PAI-1/tPA complexes in plasma (P = 0.015) were lower after transgenic xenoperfusion as compared to wild-type xenoperfusions. CONCLUSIONS: In this human-to-pig xenoperfusion model, complement inhibition by transgenic hCD46 expression led to a significant inhibition of procoagulant and antifibrinolytic pathways.

A comparison of complications in 400 patients after native nail versus silicone nail splints for fingernail splinting after injuries.

BACKGROUND: The fingertip is the most commonly injured part of the hand and is an important aesthetic part of the hand. METHODS: In this retrospective study we analyzed data from 700 patients operated on between 1997 and 2008 for complications after nail splinting with native nail or silicone nail. Inclusion criteria were patients living in Bern/Berner Land, complete documentation, same surgical team, standard antibiotics, acute trauma, no nail bed transplantation, and no systemic diseases. Groups were analyzed for differences in age, gender, cause and extension of trauma, bony injury and extent, infection, infectious agent, and nail deformities. Statistical analysis was done using the χ (2) test, Fisher's exact test, and Pearson correlation coefficients. RESULTS: A total of 401 patients, with a median age of 39.5 years, were included. There were more men with injured nails. Two hundred forty native nails and 161 silicone splints were used. There were 344 compression injuries, 44 amputations, and 13 avulsion injuries. Forty-three patients had an infection, with gram-positive bacteria (Staphylococcus aureus) causing most infections. A total of 157 nail dystrophies were observed, split nails most often. The native nail splint group showed significantly (p < 0.015) fewer nail deformities than the silicone nail splint group; otherwise, there were no statistical differences. However, there were twice as many infections in the silicone nail group. CONCLUSION: It seems to be advantageous to use the native nail for splinting after trauma, when possible. In case of a destroyed and unusable nail plate, a nail substitute has to be used.

Complement dependent early immunological responses during ex vivo xenoperfusion of hCD46/HLA-E double transgenic pig forelimbs with human blood.

BACKGROUND: Besides α1,3-galactosyltransferase gene (GGTA1) knockout, several transgene combinations to prevent pig-to-human xenograft rejection are currently being investigated. In this study, the potential of combined overexpression of human CD46 and HLA-E to prevent complement- and NK-cell-mediated xenograft rejection was tested in an ex vivo pig-to-human xenoperfusion model. METHODS: α1,3-Galactosyltransferase knockout heterozygous, hCD46/HLA-E double transgenic (transgenic) as well as wild-type pig forelimbs were ex vivo perfused with whole, heparinized human and autologous pig blood, respectively. Blood samples were analyzed for the production of porcine and/or human inflammatory cytokines as well as complement activation products. Biopsy samples were examined for deposition of human and porcine C3b/c, C4b/c, and C6 as well as CD62E (E-selectin) and CD106 (VCAM-1) expression. Apoptosis was measured in the porcine muscle tissue using TUNEL assays. Finally, the formation of thrombin-antithrombin (TAT) complexes was measured in EDTA plasma samples. RESULTS: No hyperacute rejection was seen in this model. Extremity perfusions lasted for up to 12 h without increase in vascular resistance and were terminated due to continuous small blood losses. Plasma levels of porcine cytokines IL1ß, IL-6, IL-8, IL-10, TNF-α, and MCP-1 as well as human complement activation markers C3a (P = 0.0002), C5a (P = 0.004), and soluble C5b-9 (P = 0.03) were lower in blood perfused through transgenic as compared to wild-type limbs. Human C3b/c, C4b/c, and C6 as well as CD62E and CD106 were deposited in tissue of wild-type limbs, but significantly lower levels (P < 0.0001) of C3b/c, C4b/c, and C6 deposition as well as CD62E and CD106 expression were detected in transgenic limbs perfused with human blood. Transgenic porcine tissue was protected from xenoperfusion-induced apoptosis (P < 0.0001). Finally, TAT levels were significantly lower (P < 0.0001) in transgenic limb as compared to wild-type limb xenoperfusions. CONCLUSION: Transgenic hCD46/HLA-E expression clearly reduced humoral xenoresponses since all, the terminal pathway of complement activation, endothelial cell activation, muscle cell apoptosis, inflammatory cytokine production, as well as coagulation activation, were all downregulated. Overall, this model represents a useful tool to study early immunological responses during pig-to-human vascularized xenotransplantation in the absence of hyperacute rejection.

MRI for differentiating ganglion and synovitis in the chronic painful wrist.

OBJECTIVE: The purpose of our study was to determine if preoperative MRI can differentiate between occult ganglion and synovitis in the chronic painful wrist. CONCLUSION: MRI is accurate in preoperatively distinguishing between ganglion and synovitis in the setting of chronic dorsal wrist pain. Four main criteria were useful: margin, shape, internal structure, and enhancement after administration of contrast material, with shape and internal structure being most helpful.

"Baby wrist": MRI of an overuse syndrome in mothers.

OBJECTIVE: The purpose of our study was to describe the MRI findings and interesting clinical aspects of a postpartum overuse syndrome of the wrist and thumb, de Quervain's tenosynovitis, or "baby wrist." CONCLUSION: Mothers may experience a wrist and thumb overuse syndrome, which can be diagnosed by MRI with an increase in size and low signal intensity on both T1- and T2-weighting, in and around the first dorsal tendon sheath compartment of the wrist.