Graft vasculopathy in the skin of a human hand allograft: implications for diagnosis of rejection of vascularized composite allografts.

Whereas vascularized composite allografts often undergo acute rejections early in the postgraft period, rejection manifesting with severe vascular changes (graft vasculopathy) has only been observed on three occasions in humans. We report a hand-allografted patient who developed severe rejection following discontinuation of the immunosuppressive treatment. It manifested clinically with erythematous maculopapules on the skin and pathologically with graft vasculopathy that affected both large vessels and smaller cutaneous ones. The observation that graft vasculopathy can affect skin vessels shows that it is amenable to diagnosis with usual skin biopsy as recommended for the follow-up of these allografts. Graft vasculopathy developing in the setting of vascularized composite allografts likely represents chronic rejection due to under-immunosuppression and, if confirmed, should be included in a future update of the Banff classification of vascularized composite allograft rejection.

Outcomes with respect to disabilities of the upper limb after hand allograft transplantation: a systematic review.

The aim of this work is to compare disabilities of the upper limb before and after hand allograft transplantation (HAT), and to describe the side effects of immunosuppressive (IS) agents given to recipients of hand allografts. Clinical cases of HAT published between 1999 and 2011 in English, French, or German were reviewed systematically, with emphasis on comparing disabilities of the arm, shoulder and hand (DASH) scores before and after transplantation. Duration of ischemia, extent of amputation, and time since amputation were evaluated for their effect on intrinsic musculature function. Infectious, metabolic, and oncological complications because of IS therapy were recorded. Twenty-eight patients were reported in 56 clinical manuscripts. Among these patients, disabilities of the upper limb dropped by a mean of 27.6 (±19.04) points on the DASH score after HAT (P = 0.005). Lower DASH scores (P = 0.036) were recorded after secondary surgery on hand allografts. The presence of intrinsic muscle function was observed in 57% of the recipients. Duration of ischemia, extent of transplantation, and time since amputation were not associated statistically with the return of intrinsic musculature function. Three grafts were lost to follow-up because of noncompliance with immunosuppression, rejection, and arterial thrombosis, respectively. Fifty-two complications caused by IS agents were reported, and they were successfully managed medically or surgically. HAT recipients showed notable functional gains, but most complications resulted from the IS protocols.

Assessment of the electrophysiological properties of the muscle fibers of a transplanted hand.

BACKGROUND: The muscle fibers in a transplanted hand remain denervated for a long period of time after the transplant. This prolonged inactivity may change the electrophysiological membrane properties of muscle fibers, as observed in long-term denervation. We investigated whether electrophysiological properties of the muscle fibers are preserved in a transplanted hand even after several months of denervation. Specifically, we assessed the dependence of muscle fiber conduction velocity (CV) on discharge rate in motor units of the abductor digiti minimi muscle. METHODS: Surface electromyography signals were recorded from the transplanted hand of a patient who was 35 years of age at the time of the transplant. In each of 11 experimental sessions performed over a period of 23 months after the transplant, the subject was asked to linearly increase the activation or to maintain a maximum activation of the abductor digiti minimi muscle for 60 sec. Individual motor unit action potentials were identified from the electromyography recordings and muscle fiber CV was estimated for each action potential as a function of the time interval separating the action potential from the preceding discharge (interspike interval [ISI]). RESULTS: The baseline (ISI >1000 msec) CV was 3.8±0.3 m/sec. CV decreased monotonically with increasing ISI (R=0.95). For ISI in the range 0 to 10 msec, muscle fiber CV was 24.9%±16.3% higher than the baseline value (P<0.05). CONCLUSIONS: The results indicate that in the investigated muscle, the baseline value of CV and its dependency on discharge rate were similar as in able-bodied individuals, despite a period of several months of denervation.

The International Registry on Hand and Composite Tissue Transplantation.

BACKGROUND: The International Registry on Hand and Composite Tissue Transplantation was founded in May 2002, and the analysis of all cases with follow-up information up to July 2010 is presented here. METHODS: From September 1998 to July 2010, 49 hands (17 unilateral and 16 bilateral hand transplantations, including 1 case of bilateral arm transplantation) have been reported, for a total of 33 patients. They were 31 men and 2 women (median age 32 years). Time since hand loss ranged from 2 months to 34 years, and in 46% of cases, the level of amputation was at wrist. Immunosuppressive therapy included tacrolimus, mycophenolate mofetil, sirolimus, and steroids; polyclonal or monoclonal antibodies were used for induction. Topical immunosuppression was also used in several cases. Follow-up ranges from 1 month to 11 years. RESULTS: One patient died on day 65. Three patients transplanted in the Western countries have lost their graft, whereas until September 2009, seven hand grafts were removed for noncompliance to the immunosuppressive therapy in China. Eighty-five percent of recipients experienced at least one episode of acute rejection within the first year, and they were reversible when promptly treated. Side effects included opportunistic infections, metabolic complications, and malignancies. All patients developed protective sensibility, 90% of them developed tactile sensibility, and 82.3% also developed a discriminative sensibility. Motor recovery enabled patients to perform most daily activities. CONCLUSIONS: Hand transplantation is a complex procedure, and its success is based on patient's compliance and his or her careful evaluation before and after transplantation.

Scaphoid reconstruction by a free vascularized osteochondral graft from the rib: a case report.

Nonunion and avascular necrosis of the proximal pole of the scaphoid remain one of the most difficult problems in wrist reconstructive surgery. A number of interpositional vascularized bone grafts have been proposed for scaphoid nonunion, in order to promote faster union at the contact site with both fragments. However, once the proximal pole has undergone avascular changes and is completely necrotic, there is no alternative other than to remove it. At present, more radical operations have been advocated in these cases, such as proximal row carpectomy or intercarpal arthrodesis. We present a case where the necrotic proximal pole of the scaphoid was removed and replaced with a remodeled osteochondral-free vascularized graft from the rib based on the inferior and superior intercostal arteries, branches of the internal mammary artery.

A home-care system for the telemonitoring and telerehabilitation of the hand incorporating interactive biofeedback.

We have designed and constructed force measurement equipment to assess hand-finger function in pressing tasks. The equipment was used for monitoring the follow-up of five hand-transplanted subjects. Interactive software was integrated into the instrument to monitor the functionality of the hand and fingers during exercises in realtime. The interactive software included biofeedback to provide realtime quantitative responses for the patient and the therapist. Acceptance of the system was investigated with patients and therapists: the system was found to be user-friendly and effective; it was practical both for patients and therapists. The system could be used in a telerehabilitation centre or in a patient's home.

Sustained impairment of human cytomegalovirus (HCMV)-specific CD4+ and CD8+ T cell response is responsible for recurrent episodes of disseminated HCMV infection in a D+R- hand transplant recipient.

Human cytomegalovirus (HCMV) infection is the major viral complication in solid organ transplant recipients. Seronegative recipents (R-) of organs from seropositive donors (D+) appear to be at higher risk of developing symptomatic HCMV infection. To what extent systemic life-threatening complications can be risked for non-life-saving transplant procedures? A case report describing successful treatment of repeated episodes of active HCMV infection in a D+R- hand recipient in the absence of HCMV-specific T-cell immunity is presented. In the attempt to save both the patient and the transplanted hand, a preemptive treatment strategy was adopted with the aim to boost the constitution of the virus-specific T-cell immune response and simultaneously avoid onset of disease. Careful monitoring of HCMV load in blood and HCMV-specific T-cell immunity guided administration of repeated courses of antiviral treatment and avoided emergence of HCMV-related symptoms. Following establishment of HCMV-specific CD4+ and CD8+ T-cell response, preemptive treatment was no longer required due to sustained HCMV disappearance from blood. The patient is now well, and his hand too. In conclusion, evaluation of virus-specific T-cell immunity is of crucial importance in D+R- transplant recipients and careful monitoring of HCMV-specific T cell mediated response should always parallel monitoring of HCMV load in transplant recipients.

The international registry on hand and composite tissue transplantation.

BACKGROUND: Since May 2002, all groups performing hand transplantations have supplied information to the International Registry on Hand and Composite Tissue Transplantation (IRHCCT). The analysis of all cases with follow-up information up to September 2007 is presented here. METHODS: From September 1998 to September 2007, 38 hands (18 unilateral and 10 bilateral hand transplantations) and two digits have been reported to IRHCTT, for a total of 30 patients with a follow-up period ranging from 6 months to 9 years. They were 28 males and two females, median age of 34 years. Time since hand loss ranged from 2 months to 34 years and in 50% of cases the level of amputation was at wrist level. Immunosuppressive therapy included tacrolimus, mycophenolate mofetil, rapamycin, and steroids; polyclonal or monoclonal antibodies were used for induction. Topical immunosuppression was also employed in several cases. RESULTS: Patient survival was 100%. Graft survival was 100% at 1 and 2 years. Except for the Chinese patients, current graft survival is 95.6% and cause of graft loss was always no compliance to the treatment. Acute rejection episodes occurred in 85% of the patients within the first year and they were reversible in all compliant patients. Side-effects included opportunistic infections and metabolic complications. All patients developed protective sensibility, 90% of them developed tactile sensibility, and 72% also a discriminative sensibility. Motor recovery enabled patients to perform most daily activities. CONCLUSIONS: Hand transplantation is feasible with a high success rate and satisfactory functional outcome.

Discharge variability of motor units in an intrinsic muscle of transplanted hand.

The study analyzed the discharge characteristics of the motor units in an intrinsic muscle of a transplanted hand. Multichannel electromyographic (EMG) recordings were obtained in 11 experimental sessions over 16 mo starting from day 205 after a hand was transplanted in a 35-yr-old man who had lost his right hand 22 yr earlier. The action potentials discharged by single motor units were identified from the surface EMG signals of the abductor digiti minimi muscle in the transplanted hand as the individual performed 60-s maximal and linearly increasing (ramp) contractions. Discharge rate decreased from 27.1 +/- 8.4 pulses per second (pps) at the start of the maximal contractions to 17.2 +/- 2.9 pps at the end (P < 0.001) and increased from 17.4 +/- 4.3 to 22.1 +/- 5.0 pps during the ramp contractions (P < 0.05). The SD of the interspike interval (ISI) nearly related to the mean ISI with a similar regression slope for the maximal (0.49 +/- 0.09) and ramp contractions (0.43 +/- 0.10). The coefficient of variation for ISI was higher than values in able-bodied persons and did not change during either the maximal (36.8 +/- 10.8%) or the ramp contractions (35.9 +/- 7.4%). High-frequency bursts of activity with <20 ms between two and six action potentials occurred during both maximal and ramp contractions. In conclusion, motor neurons that reinnervated a muscle in a transplanted hand discharged action potentials with a high degree of variability that suggested greater synaptic noise during the voluntary contractions.

Second report (1998-2006) of the International Registry of Hand and Composite Tissue Transplantation.

Since May 2002 all groups performing hand transplantations have supplied detailed information to the International Registry on Hand and Composite Tissue Transplantation. This report provides a review of all hand transplants performed to date. From September 1998 to February 2006 eighteen male patients underwent 24 hand/forearm/digit transplantations (eleven unilateral and four bilateral hand transplantations, two bilateral forearm transplantations, one thumb transplantation). The level of amputation was mostly at the distal forearm or wrist. Patient average age was 32. Time since hand loss ranged from 2 months to 22 years. Immunosuppressive therapy included tacrolimus, mycophenolate mofetil, rapamycin and steroids; polyclonal or monoclonal antibodies were used for induction. Topical immunosuppression was administered in some patients. Follow-up period ranged from 34 to 85 months. Patient survival was 100%. Graft survival was 100% at 1 and 2 years. Two cases of graft failure at a later date occurred and were caused by severe inflammation and progressive rejection in a non-compliant patient. In addition, 6 hands were lost due to a rejection process as the Chinese recipients did not take their immunosuppressive treatment. These failures were communicated in January 2006. Acute rejection episodes occurred in 12 patients within the first year. Rejection was completely reversible in all compliant patients. Side-effects included opportunistic infections and metabolic complications. No life-threatening complications or malignancies were reported. As it would have been very difficult to analyse transplantation functional results in a standardized way, the Registry has performed a functional score system. All patients had achieved protective sensation and in 17 of them also discriminative sensation. Extrinsic and intrinsic muscle recovery enabled patients to perform most daily activities and 90% of the recipients returned to work, and improved manual skills allowed them not only to resume their previous jobs but also, in some cases, to find more suitable employment. Fifteen recipients reported an improvement of their quality of life and we evaluated as a very important point as patient satisfaction and well-being are mandatory goals of hand transplantation.

Simultaneous vascularized bone marrow transplantation to promote acceptance of hind limb allografts and its effects on central and peripheral chimerism.

Administration of donor bone marrow (BM) cells can improve the outcome of transplantation. The ability of donor vascularized bone marrow transplantation (VBM) to provide an ongoing source of donor cells and improve survival in a rigorous rat model of hind limb transplantation (HLTX) was investigated. HLTX were performed between Brown Norway (BN) donors and Lewis recipients in three groups: HLTX; HLTX plus intravenous donor BM cells and HLTX plus simultaneous VBM transplantation. Animals received 12 weeks triple immunosuppression. Survival was compared at 4 months and donor chimerism was evaluated. Simultaneous VBM transplantation led to slight but nonsignificant prolongation of survival (P=0.056). Donor cells in the VBM were eventually replaced by recipient and there was no long-term increase in chimerism. Few donor cells were observed in thymus. Simultaneous VBM transplantation showed a trend for improved survival of HLTX however the VBM failed to provide a sustained increase in chimerism.

Osmic acid staining of myelin sheath in normal and regenerated peripheral nerves.

OBJECTIVE: To introduce a practical, economical, and time-saving method to stain (with osmic acid) the myelin sheath in normal and regenerated peripheral nerves. METHODS: A total of 12 Sprague Dawley rats, weighing 250-320 g (mean equal to 276 g+/-38 g), were divided into two groups: a normal nerve group (n equal to 6) and a regenerated nerve group (n equal to 6). In the normal nerve group, the ventral and dorsal roots of L(4) to L(6) and their sciatic nerves were harvested for histological analysis. While in the regenerated nerve group, the right sciatic nerves were severed and then repaired with an epineurial microsuture method. The repaired nerves were harvested 12 weeks postoperatively. All the specimens were fixed in 4% paraformaldehyde and transferred to 2% osmic acid for 3-5 days. Then the specimens were kept in 75% alcohol before being embedded in paraffin. The tissues were cut into sections of 3 micromolar in thickness with a conventional microtome. RESULTS: Under a light microscope, myelin sheaths were clearly visible at all magnifications in both groups. They were stained in clear dark colour with a light yellow or colorless background, which provided high contrast images to allow reliable morphometric measurements. Morphological assessment was made in both normal and regenerated sciatic nerves. The ratios of the myelin area to the fibre area were 60.28%+/-7.66% in the normal nerve group and 51.67%+/-6.85% in the regenerated nerve group, respectively (P less than 0.01). CONCLUSIONS: Osmic acid staining is easy to perform and a very clear image for morphometrical assessment is easy to obtain. Therefore, it is a reliable technique for quantitative evaluation of nerve morphology.

Cytomegalovirus-related complications in human hand transplantation.

BACKGROUND: Up to date, 24 hands/thumbs have been transplanted in 18 patients. We herein report on cytomegalovirus (CMV) infection, disease, and the adopted treatment. METHODS: Immunosuppression consisted of tacrolimus-based triple-drug therapy with antithymocyte globuline or CD25-receptor antagonist induction. Donor/recipient CMV match was negative/negative (n=8), negative/positive (n=3), positive/positive (n=3), positive/negative (n=3) and unknown in one case. Six patients (three +/-, two +/+, and one -/+) received gancyclovir i.v. followed by oral gancyclovir or valgancyclovir for prophylaxis. RESULTS: Patient and graft survival at a mean follow-up of 42.9 months were 100% and 91%, respectively. Of all patients tested for CMV, 45.5% developed CMV infection or disease. Two patients that were given a CMV-positive graft showed very high viral loads (550 and 1200/200000 leukocytes) after transplantation. Gancyclovir treatment failed to permanently control CMV in 80% of the patients experiencing CMV infection. Those patients requiring more toxic second-line therapies (foscarnet/cidofovir) suffered from side effects such as nephrotoxicity, nausea, vomiting, and diarrhea. CONCLUSIONS: CMV infection/disease complicated the postoperative course after composite tissue allograft (CTA) transplantation in five of nine recipients challenged with the virus. The close time correlation suggests an association between virus replication and rejection in some cases. CMV represents the major infectious threat in CTA transplantation. Therefore, CMV-mismatch should be avoided and prophylaxis with valgancyclovir and anti-CMV hyperimmunoglobulin should be mandatory.

Pathological score for the evaluation of allograft rejection in human hand (composite tissue) allotransplantation.

The experience obtained from the human hand allografts (HHA) performed to date suggests that the skin is a priviledged target of allograft rejection in this setting. The aim of this study was to establish a pathological score for assessing the severity of HHA rejection. The pathological slides of 89 skin biopsies obtained from the allografted limbs of six HHA recipients from day 0 to 5 years post-graft were retrospectively examined. According to the severity of the pathological changes observed, the following grades of rejection are proposed: 0: no rejection, I: mild rejection, II: moderate rejection, III: severe rejection, IV: very severe rejection. This grading system can be used as a basis for monitoring allograft rejection and for assessing the effects of the immunosuppressive treatment aiming at reversing HHA rejection; it can also be used for monitoring rejection of other skin-containing CTA.

The International Registry on Hand and Composite Tissue Transplantation.

BACKGROUND: Since May 2002 all groups performing hand transplantations have supplied detailed information to the International Registry on Hand and Composite Tissue Transplantation. This inaugural report provides a review of all hand transplants performed to date. METHODS: Between September 1998 and September 2004, 18 male patients underwent 24 hand/forearm/digit transplantations (11 monolateral and 4 bilateral hand transplantations, 2 bilateral forearm transplantations, and 1 thumb transplantation). The level of amputation was mostly at the distal forearm or wrist. The average age of the patient was 32 years. Time since hand loss ranged from 2 months to 22 years. Immunosuppressive therapy included tacrolimus, mycophenolate mofetil, rapamycin, and steroids; polyclonal or monoclonal antibodies were used for induction. Topical immunosuppression was administered in some patients. Follow-up period ranged from 17 to 70 months. RESULTS: Patient survival was 100%. Graft survival was 100% at 1 and 2 years. Two cases of graft failure at a later date were caused by severe inflammation and progressive rejection in a noncompliant patient. Acute rejection episodes occurred in 12 patients within the first year. Rejection was reversible in all compliant patients. Side effects included opportunistic infections and metabolic complications. No life-threatening complications or malignancies were reported. All patients had achieved protective sensation, and 17 patients also achieved discriminative sensation. Extrinsic and intrinsic muscle recovery enabled patients to perform most daily activities. CONCLUSIONS: Despite the enormous antigen load associated with composite tissue allograft, hand transplantation became a clinical reality with immunosuppression comparable to transplantation of solid organs.

Arterial stiffening influence of sympathetic nerve activity: evidence from hand transplantation in humans.

Studies in animals and humans suggest that sympathetic activity exerts a stiffening influence on large and middle-sized artery walls. We sought to obtain further evidence on this issue by measuring radial artery distensibility in an allotransplanted and thus denervated hand using the contralateral artery as control. In 2 men, blood pressure was measured by a semiautomatic device (Dinamap). Diastolic diameter, systo-diastolic diameter excursion (ultrasound Wall Track system), and distensibility (Reneman formula) of both radial arteries were measured at a level corresponding to 4 cm below the suture of the transplanted hand 40 days after surgery and every 4 weeks for the next 6 months. After surgery, systo-diastolic diameter excursion and distensibility were much greater in the transplanted radial artery than in the contralateral vessel, reaching values similar to the contralateral ones after 4 months, when signs of reinnervation of the transplanted hands had appeared. Radial deinnervation was accompanied by an increased arterial distensibility, which provides further evidence of the sympathetic stiffening effect on arterial wall in humans.

Reinnervation of motor units in intrinsic muscles of a transplanted hand.

Functional recovery of transplanted hand can be evaluated clinically but until now there has been no direct assessment of muscle control. In October 2000 we transplanted the right hand of a brain-dead man aged 43 onto a man aged 35 who had lost his right dominant hand 22 years before. Starting from day 205 after the transplant, multi-channel surface electromyographic (EMG) signals were recorded from intrinsic muscles of the transplanted hand in order to assess their degree of reinnervation. Eleven months post-operatively, the first motor unit action potential train was detected from the abductor digiti minimi. One month later, also the abductor pollicis brevis and the opponens pollicis muscles showed motor unit activity, while, after 15 and 24 months, the first dorsal interosseous and the first lumbricalis muscles, respectively, showed activation of their first motor units. An increase in the number of active motor units was observed after the first signs of reinnervation, although the process was rather slow. In sustained maximal contractions, the motor unit discharge rate decreased from (mean +/- S.D.) 34.0+/-6.7 pps to 23.4+/-5.1 pps in 60 s for the abductor digiti minimi, although the subject was verbally encouraged to maintain a maximal activation. Moreover, the subject was able to perform basic control tasks involving voluntary modulation of motor unit discharge rate. With a visual feedback, he could increase discharge rate of the abductor digiti minimi approximately linearly over time, from 13.4+/-6.7 pps to 32.5+/-11.2 pps in 60 s. In conclusion, we showed reinnervation of single motor units in a transplanted hand after 22 years of denervation. Moreover, voluntary modulation of discharge rates of these motor units could be performed since the first sign of reinnervation.