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Biomedical subjects

Keiichi Muramatsu

Publications and source records attributed to Keiichi Muramatsu.

At least 19 recordsLinked to original sources

The role of cyclophosphamide and granulocyte colony-stimulation factor in achieving high-level chimerism in allotransplanted limbs.

The establishment of a high-level of chimerism may be the most stable strategy for donor-specific tolerance. The purpose of this study was to evaluate the efficacy of a new protocol using cyclophosphamide (CYP) and granulocyte colony-stimulation factor (G-CSF) to induce high-level chimerism following rat whole-limb allotransplantation. Seventy-three whole-limb allotransplants from LacZ transgenic rats to LEW rats were performed. CYP was injected at day 2, and G-CSF was given from day 0 to 3. Nontreated limb allografts were rejected after 4.2 days. In FK506-treated group for 28 days, the survival time was prolonged to 64 days. In the group treated with CYP/G-CSF, limb allografts were rejected after 5.4 days and 5 of 15 recipients showed acute lethal graft-versus-host disease (GVHD). Polymerase chain reaction (PCR) study showed a high level of chimerism even within 1 week after transplantation. Fourteen of 30 recipients given CYP/G-CSF/FK506 died within 2 weeks. The limb survival was significantly prolonged, however, with three grafts surviving more than 300 days. Seven recipients (24%) showed chronic GVHD. A high-level of chimerism was maintained when limb allografts were not rejected by recipients. Limb allografting could function as a vascularized carrier for bone marrow transplantation, provide a continuous source of donor cells and contribute to a high level of chimerism in the recipient. Pretransplant CYP followed by G-CSF and FK506 treatment significantly prolonged the survival of limb allografts but frequently caused chronic GVHD in the recipients.

Animals↗

Peripheral neuropathies associated with rheumatoid synovial cysts of the elbow joint: three case reports.

We present 3 cases with peripheral neuropathies of the median, posterior interosseous, and ulnar nerves associated with rheumatoid synovial cysts of the elbow joint. Magnetic resonance imaging and electrophysiological examination led to the final diagnosis of the relation to synovial cysts of the elbow joint. All patients underwent surgical decompression of these nerves and synovectomy of the elbow joint. Functional recovery was excellent in the 2 patients with median and posterior interosseous nerve palsy but incomplete in the patient with ulnar nerve palsy. Such neuropathies require consideration in the differential diagnosis of wrist and hand disability in patients with rheumatoid arthritis. This awareness should enhance early diagnosis and treatment by surgical decompression.

Aged↗

Prolonged survival of rat whole-limb allografts treated with cyclophosphamide, granulocyte colony-stimulation factor and FK506.

We evaluated the efficacy of a new protocol using cyclophosphamide (CYP), granulocyte colony-stimulation factor (G-CSF) and FK506 to induce high level chimerism following rat whole-limb allotransplantation. The present study investigated the dose requirement and toxicity of CYP monotherapy in inducing stable bone marrow chimerism. Fifty-six whole-limb allotransplants from LacZ transgenic rats to LEW rats were performed. CYP at a dose of 100 mg to 200 mg/kg was injected 2 days before transplantation and G-CSF of 25 microg/kg/day was given for 4 days. FK506 was used for 28 days at 1 mg/kg/day. The level of chimerism was evaluated by semi-quantitative polymerase chain reaction. The survival of limb allografts in recipients treated with CYP of 150 mg/kg was significantly prolonged to 107 days. The onset of rejection was more prolonged to 158 days in recipients with CYP of 200 mg/kg, with two of eight grafts surviving >1 year and three recipients (38%) showed chronic, nonlethal GVHD with a high level of bone marrow chimerism. Limb allografting could contribute to chimerism in the recipient. Pretreatment with CYP had the dose-dependent effects of prolonging the survival of limb allografts. A CYP dose of 200 mg/kg appears to significantly prolong limb graft survival but frequently causes chronic nonlethal GVHD in the longer surviving recipients.

Animals↗

Free vascularized fibula grafting for reconstruction of the wrist following wide tumor excision.

Free vascularized fibula transfer is an established method for reconstruction of the wrist following tumor resection. In cases of resection of the radial articular surface, three reconstructive options are possible: fibular head transfer along with the shaft to replace the radial joint surface, fixation of the fibula to the scaphoid and lunate, or a complete wrist fusion. Three patients with a tumor involving the distal end of the radius were treated with wide resection, and subsequent wrist reconstruction was performed, using the above-mentioned procedures. Although our experience included only a small number of patients, both radio-carpal hemiarthroplasty and fibulo-scapho-lunate fusion similarly provided successful wrist stability and functional range of motion in these cases. Even when the wrist was totally fused with the fibula, its function was still acceptable.

Adolescent↗

Prolonged survival of rat hindlimb allografts following short-course FK506 and mycophenolate mofetil combination therapy.

The immunosuppressive effect of combined therapy using FK506 and mycophenolate mofetil (MMF) was studied in rat limb allotransplantation. Dark Agouti rat donor hindlimbs were orthotopically transplanted into Lewis rat recipients. In total, 38 models of transplantation were performed and divided into 8 groups that were treated individually or in combination with FK506 + MMF therapy. Animals were immunosuppressed for 28 days and then observed for up to 140 days. Graft rejection was evaluated both macroscopically and histologically. Survival times for rat limb allotransplants receiving combination FK506 + MMF therapy were significantly longer than with FK506 or MMF monotherapy, and this was achieved without serious side effects. A histopathological study demonstrated a significantly lower level of rejection with FK506 + MMF combination treatment compared to groups receiving FK506 or MMF monotherapy. Combined FK506 + MMF treatment can prolong the survival of rat limb allografts.

Animals↗

Gradual graft-cell repopulation with recipient cells following vascularized bone and limb allotransplantation.

Little is known about the fate of graft cells following vascularized bone allografting. This study was conducted to define the process of graft-cell repopulation with recipient cells. Sixty-five vascularized tibial bone and 50 limb allotransplantations were performed in rat sex-mismatched pairs. FK 506 was used for immunosuppression. The ratio of donor and recipient cells in the graft was evaluated by semiquantitative polymerase chain reaction, using the Y-chromosome primers. Allografted bones had no rejection episodes. In the vascularized bone allograft model, donor-derived cells were gradually replaced by cells of recipient origin, such that by 24 weeks, they comprised only 10% of total cells. In the limb allograft model, male recipient cells were detected in female grafts not at 1 week but at 48 weeks posttransplantation. The ratio of recipient cells was more than 10% in the femur and tibia. Recipient-derived cells gradually migrated into the grafted bone cells with the passage of time.

Animals↗

Donor cell engraftment in recipient lymphoid tissues after rat limb allograft.

BACKGROUND: The movement of cells from a transplanted tissue into the host organs, the so-called systemic chimerism, is a phenomenon known to occur and be associated with the development of immunologic tolerance in allotransplantation cases. The purpose of this study was to identify donor cell engraftment in recipient lymphoid tissues after performing rat hind limb allograft. MATERIALS AND METHODS: Fifty-five whole-limb allotransplantations were performed in sex-mismatched pairs of rats. Syngeneic male Lewis and allogeneic Dark Agouti donors were transplanted to female Lewis recipients. FK506 was used for immunosuppression. Donor male cells could be identified in the recipient female tissues by semiquantitative polymerase chain reaction analysis for a Y chromosome-specific DNA sequence. Chimerism was assessed at 1, 24, and 48 weeks after transplantation. RESULTS: There was no rejection episode in any of the limb grafts. Although levels of chimerism were highly variable in each lymphoid tissue, a gradual increase was noticed in all during the course of time. At 1 week after the transplant period, only intrasplenic chimerism was at high level (1%) in three groups. At 48 weeks after the transplant, all recipients with allografts showed very high level (10%) of chimerism in the bone marrow. Two, two, and two of six recipients showed very high levels in the spleen, lymph node, and liver, respectively, at 48 weeks. Intrathymic chimerism was higher at 24 weeks after transplant rather than at 48 weeks. CONCLUSION: We demonstrated donor cell engraftment into recipient lymphoid tissues after successful whole limb transplantation. We conclude that limb allograft can work as a vascularized carrier for the bone marrow transplantation, provide a continuous source of donor cells and contribute to chimerism in the recipient.

Animals↗

Cell traffic between donor and recipient following rat limb allograft.

Although cell traffic from the graft into the recipient and from the recipient into the graft had been noticed in allogeneic organ transplantation, little is known following whole-limb allografting. This study was conducted to define cell migration between donor and recipient. Sixty-seven vascularized hind limb allotransplantations were performed in rat sex-mismatched pairs and the recipient animals were treated with FK506 immunosuppression. The ratio of donor and recipient cells was evaluated by semi-quantitative PCR using the specific primers of the Y-chromosome. Allografted limbs had no rejection episode until the final assessment. The male recipient cells were detected in female limb grafts not at 1 week but at 48 weeks after transplantation. The male donor cells were detected in the humerus and tibia in the female recipient but not in the gastrocnemius muscle and leg skin. Our results demonstrated that recipient-derived cells gradually migrated into the grafted bone, muscle and skin cells with the duration of time. Donor-derived cells migrated into the healthy bones but not into the healthy muscle and skin. Because active regeneration occurs in the grafted limb to compensate graft damage secondary to ischemia and operative intervention, recipient-derived cells may mediate a muscular and dermo-epidermal renewal.

Animals↗

Functional recovery of rat hind-limb allografts.

Few papers have assessed the long-term functional recovery of animal limb allografts. In this study, the functional recovery of rat limb allografts was serially and quantitatively investigated for a period of 1 year. The donor's hind limb was orthotopically transplanted into the recipient. Fifteen recipients with allografts were treated with FK506. Functional recovery of the grafted limb was assessed serially by cutaneous reaction test, walking track analysis, and electrophysiologic evaluation. Sensibility improved to a similar extent in both isografts and allografts, and the recovery rate at 1 year was 68 percent, compared to the normal side. Sciatic function index significantly improved to - 70 points after 1 year. The amplitude recorded from the gastrocnemius muscle significantly improved, and the ratio compared to the normal side was 43 percent. Limb isografts and allografts treated with FK506 showed no significant differences in functional recovery. The data can be used as a reference standard for future investigations.

Animals↗

Neurospheres induced from bone marrow stromal cells are multipotent for differentiation into neuron, astrocyte, and oligodendrocyte phenotypes.

Bone marrow stromal cells (MSCs) can be expanded rapidly in vitro and have the potential to be differentiated into neuronal, glial and endodermal cell types. However, induction for differentiation does not always have stable result. We present a new method for efficient induction and acquisition of neural progenitors, neuronal- and glial-like cells from MSCs. We demonstrate that rat MSCs can be induced to neurospheres and most cells are positive for nestin, which is an early marker of neuronal progenitors. In addition, we had success in proliferation of these neurospheres with undifferentiated characteristics and finally we could obtain large numbers of neuronal and glial phenotypes. Many of the cells expressed beta-tubulin III when they were cultivated with our method. MSCs can become a valuable cell source as an autograft for clinical application involving regeneration of the central nervous system.

Animals↗

Microvascular anastomosis through the tibial tunnel: a new technique in free-tissue transfer to the leg.

Free-tissue transfer to a severely traumatized leg has a high rate of vascular complications. We present three successful cases using a new technique of microvascular anastomosis through the tibial tunnel. Because of the unavailability of anterior tibial artery due to posttraumatic vascular disease, donor vascular pedicles were passed posterior to the tibia through the tibial tunnel and anastomosed to the posterior tibial artery or its branch in an end-to-end fashion. The flaps survived perfectly, without any vascular complication. This technique represents a safe route, and the shortest route, to an expected anastomosis point. Our technique is indicated especially in cases with a single-vessel leg.

Adult↗

Role of vascularised tissue transfer after oncological resection in the upper extremity.

We reviewed 12 patients who had had vascularised tissue transfer after oncological resection in the upper extremity. All patients had immediate reconstruction, and one had a double tissue transfer. Tissues used were skin flaps, free muscles, and vascularised fibulas, for resurfacing the wound, motor recovery, and reconstruction of large bony defects, respectively. All the patients returned early to their daily activities. Although local recurrences were encountered in two patients, they were again rendered disease-free by salvage operations. Two patients had secondary reconstructions using pedicle latissimus dorsi flaps for late problems, including one of the two patients who developed a local recurrence. The mean functional score using the system devised by the Musculoskeletal Tumor Society was 84%. All the upper extremities were successfully rescued with satisfactory function. Vascularised tissue transfer was invaluable for achieving both curative resection of the tumour and a useful upper extremity.

Adolescent↗

Posttraumatic radioulnar synostosis treated with a free vascularized fat transplant and dynamic splint: a report of two cases.

Two cases of posttraumatic radioulnar synostosis are presented. The patients were treated with excision of the cross-union and interposition of a free vascularized fat transplant. A newly devised pronation-supination dynamic splint was employed for 3 months postoperatively in both patients. After a 1-year postoperative follow-up, an increased range of motion was restored in both cases, and there was no evidence of recurrent synostosis formation in subsequent radiographs. We suggest that an interposed vascularized fat graft may be an ideal biologic barrier to fill the space created by cross-union excision.

Adult↗

Ulnar variance and the role of joint levelling procedure for Kienböck's disease.

Forty patients with Kienböck's disease were reviewed to determine the relationship between ulnar variance and the development of Kienböck's disease in a Japanese cohort. The joint levelling procedures designed to correct ulnar variance were evaluated in 11 patients after a mean of 9.7 years following surgery. The mean ulnar variance was 0.50+/-1.67 in patients with Kienböck's disease and 0.56+/-1.76 in a control group. The onset of wrist symptoms in a younger age group was noticed after sports injuries, whereas older patients had no distinct history of trauma. Joint levelling procedures produced good clinical results, but carpal height ratio, Stahl's index and radioscaphoid angle were not improved, as evidenced radiographically. Our study suggests that in a Japanese cohort ulnar minus variance is not an important factor in the aetiology of Kienböck's disease. Although joint levelling procedures improved the clinical results, they did not reverse lunate collapse or carpal alignment.

Adolescent↗

Carpal instability in rheumatoid wrists.

This study was conducted to identify patterns of carpal instability in the rheumatoid wrist. One hundred patients with proven rheumatoid arthritis were randomly chosen, and posteroanterior and lateral radiographs of right wrists were taken. The most common isolated pattern was volar intercalated segmental instability, apparent in 16 wrists, while nine wrists had scapholunate dissociations. Larsen grading scores revealed a correlation between disease duration, carpal height ratio, and ulnar translocation index. In the early stages of disease, the scapholunate angle was significantly increased and the lunate rotated volarly due to loosening of the intrinsic carpal ligament. In later phases, the capitolunate angle significantly increased and the capitate rotated dorsally. Patterns of carpal instability in rheumatoid wrists seem to be complicated, involving combined laxity of the intrinsic and extrinsic carpal ligaments.

Adult↗