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Biomedical subjects

K Shino

Publications and source records attributed to K Shino.

At least 37 records · Page 2Linked to original sources

Effect of freeze-drying or gamma-irradiation on remodeling of tendon allograft in a rat model.

Freezing or freeze-drying and gamma-irradiation are techniques currently used for processing tendon allografts. However, it is still unknown how these processing methods affect graft remodeling. In this study, we used a rat patellar tendon transplantation model to investigate the effect of various processing methods on remodeling by quantifying loss of collagen labeled with a radioactive isotope. The grafts were divided into the following four groups according to the processing method: fresh-frozen, freeze-dried, fresh-frozen and gamma-irradiated, or freeze-dried and gamma-irradiated. The percentage of donor collagen, calculated from hydroxyproline content and radioactivity level, was used as an indicator of graft remodeling. At 2 weeks, the level of donor collagen in the fresh-frozen group was 62%; in the freeze-dried group, 59%; in the fresh-frozen and irradiated group, 57%; and in the freeze-dried and irradiated group, 44%. At 4 weeks, the percentage of donor collagen remaining in grafts decreased to 38% in the fresh-frozen group, 19% in the freeze-dried group, 27% in the fresh-frozen and irradiated group, and 12% in the freeze-dried and irradiated group. Finally, at 12 weeks, the levels were 19% in the fresh-frozen group, 20% in the freeze-dried group, 15% in the fresh-frozen and irradiated group, and 6% in the freeze-dried and irradiated group. The percentages of donor collagen in the freeze-dried and the fresh-frozen and irradiated groups were significantly lower than that in the fresh-frozen group at 4 weeks. The values for the freeze-dried and irradiated group were significantly lower than those for the fresh-frozen and irradiated group at 4 and 12 weeks. These data suggest that freeze-drying, freeze-drying followed by gamma-irradiation, and fresh-freezing followed by gamma-irradiation temporarily accelerate graft remodeling.

Animals↗

Temporal and spatial expression of transforming growth factor-beta in the healing patellar ligament of the rat.

We investigated the temporal and spatial expression of transforming growth factor-beta in the healing patellar ligament of the rat by immunohistochemistry. The mid-portion of the medial half of the patellar ligament in 14-week-old male Wistar rats was cut transversely with a scalpel. On day 1 after ligament injury, diffuse staining for transforming growth factor-beta was observed in the extracellular matrix filling the wound, and the staining in the adjacent ligament tissue was as weak as it was in the normal ligament. On day 3, the intensity of the diffuse extracellular staining decreased, and the staining was observed in correspondence with the cellular distribution in the wound site and in the adjacent uninjured ligament tissue. On day 7, the intense staining was widely distributed over the whole length of the ligament tissue. On day 28, the staining for transforming growth factor-beta was still observed at the wound site and in the adjacent uninjured ligament tissue, where the staining was reduced in intensity but still stronger than it was in the normal ligament. On day 56, the expression of transforming growth factor-beta was still detectable at the wound site; however, in the adjacent uninjured ligament tissue, it had almost subsided to the normal level. The results of the present study suggest that ligament healing may be accompanied by extensive changes in the expression of transforming growth factor-beta over the whole length of ligament tissue.

Animals↗

Avulsion fracture of the anteromedial bundle of the anterior cruciate ligament.

The avulsion fracture of the tibial attachment site of the anterior cruciate ligament is relatively rare among adults. Although partial avulsion fracture of this site is very rare, complex forces exerted on the knee joint could produce this type of fracture. We report on an adult patient with an avulsion fracture of the attachment site of anteromedial bundle of the anterior cruciate ligament associated with injuries of lateral and medial collateral ligament and fibular fracture. We speculate varus stress in a flexed knee coupled with rotation between the femur and tibia caused this type of injury.

Adult↗

Remodeling of allogeneic and autogenous patellar tendon grafts in rats.

The differences in remodeling among frozen allografts, frozen autografts, and fresh autografts were investigated using a patellar tendon transplantation model in 2 different strains of rats. In this study, isografting (transplantation among the same strain inbred animals) was adopted as an autograft model. This method makes it possible to set up a frozen autograft model. Evaluation included mechanical properties determined during tensile failure tests, cross sectional area, histologic findings, and collagen fibril distribution. Recipient rats were euthanized at 4, 8, 12, and 24 weeks after transplantation (n = 5, in each group and in each period). The frozen allografts showed significantly less tensile strength than did the frozen autografts at 4 weeks, but this difference was transient and diminished by 8 weeks. The tensile strength of the fresh autografts was significantly greater than that of the frozen autografts at 8 weeks or the frozen allografts at 8 and 12 weeks. It was suggested that freeze thawing has adverse effects on the remodeling of fresh autografts in this extraarticular graft model. The cross sectional area for each type of graft at each posttransplantation period was about twice as large as that before transplantation. The collagen fibril profile of the frozen allografts was similar to that of the frozen autografts. However, the fresh autografts showed a greater number of large diameter fibrils and a lesser number of small diameter fibrils than did the other 2 frozen groups.

Animals↗

Osteoclastogenesis in iliac bone marrow of patients with rheumatoid arthritis.

OBJECTIVE: To investigate osteoclastogenesis in bone marrow cells from patients with various pathogenic backgrounds known to induce osteoporosis, to identify specific factors that may cause generalized osteoporosis in patients with rheumatoid arthritis (RA). METHODS: Bone marrow blood was obtained from 59 women, 36 with RA and 23 without RA. Patients with RA were classified as severe (26) and mild RA (10: 5 patients with and 5 without corticosteroid therapy). The non-RA subjects were divided into 3 groups, premenopausal (7), menopausal (8), and elderly (8). As a marker of bone resorption, the pyridinoline crosslinked telopeptide domain of type I collagen (ICTP) concentration in the bone marrow supernatant was measured by radioimmunoassay. The bone marrow cells were cultured 14 days in the presence or absence of autologous bone marrow supernatant; then the number of tartrate resistant acid phosphatase positive multinucleated cells (TRAP positive MNC) was counted as an indicator of osteoclastogenesis. RESULTS: ICTP concentration of the bone marrow supernatant and the number of TRAP positive MNC showed remarkable enhancement in some patients with severe RA, but these features were not observed in the 3 control groups. CONCLUSION: Increased bone resorption and enhanced osteoclastogenesis were specifically observed in the iliac bone marrow of patients with RA, especially those with severe RA. These phenomena can be considered to accompany generalized osteoporosis in RA.

Adult↗

Transient introduction of a foreign gene into healing rat patellar ligament.

We investigated the in vivo introduction of a reporter gene into healing rat patellar ligaments using the hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer method. The mid-portion of the medial half of the patellar ligament was cut transversely with a scalpel in 14-wk-old male Wistar rats. A HVJ-liposome suspension containing beta-galactosidase (beta-gal) cDNA was injected directly into the injured site and pooled in the fascial pocket covering the injured site 3 d postoperatively. Thereafter, beta-gal-labeled cells were observed in the wound site accounting for 3% of the wound cells on the first day, 2% on the third, 7% on the seventh, 6% on the 14th, 2% on the 28th, and 0.2% on the 56th day after injection. The beta-gal-labeled cells were initially localized in and adjacent to the wound site, but they were observed spreading into the ligament substance away from the wound on the seventh day after injection. On day 28, beta-gal-labeled cells were observed throughout the length of the ligament substance. With double-labeling for marker antigens for monocyte/macrophage (ED-1) and for collagen I aminopropeptide (pN collagen I), it was revealed that fibroblastic (pN collagen I-positive) cells accounted for 63% and monocyte/macrophage lineage cells for 32% of the beta-gal-labeled cells in the day 7 wound. On day 28, they formed 58 and 35% of the beta-gal-labeled cells in the wound, respectively. Thus, we succeeded in introducing the beta-gal gene into healing rat patellar ligament. Moreover, labeling of the transfected cells made it possible to identify a biological event, namely that the cells in and around the wound site infiltrate into the uninjured ligament substance and come to populate the whole length of the ligament substance as repair progresses. These results suggest that ligament healing may involve not only the repair of the wound site itself but also extensive cellular infiltration of ligament substance adjacent to the wound.

Animals↗

The remodelling process of allogeneic and autogenous patellar tendon grafts in rats: a radiochemical study.

In order to study the remodelling of collagen fibres of transplanted tendons, one-half of the patellar tendon of the knee in 54 rats was replaced with a radioisotope (3H-proline)-labelled patellar tendon procured from a donor rat. Three transplantation models were used in this study: fresh-frozen allograft, fresh-frozen autograft (fresh-frozen isograft), and fresh autograft (fresh isograft). The percentage of old collagen was calculated as an indicator of collagen turnover from the amount of hydroxyproline and the radioactivity level of 3H-hydroxyproline in the transplanted tendons at 4, 12 and 24 weeks postoperatively. Histological evaluation was also performed at 2, 4, 12 and 24 weeks. At 4 weeks, the percentage of old collagen in the grafts from the fresh-frozen allograft group was significantly lower than in the autograft groups (20% vs. 48%). Although the percentage of old collagen in the fresh-frozen autograft group had decreased to the same level as in the fresh-frozen allograft group by 12 weeks (approximately 10%), the value was still high in the fresh autograft group at 12 weeks and remained higher (38%). Histologically, at 2 weeks, the cellularity in the fresh-frozen allograft was higher than in the fresh-frozen autograft. After 4 weeks, however, no significant difference was found between these two groups. In the fresh autograft group, the cellularity was lower than in the fresh-frozen groups at all times. In conclusion, the collagen turnover rate in the fresh-frozen allograft was the most rapid of the three transplantation models based on hydroxyproline turnover.

Animals↗

Results of isolated meniscal repair evaluated by second-look arthroscopy.

Thirty-six isolated torn menisci in 35 patients (average age, 24 years) which had been repaired arthroscopically using an inside-out technique were evaluated by second-look arthroscopy. The time from meniscal repair to second-look arthroscopy ranged from 2 to 10 months with a mean of 5 months. The indications for meniscal repair were a longitudinal or oblique tear located at the outer half of the meniscus. Twenty (56%) were graded as excellent, 10 (28%) as good, and 6 (16%) were graded as poor. Neither age nor length of time between injury and repair affected meniscal healing. The medial meniscal repairs showed better results than the lateral repairs (rate of excellent results: medial, 82%; lateral, 44%; P < .01, chi-squared test). The rate of excellent results for those with normal meniscal bodies at the time of repair was 79%, which was significantly higher than that seen in the cases with deformed and/or superficial damage to the meniscal body (36%; P < .05, chi-squared test).

Adolescent↗

A simple and secure anchoring system for Caspari's transglenoid multiple suture technique using a biodegradable poly-l-lactic acid button.

To manage a difficulty in tying sutures over the infraspinatus fascia when using Caspari's transglenoid multiple suture technique, we developed a new anchoring system using a biodegradable poly-L-lactic acid (PLLA) button and investigated its clinical efficacy in 28 patients who were followed-up for more than 2 years postoperatively (mean, 26.5 months). Twenty-four patients had Bankart lesions and 4 had detachment of the superior glenoid labrum. The mean age at operation was 22.1 years. The PLLA button measured 8 x 8 x 1.2 mm and had two holes. After multiple sutures were inserted by the routine Caspari technique (mean, 7.3 sutures), the sutures were divided into 2 bundles, passed through the holes in the button, and tied over it on the posterior scapular neck under traction. The arm was immobilized in a Velpeau bandage for 3 weeks after Bankart repair and for 1 to 2 weeks after superior labral repair. The results of Bankart repair were excellent in 13 patients, good in 7, and poor in 4 according to Rowe's rating scale (success rate, 83%), while the outcome of superior labral repair was excellent in 3 and good in 1 according to our own criteria. All 4 patients who showed a poor outcome were contact athletes who developed resubluxation postoperatively. There were no complications, but transient damages to the suprascapular nerve occurred in 2 patients. In conclusion, the PLLA button provided simple and secure suture fixation for the Caspari technique.

Adolescent↗

Arthroscopic posterior cruciate ligament reconstruction using hamstring tendons: one-incision technique with Endobutton.

The one-incision surgical technique using autogenous hamstring tendons with Endobutton (Acufex Microsurgical Inc. Mansfield, MA) femoral end fixation for posterior cruciate ligament reconstruction is described. This technique avoids a second incision on the femoral side, and is adaptable to various graft materials (autogenous or allogeneic bone-patellar tendon-bone, hamstring, or Achilles tendon.

Arthroplasty↗

Bone marrow stromal cells contribute to synovial cell proliferation in rats with collagen induced arthritis.

OBJECTIVE: To observe the migration of bone marrow stromal cells into the joint cavity and the contribution of such cells to synovial proliferation in rats with collagen induced arthritis (CIA). METHODS: After bone marrow stromal cells preliminarily labeled with fluorescent dye or 3H thymidine accumulated in the bone marrow of splenectomized rats by intraperitoneal injection, the migration of labeled stromal cells in rats with CIA was analyzed by liquid scintillation counting, autoradiography, and fluorography. RESULTS: In splenectomized control rats, labeled bone marrow stromal cells were mostly found in the bone marrow and not in the synovium. Over 2 weeks after immunization, labeled stromal cells were microscopically found migrating directly into the joint cavity through the area between the articular margin and the synovial insertion (the bare zone). Labeled stromal cells were mainly found in the sublining layers of proliferating synovial tissue. CONCLUSION: At the onset of CIA, bone marrow stromal cells migrated from the bone marrow into the affected joint cavity and seemed to contribute to synovial proliferation.

Animals↗

Large molecule fraction of plasma separated with polyethylene glycol in treatment of chronic relapsing thrombotic thrombocytopenic purpura.

Plasma exchange and plasma infusion are effective in the treatment of patients with thrombotic thrombocytopenic purpura (TTP). It has been proposed that some component in normal plasma may be deficient in TTP patients. In this experiment normal plasma was fractionated and an effective fraction for the treatment of TTP was sought. Plasma was fractionated with 5% polyethylene glycol and a large molecule fraction was obtained which was composed mainly of proteins and lipoproteins with molecules larger than that of IgG. The fraction was dissolved in physiological saline and infused in two patients with chronic relapsing TTP who had been successfully treated periodically with plasma infusion. Peripheral platelet counts and serum LDH levels were measured, and the efficacy of the treatment was assessed. Infusion of the large molecule fraction elevated platelet counts and lowered the LDH level. However, the efficacy of the large molecule fraction was 25 to 50% that of plasma. Components in the fraction may have been inactivated partially during preparation. We propose that the large molecule fraction contains factors which are effective in some patients with TTP.

Adolescent↗

Arthroscopic repair for a flap tear of the posterior horn of the lateral meniscus adjacent to its tibial insertion.

A flap tear of the posterior horn of the lateral meniscus adjacent to its tibial insertion combined with acute rupture of the anterior cruciate ligament was successfully repaired arthroscopically by a combination of advancement of the tip of the flap into a drill hole created in the tibia and an inside-out stacked suture technique using Henning instrumentation in two cases. However, the long-term function of the repaired menisci, which was reduced several millimeters in circumferential length, is still unknown.

Adolescent↗

Conservative treatment of isolated injuries to the posterior cruciate ligament in athletes.

We assessed arthroscopically 22 young athletes with an isolated acute posterior cruciate ligament (PCL) injury. Four had significant damage to the articular cartilage of the medial femorotibial compartment and were advised not to resume sports. Three underwent PCL reconstruction because of a reparable meniscal tear or instability. The other 15 were treated conservatively and resumed sport. At an average follow-up of 51 months, one had developed arthritic symptoms due to newly-developed severe chondral damage to the medial femoral condyle, but none of the other 14 had developed arthritic symptoms and most remained athletically active. Severe chondral damage should be seen at an early arthroscopy. Knees with an isolated injury to the PCL with concomitant articular damage may be successfully managed by conservative treatment.

Activities of Daily Living↗

Second-look arthroscopy after meniscal repair. Review of 132 menisci repaired by an arthroscopic inside-out technique.

From 1986 to 1993, we repaired 278 torn menisci in 264 patients using an arthroscopically assisted inside-out technique. A total of 132 meniscal repairs in 122 patients were evaluated by second-look arthroscopy. At review, only nine patients had meniscal symptoms, such as locking, swelling or pain. Ninety-seven menisci (73%) had healed completely at the repair site, but there were new tears in different areas of 21 menisci, some of which had complete healing at the repair site. Incomplete healing, seen in 23 menisci (17%), was frequently near the popliteus tendon, most commonly where there had been an associated anterior-cruciate-ligament injury. Arthroscopically-assisted meniscal repair seems to be a reliable procedure, but some clinically successful cases had incomplete healing at the repair site or a newly-formed tear in the meniscal body or both. These lesions may cause meniscal symptoms to appear at a later date.

Adolescent↗

Usefulness of magnetic resonance imaging for detecting intrasubstance tear and/or degeneration of lateral discoid meniscus.

In order to determine the usefulness of magnetic resonance imaging (MRI) for detection of intrasubstance tears or degeneration of the lateral discoid meniscus, we compared findings obtained with MRI with those of histological examination. Sixty-four symptomatic lateral discoid menisci were studied using MRI before surgical treatment. Of these, 18 (28%) met the criteria that MRI signals did not extend to the surface and that no visible tears were observable at arthroscopy. Findings of intrameniscal regions of high signal intensity and flattening in shape on MR images were found to represent intrasubstance tears or degeneration not detected at arthroscopy. Our findings thus demonstrate that MRI is more sensitive than arthroscopy in the detection of intrasubstance pathologies of the lateral discoid menisci.

Adolescent↗

A congenital variant of thrombotic thrombocytopenic purpura in two siblings.

We describe two siblings affected by chronic relapsing thrombotic thrombocytopenic purpura from infancy. The elder brother, a 12-year-old boy had 50 such episodes characterized by acute onset of fever, headache, drowsiness, vomiting, dark urine, thrombocytopenia and anemia. The younger sister, a 6-year-old girl, had 8 episodes with the same clinical manifestations. Petechiae and ecchymoses on the extremities were present throughout their lives. Furthermore, anemia with evidence of red blood cell fragmentation and thrombocytopenia were present chronically. Periodical transfusion of frozen fresh plasma prevented recurrent episodes. These cases suggest that there is a congenital variant of thrombotic thrombocytopenic purpura.

Child↗

Effects of solvent preservation with or without gamma irradiation on the material properties of canine tendon allografts.

We studied the effects of solvent preservation with and without gamma irradiation on the material properties and morphology of canine tendons. Twenty-four paired tendons were harvested from both hind limbs of eight dogs. The tendons from the right legs were divided into three treatment groups of eight tendons each. In the first group, the tendons were subjected to solvent preservation for sterilization and drying (solvent group). In the second group, the tendons were treated in the same manner and then were sterilized by gamma irradiation (solvent/gamma group). The same treatments were applied to the tendons in the third group but in reverse order (gamma/solvent group). Tendons from the contralateral left legs were frozen immediately for use as controls for each corresponding treatment group. Histologically, the tendons treated with solvent, with or without gamma irradiation, had a more prominent wavy pattern in the collagen fibers than the control tendons. Mechanically, the tangent moduli for the solvent, solvent/gamma, and gamma/solvent groups were 58, 53, and 99%, respectively, of each contralateral control. The respective tensile strengths were 85, 39, and 86% of that of the contralateral control. The tendons in the solvent/gamma group underwent the most severe changes in material properties; the tendons in the gamma/solvent group changed the least. This suggests that gamma irradiation followed by solvent drying is the procedure of choice for the preservation of tendon allografts.

Animals↗