Objective To investigate the clinical feasibility of different types of the saphenous neuro-veno-fascial cutaneous flaps. Methods From June 1996 to October 2002, 18 cases of skin defects in the knee and the lower part of the limb were treated with proximally(4 cases) or distally(11 cases) based pedicles of saphenous neuro-venofascial cutaneous flap or crossleg flap (3 cases)according to the site of defects . The sizes of the flaps ranged from 4 cm×5 cm to 9 cm×20 cm. Results The flaps survived completely in 17 cases, distal 1/5 of the flap necrosed partially in 1 case because of vein drainage disturbance. The colour and texture of flaps were excellent, the appearance and function were satisfactory after a follow up of 6-24 months.Conclusion The saphenous neuro-veno-fascial cutaneous flap is an idea flap in repairing skin defects of the knee, the leg, the ankle and the foot because it is easy to be designed and dissected and it has reliable blood supply and preserved main artery. The relationship between the septal perforating branches of the tibial posteriorartery and survival size of flap need to be investigated further.
Objective To explore the expression characteristics of chaperone interacting protein (CHIP) in normal, scar and chronic ulcer tissues and its relationship with wound healing. Methods Twenty biopsies including scar tissues(n=8), chronic ulcer tissues(n=4) and normal tissues(n=8)were used in this study. The immunohistochemical staining (power visionTMtwo-step histostaining reagent) was used to explore the amount and expression characteristics of such protein.Results The positive expression of CHIP was observed in fibroblasts, endothelial cells and epidermal cells in dermis and epidermis. It was not seen ininflammatory cells. The expression amount of CHIP in scar tissues, chronic ulcer tissues and normal tissues was 89%, 83% and 17% respectively. Conclusion Although the function of CHIP is not fully understood at present, the fact that this protein is expressed only at the mitogenic cells indicates that it may be involved in mitogenic regulation during wound healing.
Objective To investigate the effect of homograft of marrow mesenchymal stem cells (MSCs) seeded onto poly-L-lactic acid (PLLA)/gelatin on repair of articular cartilage defects. Methods The MSCs derived from36 Qingzilan rabbits, aging 4 to 6 months and weighed 2.5-3.5 kg were cultured in vitroand seeded onto PLLA/gelatin. The MSCs/ PLLA/gelatin composite was cultured and transplanted into full thickness defects on intercondylar fossa. Thirty-six healthy Qingzilan rabbits were made models of cartilage defects in the intercondylar fossa. These rabbits were divided into 3 groups according to the repair materials with 12 in each group: group A, MSCs and PLLA/gelatin complex(MSCs/ PLLA/gelatin); group B, only PLLA/gelatin; and group C, nothing. At 4,8 and 12 weeks after operation, the gross, histological and immunohistochemical observations were made, and grading scales were evaluated. Results At 12 weeks after transplantation, defect was repaired and the structures of the cartilage surface and normal cartilage was in integrity. The defects in group A were repaired by the hylinelike tissue and defects in groups B and C were repaired by the fibrous tissues. Immunohistochemical staining showed that cells in the zones of repaired tissues were larger in size, arranged columnedly, riched in collagen Ⅱ matrix and integrated satisfactorily with native adjacent cartilages and subchondral bones in group A at 12 weeks postoperatively. In gross score, group A(2.75±0.89) was significantly better than group B (4.88±1.25) and group C (7.38±1.18) 12 weeks afteroperation, showing significant differences (P＜0.05); in histological score, group A (3.88±1.36) was better than group B (8.38±1.06) and group C (13.13±1.96), and group B was better than group C, showing significant differences (P＜0.05). Conclusion Transplantation of mesenchymal stem cells seeded onto PLLA/gelatin is a promising way for the treatment of cartilage defects.
Objective To know the possibility of nerveregeneration after artery sleeve anastomosis and end-to-side suture Methods Seventy-five SD rats were divided into 5 groups. First, the distal end ofsevered peroneal nerve was sutured end-to -side with artery sleeve anastomosis withnormal nerve tibial trunk in groups A, B, C and D. Second, the tibial epineurium at the suture site was not removed in group A; the epineurium at the suturesite was removed(windowing) in group B; the distal end of pre-injured peroneal nerve was sutured after 14 days and windowing was done in group C; and the neural growth factor was injected into artery sleeve and windowing was done in group D. While the distal end of severed peroneal nerve was sutured end to side directly with normal nerve tibial trunk and windowing was done in group E. The histological observation was made and the number of nerve fibers was recorded after 4, 8 and 12 weeks of operation.Results After 4 weeks, there existed the regeneration of axons and myeline sheaths in groups C, D, E, and no nerve fiber regeneration was seen in group A. After 8 weeks, the regenerating nerve fibers were significantly more in groups C, D and E than in group B and ingroup E than groups C and D(Plt;0.05). After 12 weeks, the regenerating nervefibers were significantly more in groups C，D and E than in group B(Plt;0.05).Conclusion End-to-side coaptation with artery sleeve anastomosis is a new valuable method in repair of peripheral nerve injuries.
Objective To study the biological activities ofthe nerve regeneration conditioned fluid (NRCF), especially to further separateand identify the protein bands of the relative molecular mass of (232-440)×103. Methods The silicone nerve regeneration chambers were implanted between the cut ends of the sciatic nerve in 6 New Zealand white rabbits (weight, 1.8-2.5 kg). The proteins in NRCF were separated by the native-polycrylamide gel electrophoresis (Native-PAGE), the protein bands of the relative molecular mass of (232-440)×103 were analyzed by the Shotgun technique, liquid chromatography, and mass spectrometry. Results The Native-PAGE result showed that there was 1 protein band of the relative molecular mass over 669×103, (232-440)×103 and (140-232)×103,respectively, and 6 bands of the relative molecular mass of (67-140)×103.Besides, 54 proteins were identified with at least 2 distinct peptides in 1 protein band of the relative molecular mass of (232-440)×103, including 4 unnamed protein products, mainly at the isoelectric points of 5.5-8.0 and of the relative molecular mass of (10-40)×103. Based on their functions in the protein database, allthe identified proteins in this study were classified into the following 5 groups: conjugated protein (43%), transport protein (30%), enzyme (6%), signal transducer (4%), and molecular function-unknown protein (17%). At the subcellular localization of the identified proteins, there was mainly a secreted protein (63%), and the remaining proteins were localized in the membrane and cytoplasm. Conclusion Native-PAGE and the Shotgun technique can effectively separate and identify proteins from NRCF, and can identify the components of the protein band of the relative molecular mass of (232-440)×103 and provide basicinformation on the unnamed protein products in NRCF.
Objective To investigate the morphology and biomechanics of in vivo osteogensis after repairing rabbit skull defects with plastic engineered bone which was prefabricated with alginate gel, osteoblasts and bone granules. Methods Twenty-eight rabbits were divided into group A (n=16), group B(n=8) and group C(n=4).The bilateral skull defects of 1 cm in diameter were made. Left skull defects filled with alginate gel-osteoblasts-bone granules(group A1) and right skull defects filled withalginate gel-bone granules(group A2).The defects of group B was left, as blank control and group C had no defect as normal control. The morphological change and bone formation were observed by methods of gross, histology and biomechanics. Results In group A1, the skull defects were almost entirely repaired by hard tissue 12 weeks after operation. The alginate gel-osteoblasts-bone granule material had changed into bone tissue with fewbone granules and some residuary alginate gel. The percentage of bone formation area was 40.92%±19.36%. The maximum compression loading on repairing tissue ofdefects was 37.33±2.95 N/mm; the maximum strain was 1.05±0.20 mm; andloading/strain ratio was 35.82±6.48 N/mm. In group A2, the alginate and bone granules material partially changed into bone tissue 12 weeks after operation. The percentage of bone formation area was 18.51%±6.01%. The maximum compression loading was 30.59±4.65 N; the maximum strain was 1.35±0.44 mm; and the loading/strainratio was 24.95±12.40 N/mm. In group B, the skull defects were mainly repaired bymembrane-like soft tissue with only few bone in marginal area；the percentage of bone formation area was 12.72%±9.46%. The maximum compression loading was 29.5±2.05 N; the maximum strain was 1.57±0.31mm；and the loading/strainratio was 19.90±5.47 N/mm.In group C, the maximum compression loading was 41.55±2.52 N; the maximum strain was 095±017 mm; and the l oading/strain ratio was 47.57±11.22 N/mm. 〖 WTHZ〗Conclusion〓〖WTBZ〗The plastic engineered bone prefabricated with algina te gelosteoblastsbone granule may shape according to the bone defects and ha s good ability to form bone tissue, whose maximum compression loading can reach 89 % of normal skull and the hardness at 12 weeks after operation is similar to that of normal skull.
Objective To investigate the origin of small saphenous vein of distally-based of sural nerve nutrient vessels flap and its clinical application. Methods The origins of nutrient vessels of small saphenousvein and communicating branches of superficial-deep vein were observed on specimens of 30 adult cadaveric low limbs by perfusing red gelatin to dissect the artery. Results The nutrient vessels of small saphenous vein originated from the heel lateral artery, the terminal perforator branches of peroneal artery and intermuscular septum perforating branches of peroneal artery. There were 2 to 5 branches ofsuch distally-based perforating branches whose diameters ranged from 0.6 to 1.0 mm. Those perforating branches included fascia branches, cutaneous branches nerve and vein nutrient branches. Those nutrient vessels formed a longitudinalvessel chain of sural nerve shaft, vessel chain of vein side and vessel networkof deep superficial fascia. The small saphenous vein had 1 to 2 communicating branches of superficial-deep vein whose diameter was 1.7±0.5 mm， 3.4±0.9 cm to the level of cusp of lateral malleolus, and converged into the fibular vein. Conclusion Distally-based sural nerve, small saphenous vein, and nutrient vessles of fascia skin have the same region. The communicating branches of superficial-deep vein is 3 to 4 cm to the level of cusp lateral malleolus. These communicating branches could improve the venousdrainage of the flap.
Objective To resolve the tough problem of how to observe the growing cells in an opaque vector. Methods The urethral epithelial cells from a young male New Zealand rabbit were inoculated, and were primarily cultured in vitro and subcultured for 3 passages. Then, the urethralepithelial cells were cultured in the collagen chitosan complex for 3, 7, 14 and 21 days. The cells were dyed with 6-carboxyfluorescein diacetateacetoxymethyl ester and propidium iodine, respectively. Then, Interactive Laser Cytometer was used to detect the growing cells. Results The urethral epithelial cells grew and proliferated very well in the collagen chitosan complex vector. After the urethral epithelial cells grew in the collagen-chitosan complex vector for 3 and 7 days, the fluorescent density amount of the surviving cells were(1.09±0.13)×10.8 and (2.04±0.13)×10.8, respectively. However, after 14and 21 days, the fluorescent density amount of the surviving cells was (0.55± 0.09)×10.8 and (0.47±0.03)×108, respectively. There was a significant difference when compared with the amount of the surviving cells at 3 and 7 days（P＜0.05）.Conclusion Using Interactive Laser Cytometer for measurement of the green and red fluorescent densities of different waves, the activity of the cultured urethral epithelial cells in vitro can be rapidlymeasured with the in situ quantitation method. This method solves a difficult problem of observing the growing cells in an opaque vector. The dynamic growing state of the engineering tissues can be observed.
Objective To investigate the feasibility of repairing goat tibia defect with marrow stromal cells (MSCs).Methods MSCs were cocultured with the bio-derived bone in vitro, and the 20 mm tibia defectswere made and fixed with plate in 35 goats, and they were divided into the experimental group, control group and blank group. The defects on the right side were filled with tissue engineering bone as the experimental group, the defects onthe left side with bio-derived bone as the control group in 33 goats, and the defect on the both sides were not filled with any materials as the blank group in 2 goats. Threpair capability was assessed physically, histopathologically and biomechanically at 2, 4, 6, 8, 12,16 and 24 weeks after operation in 3 groups.Results By physical, histopathological and biomechanical examinations, the bio-derived bone was partially absorbed in the experimental group and was rarely absorbed in the control group in the 4th week; the defects were partially repaired in the experimental group, and in the control group, few new bones were observed in the two ends of the implants, in which there was fibrous tissue. The effects of biomechanics had no statistically significant difference between the experimental group and the control group(P＞0.05) in the 8th week; the defects were perfectly repaired in the experimental group and the effects of biomechanics had statistically significant difference between two groups (P＜0.05) in the 12th weeks. The defects were not repaired in the 24th week in the blank group.Conclusion The tissue engineering bone can efficiently repair bone defect, and itsrepair capability is better than that of bio-derived bone alone both in quantity and in quality of bone formation.
Objective To introduce the experience about thereconstruction of median sternotomy wound dehiscence. Methods From February 2002 to October 2004, 10 patients with median sternotomy wound dehiscence due to coronary artery revascularization were treated. There were 7 males and 3 females, aging from 68 to 76 years. The sizes of defects ranged from3 cm×5 cm to 5 cm×15 cm. After debridement of necrotic soft tissue, sternum and rib, infected median sternotomy wound was reconstructed with rectus abdominis myocutanous flap, pectoralis major myocutanous flap and latissimus dorsi flap or single muscle flap. The sizes of flaps ranged from 3 cm×5 cm to 5 cm×16 cm.Results Allpatients were followed up from 3 to 11 months with anaverage of 6 months. All the patients achieved healing by first intention with normal respiration and normal function of upper limbs. The wound of donor site healed well.No abdominal hernia and other complications occurred. The wound of donor site healed well.The results were satisfactory.Conclusion According to different stages of the disease and different conditions of an operation, the surgical management should vary with each individual.