Objective To construct a replicationdefective recombinant adinovirus including the target gene human bone morphogenetic protein 4(fragment hBMP-4). Methods The hBMP-4 gene fragment was cut down from pCS2(+)/hBMP-4, cloned into the eukaryotic expressive vector pcDNA 3.1(+), then subcloned into pShuttle-CMV and transformed into the competent E. coli BJ5183/p by electroporation. The resulting recombinant plasmid pAdE/hBMP-4 was transformed into the packaging of thecell lines HEK293 to produce the replication-defective recombinant adenovirusescontaining the hBMP-4 gene. These replication-defective recombinant adinoviruses were transfected into HEK293 and HeLa cells. Then, total RNA and total protein were detected by RT-PCR and the Western-blot assay. Results The pAdE/hBMP-4 was confirmed by the restrictional endonuclease digestion. In HEK293 and HeLa cells, the specific transcription of the hBMP-4 gene was confirmed by RT-PCR, and the expression of the hBMP-4 protein was confirmed by theWestern-blot assay. Conclusion The replication-defective recombinant adinovirus expression vector containing the hBMP-4 gene can be constructed and expressed successfully, which has laid a foundation for the further research on the genetherapy of hBMP-4.
OBJECTIVE: To explore the possibility to bridge peripheral nerve defects by xenogeneic acellular nerve basal lamina scaffolds. METHODS: Thirty SD rats were randomly divided into 5 groups; in each group, the left sciatic nerves were bridged respectively by predegenerated or fresh xenogeneic acellular nerve basal lamina scaffolds, autogenous nerve grafting, fresh xenogeneic nerve grafting or without bridging. Two kinds of acellular nerve basal lamina scaffolds, extracted by 3% Triton X-100 and 4% deoxycholate sodium from either fresh rabbit tibial nerves or predegenerated ones for 2 weeks, were transplanted to bridge 15 mm rat sciatic nerve gaps. Six months after the grafting, the recovery of function was evaluated by gait analysis, pinch test, morphological and morphometric analysis. RESULTS: The sciatic nerve function indexes (SFI) were -30.7% +/- 6.8% in rats treated with xenogeneic acellular nerve, -36.2% +/- 9.7% with xenogeneic predegenerated acellular nerve, and -33.9% +/- 11.3% with autograft respectively (P gt; 0.05). The number of regenerative myelinated axons, diameter of myelinated fibers and thickness of myelin sheath in acellular xenograft were satisfactory when compared with that in autograft. Regenerated microfascicles distributed in the center of degenerated and acellular nerve group. The regenerated nerve fibers had normal morphological and structural characters under transmission electron microscope. The number and diameter of myelinated fibers in degenerated accellular nerve group was similar to that of autograft group (P gt; 0.05). Whereas the thickness of myelin sheath in degenerated accellular nerve group was significantly less than that of autograft group (P lt; 0.05). CONCLUSION: The above results indicate that xenogeneic acellular nerve basal lamina scaffolds extracted by chemical procedure can be successfully used to repair nerve defects without any immunosuppressants.
Objective To investigate the effect of simvastatin on inducing endothel ial progenitor cells (EPCs) homing and promoting bone defect repair, and to explore the mechanism of local implanting simvastatin in promoting bone formation. Methods Simvastatin (50 mg) compounded with polylactic acid (PLA, 200 mg) or only PLA (200 mg) was dissolved in acetone (1 mL) to prepare implanted materials (Simvastatin-PLA material, PLA material). EPCs were harvested from bone marrow of 2 male rabbits and cultured with M199; after identified by immunohistochemistry, the cell suspension of EPCs at the 3rd generation (2 × 106 cells/mL) was prepared and transplanted into 12 female rabbits through auricular veins(2 mL). After 3 days, the models of cranial defect with 15 cm diameter were made in the 12 female rabbits. And the defects were repaired with Simvastatin-PLA materials (experimental group, n=6) and PLA materials (control group, n=6), respectively. The bone repair was observed after 8 weeks of operation by gross appearance, X-ray film, and histology; gelatin-ink perfusion and HE staining were used to show the new vessels formation in the defect. Fluorescence in situ hybridization (FISH) was performed to show the EPCs homing at the defect site. Results All experimental animals of 2 groups survived to the end of the experiment. After 8 weeks in experimental group, new bone formation was observed in the bone defect by gross and histology, and an irregular, hyperdense shadow by X-ray film; no similar changes were observed in control group. FISH showed that the male EPC containing Y chromosome was found in the wall of new vessels in the defect of experimental group, while no male EPC containing Y chromosome was found in control group. The percentage of new bone formation in defect area was 91.63% ± 4.07% in experimental group and 59.45% ± 5.43% in control group, showing significant difference (P lt; 0.05). Conclusion Simvastatin can promote bone defect repair, and its mechanism is probably associated with inducing EPCs homing and enhancing vasculogenesis.
Objective To investigate the clinical results of allograft and sural neurovascular flap in repairing calcaneus and skin defects.Methods From February 1996 to December 2002, allograft and sural neurovascular flap were used to repair calcaneusand skin defects in 6 cases. The causes included road accident in 3 cases, strangulation in 2 cases and crashing object in 1 case. The defect locations were at theback of the calcaneus( 1/3, 1/2 and 2/3 of calcaneus in 3 cases, 2 cases and 1case respectively). The flap area ranged from 6 cm×7 cm to 12 cm×17 cm. Results The flaps survived completely in 4 cases; the distal flaps necrosed partly in 2 cases and the wound healed by dressing. The postoperative X-ray films showed that the repaired bone and joint had normal position and the arcus plantaris recovered. After a follow upof 6 months to 3 years all the patients were achieved bone union in allograft and had no complications of absorption, infection and repulsion. The weightbearing and walking functions were restored and the injured foot obtained a satisfactory contour. After 36 months of operation, the sensory recovery of foot occurred. Conclusion The used-allograft iseasy to be obtained and arcus plantaris is easy to recover. The reversesural neurovascular- flap in repairing calcaneus and skin defects has the following advantages: the maintenance of blood supply for injured foot, the less dangerous operation, the simple procedure, the recovery of walking function, and the good appearance and sensation.
OBJECTIVE: To investigate the method and clinical effect of temporal fascia flap, free forearm flap, free iliac bone transfer and immediate implant on reconstruction of maxillary defect. METHODS: From February 1999 to July 2002, 8 cases of maxillary defects due to excision of cancer were repaired immediately with temporal fascia flap, free forearm flap, free iliac bone transfer and implant. Out of 8 patients, there were 6 males and 2 females, aged 32-49 years, with a disease course of 3 months to 2 years. RESULTS: Free iliac bone and forearm flap survived in all 8 cases. Osseo-integration could be seen and the implants could be used for denture repair and chew function. After 6-12 months, X-ray examination showed iliac bone healing; facial shape and functional restoration were satisfactory. CONCLUSION: Temporal fascia flap, free forearm flap, free iliac bone transfer and immediate implant is an ideal method to repair maxillary defect immediately and reconstruct its function.
Objective To explore the clinical effect of latissimus dorsi musculocutaneous flap with a few muscle in repairing the soft tissue defect of lower limbs. Methods From June 2000 to December 2006, 8 patients with soft tissue defects of lower limbs were repaired with the latissimus dorsi musculocutaneous flaps. There were 6 males and 2 females, aged from 2569 years. The locations were heel in 3 cases, dorsum pedis in 2 cases, anticnemion in 2 cases, and the right leg (squamous carcinoma) in 1 case. The area of soft defect ranged from 10 cm×7 cm～18 cm×12 cm. The flap in size ranged from 15 cm×8 cm to 22 cm×15 cm. Results Of all the flaps,6 survived,1 had vascular necrosis 2 hours after operation and survived by skin grafts, 1 had delayed healing because of infection. The wound and donor site achieved primary healing. The followup for 3 to 12 months revealed that all the flaps had a good appearance. The function of donor site was as normal. Conclusion It is an ideal method to repair the softtissue defect of lower limbs with latissimus dorsi musculocutaneous flap.
Objective
To summarize the effectiveness of radial dorsal fasciocutaneous flap of thumb for repairing severe nail bed defects.
Methods
Between May 2009 and January 2012, 16 patients with severe nail bed defect were treated with radial dorsal fasciocutaneous flap of the thumb. There were 10 males and 6 females, aged 16-54 years (mean, 36 years). The causes of injury included crush injury in 10 cases, chainsaw injury in 4 cases, and scald in 2 cases; injured fingers were thumb in 3 cases, index finger in 4 cases, middle finger in 5 cases, ring finger in 3 cases, and little finger in 1 case. The time between injury and operation was 2 hours to 8 days (mean, 19.3 hours). Of 16 patients, 9 complicated by distal phalanx fracture. The area of defect ranged from 0.9 cm × 0.6 cm to 2.3 cm × 2.1 cm. According to ZHOU Qingwen’s grading system for nail bed defects, 6 cases were rated as degree III and 10 cases as degree IV. The area of flap ranged from 1.0 cm × 0.6 cm to 2.5 cm × 2.2 cm. Retrograde transposition was performed to repair the thumb defect, and pedicled transposition to repair the 2nd-5th fingers defects. The donor sites were directly sutured or were repaired with skin graft.
Results
All flaps and skin grafts survived, and wounds healed by first intention. All patients were followed up 6-12 months (mean, 8 months). The color, texture, and contour of the flaps were good. According to total active motion standard, the finger function was assessed as excellent in 10 cases, good in 4 cases, and fair in 2 cases, and the excellent and good rate was 87.5%.
Conclusion
Radial dorsal fasciocutaneous flap of thumb is a reliable flap with easy dissection and less trauma in repair of severe nail bed defects.
From 1984 to 1994, 236 different types of traumatic defects of foot were repaired by microsurgical tissue grafting. They included simple cutaneous flap in 187 and composite flap in 49. Among the 236 different tissue flaps, vascularized flap was used in 97 and pedicled flap in 139. The 4 fore-foot and 6 heel defects were repaired by composite skeleted cutaneous grafts with scapula and vascularized febula respectively. After the follow-up from 1 to 10 years, all the grafted tissues were survived and healed well. The functions were satisfactory, and 186 patients had resumed their original works. The key to good function following repair was to maintaion the integrity of foot structures and the sensation of the sole and heel.
Objective
To investigate the operative method and effectiveness of repairing defects at medial malleolus in children with vascularized fibular head composite flap.
Methods
Between November 2008 and January 2011, 8 children with bone and soft tissue defects at the medial malleolus were treated. There were 5 boys and 3 girls, aged 2-9 years (mean, 4.6 years). Injuries were caused by machine twisting in 2 cases and by wheel twisting in 6 cases. Soft tissue defect area ranged from 3.5 cm × 3.0 cm to 7.0 cm × 4.5 cm; defect was total in all medial malleolus. The disease duration from injury to admission was 2-8 hours (mean, 4.5 hours). Defects were repaired with vascularized fibular head composite flap carrying the skin around the head of the fibula in 5 cases, and with vascularized fibular head composite flap and skin flap above the medial malleolus in 3 cases having too large defect (gt; 5 cm × 4 cm). The donor sites were repaired with direct suture in 2 cases and with skin graft in 6 cases.
Results
All 8 fibular head composite flaps and 3 skin flaps above the medial malleolus survived completely. Wounds healed by first intention; the skin grafts at donor sites survived in the other cases except 1 case having local necrosis, with healing of incision by first intention. The patients were followed up 10 months to 3 years (mean, 22 months). The color and elasticity of the flaps were good. All the children had equal leg length. Of 8 cases, 6 had no joint valgus; 2 cases had progressive ankle varus after 1 year of operation. The ankle flexion and extension function returned to normal in 5 cases, and was slightly limited in 3 cases; horizontal side, forward and backward movements had no difference compared with normal side. According to American Orthopaedic Foot and Ankle Society (AOFAS) ankle function evaluation criteria, the results were excellent in 5 cases, and good in 3 cases at 10 months after operation. X-ray film showed that the ankle hole gap development of both sides was similar; no premature closure of the epiphysis or bone bridge formation of the medial malleolus was observed in 6 cases, and bone bridge formed in 2 cases after 1 year of operation.
Conclusion
The satisfactory short-term effectiveness can be obtained in repairing children medial malleolus and soft tissue defects by vascularized fibular head composite flap, and the reconstructed medial malleolus can develop with the growth of children. Long-term effectiveness still need more follow-up study.
Objective?To investigate the surgical methods and clinical results of repairing soft tissue defects in the thumb with distally-based dorsal thumb neurocutaneous vascular flap.?Methods?From January 2006 to October 2007, 23 patients with soft tissue defect in the thumb were treated, including 20 males and 3 females aged 19-46 years old (average 27.5 years old). The defect was caused by crush injury in 1 case, electric planer accident in 6 cases, incised injury in 8 cases, and avulsion injury in 8 cases. The defect was located on the palmar aspect of the thumb distal phalanx in 3 cases, the dorsal-radial aspect of the thumb distal phalanx in 3 cases, and ulnar or dorsal aspect in 17 cases. The defect size ranged from 3.3 cm × 1.2 cm to 4.2 cm × 1.2 cm. Among them, 18 cases were complicated with distal 1/2 nail bed defect or injury. The time between injury and hospital admission was 1- 72 hours (average 22 hours). During operation, the defect was repaired with distally-based dorsal-radial neurovenocutaneous vascular flap of the thumb in 3 cases and distally-based dorsal-ulnar neurovenocutaneous vascular flap of the thumb in 20 cases. The size of those flaps was 4.0 cm × 1.6 cm-5.0 cm × 3.0 cm. The donor site underwent direct suture or split thickness skin graft repair.?Results?At 10 days after operation, 3 cases suffered from the epidermal necrosis in the distal part of the flap, 2 of them experienced the exfoliation of dark scab 14 days later and the flap survived, and the flap of the rest one survived after dressing change. The other flaps and the skin graft at the donor site all survived uneventfully. The wounds healed by first intention. All the patients were followed up for 10-16 months (average 12.6 months). The flaps were soft in texture and full in appearance. The two-point discrimination value 6 months after operation was 8-10 mm. At 12 months after operation, the growth of the residual fingernail was evident in 18 cases, including 4 cases of curved or hook fingernail. Active flexion and extension of the thumb were normal. The abduction of the first web space reached or surpassed 80 percent of the normal side in 20 cases and was below 80 percent of the normal side in 3 cases. The clinical outcomes were satisfactory in 11 cases, approximately satisfactory in 8 cases, and unsatisfactory in 4 cases according to self-designed evaluation system.?Conclusion?The operative method of repairing the soft tissue defects in the thumb with the distally-based dorsal thumb neurocutaneous vascular flap is simple, stable in anatomy, in line with the principle of proximity, and suitable for repairing thumb tip defect 3 cm in size. It can bring a good postoperative appearance of the thumb and little influence on the hand function.
