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Objective:To explore effects of trehalose as a cryopreserve agent on survival rate of fatty tissue after cryopreservation.Methods:The liposuction was used on the abdomen of adult female.After centrifugation and purification,adipose was randomized into the following three groups,the trehalose group,the fetal bovine serum (FBS)+ 10％DMSO group and the physiological saline group.The specimens were cryopreserved at-196 ℃ for 3 months and then the HE staining,glucose transfer method and CK method were used to detect the cell survival rate in each group.Results:The activity of adipose in the trehalose group and FBS+10％DMSO group adipose was higher than that in the physiological saline group (P＜0.05);while there was no significant difference between the trehalose group and FBS+10％DMSO group (P＞0.05).Conclusion:As cryoprotectant,trehalose could keep fat cell viability,and adipose tissue can be used for clinical transplantation after 3 months' freezing.
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Objective:To evaluate the clinical effect of combination of autologous tissue reconstruction of tarsal plate with temporal flap on repair of full-thickness lower eyelid defect.Methods:Eleven patients (11 eyes) underwent hard palate mucosa or ear cartilage combined the emporal flap with the orbicularis oculi muscle to repair full-thickness defect ofpalpebra inferior.Of the 11 patients,6 had more than 75％ eyelid defect area,and 5 had more than 50％ eyelid defect area.Results:All 11 eyes closed completely,with no entropion or ectropion,and returned to normal basically.Postoperative follow-up was performed for 6 months to 5 years,3 years and 4 months on average.The function and form of eyelid remained stable.Infection,leakage or contracture was not found on reconstruction tarsus.Conclusion:Reconstruction of eyelid with autogenous hard palate mucosa or ear cartilage combined the emporal flap with the orbicularis oculi muscle is a simple,convenient and effective method.
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BACKGROUND: The soft tissues would shrink with nasal framework collapse following surgical trauma, which cause aseptic inflammation, lead to parts of cartilage resorption. Accordingly, long-term saddle nose deformity usually accompanied by short nasal columella. Complications such as skin perforation or ulceration would appear if corrected the deformity using medical silicone rubber with large tension.OBJECTIVE: To explore the effectiveness of repairing post-traumatic saddle nose deformity with autologous costal cartilage transplantation.METHODS: A total of 21 cases with post-traumatic saddle nose deformity accompanied by short nasal columella were selected, including 6 males and 15 females, aged 16 45 years. All of the cases had trauma history and agreed with the treatment. The costal cartilage was obtained from the seventh rib and formed to babylon weeping willow leaf shape and columella nasi stent to repair post-traumatic saddle nose deformity. The "V-Y" progradation suture was used in the philtrum introcession and the botton of nasal columella to extend the nasal columella. The recovery of saddle nose deformity after transplantation, discharge of transplanted cartilage, as well as the incision scar status was observed.RESULTS AND CONCLUSION: The results were satisfactory and there were no complications after transplantation. All the cases were followed up from 6 months to 2 years. No case suffered costal cartilage grafts discharge or chondral deformation. The scar was little at the bottom of nasal columella. It is an ideal method for repairing post-traumatic saddle nose deformity using transplantation of autologous costal cartilage with "V-Y" progradation suture.
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Objective To conduct an empirical study for quantifying the anastomosis between two vessels after skin expanded technique by the method of angiography and to provide a precise basis for vascular study in skin flap.Methods Bilateral skin flaps based on deep iliac circumflex vessels were elevated from the abdominal wall including deep superior epigastric vessels.One was expanded at the boundary between two vessels and the other unexpanded.An X-ray image was obtained by carotid arterial injection of gelatin-lead oxide mixture.Three parallel lines with equal intervals perpendicular to long axis of the two vessels were designed at the communication area.Vessel anastomosis quantity was determined by counting the number of marks derived from the intersections of the lines and the vessels and statistical analysis was carried out.Results The marks of intersection in expanded group were more than unexpanded group with statistical significance.Conclusions The method for quantifying vessel anastomosis in skin flap is reliable.The principles of this procedure may also be applied to other experimental and elinical studies.
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BACKGROUND: Compared with normal physiological flap, main advantage of venous skin flap is that it throws off the limitation of arterial vascular territory on donor site and recipient site of traditional axial skin flap. However, its survival rate is unstable. OBJECTIVE: To explore effects of basic fibroblast growth factor (bFGF)-poly(lactic-co-glycolic-acid)] (PLGA)-sustained release microspheres on the survival of rabbit venous flaps.DESIGN, TIME AND SETTING: A randomized controlled animal study was performed at the University of South China from May to October 2008.MATERIALS: A total of 24 healthy New Zealand rabbits were equally randomly assigned to bFGF-PLGA sustained release microsphere, blank microsphere and blank control groups.METHODS: The formulation of bFGF microspheres was optimized by orthogonal design. bFGF-PLGA microspheres were prepared by optimized method. Lateral abdominal wall skin flap was created in rabbits from 3 groups. Five days before operation, 28.85 g/L bFGF-PLGA microspheres 3 mL (containing bFGF 20 μg) was intradermally injected into rabbits from the bFGF-PLGA sustained release microsphere group. An equal volume of blank microsphere + bFGF was injected in the rabbits of the blank microsphere group. Rabbits from the blank control group were infused with the same volume of saline. MAIN OUTCOME MEASURES: Morphology and particle distribution of bFGF-PLGA microspheres, drug loading volume, encapsulation efficiency, in vitro drug release characteristics were measured. After seven days, the survival area of skin was determined. Rabbit skin samples received CD34~+ immunohistochemical staining to detect the expression of CD34~+ and average number of blood vessels. RESULTS: The bFGF microsphere prepared based on optimized formulation exhibited well-defined properties, with the even and uniform sphere in appearance, regular particles without adhesion, about 98% of particles with a size distribution between 12.50 to 43.49 μm, with a mean particle size of 26.93μm and size span of (0.611 ± 6.60). The drug loading volume and encapsulation efficiency of bFGF microsphere reached [(23.11 ±0.44 )x10~3]% and (86.51±0.83)%, respectively. In the burst release phase, the rate of in vitro drug release amounted to 27.78%, but rose to 81.56% accumulatively 30 days later. The in vitro drug release of bFGF microsphere corresponded with Higuichi equation (r= 0.997). The sustained-release microspheres, blank microspheres and normal saline group, the average survival of the flap and the average number of blood vessels were similar (P=0.597, P=0.336), but still significantly lower than the bFGF-PLGA sustained release microsphere group (P=0.000). Results of immunohistochemical staining revealed that bFGF-PLGA promoted blood supple between flap and surroundings, improved flap survival and abundant CD34~+ expression. CONCLUSION: bFGF microsphere with good morphology, high drug loading volume and encapsulation efficiency can be obtained using W/O/W multiple emulsion evaporation method. The bFGF microsphere can promote the survival of rabbit venous flaps through a long period due to sustained release of bFGF.
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BACKGROUND:Compared with normal physiological flap,main advantage of venous skin flap is that it throws off the limitation of arterial vascular territory on donor site and recipient site of traditional axial skin flap.However,its survival rate is unstable.OBJECTIVE:To explore effects of basic fibroblast growth factor (bFGF)-[poly(lactic-co-glycolic-acid)] (PLGA)-sustained release microspheres on the survival of rabbit venous flaps.DESIGN,TIME AND SETTING:A randomized controlled animal study was performed at the University of South China from May to October 2008.MATERIALS:A total of 24 healthy New Zealand rabbits were equally randomly assigned to bFGF-PLGA sustained release microsphere,blank microsphere and blank control groups.METHODS:The formulation of bFGF microspheres was optimized by orthogonal design.bFGF-PLGA microspheres were prepared by optimized method.Lateral abdominal wall skin flap was created in rabbits from 3 groups.Five days before operation,28.85 g/L bFGF-PLGA microspheres 3 mL (containing bFGF 20 ?g) was intradermally injected into rabbits from the bFGF-PLGA sustained release microsphere group.An equal volume of blank microsphere + bFGF was injected in the rabbits of the blank microsphere group.Rabbits from the blank control group were infused with the same volume of saline.MAIN OUTCOME MEASURES:Morphology and particle distribution of bFGF-PLGA microspheres,drug loading volume,encapsulation efficiency,in vitro drug release characteristics were measured.After seven days,the survival area of skin was determined.Rabbit skin samples received CD34+ immunohistochemical staining to detect the expression of CD34+ and average number of blood vessels.RESULTS:The bFGF microsphere prepared based on optimized formulation exhibited well-defined properties,with the even and uniform sphere in appearance,regular particles without adhesion,about 98% of particles with a size distribution between 12.50 to 43.49 ?m,with a mean particle size of 26.93 ?m and size span of (0.611 ? 6.60).The drug loading volume and encapsulation efficiency of bFGF microsphere reached [(23.11?0.44)?10-3]% and (86.51?0.83)%,respectively.In the burst release phase,the rate of in vitro drug release amounted to 27.78%,but rose to 81.56% accumulatively 30 days later.The in vitro drug release of bFGF microsphere corresponded with Higuichi equation (r=0.997).The sustained-release microspheres,blank microspheres and normal saline group,the average survival of the flap and the average number of blood vessels were similar (P=0.597,P=0.336),but still significantly lower than the bFGF-PLGA sustained release microsphere group (P=0.000).Results of immunohistochemical staining revealed that bFGF-PLGA promoted blood supple between flap and surroundings,improved flap survival and abundant CD34+ expression.CONCLUSION:bFGF microsphere with good morphology,high drug loading volume and encapsulation efficiency can be obtained using W/O/W multiple emulsion evaporation method.The bFGF microsphere can promote the survival of rabbit venous flaps through a long period due to sustained release of bFGF.
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BACKGROUND: Alginic sodium diester (ASD) possesses neuroprotective function because of its selective calcium antagonist effects.OBJECTIVE: To compare the influences of ASD on intraneuronal Ca2+content and nerve cell apoptosis before and after reperfusion in focal cerebral ischemic rats.DESIGN: Randomized controlled observation.SETTING: Neurological Department of Xiangya Hospital Affiliated to South China University; Laser Orthopedic Surgery of the First Hospital Affiliated to Southern China University.MATERIALS: This experiment was carried out between November 2003and April 2004 at the Neurological Department of Xiangya Hospital Affiliated to South China University. A total of 65 male SD rats were recruited and randomized into 6 groups; 17 got lost during the experiment, and the other 48 rats completed the experiment with 8 rats in each group.METHODS: In sham operation group, an incision was made on rats' cervical skin and sutured. Right cerebral middle artery was occluded in rats of ischemic group, ASD 5 mg/kg preischemic group, ASD 5 mg/kg postischemic group, ASD 10 mg/kg preischemic group, and ASD 10 mg/kg postischemic group. After that, rats in all but ischemic group were subjected to intraperitoneal injection of various dosage of ASD or excipient 30minutes before reperfusion and 5 hours after reperfusion. FCM was used to determine intraneuronal Ca2+ content and rate of nerve cell apoptosis;meanwhile, neurological dysfunction was scored.MAIN OUTCOME MEASURES: [1] Influence of ASD on the score for neurological dysfunction, intraneuronal Ca2+ fluorescence intensity, and neuronal apoptosis in rats with right cerebral middle artery ischemia. [2]Correlation of behavioral obstacle score with intraneuronal Ca2+ fluorescence intensity and neuronal apoptosis in rats with right cerebral middle artery ischemia.RESULTS: Totally 65 rats were enrolled in this study, 17 of which got lost and the other 48 rats entered the result analysis. [1] Influence of ASD on the score for neurological dysfunction, intraneuronal Ca2+ fluorescence intensity, and neuronal apoptosis in rats with right cerebral middle artery ischemia: The score was obviously reduced in ASD 5 mg/kg preischemic group, ASD 5 mg/kg postischemic group, ASD 10 mg/kg preischemic group and ASD 10 mg/kg postischemic group as compared with ischemic group (1.80±0.21, 2.20±0.23, 1.20±0.11, 2.00±0.22, 3.40±0.65); moreover,functional improvement was more obvious due to pre-reperfusional administration than post-reperfusional administration. Intraneuronal Ca2+ concentration was reduced after ASD administration at different degrees and lower than that of ischemic group. Decrement of intraneuronal Ca2+ concentration was found most obvious due to 10 mg/kg ASD administration 30 minutes before reperfusion, approximately reduced by 70%; moreover, neuronal apoptosis rate on the ischemic side was obviously suppressed by ASD administration, displaying time-dependent manner, with apoptotic suppression effect more obvious in pre-reperfusional group than in post-reperfusional group (5.68%, 10.03%; 4.00%, 9.91%). [2] Correlation of behavioral obstacle score of right cerebral middle artery ischemic rats with intraneuronal Ca2+ fluorescence intensity and membrane associated protein/propidium iodide apoptosis: Obvious positive correlation was found between behavioral obstacle score and intraneuronal Ca2+ fluorescence intensity and detection rate of membrane associated protein/propidium iodide apoptosis (r=0.51,0.62, P ＜ 0.05); intraneuronal Ca2+ fluorescence intensity was also positively correlated with the detection rate of membrane associated protein/propidium iodide apoptosis (r=0.84, P ＜ 0.05).CONCLUSION: [1] ASD can exert anti-apoptosis effect by suppressing the increment of intraneuronal Ca2+ concentration, thus having neuroprotective function and ultimately improving neurological dysfunction. [2] Its effect displays time-dependent manner, and neurological functional improvement is more obvious by pre-reperfusional administration than by post-operational administration.