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Objective@#To investigate the effect of local autologous dermal fibroblasts transplantation on hypertrophic scar formation and wound healing quality in early scar formation. To explore the feasibility of fibroblasts for prevention and treatment of hypertrophic scar.@*Methods@#Dermal fibroblasts were isolated from the dorsal skin tissue of New Zealand white rabbits by mechanical method combined with enzyme digestion. Passage 3 cells were induced to differentiate into osteoblasts and adipocytes. The complete epithelialization time and hypertrophic scar formation after full-thickness skin defect were confirmed by pre-experiment study. In the experiment, 6 rabbits were used, left ear as experimental group and right ear as control group. In the experimental group, the passage 4 dermal fibroblasts labeled with 5-bromodeoxyuridine (5-BrdU) were injected subcutaneously around the wound and hypertrophic scar on 20 d (day 2 after epithelialization) and 30 d (most obvious scar hyperplasia) after surgery. As a control group, physiological saline was injected following the same protocol. On 37 days after surgery, the hypertrophic scar tissue were harvested and assessed by gross view and histological examination. The transplanted cells were detected by immunofluorescence staining, transforming growth factor beta 1 (TGF-β1) and decorin(DCN) mRNA expression were assayed by real-time fluorescence polymerase chain reaction (RT-PCR), and the protein expression of TGF-β1、DCN、Collagen type Ⅰ and type Ⅲ were tested by enzyme linked immunosorbent assay(ELISA).@*Results@#Compared with the control group, the scar in the experimental group was flatter and softer, the color was slightly lighter, and the volume was reduced. The histological results showed that compared with the control group, the number of fibroblasts and inflammatory cells in the superficial dermis was reduced, the proliferation of connective tissue and collagen deposition were reduced, and the basal cells and collagen fibers were arranged in order in the experimental group. The results of RT-PCR showed that TGF-β1 mRNA expression level in the hypertrophic scar tissue reduced significantly and DCN increased significantly in the experimental group, compared with the control group (P<0.01). ELISA results showed that TGF-β1, type Ⅰ and Ⅲ collagen contents in hypertrophic scar tissue were significantly lower than that in the control group, while the DCN expression was significantly increased (P<0.01).@*Conclusions@#The rabbit dorsal dermal fibroblasts significantly inhibited the hypertrophic scar in rabbit ears. The mechanism may be related to the up-regulation of DCN expression and down-regulation of TGF-β1 expression, reduction of type Ⅰ and Ⅲ collagen synthesis and extracellular matrix deposition, in the hypertrophic scar tissue.
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Objective To establish a stable experimental model of vascularized composite allograft (VCA),which would facilitate us to study of the reaction and intervening measure regarding rejection reaction in the future.Methods From September,2016 to July,2017,30 healthy male New Zealand rabbits,weighted 2.5-3.0 kg each,were chosen.Their ears should be intact without defect or necrosis.All of them were randomly and eaqually divided into 2 groups:transverse amputated group and V-shaped amputated group.In situ ear replantation after the amputation was performed.Histology analysis of skin and cartilage were done through HE and TUNEL staining,in order to compare vital rate of these ears.Results Thirty rabbits underwent ear replantation,including 13 via transverse incision and 17 via V-shaped incision.In transverse group,no ear survived,and some of them encountered vein crisis gradually after operation.The survival time ranged from 1 day to 10 days.There were 2 ears survived in V-shaped group.From HE staining,it was found certain vacuolar degenerated cells within skin and cartilage in failure ears.The rates of cell necrosis and apoptosis were higher than the survived ears.Conclusion Rabbit ear replantation model is viable.However,the rabbit ear replantation model is not suitable to be used in large samples.
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Objective To study whether ferulic acid can promote healing on chronic ischemic wounds and its possible mechanisms.Methods 40 male New Zealand white rabbits were randomly divided into 4 groups:vaseline group,ischemic control group,5％ ferulic acid group and recombinant bovine basic fibroblast growth factor for external use (rb-bFGF) group.Gross wounds were carefully observed and HE staining was used to observe the wound healing and immumohistochemical staining to observe the expression of the VEGF and CD31.The RNA was extracted to detect the expression of VEGF and HIF-1a by real-time PCR.Results The general observation and the HE staining of each specimen 11 days after operation all indicated that the duration of wound healing of the 5 ％ ferulic acid group was similar to that of the rb-bFGF group and markedly shorter than the ischemic control group and the vaseline smear group.The result of the immunohistochemical staining indicated that the content of the VEGF and CD31 expression of the 5 ％ ferulic acid groups and the rb-bFGF group made lit tle difference,but there was markedly less VEGF and CD31 in ischemic control group and the vaseline smear group.The result of the PCR showed that expression level of VEGF and HIF-1α in the 5 ％ fer ulic acid group was similar to that in the rb bFGF group and the vaseline smear group,but was obviously more than that of the ischemic control group and the vaseline smear group (P ＜ 0.05).Conclusions Ferulic acid can promote angiogenesis by increasing VEGF and HIF-1α which are closely related to angiogenesis and then promote the healing of chronic wounds.
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Objective To investigate the feasibility and effectiveness of focused ultrasound on the acne of rabbit ear.Methods Eighteen male rabbits were randomly divided into blank control group,model control group and ultrasound irradiation group.Acne rabbit ear model was established by Kligman's method in model control group and ultrasound irradiation group.Then,continuous ultrasonic irradiation was given in ultrasound irradiation group.Pathological changes were observed at the time of before and after ultrasound irradiation and 14 days after ultrasound irradiation.Results After modeling,hair follicle expansion,hyperkeratosis,acupoint hypertrophy were observed in blank control group.Compared with blank control group,the thickness of horn layer,the sebaceous gland diameter and other index in model controlgroup were statistically significant (P＜0.01);After irradiation,there were hyperemia and edema in epidermis,angiotelectasis and inflammatory infiltrating,but coagulative necrosis in slice was not observed,14 days later,hair follicle mild expansion,mild keratosis,spiny thinning were observed in ultrasound irradiation group,when compared with model control group,the thickness of horn layer,sebaceous gland diameter and other index were decreased in ultrasound irradiation group,and the differences were statistically significant(P＜0.01).Conclusion The ultrasonic focusing treatment on rabbit ear acne is feasible and effective.
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Objective To observe the morphological changes by different electron beam radiation methods and different triamcinolone acetonide treatments on the role of hypertrophic scar of rabbit ear,and to explore the optimal treatment for the rabbit ear scar.Methods The ventral rabbit ear hypertrophic scar model was established,and intervention began at 28 days after surgery.The animals were randomly divided into control group (group A),saline group (group B),surgery plus saline group (group C),triamcinolone acetonide group (group D),surgery plus triamcinolone group (group E),and electron beam group (group F),2 days after surgical electron beam group (group G),and 7 days after surgical electron beam group (group H).HE stain,VG stain,toluidine blue stain,CD34 and Alpha-SMA immunohistochemical staining were performed,and comparied on histomorphology of each group.Results Collagen fibers observed showed that there was no statistically significance between groups A and B,groups D and G and groups F and H (P＞0.05),and the difference was statistically significant between other groups (P＜0.05).Mast cells were not typical by toluidine blue staining.The microvascular observation showed there was no statistically significance between groups A and B,groups F and H,and groups G and H (P＞0.05),and the difference was statistically significant between other groups (P＜0.05).Myofibroblasts showed no statistically significance between the groups A and B (P＞0.05),but the difference was statistically significant between other groups (P＜0.05).Conclusions Compared with electron beam radiation methods,triamcinolone acetonide injection is a better treatment.Early postoperative radiotherapy is better than late.Triamcinolone acetonide or electron beam radiation combined with surgery is better than simple treatment.
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OBJECTIVE: To explore the therapeutic effect of compound Lingzhi cream for mild frostbite on rabbit ears. METHODS: Rabbit ears were immersed into alcoholic freezing chamber at -24.5°C to cause mild frostbite, then the therapeutic effect of compound Lingzhi cream of different formula was explored after smearing the cream on the injured site, using appearance change, swelling and distension degree, survival area and pathological change of the ears as the observation index. RESULTS: Compound Lingzhi cream formula 2 was effective on mild frostbite of rabbit ears to some extend, but the effect was not as obvious as those of compound Lingzhi cream formula 3 and 4. CONCLUSION: The effective dosage of compound Lingzhi cream on mild frostbite of rabbit ears is 0.5 g crude Lucid Ganoderma and 0.25 g crude Cowherb Seed in 1 g cream. The optimal dosage is 1 g cream containing 1 g crude Lucid Ganoderma and 0.5 g crude Cowherb Seed. The therapeutic effect would not further increase when increasing the drug dosage. Copyright 2012 by the Chinese Pharmaceutical Association.
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Objective To investigate a medicine which can inhibit the expression of connective tissue growth factor (CTGF) in hypertrophic scar of rabbit ears. Methods 24 bigears white rabbits were used to establish a model of hypertrophic scar of rabbit ears, which was randomly divided into four groups. The hypertrophic scar was injected intralesionally with antisense CTGF (group A), betamethason (group B), triamcinolone acetonide (group C) and physiological saline (group D). Some scar tissue samples were sectioned in every group when the scar was treated after 7, 14, 30, and 60 days, respectively. The expression of CTGF mRNA in the scar was assessed by in situ hybridization and hematoxylin and eosin stain (HE stain) in every samples. Results The expression of CTGF mRNA and the counting of fibroblasts decreased in group A, which showed statistical difference as compared with groups B, C and D. Conclusions The results suggesz that antisense CTGF is able to inhibit the proliferation process of hypertrophic scar in rabbit ears and to remarkably decrease the degree of fiborsis in the scar.
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Objective To observe the novel endothelial like cells of the marginal ear vein intima of rabbit by scanning electron microscope.Methods The marginal ear veins were obtained from male Japanese big ear white rabbits,and prepared for scanning electron microscope(SEM).The novel endothelial like cells of the marginal ear vein intima were observed.Results Between endothelial cells and basement membrane of the marginal ear vein intima of rabbit,some oval-shaped endothelial like cells were found.This kind of cells was measured,the length was(8.32?1.04)?m,width was(5.79?0.68)?m,and length/width is 1.45?0.22.The cells were connected each other by some filament structure,and the average length between two adjacent cells was(6.41?2.45)?m.Conclusion There are novel endothelial like cells in the marginal ear vein intima of rabbit.The filament structure,connected between endothelial like cells,might be a new type of cell junction.