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2003 Vol. 19, No. Z1

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Original Article
Experimental studies on the biocompatibility of spongy PLGA-collagen membrane as a tissue-engineering dermal scaffold
TIAN Jian-guang, BAI Dong-hai, LIU Zhi-guo, TANG Hong-tai, XIA Zhao-fan
2003, 19(Z1): 1-4. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.101
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Objective To evaluate the biocompatibility of spongy PLGA-collagen membrane as a dermal scaffold. Methods Adult SD rats, cultured human keratinocytes and fibroblasts were employed in the study. The in vivo degradation process and the in vitro cytotoxicity and hemolytic response of the PLGA-collagen membrane, and the cell affinity of the PLGA membrane with the human cells were investigate. Results The PLGA-collagen membrane degraded completely in vivo at the 9th week with an acceptable speed, It was found that new tissues gradually grew into the pores of the matrix, and blood vessels could be the matrix at the 3rd week of implantation subcutaneously without obvious inflammatory reaction. The lixivium of the PLGA-collagen membrane did not induce apparent cytotoxicity and hemolytic response, and it could induce rapid adherence and proliferation of fibroblasts and keratinocytes on it. Conclusion The PLGA-collagen membrane posessed good biocompatibility cell affinity, with appropriate degradation speed, These properties seemed to meet the regairemeat of a tissue-engineering dermal scaffold.
Effects of thrombin peptides and thrombin on the release of TGF-β and VEGF from human fibroblasts
HUANG Yue-sheng, YANG Zhong-cheng
2003, 19(Z1): 5-7. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.102
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Objective To explore the effect of thrombin peptide 508 (TP508) and thrombin on the release of TGF-β and VEGF from cultured human fibroblasts( Lu13 ) in vitro. Methods TP508 and thrombin in different concentrations were added to the confluent cultured Lu13 and incubated for 8 to 24 hours. Then the supernatant was obtained by centrifugation, and the TGF-β 1, TGF-β 2 and VEGF contents were measured by ELISA. Results TGF-β 1 release from Lu13 was markedly increased by thrombin, but decreased obviously by TP508 ( P <0.05). A combinedation application of TP508 and thrombin showed a synergistic effect to the release of TGF-β1, from Lu13. The release of TGF-β2 was also increased( P <0.05) after thrombin treatment, while slightly decreased when incubated with 100ug/ml of TP508 ( P > 0.05). VEGF release was also stimulated by thrombin and inhibited by TP508. Conclusion The release of TGF-β 1, TGF-β 2 and VEGF from Lu13 could be stimulated by thrombin, and inhibited by TP508, which might be beneficial to reducing scar formation and enhancing the healing quality of the wound.
A preliminary report of therapeutic effects of topical capsaicin in postburn hypertrophic scar
XU Ming-huo, CHAI Jia-ke, LU Jun-ling, CHEN Bao-ju, SHENG Zhi-yong, WANG Shi-ling
2003, 19(Z1): 8-10. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.103
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Objective To observe the therapeutic effects of capsaicin on hypertrophic scar ( HS). Methods 12 patients with hypertrophic scar within 3 months were randomly selected for a double-blind study. Each patient received two treatments, 0.05% capsaicin cream for one area of HS, cream vehicle for adjacent HS. The scores of pruritus and side effects of capsaicin were evaluated. Skin biopsies were collected, routine pathological examination, immunohistochemical staining for substance P and special staining for collagen fiber were also performed. Results Capsaicin definitely has an anti-pruritus effect on HS, which is correlated with the decline of content of substance P in scar tissue. The color, texture, thickness of hypertrophic scar and distribution of collagen fiber in HS tissue seemed to be similar between two groups. Conclusion Capsaicin is a good anti-pruritus drug for HS with no severe side effect, but it does not inhibit hypertrophy of scars.
A study on the promoting effect of L-arginine on the burn wound healing of the rats with diabetes
GE Kui, LU Shu-liang, QING Chun, XIE Ting, NIU Yi-wen, RONG Liu, WANG Min-jun, LIN Wei-dong, JIN Shu-wen, HUA Lan-nv
2003, 19(Z1): 11-14. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.105
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Objective To explore the effect and possible mechanism of L-arginine on burn wound healing of the rats with diabetie. Methods One hundred and twenty three Spraque-Dawley( SD) rats were enrolled in the study and randomly divided into four groups. i. e. group A (thirty rats as normal control),group B( diabetic control), group C( diabetes with L-arginine administration) and group D( diabetes with L-glycine administration). Diabetes was induced in rats in B, C and D groups with streptozotocin ( STZ), and they were subjected to 20% TBSA of deep partial thickness scalding on the back 8 weeks later. The percentage of healing areas, the histological and morphological changes, the levels of TGF-b1 released from wound site, the content of hydroxyproline and the glucose content in local wound tissues were observed and measured on the 0, 1 , 3 , 7, 14, and 21 post-burn days ( PBD). Results With the supplementation of L-arginine , the inflammatory reaction , the shedding of necrotic tissue and the advancement of the epithelial cells in the wound tissue appeared earlier and more markedly, with glucose content in the skin decreased and OHP and TGF-β1, contents and the percentage of healing areas increased, compared with those in the group B( P< 0.05). Conclusion The healing of burn wounds in diabetic rats could be promoted by the use of L-arginine by reducing the glucose content in the skin and increasing TGF-β1, synthesis and release.
Expression of microtubule destabilizer-oncoprotein18/stathmin gene in the hypertrophic scar before and after the healing of the deep partial thickness burn wounds
YAO Qing-jun, JIA Chi-yu, CHEN Bi, LU Ning, XU Ming-da, LIU Xin-ping, YAO LI-bo, WANG Shu-sen
2003, 19(Z1): 15-17. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.107
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Objective To study the expression of microtubule destabilizer-oncoprotein18/stathmin (Op18) gene in the hypertrophic scar before and after the healing of the deep partial thickness burn wound. Methods Total RNA was extracted from the hypertrophic scar of the patients with deep partial thickness burn at different time before and after the wound healing. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of Op18 gene with GAPDH as an internal index. Results The expression of Op18 gene in the hypertrophic scar tissue decreased significantly before wound healing, but increased rapidly after wound healing when compared with that in normal tissue( P <0.05). A sleady over-expression of Op18 gene was found in the hypertrophic scar tissue during 4 to 32 months after wound healing. Conclusion The lower expression of Op18 gene during early postburn stage met the need of cell proliferation and wound tissue repair. The scar hypertrophy could be inhibited by the over-expression of Op18gene in the hypertrophic scar tissue.
Effects of Ulinastatin on the intestinal mucosal barrier of the mini - pigs after severe burn injury
HU Xiaohua, GE Yan-ling, XIN Dong, LI Hai-bo, ZHANG Hui-ying, SUN Yong-hua
2003, 19(Z1): 18-21. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.108
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Objective To explore the protective effect of a human proteinkinase inhibitor - Ulinastatin ( UTI) on the prevention of inflammatory mediators oxidative free radical production and changes in intestinal mucosa permeability in mini - pigs at early postburn stages. Methods Twelve Guizhou male mini - pigs were randomly divided into A (control, n =6) and B( with UTI treatment, n = 6) groups. The mini-pigs were inflicted with 35% TBSA full thickness burn on the back and received fluid resuscitation immediately after injury. UTI(5 000 U/kg, 3 times a day) were administered to group B one hour after injury, while an equal amount of normal saline were administered to group A untill the pigs were sacrified. Blood samples were harvested for the determination of serum levels of TNFα , IL-6 , MDA , SOD, DAO, D-lactate at pre- (as normal control) and 6,24,48 ,72 postburn hours( PBHs). Results The serum levels of TNFα and IL-6 in each group at 6 PBH were increased dramatically, especially in group A, peaked at 24 PBH, and seemed to decrease thereafter. The serum levels of MDA and the SOD consumption in group A at 6 - 72 PBH increased obviously than that in normal control. The MDA content in each time points after burn in group B decreased and the SOD consumption increased dramatically when compared with those in group A( P <0.05). The serum DAO activity and the D - lactate content of the rats were significantly increased after burns with barrier function of the intestinal mucosa being severely damaged. The intestinal functions of the rats in group B was markedly improved with functional enzymes obviously decreased. Conclusion UTI can significantly lessen the production of inflammatory.
Observation of the effect of hyperoxic sodium chloride solution on hypoxic injury
SUN Yong-hua, HU Xiaohua, CHEN Zhong, CHENG An-yuan, ZHONG Sui-hang
2003, 19(Z1): 22-26. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.110
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Objective To observe and explore the effects of compound hyperoxic sodium chloride solution on the management of burn shock. Methods (1) Wistar rats were randomly divided into 6 groups, i. e. A group (normal rats as control), B and C groups(the rats of 30% TBSA full thickness burn with compound sodium chloride solution at 1 and 6 postburn hours ( PBHs) respectively) ; D and E groups (with hyperoxic sodium chloride solution at 1 and 6 PBHs respectively), and F group( without treatment). The changes in the serum contents of LPS, IL-6, DAO, D-lactate and M DA in the rats of each group were observed dynamically. (2) The burn patients admitted within 3 PBHs with 50% -69% TBSA burns were enrolled in the study and were randomly divided into the treatment (with hyperoxic sodium chloride solution) and the control (with sodium chloride solution) groups, The changes during shock stage, the postburn complications and the changes in the PtcO2 and the serum contents of Hb, Hct were observed. Results All indices examined in B, C, D and E groups were evidently higher than those in A group in a progressive manner, especially in F group. The sequence was as follows : F group > C group > B group > E group > D group ( P < 0.05). The patients in treatment group could undergo the shock stage more steadily with decreased ammount of fluid infusion and increased PO2 and less complications when compared with those in control group. But there was no obvious difference of the content of blood oxygen saturation between the two groups. Conclusion Application of hyperoxic sodium chloride solution during the early postburn stage could be beneficial to the management of burn shock.
Experimental study on the proportion of three nutrients in burn rats with enteral nutrition
ZANG Jian, LU Shu-liang, LIAO Zhen-jiang, JIN Shu-wen, WANG Xin, IKEDA Shigeo, SAITO Hideaki, SHI Ji-xiang
2003, 19(Z1): 27-30. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.112
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Objective To observe the effect of alteration of proportion of carbohydrate, protein and fat intake on the metabolic level, immunological function and healing of the burn wound. Methods Twenty-one SD rats were inflicted with 30% TBSA scalding on the back after the gastrostomy was performed on them. The rats were divided into three groups: i. e. group A (with low-fat and low-protein diet) , group B (with high-fat and low-protein diet) and group C ( with low-fat and high-protein diet) . The intake caloric value was set at 732 kJ • kg -1 • d -1. Equal amounts of calories and volume for three groups were given in aconstant speed evenly with a pump within 24 hours through the tube inserted through the gastrostomy stoma. The response of splenic lymphocyte to ConA, the level of PGE2, IL-2, serum albumin, plasma transfferin, glucagon, and cortisol in blood, the content of hydroxyproline and the ratio of type I to type III collagen in burn wounds, and the nitrogen content in the liver and the jejunal mucosa were measured ten days after injury. Results In group C, the level of serum albumin, the nitrogen content of the liver and the secretion of PGE2 from peritoneal macrophages were significantly higher than those in group A and B. The level of IL-2in plasma and the content of hydroxyproline in burn wound of Group C was also significantly higher than that in group A, while the ratio of type 1 to type Ⅲ collagen in group B and C was significantly lower than that in group A. There were no significant differences among the three groups in the levels of plasma cortisol, glucagon and urine vanillylmandelic acid( VAM). Conclusion The low-fat and high-protein diet might be beneficial to the synthesis of protein, immunological function and the burn wound healing.
Preliminary observation of the effect of hyperoxic Ringer's solution on the patients with inhalation injury at early post-burn stage
JIA Chi-yu, HUANG Li-feng, CHEN Jian
2003, 19(Z1): 31-32. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.114
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Objective To investigate the clinical application of hyperoxic Ringers solution in the treatment of the burn patients with inhalation injury at the early postburn stage. Methods Eight patients with over 30% TBSA and moderate inhalation injury were randomly enrolled in the study ( + ), while other 4 cases with similar conditions as control( - ). Intravenous infusion of 1500ml hyperoxic Ringers solution were given to the patients every day and for 7 days in the former group, while equal amount of Ringer's solution was given to the patients in the control group. The vital signs, the blood biochemical indices and the blood gas analysis of the two groups were monitored dynamically. Results The vital signs of the patients in group T were steady with slight changes in the blood biochemical indices when compared with those in group C, while the level in PaO2 was significantly higher than that in group C( P <0.05). Conclusion The infusion of hyperoxic Ringer's solution is beneficial in the early treatment of burn patients with inhalation injury.
The therapeutic effect of early enteral nutrition togather with recombinant human growth hormone on severely scalded rats
GUO Guang-hua, WU Chou-lang, LI Guo-hui, YU Yu-rong, Cai Chen
2003, 19(Z1): 33-36. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.115
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Objective To evaluak the therapeutic effects of early enteral nutrition ( EEN ) togather with recombinant human growth hormone ( rhGH) on severely scalded rats. Methods One hundred and thirty Wistar rats were randomly divided into four groups, i. e. normal ( group N , n = 10) ,control( group C,n =40) , early enteral nutrition(group EEN , n =40) and early enteral nutrition with rhGH ( group EEN + rhGH ,n =40). The rats in C, EEN and EEN + rhGH groups were subgected to 30% TBSA Ⅲ degree scald on the back. The changes in body weight, hemoglobin, white blood cell count, total protein, albumin, morphology of ileal mucosa, serum levels of endotoxin and TNFα were observed on 1 ,3,5,9 postburn day ( PBD). Results (1 ) The rat body weight in C group decreased significantly ( P <0.05~0.01 ), while that in EEN+ rhGH group recovered more rapidly than that in other groups. ( 2 ) The serum endotoxin levels in EEN and EEN + rhGH groups were lower than that in C group( P <0.01), while that in EEN + rhGH groups was slightly lower than that in EEN group. (3) The serum TNFa levels in EEN group and EEN + rhGH group were lower than that in C group ( P <0.01) , while that in EEN + rhGH group was lower than that in EEN group on 3 and 9 postburn days( PBDs) ( P <0.05 ~0.01) . (4) The serum levels of total protein in EEN+ rhGH group was higher than that in EEN group on 9 PBD( P < 0.05). (5) The serum levels of albumin and blood hemoglobin levels exhibited no obvious differences among all the groups. (6) Intestinal mucosal injury and necrosis were only observed in C group. Imflammatory reaction was observed in EEN group, and the villous height in EEN + rhGH group was obviously higher than that in the other groups. Conclusion The structure and function of intestine in severely scalded rats could be maintained and the hypermetabolism reaction could be alleviated by early enteral nutrition, and EEN togather with rhGH seemed fohave better resulls than that of EEN alone.
A reprot of 2-phase clinical trioe of Bing Shi Yu Shang ointment for the treatment of partial thickness burn -
HUANG Yue-sheng, SUN Yong-hua, XU Ming-da, ZHANG Zhen-xin, LI Xue-cheng, WANG Zhang-yang, PEN Yi-zhi, JIA Chi-yu, LUO Qi-zhi
2003, 19(Z1): 37-39. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.116
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Objective To observe the clinical effect and the safety of the Bing Shi Yu Shang ointment on the patients with partial thickness burn and residual wounds. Methods Two hundred patients were enrolled in the study and randomly divided into C ( 100 cases) and T groups( 100 cases) by the paralleled and single placebo methods. The patients in group C were treated with Jing Wan Hong ointment, while those in T group were treated with Bing Shi Yu Shang ointment and the indices reflecting the effect and safety of the latter were observed. Results (1) The healing rate and times of healing : the healing rate and time of healing of the partial thickness burn in group T were 67.35% on the 7 PBD and 10.08 PBD respectively, while those in group C were 45.01% and 12.66 PBD respectively. (2) The healing rate on 15 PBD and time of the patients in group T with the partial thickness burn were 67.35% and 10.08PBD respectively, while those in group C were 70.90% and 18.81 PBD respectively. The average healing time of the patients in group T and C were 12.53 and 15.11 PBD respectively, and no toxic and side effect were found in the group T. Conclusion The application of Bing Shi Yu Shang ointment for the treatment of burn wounds of the patients with partial thickness burn was safe and effective.
The effects of NO inhalation on cardice function in the burn patients with inhalation injury
Qi Shun-zhen, ZHAO Hong-liang, ZHAO Lian-kui, SUN Wei-hai
2003, 19(Z1): 40-43. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.117
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Objective To observe the effects of nitric oxide ( NO) inhalation on the cardiac function of the burn patients with inhalation injury. Methods Twelve burn patients with moderate degree of inhalation injury were enrolled in the study and randomly divided into control group ( C, six cases with conventional therapy) and treatment group( T, six cases with conventional therapy and NO inhalation of 10 ppm). Swan-Ganz catheters were placed in all of the patients. The changes in the indices of the cardiac function and plasma contents of endothelin (ET) and NO in the patients in both groups were measured before treatment time and 6, 12, 24, 48 and 72 hours after treatment ( HAT). The changes in the plasma contents of ET and NO in the T group were analyzed correlatively. Results Both the cardiac output (CO) and stroke volume (SV) in the T group were significantly higher than the normal values at 24 HAT ( P < 0.05 , P < 0.01),and the ineredse occurred earlier than those in control group ( P < 0.05). The improvement in LVSWI and RVSWI in T group was observed earlier than those in C group. There was evident difference of RVSWI between T and C groups( P <0.01). The plasma content of ET decreased significantly, while the NO content increased dramatically in T group compared with those in C group ( P <0.01). Conclusion The cardiac function of the patients with inhalation injury could be improved by NO inhalation of 10ppm, which might be related to the regulation of the tension of the pulmonary vascular smooth muscle by NO.
Expression of some active peptides and extracellular matrix in the in vitro cultured tissue engineering skin
XIAO Shi-chu, XIA Zhao-fan, YANG Jun, LIU Zhi-guo, BEN Dao-feng, YU Wei-rong, YANG Yong, ZHU Shi-hui
2003, 19(Z1): 44-46. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.119
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Objective To investigate the bioactivity in the in vitro cultured tissue engineering skin. Methods Epithelial cells and fibroblasts were seeded onto the surface of an acellular dermal matrix to form a skin substitute, and the culture supernatants were collected on the 3rd and the 7th day. The changes in the content of interleukin-6 and interleukin-8 (IL-6,8) and transforming growth factor 1 (TGF-β1) in the supernatants were measured by enzyme linked immunosorbent assay ( ELISA), while the changes in the content of laminin, hyaluronic acid, type Ⅰ and type Ⅲ collagen were quantified by radioactive immunoassay (RIA). Results A certain amounts of cytokines, growth factors and extracellular matrix could be detected in the supernatant of the skin substitute, and the contents of these substances on the 7th day were significantly higher than those on the 3rd day( P <0.01) . The contents of type I and type Ⅲ collagen in the skin substitute containing both epidermis and fibroblast were dramatically lower compared with those in the skin substitute only containing fibroblast( P <0.01). Conclusion The in vitro cultured skin substitute possessed relatively powerful bioactivity, and the fibroblast function could be modulated to some extent by seeding epidermis on it.
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