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[不同人工真皮支架对猪全层烧伤创面血管化及瘢痕形成的影响]

[Effects of different artificial dermal scaffolds on vascularization and scar formation of wounds in pigs with full-thickness burn].

作者信息

Teng Jian-ying, Guo Rui, Xie Jing, Sun Dong-jie, Shen Ming-qiang, Xu Shao-jun

机构信息

Clinical Medical College of Hangzhou Normal University, Hangzhou, China.

出版信息

Zhonghua Shao Shang Za Zhi. 2012 Feb;28(1):13-8.

Abstract

OBJECTIVE

To investigate the effects of three kinds of artificial dermal scaffolds on vascularization and scar formation of wounds in pigs with full-thickness burn.

METHODS

Eighteen Bama miniature pigs were divided into chitosan scaffold (CS) group, sulfonated carboxymethyl chitosan scaffold (SCCS) group, and acellular dermal matrix (ADM) scaffold group according to the random number table, with 6 pigs in each group. Every pig in all groups was inflicted with 4 or 8 full-thickness scald wounds on the back (totally 96 wounds). Forty-eight hours after injury, eschars of all wounds were excised. Twenty-four wounds in CS group were transplanted with double-layer artificial dermis of collagen-chitosan and silicone rubber, those in SCCS group with double-layer artificial dermis of collagen-sulfonated carboxymethyl chitosan and silicone rubber, and those in ADM scaffold group with ADM. The rest 24 wounds in the three groups were dressed with vaseline gauze as control group. After 2 weeks of treatment, all wounds of every group were covered with skin. In post treatment (scaffold transplantation or gauze covering) week (PTW) 1, 2, 3, and 4, gross condition of wound was observed, and specimens from central parts of wounds were harvested for observation and assessment of vessels or cells with positive expression of CD31, α smooth muscle actin (α-SMA), TGF-β(1) and TGF-β(3) with SP staining. Data were processed with one-way analysis of variance and LSD test.

RESULTS

(1) Degree of vascularization in SCCS group was better than that in the other three groups. (2) The number of vessels with positive expression of CD31 in CS, SCCS, ADM scaffold, and control groups increased gradually from PTW 1 to PTW 3, and decreased in PTW 4. There were statistical differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 24.005, 38.822, 25.274, 3.856, P < 0.05 or P < 0.01). The numbers of vessels that expressed CD31 in SCCS group from PTW 1 to PTW 3 were more than those in the other three groups (with P values all below 0.05). (3) The numbers of vessels that expressed α-SMA in CS, SCCS, and ADM scaffold groups from PTW 1 to PTW 3 showed the similar trend of change to those of vessels that expressed CD31, which increased gradually in control group from PTW 1 to PTW 4. There were obvious differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 22.637, 28.087, 62.651, 18.055, P values all below 0.01). The number of vessels that expressed α-SMA in SCCS group from PTW 1 to PTW 4 was more than that in the other three groups (with P values all below 0.05). (4) From PTW 1 to PTW 4, the number of cells with expression of TGF-β(1) in CS group was respectively (127 ± 8), (167 ± 19), (170 ± 18), (144 ± 10) per 400 times visual field, that in SCCS group was respectively (171 ± 17), (207 ± 25), (130 ± 30), (69 ± 16) per 400 times visual field, that in ADM scaffold group was respectively (106 ± 8), (159 ± 17), (171 ± 11), (145 ± 11) per 400 times visual field, and that in control group was respectively (100 ± 20), (150 ± 18), (200 ± 14), (172 ± 20) per 400 times visual field. There were statistical differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 29.675, 9.503, 13.107, 54.515, P values all below 0.01). Compared with those in SCCS group, the number of cells that expressed TGF-β(1) in the other three groups was decreased in PTW 1, 2 but increased in PTW 3, 4 (with P values all below 0.05). (5) The number of cells that expressed TGF-β(3) in 4 groups increased gradually from PTW 1 to PTW 3, and decreased or increased continually in PTW 4. There were statistical differences among 4 groups from PTW 1 to PTW 4 (with F value respectively 140.612, 945.850, 714.037, 119.147, P values all below 0.01). The number of cells with positive expression of TGF-β(3) in SCCS group from PTW 1 to PTW 4 was more than that in the other three groups (with P values all below 0.05).

CONCLUSIONS

The collagen-sulfonated carboxymethyl chitosan dermal scaffold can rapidly induce growth and maturation of blood vessels during wound healing after burn. It is beneficial for wound repair at early stage with inhibition of scar proliferation.

摘要

目的

探讨三种人工真皮支架对猪全层烧伤创面血管化及瘢痕形成的影响。

方法

将18只巴马小型猪按随机数字表法分为壳聚糖支架(CS)组、磺化羧甲基壳聚糖支架(SCCS)组和脱细胞真皮基质(ADM)支架组,每组6只。各组每只猪背部造成4处或8处全层烫伤创面(共96处创面)。伤后48小时,切除所有创面的焦痂。CS组24处创面移植胶原 - 壳聚糖双层人工真皮及硅橡胶,SCCS组移植胶原 - 磺化羧甲基壳聚糖双层人工真皮及硅橡胶,ADM支架组移植ADM。三组其余24处创面用凡士林纱布包扎作为对照组。治疗2周后,各组所有创面均被皮肤覆盖。在治疗后(支架移植或纱布覆盖)第1、2、3和4周(PTW),观察创面大体情况,取创面中央部位标本,采用SP染色观察和评估CD31、α平滑肌肌动蛋白(α - SMA)、转化生长因子 - β(1)(TGF - β(1))和转化生长因子 - β(3)(TGF - β(3))阳性表达的血管或细胞。数据采用单因素方差分析和LSD检验进行处理。

结果

(1)SCCS组血管化程度优于其他三组。(2)CS组、SCCS组、ADM支架组和对照组中CD31阳性表达血管数量从PTW 1至PTW 3逐渐增加,PTW 4时减少。PTW 1至PTW 4四组间有统计学差异(F值分别为24.005、38.822、25.274、3.856,P < 0.05或P < 0.01)。SCCS组PTW 1至PTW 3表达CD31的血管数量多于其他三组(P值均< 0.05)。(3)CS组、SCCS组和ADM支架组PTW 1至PTW 3表达α - SMA的血管数量变化趋势与表达CD31的血管相似,对照组PTW 1至PTW 4逐渐增加。PTW 1至PTW 4四组间有明显差异(F值分别为22.637、28.087、62.651、18.055,P值均< 0.01)。SCCS组PTW 1至PTW 4表达α - SMA的血管数量多于其他三组(P值均< 0.05)。(4)PTW 1至PTW 4,CS组每400倍视野中表达TGF - β(1)的细胞数分别为(127 ± 8)、(167 ± 19)、(170 ± 18)、(144 ± 10),SCCS组分别为(171 ± 17)、(207 ± 25)、(130 ± 30)、(69 ± 16),ADM支架组分别为(106 ± 8)、(159 ± 17)、(171 ± 11)、(145 ± 11),对照组分别为(100 ± 20)、(150 ± 18)、(200 ± 14)、(172 ± 20)。PTW 1至PTW 4四组间有统计学差异(F值分别为29.675、9.503、13.107、54.515,P值均< 0.01)。与SCCS组相比,其他三组PTW 1、2时表达TGF - β(1)的细胞数减少,PTW 3、4时增加(P值均< 0.05)。(5)四组中表达TGF - β(3)的细胞数从PTW 1至PTW 3逐渐增加,PTW 4时持续减少或增加。PTW 1至PTW 4四组间有统计学差异(F值分别为140.612、945.850、714.037、119.147,P值均< 0.01)。SCCS组PTW 1至PTW 4 TGF - β(3)阳性表达细胞数多于其他三组(P值均< 0.05)。

结论

胶原 - 磺化羧甲基壳聚糖真皮支架可在烧伤创面愈合过程中快速诱导血管生长和成熟,有利于早期创面修复并抑制瘢痕增生。

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