烧伤%皮肤,人工%新生血管化,生理性%瘢痕%真皮支架
燒傷%皮膚,人工%新生血管化,生理性%瘢痕%真皮支架
소상%피부,인공%신생혈관화,생이성%반흔%진피지가
Burns%Skin,artificial%Neovascularization,physiologic%Cicatrix%Dermal scaffold
目的 了解移植3种人工真皮支架对Ⅲ度烧伤创面血管化及其瘢痕形成的影响.方法 采用随机数字表法将18只广西巴马小型猪分成壳聚糖支架组、磺化羧甲基壳聚糖支架组、ADM支架组,每组6只,在每只猪背部制作Ⅲ度烫伤(下称烧伤)创面4或8个,共96个.伤后48 h切痂后,前2组创面分别移植胶原-壳聚糖/硅橡胶双层人工皮、胶原-磺化羧甲基壳聚糖/硅橡胶双层人工皮,ADM支架组创面移植ADM,每组创面24个；剩余24个创面作为油纱对照组,直接覆盖油纱.于支架移植或油纱覆盖后l、2、3、4周(“4周”指植入支架或覆盖油纱后2周+移植表皮后2周),观察各组创面的大体情况；取创面中央部位组织,采用生物素-链霉亲和素-过氧化物酶染色法观察CD31、α平滑肌肌动蛋白(α-SMA)、TGF-β1、TGF-β3表达阳性的血管或细胞,并计数.对数据行单因素方差分析和LSD检验. 结果(1)大体观察可见,磺化羧甲基壳聚糖支架组创面血管化程度明显优于其余3组.(2)移植或覆盖后1～3周各组创面组织CD31阳性血管数持续增多,第4周时3个支架组均减少.各时相点4组间比较,F值分别为24.005、38.822、25.274、3.856,P＜0.05或P＜0.01；磺化羧甲基壳聚糖支架组l～3周明显多于其余3组(P值均小于0.05).(3)移植后1～3周,3个支架组创面组织α-SMA阳性血管数均持续增多,4周时下降；油纱对照组覆盖后1～4周持续增多.各时相点4组间比较,F值分别为22 637、28.087、62.651、18.055,P值均小于0.01.磺化羧甲基壳聚糖支架组各时相点均明显多于其余3组(P值均小于0.05).(4)移植或覆盖后1～4周每400倍视野下TGF-β1阳性细胞数:壳聚糖支架组分别为(127±8)、(167±19)、(170±18)、(144±10)个,磺化羧甲基壳聚糖支架组为(171±17)、(207±25)、(130±30)、(69±16)个,ADM支架组为(106±8)、(159±17)、(171±11)、(145±11)个,油纱对照组为(100±20)、(150±18)、(200±14)、(172±20)个；各时相点4组间比较,F值分别为29 675、9.503、13 107、54.515,P值均小于0.01.1、2周时磺化羧甲基壳聚糖支架组明显高于其余3组(P值均小于0.05),3、4周时其表达量下降,明显低于其余3组(P值均小于0 05).(5)移植或覆盖后1～3周,各组创面组织每400倍视野下TGF-β3阳性细胞数持续增多,4周时下降或继续升高；各时相点4组间比较,F值分别为140.612、945 850、714.037、119.147,P值均小于0.01.磺化羧甲基壳聚糖支架组各时相点明显高于其余3组(P值均小于0 05). 结论 胶原-磺化羧甲基壳聚糖真皮支架在烧伤创面愈合过程中可快速诱导血管生成和成熟,有利于创面早期修复和抑制瘢痕增生.
目的 瞭解移植3種人工真皮支架對Ⅲ度燒傷創麵血管化及其瘢痕形成的影響.方法 採用隨機數字錶法將18隻廣西巴馬小型豬分成殼聚糖支架組、磺化羧甲基殼聚糖支架組、ADM支架組,每組6隻,在每隻豬揹部製作Ⅲ度燙傷(下稱燒傷)創麵4或8箇,共96箇.傷後48 h切痂後,前2組創麵分彆移植膠原-殼聚糖/硅橡膠雙層人工皮、膠原-磺化羧甲基殼聚糖/硅橡膠雙層人工皮,ADM支架組創麵移植ADM,每組創麵24箇；剩餘24箇創麵作為油紗對照組,直接覆蓋油紗.于支架移植或油紗覆蓋後l、2、3、4週(“4週”指植入支架或覆蓋油紗後2週+移植錶皮後2週),觀察各組創麵的大體情況；取創麵中央部位組織,採用生物素-鏈黴親和素-過氧化物酶染色法觀察CD31、α平滑肌肌動蛋白(α-SMA)、TGF-β1、TGF-β3錶達暘性的血管或細胞,併計數.對數據行單因素方差分析和LSD檢驗. 結果(1)大體觀察可見,磺化羧甲基殼聚糖支架組創麵血管化程度明顯優于其餘3組.(2)移植或覆蓋後1～3週各組創麵組織CD31暘性血管數持續增多,第4週時3箇支架組均減少.各時相點4組間比較,F值分彆為24.005、38.822、25.274、3.856,P＜0.05或P＜0.01；磺化羧甲基殼聚糖支架組l～3週明顯多于其餘3組(P值均小于0.05).(3)移植後1～3週,3箇支架組創麵組織α-SMA暘性血管數均持續增多,4週時下降；油紗對照組覆蓋後1～4週持續增多.各時相點4組間比較,F值分彆為22 637、28.087、62.651、18.055,P值均小于0.01.磺化羧甲基殼聚糖支架組各時相點均明顯多于其餘3組(P值均小于0.05).(4)移植或覆蓋後1～4週每400倍視野下TGF-β1暘性細胞數:殼聚糖支架組分彆為(127±8)、(167±19)、(170±18)、(144±10)箇,磺化羧甲基殼聚糖支架組為(171±17)、(207±25)、(130±30)、(69±16)箇,ADM支架組為(106±8)、(159±17)、(171±11)、(145±11)箇,油紗對照組為(100±20)、(150±18)、(200±14)、(172±20)箇；各時相點4組間比較,F值分彆為29 675、9.503、13 107、54.515,P值均小于0.01.1、2週時磺化羧甲基殼聚糖支架組明顯高于其餘3組(P值均小于0.05),3、4週時其錶達量下降,明顯低于其餘3組(P值均小于0 05).(5)移植或覆蓋後1～3週,各組創麵組織每400倍視野下TGF-β3暘性細胞數持續增多,4週時下降或繼續升高；各時相點4組間比較,F值分彆為140.612、945 850、714.037、119.147,P值均小于0.01.磺化羧甲基殼聚糖支架組各時相點明顯高于其餘3組(P值均小于0 05). 結論 膠原-磺化羧甲基殼聚糖真皮支架在燒傷創麵愈閤過程中可快速誘導血管生成和成熟,有利于創麵早期脩複和抑製瘢痕增生.
목적 료해이식3충인공진피지가대Ⅲ도소상창면혈관화급기반흔형성적영향.방법 채용수궤수자표법장18지엄서파마소형저분성각취당지가조、광화최갑기각취당지가조、ADM지가조,매조6지,재매지저배부제작Ⅲ도탕상(하칭소상)창면4혹8개,공96개.상후48 h절가후,전2조창면분별이식효원-각취당/규상효쌍층인공피、효원-광화최갑기각취당/규상효쌍층인공피,ADM지가조창면이식ADM,매조창면24개；잉여24개창면작위유사대조조,직접복개유사.우지가이식혹유사복개후l、2、3、4주(“4주”지식입지가혹복개유사후2주+이식표피후2주),관찰각조창면적대체정황；취창면중앙부위조직,채용생물소-련매친화소-과양화물매염색법관찰CD31、α평활기기동단백(α-SMA)、TGF-β1、TGF-β3표체양성적혈관혹세포,병계수.대수거행단인소방차분석화LSD검험. 결과(1)대체관찰가견,광화최갑기각취당지가조창면혈관화정도명현우우기여3조.(2)이식혹복개후1～3주각조창면조직CD31양성혈관수지속증다,제4주시3개지가조균감소.각시상점4조간비교,F치분별위24.005、38.822、25.274、3.856,P＜0.05혹P＜0.01；광화최갑기각취당지가조l～3주명현다우기여3조(P치균소우0.05).(3)이식후1～3주,3개지가조창면조직α-SMA양성혈관수균지속증다,4주시하강；유사대조조복개후1～4주지속증다.각시상점4조간비교,F치분별위22 637、28.087、62.651、18.055,P치균소우0.01.광화최갑기각취당지가조각시상점균명현다우기여3조(P치균소우0.05).(4)이식혹복개후1～4주매400배시야하TGF-β1양성세포수:각취당지가조분별위(127±8)、(167±19)、(170±18)、(144±10)개,광화최갑기각취당지가조위(171±17)、(207±25)、(130±30)、(69±16)개,ADM지가조위(106±8)、(159±17)、(171±11)、(145±11)개,유사대조조위(100±20)、(150±18)、(200±14)、(172±20)개；각시상점4조간비교,F치분별위29 675、9.503、13 107、54.515,P치균소우0.01.1、2주시광화최갑기각취당지가조명현고우기여3조(P치균소우0.05),3、4주시기표체량하강,명현저우기여3조(P치균소우0 05).(5)이식혹복개후1～3주,각조창면조직매400배시야하TGF-β3양성세포수지속증다,4주시하강혹계속승고；각시상점4조간비교,F치분별위140.612、945 850、714.037、119.147,P치균소우0.01.광화최갑기각취당지가조각시상점명현고우기여3조(P치균소우0 05). 결론 효원-광화최갑기각취당진피지가재소상창면유합과정중가쾌속유도혈관생성화성숙,유리우창면조기수복화억제반흔증생.
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-β1 and TGF-β3 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-β1 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 inADM 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-β1 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-β3 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-β3 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.
