目的 探讨烧伤变性脱细胞真皮基质(DADM)作为真皮替代物用于创面修复的可行性. 方法 (1)取12只Wistar大鼠,其中9只背部造成深Ⅱ度烫伤(以下称烧伤)创面,分别于伤后1、2、3d(每时相点3只)取创面全厚皮肤,用2.5 g/L胰蛋白酶-体积分数0.5％ Triton X-100行脱细胞处理制成DADM,相应称为DADM-1 d、DADM-2 d、DADM-3 d；余下3只大鼠不致伤,同法制备ADM作为对照.对ADM和各DADM行大体、组织学观察及微生物学和生物力学检测(极限抗拉强度、最大拉力、断裂伸长率、应力-应变关系).(2)另取64只Wistar大鼠,按随机数字表法分为ADM组、DADM-1d组、DADM-2 d组、DADM-3d组,每组16只.于各大鼠背部制作一2.0 cm×1.8 cm大小皮瓣,将前述ADM和DADM-1 d、DADM-2d、DADM-3 d切制成1.8cm×1.5 cm大小后分别埋植于皮瓣下.术后1、3、5、9周,每组每时相点取4只大鼠,肉眼观察创面愈合情况及埋植物的变化,并对埋植物行组织学观察.对数据行单因素方差分析及t检验. 结果 (1)新鲜制备的各DADM呈乳白色,质地柔软有弹性,韧性较ADM弱.光学显微镜下见ADM和各DADM中均无上皮结构和细胞存在,DADM胶原纤维增粗程度不均匀,排列紊乱,嗜伊红性增强.各DADM微生物学检测结果均为阴性.DADM-1d、DADM-2 d、DADM-3 d之间极限抗拉强度、最大拉力、断裂伸长率和应力-应变关系比较,差异均无统计学意义(F值为0.088～3.591,P值均大于0.05)；其中DADM-3d此4项指标值最高,分别为(13.0±2.4) MPa、(61 ±4)N、(173±7)％、(45 7±2.0)％.ADM此4项指标各为(19.0±2.6) MPa、(95±4)N、(201±5)％、(62.5±2.2)％,与3种DADM两两比较均偏高(t值为6.424 ～17.125,P值均小于0.01).(2)埋植术后1周,各组大鼠创面无渗出或红肿,埋植物未收缩或卷曲；创面有炎性细胞浸润并有Fb及毛细血管长入.术后3周时,DADM-1d、DADM-2 d、ADM组埋植区毛发生长正常,DADM-3 d组埋植区毛发较稀疏；各组炎性细胞减少,Fb增多,有新生小血管长入,DADM-3 d组炎性细胞减退稍延迟.术后第5周,各组埋植区毛发恢复正常,埋植物收缩、变薄,表面纤维膜包裹紧密,有较大血管束长入；各组真皮基质与周围正常组织融合.术后第9周,ADM和DADM均为薄、软白色组织片,与皮瓣内面连接紧密；各组无炎性细胞浸润,胶原纤维排列规则、致密,与正常胶原组织融合. 结论 烧伤DADM无明显免疫原性,生物相容性好,经离体改造有望成为创面修复治疗中的真皮替代物.
目的 探討燒傷變性脫細胞真皮基質(DADM)作為真皮替代物用于創麵脩複的可行性. 方法 (1)取12隻Wistar大鼠,其中9隻揹部造成深Ⅱ度燙傷(以下稱燒傷)創麵,分彆于傷後1、2、3d(每時相點3隻)取創麵全厚皮膚,用2.5 g/L胰蛋白酶-體積分數0.5％ Triton X-100行脫細胞處理製成DADM,相應稱為DADM-1 d、DADM-2 d、DADM-3 d；餘下3隻大鼠不緻傷,同法製備ADM作為對照.對ADM和各DADM行大體、組織學觀察及微生物學和生物力學檢測(極限抗拉彊度、最大拉力、斷裂伸長率、應力-應變關繫).(2)另取64隻Wistar大鼠,按隨機數字錶法分為ADM組、DADM-1d組、DADM-2 d組、DADM-3d組,每組16隻.于各大鼠揹部製作一2.0 cm×1.8 cm大小皮瓣,將前述ADM和DADM-1 d、DADM-2d、DADM-3 d切製成1.8cm×1.5 cm大小後分彆埋植于皮瓣下.術後1、3、5、9週,每組每時相點取4隻大鼠,肉眼觀察創麵愈閤情況及埋植物的變化,併對埋植物行組織學觀察.對數據行單因素方差分析及t檢驗. 結果 (1)新鮮製備的各DADM呈乳白色,質地柔軟有彈性,韌性較ADM弱.光學顯微鏡下見ADM和各DADM中均無上皮結構和細胞存在,DADM膠原纖維增粗程度不均勻,排列紊亂,嗜伊紅性增彊.各DADM微生物學檢測結果均為陰性.DADM-1d、DADM-2 d、DADM-3 d之間極限抗拉彊度、最大拉力、斷裂伸長率和應力-應變關繫比較,差異均無統計學意義(F值為0.088～3.591,P值均大于0.05)；其中DADM-3d此4項指標值最高,分彆為(13.0±2.4) MPa、(61 ±4)N、(173±7)％、(45 7±2.0)％.ADM此4項指標各為(19.0±2.6) MPa、(95±4)N、(201±5)％、(62.5±2.2)％,與3種DADM兩兩比較均偏高(t值為6.424 ～17.125,P值均小于0.01).(2)埋植術後1週,各組大鼠創麵無滲齣或紅腫,埋植物未收縮或捲麯；創麵有炎性細胞浸潤併有Fb及毛細血管長入.術後3週時,DADM-1d、DADM-2 d、ADM組埋植區毛髮生長正常,DADM-3 d組埋植區毛髮較稀疏；各組炎性細胞減少,Fb增多,有新生小血管長入,DADM-3 d組炎性細胞減退稍延遲.術後第5週,各組埋植區毛髮恢複正常,埋植物收縮、變薄,錶麵纖維膜包裹緊密,有較大血管束長入；各組真皮基質與週圍正常組織融閤.術後第9週,ADM和DADM均為薄、軟白色組織片,與皮瓣內麵連接緊密；各組無炎性細胞浸潤,膠原纖維排列規則、緻密,與正常膠原組織融閤. 結論 燒傷DADM無明顯免疫原性,生物相容性好,經離體改造有望成為創麵脩複治療中的真皮替代物.
목적 탐토소상변성탈세포진피기질(DADM)작위진피체대물용우창면수복적가행성. 방법 (1)취12지Wistar대서,기중9지배부조성심Ⅱ도탕상(이하칭소상)창면,분별우상후1、2、3d(매시상점3지)취창면전후피부,용2.5 g/L이단백매-체적분수0.5％ Triton X-100행탈세포처리제성DADM,상응칭위DADM-1 d、DADM-2 d、DADM-3 d；여하3지대서불치상,동법제비ADM작위대조.대ADM화각DADM행대체、조직학관찰급미생물학화생물역학검측(겁한항랍강도、최대랍력、단렬신장솔、응력-응변관계).(2)령취64지Wistar대서,안수궤수자표법분위ADM조、DADM-1d조、DADM-2 d조、DADM-3d조,매조16지.우각대서배부제작일2.0 cm×1.8 cm대소피판,장전술ADM화DADM-1 d、DADM-2d、DADM-3 d절제성1.8cm×1.5 cm대소후분별매식우피판하.술후1、3、5、9주,매조매시상점취4지대서,육안관찰창면유합정황급매식물적변화,병대매식물행조직학관찰.대수거행단인소방차분석급t검험. 결과 (1)신선제비적각DADM정유백색,질지유연유탄성,인성교ADM약.광학현미경하견ADM화각DADM중균무상피결구화세포존재,DADM효원섬유증조정도불균균,배렬문란,기이홍성증강.각DADM미생물학검측결과균위음성.DADM-1d、DADM-2 d、DADM-3 d지간겁한항랍강도、최대랍력、단렬신장솔화응력-응변관계비교,차이균무통계학의의(F치위0.088～3.591,P치균대우0.05)；기중DADM-3d차4항지표치최고,분별위(13.0±2.4) MPa、(61 ±4)N、(173±7)％、(45 7±2.0)％.ADM차4항지표각위(19.0±2.6) MPa、(95±4)N、(201±5)％、(62.5±2.2)％,여3충DADM량량비교균편고(t치위6.424 ～17.125,P치균소우0.01).(2)매식술후1주,각조대서창면무삼출혹홍종,매식물미수축혹권곡；창면유염성세포침윤병유Fb급모세혈관장입.술후3주시,DADM-1d、DADM-2 d、ADM조매식구모발생장정상,DADM-3 d조매식구모발교희소；각조염성세포감소,Fb증다,유신생소혈관장입,DADM-3 d조염성세포감퇴초연지.술후제5주,각조매식구모발회복정상,매식물수축、변박,표면섬유막포과긴밀,유교대혈관속장입；각조진피기질여주위정상조직융합.술후제9주,ADM화DADM균위박、연백색조직편,여피판내면련접긴밀；각조무염성세포침윤,효원섬유배렬규칙、치밀,여정상효원조직융합. 결론 소상DADM무명현면역원성,생물상용성호,경리체개조유망성위창면수복치료중적진피체대물.
Objective To explore the feasibility of burn denatured acellular dermal matrix (DADM) as dermal substitute in repairing wounds. Methods ( 1 ) Nine Wistar rats received a deep partial-thickness scald on the back.Full-thickness wounded skin was collected on post scald day (PBD) 1,2,and 3 (with 3 rats at each time point),and it was treated with 2.5 g/L trypsin/0.5％ Triton X-100 to remove cells to prepare DADM,respectively called DADM-1 d,DADM-2 d,and DADM-3 d.Another 3 rats without scald injury were treated with the same method as above to prepare acellular dermal matrix (ADM)to serve as control.Gross and histological observations and microbiological and biomechanical tests,including ultimate tensile strength,maximum tension,stretched length at breaking,stress-strain relationship,were conducted for the resulting ADM and DADM.(2) Another 64 rats were divided into ADM group and DADM-1 d,DADM-2 d,and DADM-3 d groups according to the random number table,with 16 rats in each group.A skin flap in size of 2.0 cm× 1.8 cm was raised on the back of each rat.The above-mentioned ADM,DADM-1 d,DADM-2 d,and DADM-3 d were cut into pieces in the size of 1.8 cm × 1.5 cm,and they were respectively implanted under the skin flaps of rats in corresponding group.At post surgery week (PSW) 1,3,5,or 9,4 rats in each group were used to observe wound healing condition and change in implants with naked eye,and histological observation of the implants was conducted.Data were processed with one-way analysis of variance and t test. Results ( 1 ) The freshly prepared DADM was milky white,soft in texture with flexibility,but poor in elasticity as compared with ADM.No epithelial structure or cellular component was observed in ADM or DADM under light microscope.Collagen fibers of DADM were seen to be thickened unevenly and arranged in disorder and eosinophilic.All microbiological results of DADM were negative.There was no statistically significant difference among DADM-1 d,DADM-2 d,and DADM-3 d in levels of ultimate tensile strength,maximum tension,stretched length at breaking,and stress-strain relationship ( with F valucs from 0.088 to 3.591,P values all above 0.05 ).Values of the above-mentioned four indexes were the highest in DADM-3 d,they were respectively (13.0±2.4) MPa,(61 ±4)N,(173 ±7) ％,(45.7 ± 2.0) ％.Values of the four indexes of ADM were respectively ( 19.0 ± 2.6) MPa,(95 ±4) N,(201 ±5)％,(62.5 ±2.2)％,which were higher than those of DADM-1 d,DADM-2 d,and DADM-3 d (with t values from 6.424 to 17.125,P values all below 0.01 ).(2) No exudate or swelling in the wounds of rats,and no contraction or curling of implants were observed in every group at PSW 1,but inflammatory cells infiltration and Fbs inward migration were observed in the wound.At PSW 3,the growth of hair was normal in the wound in DADM-1 d,DADM-2 d,and ADM groups,but few and scattered hair grew in DADM-3 d group.The inflammatory cells decreased,while Fbs increased,and new capillaries were found to grow inwardly in each group.The decrease in inflammatory cells was slightly delayed in DADM-3 d group.At PSW 5,hair growth became normal,and implants shrank and thinned with fiber membrane wrapped densely and bundles of ingrowing large caliber blood vessels in all groups.The dermal matrix in each group merged with the surrounding normal tissue.At PSW 9,ADM and DADM became white,thin,and soft tissue sheet which was closely connected with the inner side of the flap.There was no infiltration of inflammatory cells in implants in either group.The collagen fibers arranged regularly and densely,and they were integrated with normal collagen tissue. Conclusions The burned DADM does not have obvious immunogenicity,but with good biocompatibility.It is prospective to become as a dermal substitute in repairing wounds.
