激光微孔脱细胞真皮基质生物力学测试

目的：研究激光微孔脱细胞基质的生物力学性质变化,制作合适规格的脱细胞基质。方法：对不同间距的脱细胞真皮基质进行应力-应变和应力-松弛实验等检测,确定最佳孔径的脱细胞真皮基质。结果：确定350μm间距的脱细胞真皮基质有良好的生物力学性能。结论：超脉冲CO2激光可以对脱细胞真皮基质进行精细的微加工,脱细胞真皮基质适用于构建组织工程支架。     

异体颗粒状脱细胞真皮基质与自体刃厚皮复合移植修复大鼠皮肤缺损创面效果观察

目的 观察异体颗粒状脱细胞真皮基质(PADM)与自体刃厚皮复合移植修复大鼠皮肤缺损创面的效果.方法 采用随机数字表法将12只SD大鼠分为实验组和对照组,每组6只.于2组大鼠背部制作全层皮肤缺损创面,实验组创面复合移植SD大鼠异体PADM(扩张比10:5)及厚度0.20 mm的自体刃厚皮,对照组创面仅移植厚度0.20 mm自体刃厚皮.术后2周起打开敷料观察大鼠创面愈合情况.术后2、3、4、6、8、12、20周计算2组创面移植皮片成活率、收缩率(或扩张率).术后20周取2组创周正常皮肤及创面皮肤标本,采用HE染色法观察胶原纤维束结构,测量胶原纤维束直径和间隙率;用天狼星红染色法观察Ⅰ、Ⅲ型胶原分布情况,测量Ⅰ、Ⅲ型胶原含量及其比值.对实验数据行独立样本t检验、Levene检验、t'检验.结果 (1)术后2周,实验组大鼠创面移植皮片成活率[(76.1±13.1)%]低于对照组[(94.5±1.3)%,t'=3.440,P=0.018].术后3周,实验组创面移植皮片收缩率[(34±8)%]明显大于对照组[(16±12)%,t=-3.211,P=0.009];术后8周,2组移植皮片扩张率接近一致.(2)HE染色和天狼星红染色显示,与大鼠创周正常皮肤比较,对照组移植皮片胶原纤维束呈均质化改变,胶原纤维纤细,排列紊乱;实验组移植皮片胶原纤维束结构、排列更接近创周正常皮肤,可见未完全降解的PADM.与对照组创面皮肤胶原纤维束直径[(7.3±1.4)μm]、间隙率[(17±4)%]、Ⅰ型胶原含量[(68.1±8.4)%]、Ⅲ型胶原含量[(32.0±8.4)%]以及Ⅰ、Ⅲ型胶原比例(2.3±1.0)比较,实验组胶原纤维束更粗[(9.6±0.8)μm,t=-3.562,P=0.005],间隙率更大[(24±5)%,t=-2.760,P=0.020],Ⅰ型胶原含量更高[(80.2±5.4)%,t=-2.981,P=0.014],Ⅲ型胶原含量更低[(19.8±5.4)%,t=2.981,P=0.014],Ⅰ、Ⅲ型胶原比例更高(4.3±1.2,t=-3.204,P=0.009).实验组创面皮肤上述胶原相关指标更接近于创周正常皮肤水平.结论 异体PADM在体内作为真皮再生模板,有助于改善自体刃厚皮所修复的大鼠皮肤缺损创面中真皮胶原纤维束的结构,提高再生真皮组织的成熟度.

Abstract：

Objective To evaluate the effects of mixed grafting of allogeneic PADM and autologous STS on wound healing of full-thickness defect in rats. Methods Full-thickness defects with size of 6 cm×4 cm were produced on the back of 12 SD rats, and they were divided into E group(n =6) and C group ( n = 6) according to the random number table. The wounds in E group were grafted with a mix of allogeneic PADM (expansion rate 10: 5) and autologous STS with thickness of 0.2 mm, while those in C group were grafted with autologous STS in the same thickness. The wound healing rate, survival rate, contraction rate,and expansion rate of transplanted skin were observed at post operation week (POW) 2, 3, 4, 6, 8, 12,20. Tissue samples form wounds and surrounding normal skin were harvested at POW 20 for histopathological observation as follows. The structure of collagen fiber bundle was observed by HE staining, the diameter and gap rate of collagen fiber bundle were also measured. The distribution of type Ⅰ and Ⅲ collagen was observed by sirsus red staining, and the contents of type Ⅰ , Ⅲ collagen and their ratio were also examined.Data were processed with independent samples t test, Levene test, and t' test. Results Survial rate of transplanted skin in E group at POW 2 [(76. 1 ± 13. 1)%] was obviously lower than that in C group [(94.5 ± 1.3)%, t' =3.440, P =0.018]. Contraction rate of transplanted skin in E, C groups at POW 3 showed significant difference [(34±8)%vs. (16 ±12)%, t = -3.211, P =0.009]. Compared with those in peri-wound normal skin, collagen fiber bundles in C group showed signs of homogenization, and collagen fibers were thin with irregular arrangement. Collagen fiber structure and arrangement of composite skin in E group were similar to those surrounding normal skin with incomplete degradation of PADM. Diameter of collagen fiber bundle [( 9.6 ± 0.8) μm] , gap rate between collagen bundle [( 24±5) %] , content of type Ⅰ collagen [( 80.2 ± 5.4) %] and the ratio of typeⅠto type Ⅲ collagen(4.3 ± 1.2) in E group were all increased as compared with those inC group [(7.3±1.4) μm (t = -3.562, P =0.005), (17±4)%( t =-2.760, P =0.020), (68.1 ±8.4)%(t = -2.981, P =0.014), 2.3±1.0(t = -3.204, P =0. 009)], while content of type Ⅲ collagen [( 19.8 ± 5.4) %] in E group was lower than that in C group [(32.0 ±8.4)% , t = 2. 981, P = 0. 014]. Above-mentioned indexes of collagen in wound of E group were similar to those of normal skin surrounding the wound. Conclusions Allogeneic PADM used as dermal regeneration template is beneficial in improving collagen fiber bundle structure in dermis layer of rats with fullthickness skin wounds when repaired with autologous STS, and it accelerates maturation of regenerative dermal tissue.     

异种(猪)脱细胞真皮基质微粒注射填充研究

目的：探讨一种新型的注射填充材料，方法：将异种（猪）脱细胞真皮基质微粒（简称ADM微粒）注入兔耳背皮下并形成皮丘，定时对植入物进行测量，光镜及电镜观察，临床应用于4例面部凹陷畸形整复。结果：ADM微粒注射后从外围向中央细胞化及血管化，12周左右体积基本稳定，48周时吸收率为26．93％左右，临床应用结果满意。结论：ADM微粒安全性好，吸收率低、使用方便，微创。     

脱细胞异体真皮基质皮下移植后胶原的动态变化

目的　探讨脱细胞异体真皮基质 (ADM)移植后胶原的变化情况。方法　将异体ADM移植于SD鼠皮下 ,测定移植后ADM中胶原的含量及Ⅰ、Ⅲ型胶原的比例。结果　异体ADM移植后胶原含量及Ⅰ、Ⅲ型胶原比例无明显变化。结论　异体ADM是一种良好的软组织填充材料。     

脱细胞真皮基质在整形美容外科中的应用

目的 为改善皮肤移植的形态和功能，提高大面积烧伤缺乏自体供皮来源患者的美容整形治疗效果。方法 应用自制脱细胞真皮基质(ADM)，植入松解或切除瘢痕后的创面，5～10d后再行自体刃厚皮移植，术后观察ADM和创面愈合、瘢痕增生及功能恢复情况。3例再次手术患者留取复合移植组织进行组织学检测。结果 15例23次复合皮移植：14例22次成活率为100％，1例1次成活率为80％，植皮区平整光滑，瘢痕增生不明显，组织学检测提示复合皮血管化明显，胶原排列整齐，表皮-真皮连接结构重建充分，未见排异反应。结论 ADM与自体刃厚皮复合移植能改善创面愈合的质量，是较理想的整形美容修复材料。     

异种脱细胞真皮基质的制作和临床应用观察

目的　降低异种 (猪 )真皮组织的抗原性 ,探讨在临床复合移植的可行性。　方法　健康小猪中厚皮 ,半数经戊二醛交联后采用胰蛋白酶和TritonX 10 0等脱细胞处理 ,制成未交联(sADM0 )和交联型 (sADM1)网状脱细胞真皮基质 ,通过动物实验和临床应用 ,观察 2种sADM组织学变化和移植效果。　结果　 (1)兔皮下埋植sADM0 宿主细胞侵入迅速 ,炎症反应和组织降解明显 ,而sADM1炎症反应轻 ,皮片降解和收缩不明显 ;(2 ) 10例烧伤患者Ⅲ度创面和 1例胸部瘢痕切除后 ,以自体薄中厚皮 (ATS)和自体刃厚皮或超薄皮 (UTS)为对照 ,进行sADM UTS复合移植 ,19块复合皮平均成活率为 78.9% ,与ATS组无显著差异。其中sADM0 UTS组早期炎症反应和创面收缩明显 ,外观效果与UTS组相当 ;在sADM1 UTS组 ,早期炎症反应和创面收缩较轻 ,外观平整 ,触软 ,与ATS组移植效果相当 ,但有 3例患者的 6块创面在术后平均 (12 .8± 6 .9)周时 ,出现表皮破溃、sADM1外露和组织学严重的异物巨噬细胞反应。　结论　临床用交联型sADM作复合皮的真皮替代物 ,能减缓早期免疫反应 ,提高UTS移植效果 ,但仍不可避免迟发性异种排异反应。     

十二烷基硫酸钠制备脱细胞真皮基质的实验研究

目的 探讨十二烷基硫酸钠制备脱细胞真皮基质的最佳浓度及作用时间.方法 取临床截肢患者废弃的皮肤,分成90块,随机分成6组,每块约1 cm×1 cm,分别置于0.1%、0.25%、0.5%、1.0%、2.5%、5.0%十二烷基硫酸钠溶液中脱细胞,每隔8 h取材一次,分别取材15次.取材标本行大体和组织学观察.结果 肉眼观察十二烷基硫酸钠作用8 h后,0.25%组可将表皮与真皮完整分离;光镜检查显示0.25%十二烷基硫酸钠在作用40 h后,能将真皮内的细胞及细胞碎片脱净,形成脱细胞真皮基质.制备的脱细胞真皮基质,主要由胶原纤维网架构成,其他组作用120 h也无法形成真正的脱细胞真皮基质.结论 十二烷基硫酸钠制备脱细胞真皮基质的最佳浓度为0.25%,最佳时间为40 h.与其他方法比较,具有简便、快捷、有效、价廉、宜推广等特点,可为临床急诊的修复争取宝贵的时间.     

十二烷基硫酸钠制备脱细胞真皮基质的实验研究

目的探讨十二烷基硫酸钠制备脱细胞真皮基质的最佳浓度及作用时间。方法取临床截肢患者废弃的皮肤,分成90块,随机分成6组,每块约1cm×1cm,分别置于0.1%、0.25%、0.5%、1.0%、2.5%、5.0%十二烷基硫酸钠溶液中脱细胞,每隔8h取材一次,分别取材15次。取材标本行大体和组织学观察。结果肉眼观察:十二烷基硫酸钠作用8h后,0.25%组可将表皮与真皮完整分离;光镜检查显示:0.25%十二烷基硫酸钠在作用40h后,能将真皮内的细胞及细胞碎片脱净,形成脱细胞真皮基质。制备的脱细胞真皮基质,主要由胶原纤维网架构成,其他组作用120h也无法形成真正的脱细胞真皮基质。结论十二烷基硫酸钠制备脱细胞真皮基质的最佳浓度为0.25%,最佳时间为40h。与其他方法比较,具有简便、快捷、有效、价廉、宜推广等特点,可为临床急诊的修复争取宝贵的时间。     

脱细胞真皮基质的研究进展

烧伤或皮肤组织缺损的创面修复多采用自体中厚或全厚皮片移植 ,供皮区损伤较重易形成疤痕 ,影响外观及功能。而早期自体薄厚皮片 (autologeus thin split- thickness skin,auto- TTS)或刃厚皮移植 ,则往往由于皮片薄、缺少有效的真皮成分和引导性组织再生的作用 ,皮片成活后常形成不同程度的瘢痕增生和挛缩畸形 ;在关节等功能部位更易造成功能障碍 ,影响患者生活质量。近年来脱细胞真皮基质(acellular dermal matrix,ADM)作为一种新型真皮替代物不但受到烧伤与整形外科界的重视 ,也引起了整个美容医学的兴趣。本文对其综述如下 :1　 ADM…     

脱细胞真皮基质与自体刃厚皮片复合移植的临床及组织学观察

目的 观察异体脱细胞真皮基质与自体刃厚皮片复合移植修复创面的中、长期临床效果,并做组织学观察.方法 选取23例接受异体脱细胞真皮基质加自体刃厚皮片复合移植的患者,于术后3、6、12、18个月进行临床随访,并作组织学观察.结果 移植后3、6、12、18个月,复合皮表面平滑,无明显瘢痕增生及色素沉着现象,柔软有弹性,关节部位活动自如.组织学观察接近正常皮肤形态.结论 异体脱细胞真皮基质与自体刃厚皮片复合移植后愈合创面表面光滑,平整柔软,无明显挛缩,关节活动自如,未见明显排汗现象,可作为修复创面的良好选择.     

Preparation of laser micropore porcine acellular dermal matrix for skin graft: an experimental study

In our previous study, we used composite grafts consisting of meshed porcine acellular dermal matrix (PADM) and thin split-thickness autologous epidermis to cover full thickness burn wounds in clinical practice. However, a certain degree of contraction might occur because the distribution of dermal matrix was not uniform in burn wound. In this study, we prepare a composite skin graft consisting of PADM with the aid of laser to improve the quality of healing of burn wound. METHODS: PADM was prepared by the trypsin/Triton X-100 method. Micropores were produced on the PADM with a laser punch. The distance between micropores varied from 0.8, 1.0, 1.2 to 1.5mm. Full thickness defect wounds were created on the back of 144 SD rats. The rats were randomly divided into six groups: micropore groups I-IV in which the wound were grafted with PADM with micropores, in four different distances, respectively and split-thickness autograft; mesh group rats received meshed PADM graft and split-thickness autograft; control group received simple split-thickness autografting. The status of wound healing was histologically observed at regular time points after surgery. The wound healing rate and contraction rate were calculated. RESULTS: The wound healing rate in micropore groups I and II was not statistically different from that in control group, but was significantly higher than that in mesh group 6 weeks after grafting. The wound healing rate in micropore groups III and IV was lower than that in mesh and control groups 4 and 6 weeks after grafting. The wound contraction rate in micropore groups I and II was remarkably lower than that in control group 4 and 6 weeks after surgery and it was significantly much lower than that in mesh group 6 weeks after surgery. Histological examination revealed good epithelization, regularly arranged collagenous fibers and integral structure of basement membrane. CONCLUSION: Laser micropore PADM (0.8 or 1.0mm in distance) grafting in combination with split-thickness autografting can improve wound healing. The PADM with laser micropores in 1.0mm distance is the better choice.     

异种(猪)脱细胞真皮基质移植后炎症免疫反应的实验研究

目的：研究异种（猪）脱细腻真皮基质与薄自体皮复合移植后炎症反应的变化规律及对Th1/Th2细胞因子的影响。方法：将异种（猪）脱细胞真皮基质与薄自体皮复合移植于大鼠，分别于移植后1，2，3，4，8，12，16周取标本，采用组织学观察及RT-PCR技术动态检测异种（猪）脱细胞真皮基质复合移植后炎症反应及Th1/Th2细胞因子的变化，同时以单纯薄自体皮移植作对照。结果：异种（猪）脱细胞真皮与薄自体皮复合移植后早期存在炎症反应，并随着创面愈合的进行而逐渐消失，Th2细胞因子的表达明显高于对照组。结论：异种（猪）脱细胞真皮基质仍具有一定的免疫原性，Th2细胞因子的高表达说明机体对异种（猪）脱细胞真皮基质的免疫反应主要以体液免疫为主，也可能是异种（猪）脱细胞真皮基质移植后未被排斥的原因之一。     

无细胞真皮基质与自体皮片复合移植的临床应用

目的：改善烧伤创面移植皮片愈合后皮肤的外形和功能,并减轻供皮区瘢痕增生。方法：应用自制的无细胞真皮基质与自体刃厚皮片复合移植于4例切（或削）痂后的烧伤创面。结果：未发现对复合移植皮片的排异反应。创面愈合后,大体观察发现移植自体网状皮片及自体小皮片的轮廓界面变得不明显或基本消失,自体大张皮片与真皮基质复合移植区出现较厚而特殊的脱屑,均反映了愈合过程的特殊性。光镜和电镜观察发现复合皮片移植区棘细胞间桥粒清晰,基底膜连续而完整等。与对照部位相比,复合皮片移植区瘢痕增生减轻,未见明显挛缩,皮肤弹性较好。结论：在3－4个月的观察期内,自体刃厚皮片与无细胞真皮基质复合移植后,功能和形态等同或优于单纯自体刃厚皮片移植。     

Influence of hyaluronic acid on wound healing using composite porcine acellular dermal matrix grafts and autologous skin in rabbits

This study aims to explore the influence of hyaluronic acid (HA) on wound healing during xenogeneic porcine acellular dermal matrix (PADM) composite skin grafting. The results will facilitate the development of methods for improving graft contracture and poor elasticity of composite transplantation. Exogenous HA was added to composite PADM grafts and to thin autologous skin grafts during rabbit full‐thickness skin wound repair. The influence of HA on wound healing was evaluated according to its contracture rate and its expression of collagen types I and III. The possible mechanism was then explored based on HA metabolism and vascularisation in the skin graft. The results show that exogenous HA relieves graft contracture on rabbit wound surfaces, increases collagen I and III expression and decreases the ratio between collagen types. HA stimulates the generation of more CD44 receptors to strengthen its enzymolysis. The resulting metabolites promote the vascularisation of the wound surface, which are conducive for mitigating graft contracture, and further improve the composite grafting effect.     

无细胞真皮基质与自体皮复合移植愈合过程的观察

目的 :为改善植皮区外观和功能 ,并同时减轻供皮区瘢痕增生。方法 :应用自制的无细胞真皮基质与薄的刃厚自体皮复合移植应用于 2例切削痂后的烧伤创面。结果 :未发现对复合移植物的排异反应。创面愈合后 ,大体观察发现移植网状自体皮的网状形式已变得不明显 ,自体小皮片的轮廓基本消失 ,均反应了愈合过程的特殊性。光镜和电镜观察发现复合移植区基底膜清晰、连续且完整等。与对照部位相比 ,复合移植区瘢痕增生减轻 ,未见明显挛缩 ,皮肤弹性较好。结论 :至少到目前为止的观察期 (3～ 4个月 )内 ,较薄的刃厚自体皮与无细胞真皮基质复合移植后外观和功能等同或优于单纯刃厚自体皮移植     

无细胞真皮基质的研制

目前治疗大面积深度烧伤的方法是刃厚自体皮移植 ,后者因缺乏足够的真皮成分 ,常常造成瘢痕和挛缩。因此 ,研制一种含有足够量真皮成分的永久性皮肤替代物将具有很大的临床意义。将异体 (或异种 )皮用高渗盐溶液处理以去除表皮 ,去污剂处理以去除真皮中细胞成分 ,得到一种无细胞真皮基质。经光镜和透射电镜观察证实 ,它去除了表皮全层和真皮中所有细胞成分 ,保留了完整的基底膜复合物 ,胶原束结构和排列保持正常 ,弹力纤维亦正常存在。与AlloDerm相比 ,无细胞真皮基质具有以下主要优点 :①由于延长去污剂的作用时间 ,真皮中残留的细胞成分被完全清除 ,因此制得的真皮基质更为理想 ;②AlloDerm用同种异体皮制备 ,而无细胞真皮基质除用同种异体皮外 ,亦可用异种猪皮制备 ,因此来源广泛 ,价格低廉     

异体脱细胞真皮基质加自体刃厚皮复合移植远期随访评价

目的 评价异体ADM+自体刃厚皮复合移植的临床远期效果.方法 选择2001年3月-2008年10月,笔者单位收治的19例行异体ADM+自体刃厚皮复合移植患者为复合移植组(34个创面),同期9例行自体刃厚皮移植患者为对照组(11个创面).患者术后均随访2年以上.随访时,在曼彻斯特瘢痕量表的基础上设计随访对象评估表,评估移植皮肤的颜色、平整度、质地、挛缩、感觉、并发症情况,分值1～4分,得分越高、情况越差;采用温哥华瘢痕量表评估供皮区瘢痕形成情况;发放问卷调查患者满意度、移植期内健康记录;组织病理学方法观察其中4例患者皮肤组织结构.采用中立位法描述术前、术后及随访时患者关节活动范围.对数据进行非参数秩和检验、t检验或x2检验.结果 (1)复合移植组皮肤平整度、挛缩、质地评分分别为(1.6±0.5)、(1.8±0.8)、(1.5±0.8)分,显著低于对照组的(2.0±0.7)、(2.2±0.9)、(2.3±0.7)分(Z值分别为-2.058、-2.220、-2.323,P值均小于0.05);2组皮肤颜色、感觉、并发症评分结果相近(Z值分别为-0.628、-0.428、-2.520,P值均大于0.05).(2)复合移植组仅1个供皮区部分区域有轻度瘢痕.(3)复合移植组和对照组患者在疼痛、瘙痒和满意度方面比较,差异均无统计学意义(x2值分别为0.187、0.019、2.628,P值均大于0.05).(4)病理结果显示,手部复合移植后2年可见神经纤维结构,ADM在受体内未引起强烈的炎症反应.(5)复合移植组11处关节部位功能得到恢复或改善,另2处需再次手术.对照组2处关节部位均明显挛缩.结论 异体ADM+自体刃厚皮复合移植在防止瘢痕挛缩,改善功能及外观方面效果明显,长期存留于成人和儿童患者体内均未出现安全问题.