异种(猪)脱细胞真皮基质与自体皮复合移植的临床应用

目的：观察基底良好的深度创面用异种（猪）脱细胞真皮基质与自体皮复合移植的临床疗效。方法：2000年1月-2002年2月用异种（猪）真皮基质与自体薄片复合移植（二步法）修复基底良好的深度创面52例。结果：37例（71.1%）植皮完全成活，15例（28.9%）植皮95%以上成活，无一例补植皮。随访3-6个月复合移植皮肤颜色、质地基本接近于正常皮肤，瘢痕增生不明显。结论：异种（猪）真皮基质可推广应用于修复基底良好的深度创面的复合移植。     

成纤维细胞-无细胞真皮替代物的生物学活性及移植实验

目的 研究含成纤维细胞的无细胞真皮替代物的生物学活性及真皮支架作用。方法 将成纤维细胞种植于无细胞真皮表面培养,形成活性真皮替代物。采用ELISA法和RIA法测定培养上清中IL－6、IL－8、TGF－β1及细胞外基质层粘连蛋白、透明质酸的分泌。并将真皮替代物植入BALB／c-nu小鼠（裸鼠）全层皮肤缺损创面,观察血管化速度、创面收缩率。结果 成纤维细胞种植于无细胞真皮表面生长良好,可形成单层细胞膜片,并分泌多种细胞因子和细胞外基质成份。活性真皮替代物植入创面后,与单纯无细胞真皮移植相比,血管化速度加快,收缩率减小。结论 无细胞真皮上种植成纤维细胞后具有较强的生物学活性,可作为较好的真皮替代物。     

The application of new biosynthetic artificial skin for long-term temporary wound coverage

Temporary dressings protect wounds from desiccation and infection. In our previous study, we used meshed acellular porcine dermis (APD) to enhance wound healing and decrease wound contraction; however, the wounds showed meshed scar [Wang HJ, Chen TM, Cheng TY. Use of a porcine dermis template to enhance widely expanded mesh autologous split-thickness skin graft growth: preliminary report. J Trauma 1997;42(2):177–82]. In this study, we produced an artificial skin composed of a cross-linked silicon sheet on the surface of APD which we have called silicone acellular porcine dermis (SAPD). This new artificial skin can protect the wound long enough to promote wound healing either by second intention or covered long enough until cultured epithelium autograft (CEA) or autologous skin graft can be harvested for permanent coverage.

We delivered 4 cm × 5 cm full-thickness wound on the back of 350 g Sprague–Dawley rats. Thirty-six rats were divided into two groups. Eighteen rats had SAPD and the other 18 were covered with Biobrane. The wounds were first examined 2 weeks after grafting and followed weekly for an additional 4 weeks to evaluate the wound and study pathological changes by using H.E. and Masson's stains. Wound size was calculated by ruler and analyzed by Student's t-test.

At the 2-week inspection, both SAPD and Biobrane showed tight adherence to the wound with no change of wound size. Both the SAPD and Biobrane dermal templates were pink. In the Biobrane-covered group, the wounds contracted soon after the tie-over dressing was removed. Its dermal layer is a layer of thin porcine dermal substance, which was promptly digested by tissue hyaluronidase and provides no real dermal template. In the SAPD-covered group however, the wound size was maintained significantly from third to sixth week after grafting (p < 0.001). SAPD was designed with thick epidermal silicone and a well-organized porcine dermis so that it incorporates into the recipient wound. Clinically the silicone layer of SAPD dislodged from APD about 6–7 weeks after grafting and was followed by dermal matrix exposure and infection. In pathological examination, much like a human skin graft, new vessels were found in APD about 1 week after grafting with minimal inflammatory cells infiltrated in the graft and wound. Six weeks after grafting, the collagen of APD incorporated into the wound, showing palisade arrangement and no sign of rejection. In the Biobrane group however, the wounds showed severe inflammation, the porcine dermal matrix was digested and disappeared 3 weeks after coverage.

In conclusion, SAPD is a thick biosynthetic artificial skin, which protects the rat wound significantly longer than Biobrane and prevents contraction. We expect that using of SAPD for temporary wound coverage will provide enough time to grow autologous-cultured epithelium or to reharvest skin grafts.     

异种脱细胞真皮基质与自体皮复合移植后基底膜重塑的实验观察

目的　观察异种 (猪 )脱细胞真皮基质与薄自体皮复合移植模型中基底膜重塑的变化规律。　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创面 ,分别于移植后 1、2、3、4、8、12、16周留取标本 ,采用免疫组化、透射电镜等方法 ,观察基底膜中层粘连蛋白的变化规律及 12周时基底膜的超微结构 ,同时以单纯薄自体皮移植进行对照。　结果　异种脱细胞真皮基质与薄自体皮复合移植后 ,层粘连蛋白的表达高于单纯薄自体皮移植。 12周时复合移植组基底膜清晰连续 ,半桥粒较多且分布均匀 ;薄自体皮组基底膜致密板模糊、不连续 ,半桥粒减少且分布不均。　结论　异种脱细胞真皮基质与薄自体皮复合移植后 ,基底膜中层粘连蛋白表达增高 ,可能有利于复合皮移植后基底膜的重塑 ,增强表皮和真皮之间的连接 ,进而改善创面愈合质量     

用含活性细胞的胶原支架复合皮修复小鼠全层皮肤缺损的研究

目的构建具有活性细胞成分的复合皮，观察其在修复小鼠全层皮肤缺损中的作用。方法将异种成纤维细胞种植于脱软骨细胞胶原支架上培养3d，形成真皮替代物。再将其放置于培养的上皮细胞膜上共同培养10d，组成含活性细胞成分的复合皮。将复合皮移植于小鼠的全层皮肤缺损处，记录其生长情况并于术后定期活检，进行组织学观察。结果胶原支架中可见生长良好的成纤维细胞和复层表皮细胞。移植术后1周，复合皮与小鼠缺损创面粘连紧密，可见明显的血管化。术后6周，创面愈合良好，移植物与创缘融合，未见明显排斥反应。结论以脱软骨细胞胶原为支架构建的复合皮，可作为皮肤替代物修复全层皮肤组织缺损。     

Partial breast reconstruction of 30 cases with peri-mammary artery perforator flaps

Self-repair of lower limb wounds has always been one of the research hotspots. Flaps and skin graft are the preferred treatment for lower extremity wound reconstruction. However, these treatments have many disadvantages, such as secondary damage, poor healing quality. In recent years, the use of acellular dermal matrix has emerged as an alternative treatment option for extremity ulcers. This study aimed to explore whether acellular dermal matrix can be used as a single treatment to promote wound healing. 7 patients with lower extremities cutaneous deficiency exposing bone or tendon, were covered by Pelnac, which was an acellular dermal matrix product approved by China Food and Drug Administration. All the wound was treated by Pelnac without flaps and skin graft. The external dressing was changed every 10 days. After a maximum of 20 weeks, all the wounds were completely healed. During the 12 months follow-up period none of the patients developed skin wear on the treatment area. All patients maintained their postoperative ambulatory ability. All patients were satisfied with the appearance and feeling after wound healing. These findings may mean acellular dermal matrix is a novel method offering opportunity for treatment of lower extremities cutaneous deficiency exposing bone or tendon. It also has the potential to close wounds of all uninfected, non-ischemic, full-thickness cutaneous deficiency.     

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.     

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。     

应用脱细胞异体真皮植入Bucks筋膜下加大阴茎

目的探讨一种加大阴茎的手术方法。方法将脱细胞异体真皮填充在阴茎Bucks筋膜与白膜之间加大阴茎。结果自2002年3月以来,我们在临床应用12例,术后自然状态下阴茎周径加大13～31cm,平均26cm,术后3个月有正常的性生活。1例因包扎过紧至阴茎皮肤部分坏死,经转移阴囊皮瓣修复愈合。结论该方法用于阴茎加大,创伤小、操作简便、效果确实,无不良反应。     

复合皮混合移植治疗深Ⅱ度烧伤患者创面疗效观察

目的观察深Ⅱ度烧伤患者创面削痂术后应用复合皮混合移植治疗的效果。方法对23例烧伤患者的30个深Ⅱ度烧伤肢体在伤后3d内分次行削痂术,削至浅筋膜后移植大张异体脱细胞真皮基质,然后切取大张自体刃厚皮(0.10～0.25mm)覆盖于其上。术后10—12d计算移植皮片的存活率,记录创面愈合时间。观察随访3—6个月时患者的肢体外观及功能恢复情况。取1例患者随访3个月时的愈合创面皮肤标本,行病理学观察。结果本组患者复合皮片成活率为93%,7%的皮片因术中固定较差,移植后自体刃厚皮与异体脱细胞真皮基质分离致皮片坏死,或因感染致皮片溶解。随访3—6个月,移植部位皮肤外观、弹性及功能恢复良好。病理学观察显示,成活皮片表皮、真皮结构正常。结论烧伤后早期削痂立即移植复合皮是治疗深Ⅱ度创面的有效方法。     

脱细胞真皮移植部位血管细胞黏附分子1的表达

目的探讨血管细胞黏附分子1（vascular cell adhesion molecule 1,VCAM-1）在脱细胞真皮移植部位的表达变化和意义。方法取近交系小型香猪15只建立猪实验模型,分别于每只猪脊背两侧制作6cm×6cm各3处6个创面。随机分为三组,每组5只。A组为对照组,猪刃厚自体皮移植;B组异体脱细胞真皮与猪刃厚自体皮复合移植;C组J-1型（人）脱细胞真皮与猪刃厚皮复合移植。分别于移植术后3、9、21及30d观察移植皮片成活情况和面积,于移植后3、6、9、12、21及30d各时间点取移植组织标本,流式细胞仪检测VCAM-1的表达变化。结果移植术后3d各组移植皮片基底分离;9dA组皮片转红润基底粘连较紧密,揭开皮片可见渗血,B、C组皮片颜色次之,基底粘连不紧密;21d,A、B、C组移植皮片成活面积分别为94%±12%、92%±9%和91%±11%;术后30d各组移植皮片均愈合良好。各组VCAM-1表达,移植术后3d无明显差别;6dA组表达明显高于B、C组有统计学意义（P〈0.05）;9、12dB、C组明显高于A组有统计学意义（P〈0.05）;30d,各组VCAM-1表达均低于3d有统计学意义（P〈0.01）。结论同种和异种真皮基质移植部位VCAM-1表达高峰迟滞于自体皮移植;提示VCAM-1表达可能与脱细胞真皮活化、血管再生有关。同种和异种真皮移植物VCAM-1表达无差别。     

PDGF-B基因表达在组织工程化皮肤移植后血管重建中的作用

目的构建含有血小板衍化生长因子B(PDGFB)基因的组织工程化皮肤,进行动物移植实验,研究PDGFB基因表达在真皮血管重建中的作用。方法构建PDGFB真核表达质粒,用脂质体LipofectAMINE介导转染成纤维细胞。分别构建3种不同类型的组织工程化皮肤角质形成细胞 脱细胞猪真皮基质(A组),角质形成细胞 脱细胞猪真皮基质 成纤维细胞(B组),角质形成细胞 脱细胞猪真皮基质 PDGFB基因转染的成纤维细胞(C组),分别移植于大鼠背部创面,观察术后2、4、6周真皮血管重建情况。结果术后2周C组真皮浅层内可见较多新生毛细血管长入,B组次之,A组毛细血管长入较少(P<005);术后4周各组真皮浅层内毛细血管数逐渐增加,但C组毛细血管数仍明显高于B、A两组(P<005);术后6周各组织工程化皮肤内毛细血管数差异无显著性意义。结论PDGFB基因在组织工程化皮肤移植后早期真皮血管重建中发挥了重要的作用,为移植后皮片成活提供了保障。     

激光微孔猪脱细胞真皮基质制备及创面移植的观察

目的 制备激光微孔猪脱细胞真皮基质（PADM），寻找其与自体刃厚皮复合移植的最佳孔间距。方法 采用胰蛋白酶法制备PADM，用激光加工机打孔，制备孔间距为0．8、1．0、1．2、1．5mm的激光微孔PADM。在144只SD大鼠背部制作全层皮肤缺损创面，分为微孔Ⅰ、Ⅱ、Ⅲ、Ⅳ组，依次采用前述4种孔间距的激光微孔 PADM＋自体刃厚皮移植；网状组采用网状PADM＋自体刃厚皮移植；对照组单纯移植自体刃厚皮。每组24只大鼠。观察移植后2、4、6周各组创面愈合情况，计算移植物成活率和创面收缩率，并行组织学观察。结果 移植后2、4周，微孔Ⅰ、Ⅱ组移植物成活率均低于对照组（P〈0．05），但与网状组比较，差异无统计学意义（P〉0．05）；移植后6周，微孔Ⅰ、Ⅱ组成活率明显高于网状组（P〈0．05）。移植后4、6周微孔Ⅰ、Ⅱ组创面收缩率较对照组显著降低（P〈0．05），移植后6周[（16．0±2．6）％、（15．1±2．4）％]明显低于网状组[（19．3±2．4）％，P〈0．05]。组织学观察见各微孔组上皮化良好，胶原纤维排列整齐，基底膜结构完整。结论 孔间距为0．8mm或1．0mm的激光微孔PADM与自体刃厚皮复合移植可提高创面修复质量，其中孔间距1．0mm效果最佳。     

Wound healing effect of acellular artificial dermis containing extracellular matrix secreted by human skin fibroblasts

In this study, an acellular artificial dermis, composed of human collagen and glycosaminoglycan (GAG) secreted by cultured human fibroblasts on a bovine collagen sponge, was developed. Much of the newly secreted extracellular matrix (ECM) remained after the cell removal process. The main theme of this study focused on the matrix, rather than the viable cell components of the skin, as the major dermal deficit in the wound. Both the acellular artificial and bioartificial dermises, containing viable cells with ECM, were significantly less soluble than the collagen sponge, and the relative GAG content in the bioartificial and acellular artificial dermises was approximately 115-120% of the chondroitin-6-sulfate (CS) content found in the collagen sponge. In the group receiving the collagen sponge, the wound area gradually decreased to approximately 10% of its original area, while in the groups receiving the bioartificial and acellular artificial dermises, the wound area also gradually decreased to approximately 60 and 50%, respectively, of the original size over the 5 weeks after grafting. Both the bioartificial and acellular artificial dermises formed thicker, denser collagen fibers; more new blood vessel formation was observed in both cases. The basement membrane of the regenerated epidermal-dermal junction was thicker and more linear in the acellular artificial dermis graft than in the collagen sponge graft. In conclusion, the wound healing effects of acellular artificial dermis are no less than those of the bioartificial dermis, and much better than the collagen sponge graft with respect to wound contraction, angiogenesis, collagen formation, and basement membrane repair.     

真皮替代物移植后的血管化过程及组织学变化的实验研究

目的了解不同种类真皮替代物移植后的血管化过程及组织学变化。方法将21只SD大鼠根据其皮下埋植不同的真皮替代物分为猪脱细胞真皮基质(sADM)组、人脱细胞真皮基质(hADM)组及人工真皮(Integra)组。于埋植后2、3、4、7、10、14、21、30、60、90、120、150、180d行移植物大体观察,采用免疫组织化学法观察移植物的血管化过程及组织学变化。结果大体观察术后各组大鼠创口周围皮肤无明显红肿及炎性反应,切口愈合良好,移植物与创面接触紧密。90d后各组移植物不易从体表触及。180d时,部分移植物面积缩小、厚度变薄甚至难以辨认。组织学观察移植术后2d起可见成纤维细胞、中性粒细胞、淋巴细胞等侵入移植物内,3d时与受床组织连接处可见长入的新生毛细血管芽。30—60d,移植物内形成丰富的血管网。150d后近似正常真皮结构。180d后部分移植物有不同程度吸收退化。结论3种真皮替代物移植后均能很快建立与受床组织的血液循环,并长时间存留于创面,但有一定程度的吸收退化。     

异体脱细胞真皮基质与自体皮片复合移植的临床应用

目的:探讨异体脱细胞真皮基质与自体皮片复合移植修复大面积深度烧伤及瘢痕切除后皮肤缺损创面及其愈合后皮肤的外形和功能。方法:应用异体脱细胞真皮基质与自体刃厚皮片组成复合皮移植,以自体刃厚皮片移植作为对照,采用一步移植法治疗切痂后大面积深度烧伤创面及瘢痕切除后皮肤缺损共56例患者60处创面,观察术后皮片的成活情况、外形及功能恢复情况并随访。结果:60处创面全部愈合,移植皮片生长良好,瘢痕增生不明显,未见明显挛缩,皮肤弹性较好。在6～12个月的观察期内,自体刃厚皮片与异体脱细胞真皮基质复合移植后,功能和形态优于单纯自体刃厚皮片移植;随访2年复合移植未发现明显的排异反应。结论:异体脱细胞真皮基质与自体皮片复合移植修复大面积深度烧伤及瘢痕切除后皮肤缺损创面愈合良好,无瘢痕增生,皮肤外观功能满意,无排异反应。     

Acellular allograft dermal matrix: immediate or delayed epidermal coverage?

In a prospective, randomized study seventeen patients received skin grafts to a freshly excised burn wound. One group was grafted with a deantigenized dermal matrix and immediately overgrafted with thin autograft. The second group was grafted with dermal matrix, which was then covered with bank allograft for protection, and autografted 1 week later. Each group also received a standard split thickness control graft. Assessment was carried out for up to 1 year. There were no statistically significant differences of graft take between any of the groups, or in the Vancouver scar score at follow-up. Thin donor sites used for dermal matrix coverage healed faster than standard control graft sites, P<0.001. Immediate grafting of acellular dermal matrix with thin autograft works well and leads to an acceptable late result, with faster donor site healing than standard split thickness grafts.     

Hautersatzverfahren in der Verbrennungschirurgie

Patients with extensive deep partial or full thickness burns require early excision of necrotic tissue, however, in many of these cases simultaneous autografting is not possible due to the general condition of the patient. In this instance temporary dressings like allogeneic or xenogeneic skin or foam dressings can be applied to minimize fluid and protein loss. In Europe glycerolized preserved allogeneic skin remains the treatment standard. Dermal replacements are considered to optimize the long-term outcome of split thickness skin grafting. Reduced contracture rates and increased pliability have been reported after additional dermal enhancement with either collagen-glycosaminoglycan matrix, acellular allogeneic dermis or collagen/elastin matrix. True regeneration of the dermis has not yet been observed. However, these materials are suitable for improvement of the wound bed and also the final result after split thickness skin transplantation.     

人表皮细胞与无细胞异种真皮复合移植的实验研究

目的 观察人表皮细胞与无细胞异种真皮复合移植全层皮肤缺损创面后的效果,寻找一种新的创面覆盖物。方法 42只裸鼠作背部全层皮肤缺损创面,分别进行复合皮移植（复合皮组）和单纯表皮细胞膜片移植（对照组）,术后定期观察创面愈合情况并进行创面愈合率及创面收缩率的计算,同时留取创面组织进行组织学检查。结果 与对照组相比较,复合皮组的创面愈合及外观情况良好,两组创面愈合率及收缩率的差异有显性意义（P＜0．05）;组织学检测提示复合皮上皮分化充分,胶原增生有序,表皮－真皮连接结构重建明显,未见明显的急性期免疫排斥反应。结论 人表皮细胞与无细胞异种真皮复合移植能改善创面愈合质量,可作为一种新的皮肤代用品。     

自体网状皮片与异种(猪)脱细胞真皮基质复合移植的临床应用

目的：探讨1：3自体网状皮片与异种（猪）脱细胞真皮基质复合移植的可行性。方法：选择9例烧伤及瘢痕患者，在Ⅲ度烧伤创面行切痂术或瘢痕切除术后的创面上移植异种（猪）脱细胞真皮基质，摊平固定后，在支架上再植1：3网状自体刃厚皮片，表面再覆盖辐照猪皮。结果：9例复合移植皮肤全部成活，临床效果满意。结论：1：3自体网状刃厚片可以与异种（猪）脱细胞真皮基质复合移植，同时将自体皮扩增3倍，成活后的复合网状皮片有相当于中厚皮片或者全厚皮片移植的效果。