{"title":"血管造影剂引导的筋膜瓣灌注脱细胞术","authors":"Liya Yang, Xueshan Bai, Yuanbo Liu, Shan Zhu, Shanshan Li, Zixiang Chen, Tinglu Han, Shengyang Jin, Mengqing Zang","doi":"10.1055/a-2404-2608","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong> Tissue engineering based on whole-organ perfusion decellularization has successfully generated small-animal organs, including the heart and limbs. Herein, we aimed to use angiosome-guided perfusion decellularization to develop an acellular fasciocutaneous flap matrix with an intact vascular network.</p><p><strong>Methods: </strong> Abdominal flaps of rats were harvested, and the vascular pedicle (iliac artery and vein) was dissected and injected with methylene blue to identify the angiosome region and determine the flap dimension for harvesting. To decellularize flaps, the iliac artery was perfused sequentially with 1% sodium dodecyl sulfate (SDS), deionized water, and 1% Triton-X100. Gross morphology, histology, and DNA quantity of flaps were then obtained. Flaps were also subjected to glycosaminoglycan (GAG) and hydroxyproline content assays and computed tomography angiography.</p><p><strong>Results: </strong> Histological assessment indicated that cellular content was completely removed in all flap layers following a 10-hour perfusion in SDS. DNA quantification confirmed 81% DNA removal. Based on biochemical assays, decellularized flaps had hydroxyproline content comparable with that of native flaps, although significantly fewer GAGs (<i>p</i> = 0.0019). Histology and computed tomography angiography illustrated the integrity and perfusability of the vascular system.</p><p><strong>Conclusion: </strong> The proposed angiosome-guided perfusion decellularization protocol could effectively remove cellular content from rat fasciocutaneous flaps and preserve the integrity of innate vascular networks.</p>","PeriodicalId":16949,"journal":{"name":"Journal of reconstructive microsurgery","volume":" ","pages":""},"PeriodicalIF":2.2000,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Angiosome-Guided Perfusion Decellularization of Fasciocutaneous Flaps.\",\"authors\":\"Liya Yang, Xueshan Bai, Yuanbo Liu, Shan Zhu, Shanshan Li, Zixiang Chen, Tinglu Han, Shengyang Jin, Mengqing Zang\",\"doi\":\"10.1055/a-2404-2608\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong> Tissue engineering based on whole-organ perfusion decellularization has successfully generated small-animal organs, including the heart and limbs. Herein, we aimed to use angiosome-guided perfusion decellularization to develop an acellular fasciocutaneous flap matrix with an intact vascular network.</p><p><strong>Methods: </strong> Abdominal flaps of rats were harvested, and the vascular pedicle (iliac artery and vein) was dissected and injected with methylene blue to identify the angiosome region and determine the flap dimension for harvesting. To decellularize flaps, the iliac artery was perfused sequentially with 1% sodium dodecyl sulfate (SDS), deionized water, and 1% Triton-X100. Gross morphology, histology, and DNA quantity of flaps were then obtained. Flaps were also subjected to glycosaminoglycan (GAG) and hydroxyproline content assays and computed tomography angiography.</p><p><strong>Results: </strong> Histological assessment indicated that cellular content was completely removed in all flap layers following a 10-hour perfusion in SDS. DNA quantification confirmed 81% DNA removal. Based on biochemical assays, decellularized flaps had hydroxyproline content comparable with that of native flaps, although significantly fewer GAGs (<i>p</i> = 0.0019). Histology and computed tomography angiography illustrated the integrity and perfusability of the vascular system.</p><p><strong>Conclusion: </strong> The proposed angiosome-guided perfusion decellularization protocol could effectively remove cellular content from rat fasciocutaneous flaps and preserve the integrity of innate vascular networks.</p>\",\"PeriodicalId\":16949,\"journal\":{\"name\":\"Journal of reconstructive microsurgery\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2024-09-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of reconstructive microsurgery\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1055/a-2404-2608\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"SURGERY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of reconstructive microsurgery","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1055/a-2404-2608","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"SURGERY","Score":null,"Total":0}
引用次数: 0
摘要
背景 基于全器官灌注脱细胞的组织工程已成功生成了包括心脏和四肢在内的小动物器官。在此,我们旨在利用血管组引导的灌注脱细胞生成具有完整血管网络的无细胞筋膜皮瓣基质。方法 采集大鼠腹部皮瓣,解剖血管蒂(髂动脉和静脉)并注射亚甲蓝,以确定血管蒂区域并确定皮瓣采集尺寸。为使皮瓣脱细胞,用1%十二烷基硫酸钠、去离子水和1% Triton-X100依次灌注髂动脉。然后获得皮瓣的大体形态、组织学和 DNA 数量。还对皮瓣进行了糖胺聚糖和羟脯氨酸含量检测以及计算机断层扫描血管造影。结果 组织学评估表明,在十二烷基硫酸钠中灌注 10 小时后,所有皮瓣层中的细胞成分都被完全清除。DNA 定量证实,81% 的 DNA 被清除。根据生化检测结果,脱细胞皮瓣的羟脯氨酸含量与原生皮瓣相当,但糖胺聚糖明显较少(p = 0.0019)。组织学和计算机断层扫描血管造影显示了血管系统的完整性和可灌注性。结论 拟议的血管造影剂引导的灌注脱细胞方案可有效去除大鼠筋膜瓣中的细胞成分,并保持先天血管网络的完整性。
Angiosome-Guided Perfusion Decellularization of Fasciocutaneous Flaps.
Background: Tissue engineering based on whole-organ perfusion decellularization has successfully generated small-animal organs, including the heart and limbs. Herein, we aimed to use angiosome-guided perfusion decellularization to develop an acellular fasciocutaneous flap matrix with an intact vascular network.
Methods: Abdominal flaps of rats were harvested, and the vascular pedicle (iliac artery and vein) was dissected and injected with methylene blue to identify the angiosome region and determine the flap dimension for harvesting. To decellularize flaps, the iliac artery was perfused sequentially with 1% sodium dodecyl sulfate (SDS), deionized water, and 1% Triton-X100. Gross morphology, histology, and DNA quantity of flaps were then obtained. Flaps were also subjected to glycosaminoglycan (GAG) and hydroxyproline content assays and computed tomography angiography.
Results: Histological assessment indicated that cellular content was completely removed in all flap layers following a 10-hour perfusion in SDS. DNA quantification confirmed 81% DNA removal. Based on biochemical assays, decellularized flaps had hydroxyproline content comparable with that of native flaps, although significantly fewer GAGs (p = 0.0019). Histology and computed tomography angiography illustrated the integrity and perfusability of the vascular system.
Conclusion: The proposed angiosome-guided perfusion decellularization protocol could effectively remove cellular content from rat fasciocutaneous flaps and preserve the integrity of innate vascular networks.
期刊介绍:
The Journal of Reconstructive Microsurgery is a peer-reviewed, indexed journal that provides an international forum for the publication of articles focusing on reconstructive microsurgery and complex reconstructive surgery. The journal was originally established in 1984 for the microsurgical community to publish and share academic papers.
The Journal of Reconstructive Microsurgery provides the latest in original research spanning basic laboratory, translational, and clinical investigations. Review papers cover current topics in complex reconstruction and microsurgery. In addition, special sections discuss new technologies, innovations, materials, and significant problem cases.
The journal welcomes controversial topics, editorial comments, book reviews, and letters to the Editor, in order to complete the balanced spectrum of information available in the Journal of Reconstructive Microsurgery. All articles undergo stringent peer review by international experts in the specialty.