Minseong Kim, MyungGu Yeo, KyoungHo Lee, Min-Jeong Park, Gyeongyeop Han, Chansong Lee, Jihyo Park, Bongsu Jung
{"title":"Extraction and Characterization of Human Adipose Tissue-Derived Collagen: Toward Xeno-Free Tissue Engineering.","authors":"Minseong Kim, MyungGu Yeo, KyoungHo Lee, Min-Jeong Park, Gyeongyeop Han, Chansong Lee, Jihyo Park, Bongsu Jung","doi":"10.1007/s13770-023-00612-2","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Collagen is a key component of connective tissue and has been frequently used in the fabrication of medical devices for tissue regeneration. Human-originated collagen is particularly appealing due to its low immune response as an allograft biomaterial compared to xenografts and its ability to accelerate the regeneration process. Ethically and economically, adipose tissues available from liposuction clinics are a good resource to obtain human collagen. However, studies are still scarce on the extraction and characterization of human collagen, which originates from adipose tissue. The aim of this study is to establish a novel and simple method to extract collagen from human adipose tissue, characterize the collagen, and compare it with commercial-grade porcine collagen for tissue engineering applications.</p><p><strong>Methods: </strong>We developed a method to extract the collagen from human adipose tissue under quasi-Good Manufacturing Practice (GMP) conditions, including freezing the tissue, blood removal, and ethanol-based purification. Various techniques, including protein quantification, decellularization assessment, SDS-PAGE, FTIR, and CD spectroscopy analysis, were used for characterization. Amino acid composition was compared with commercial collagen. Biocompatibility and cell proliferation tests were performed, and in vitro tests using collagen sponge scaffolds were conducted with statistical analysis.</p><p><strong>Results: </strong>Our results showed that this human adipose-derived collagen was equivalent in quality to commercially available porcine collagen. In vitro testing demonstrated high cell attachment and the promotion of cell proliferation.</p><p><strong>Conclusion: </strong>In conclusion, we developed a simple and novel method to extract and characterize collagen and extracellular matrix from human adipose tissue, offering a potential alternative to animal-derived collagen for xeno-free tissue engineering applications.</p>","PeriodicalId":23126,"journal":{"name":"Tissue engineering and regenerative medicine","volume":" ","pages":"97-109"},"PeriodicalIF":4.4000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10764687/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Tissue engineering and regenerative medicine","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s13770-023-00612-2","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/12/11 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"CELL & TISSUE ENGINEERING","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Collagen is a key component of connective tissue and has been frequently used in the fabrication of medical devices for tissue regeneration. Human-originated collagen is particularly appealing due to its low immune response as an allograft biomaterial compared to xenografts and its ability to accelerate the regeneration process. Ethically and economically, adipose tissues available from liposuction clinics are a good resource to obtain human collagen. However, studies are still scarce on the extraction and characterization of human collagen, which originates from adipose tissue. The aim of this study is to establish a novel and simple method to extract collagen from human adipose tissue, characterize the collagen, and compare it with commercial-grade porcine collagen for tissue engineering applications.
Methods: We developed a method to extract the collagen from human adipose tissue under quasi-Good Manufacturing Practice (GMP) conditions, including freezing the tissue, blood removal, and ethanol-based purification. Various techniques, including protein quantification, decellularization assessment, SDS-PAGE, FTIR, and CD spectroscopy analysis, were used for characterization. Amino acid composition was compared with commercial collagen. Biocompatibility and cell proliferation tests were performed, and in vitro tests using collagen sponge scaffolds were conducted with statistical analysis.
Results: Our results showed that this human adipose-derived collagen was equivalent in quality to commercially available porcine collagen. In vitro testing demonstrated high cell attachment and the promotion of cell proliferation.
Conclusion: In conclusion, we developed a simple and novel method to extract and characterize collagen and extracellular matrix from human adipose tissue, offering a potential alternative to animal-derived collagen for xeno-free tissue engineering applications.
背景:胶原蛋白是结缔组织的重要组成部分,经常被用于制造组织再生医疗器械。与异种移植物相比,人源胶原蛋白作为同种生物材料的免疫反应较低,而且能够加速再生过程,因此特别具有吸引力。从道德和经济角度来看,从吸脂诊所获得的脂肪组织是获取人体胶原蛋白的良好资源。然而,关于提取和表征源自脂肪组织的人体胶原蛋白的研究仍然很少。本研究旨在建立一种新颖而简单的方法,从人体脂肪组织中提取胶原蛋白,表征胶原蛋白,并将其与商业级猪胶原蛋白进行比较,以用于组织工程应用:我们开发了一种在准良好生产规范(GMP)条件下从人体脂肪组织中提取胶原蛋白的方法,包括冷冻组织、去除血液和基于乙醇的纯化。在表征过程中采用了多种技术,包括蛋白质定量、脱细胞评估、SDS-PAGE、傅立叶变换红外光谱和 CD 光谱分析。氨基酸组成与商用胶原蛋白进行了比较。进行了生物相容性和细胞增殖测试,并使用胶原蛋白海绵支架进行了体外测试和统计分析:结果:我们的研究结果表明,这种源自人体脂肪的胶原蛋白在质量上与市售猪胶原蛋白相当。体外测试表明,这种胶原蛋白具有很高的细胞附着性,并能促进细胞增殖:总之,我们开发了一种简单而新颖的方法来提取和表征人体脂肪组织中的胶原蛋白和细胞外基质,为无异种组织工程应用提供了一种潜在的动物源胶原蛋白替代品。
期刊介绍:
Tissue Engineering and Regenerative Medicine (Tissue Eng Regen Med, TERM), the official journal of the Korean Tissue Engineering and Regenerative Medicine Society, is a publication dedicated to providing research- based solutions to issues related to human diseases. This journal publishes articles that report substantial information and original findings on tissue engineering, medical biomaterials, cells therapy, stem cell biology and regenerative medicine.