Douglas M Gewehr, Pedro E P Carvalho, Fernanda P Izar, Alexandre G Haddad, Victor D Falkenbach Tenius, Allan F Giovanini, Luiz F Kubrusly
{"title":"在 4% 甲醛中长期储存对戊二醛处理过的牛心包中免疫表型标记物的影响","authors":"Douglas M Gewehr, Pedro E P Carvalho, Fernanda P Izar, Alexandre G Haddad, Victor D Falkenbach Tenius, Allan F Giovanini, Luiz F Kubrusly","doi":"10.1097/PAI.0000000000001221","DOIUrl":null,"url":null,"abstract":"<p><p>Bovine pericardium (BP) is widely used as a biomaterial for tissue engineering. Glutaraldehyde and formaldehyde are commonly employed in the reticulation processes to enhance the material's resistance and preservation. In this study, we assessed the impact of long-term storage in 4% formaldehyde on the quantitative expression of immunophenotypic markers in glutaraldehyde-treated BP. Histologic and immunohistochemical analyses were performed on 2 BP patches, manufactured in 2009 and 2020, respectively. Braile Biomédica provided the BP patches. Sections of BP were stained with H&E, Weigert, and picrosirius red, followed by immunolabeling for vimentin, laminin 5, collagen I, and collagen IV using a standardized protocol. Microscopic images were captured using light and polarized microscopy, and the area of the antibody signal was quantified using Image J Software. Histologic analysis showed no autolysis or significant changes in the patches. Immunohistochemical analysis revealed a diffuse distribution of collagen I and collagen IV throughout the connective tissue of the patches. The 2020 specimen exhibited higher expression levels of collagen I (21.36%) and collagen IV (24.67%) compared with the 2009 specimen (collagen I: 15.87%; collagen IV: 12.02%). Laminin did not show reactivity in either specimen. Notably, vimentin immunopositivity differed significantly between the patches, with a larger area of expression observed in the 2020 patch (54%) compared with the 2009 patch (13%). In summary, there were no substantial differences in immunophenotypic expression between the 2009 and 2020 BP patches, except for the higher vimentin expression in the 2020 BP patch.</p>","PeriodicalId":48952,"journal":{"name":"Applied Immunohistochemistry & Molecular Morphology","volume":null,"pages":null},"PeriodicalIF":1.3000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Impact of Long-term Storage in 4% Formaldehyde on Immunophenotypic Markers in Glutaraldehyde-treated Bovine Pericardium.\",\"authors\":\"Douglas M Gewehr, Pedro E P Carvalho, Fernanda P Izar, Alexandre G Haddad, Victor D Falkenbach Tenius, Allan F Giovanini, Luiz F Kubrusly\",\"doi\":\"10.1097/PAI.0000000000001221\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Bovine pericardium (BP) is widely used as a biomaterial for tissue engineering. Glutaraldehyde and formaldehyde are commonly employed in the reticulation processes to enhance the material's resistance and preservation. In this study, we assessed the impact of long-term storage in 4% formaldehyde on the quantitative expression of immunophenotypic markers in glutaraldehyde-treated BP. Histologic and immunohistochemical analyses were performed on 2 BP patches, manufactured in 2009 and 2020, respectively. Braile Biomédica provided the BP patches. Sections of BP were stained with H&E, Weigert, and picrosirius red, followed by immunolabeling for vimentin, laminin 5, collagen I, and collagen IV using a standardized protocol. Microscopic images were captured using light and polarized microscopy, and the area of the antibody signal was quantified using Image J Software. Histologic analysis showed no autolysis or significant changes in the patches. Immunohistochemical analysis revealed a diffuse distribution of collagen I and collagen IV throughout the connective tissue of the patches. The 2020 specimen exhibited higher expression levels of collagen I (21.36%) and collagen IV (24.67%) compared with the 2009 specimen (collagen I: 15.87%; collagen IV: 12.02%). Laminin did not show reactivity in either specimen. Notably, vimentin immunopositivity differed significantly between the patches, with a larger area of expression observed in the 2020 patch (54%) compared with the 2009 patch (13%). In summary, there were no substantial differences in immunophenotypic expression between the 2009 and 2020 BP patches, except for the higher vimentin expression in the 2020 BP patch.</p>\",\"PeriodicalId\":48952,\"journal\":{\"name\":\"Applied Immunohistochemistry & Molecular Morphology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.3000,\"publicationDate\":\"2024-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Applied Immunohistochemistry & Molecular Morphology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1097/PAI.0000000000001221\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/8/22 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"ANATOMY & MORPHOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Applied Immunohistochemistry & Molecular Morphology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1097/PAI.0000000000001221","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/8/22 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"ANATOMY & MORPHOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
牛心包(BP)被广泛用作组织工程的生物材料。在网状化过程中通常会使用戊二醛和甲醛来增强材料的耐受性和保存性。在本研究中,我们评估了在 4% 甲醛中长期储存对戊二醛处理 BP 免疫表型标记物定量表达的影响。我们分别对 2009 年和 2020 年生产的 2 块 BP 补丁进行了组织学和免疫组化分析。Braile Biomédica 公司提供了 BP 补丁。用 H&E、Weigert 和 picrosirius 红对 BP 切片进行染色,然后用标准化方案对波形蛋白、层粘连蛋白 5、胶原蛋白 I 和胶原蛋白 IV 进行免疫标记。使用光学显微镜和偏光显微镜采集显微图像,并使用 Image J 软件量化抗体信号的面积。组织学分析表明,斑块没有自溶或明显变化。免疫组化分析显示,胶原蛋白 I 和胶原蛋白 IV 在整个斑块的结缔组织中呈弥漫性分布。与 2009 年的标本(胶原 I:15.87%;胶原 IV:12.02%)相比,2020 年的标本显示出更高的胶原 I(21.36%)和胶原 IV(24.67%)表达水平。层粘连蛋白在两种标本中均未显示反应性。值得注意的是,不同斑块的波形蛋白免疫阳性率有显著差异,2020 年斑块(54%)的表达面积大于 2009 年斑块(13%)。总之,除了 2020 年 BP 补丁中的波形蛋白表达较高外,2009 年和 2020 年 BP 补丁的免疫表型表达没有实质性差异。
Impact of Long-term Storage in 4% Formaldehyde on Immunophenotypic Markers in Glutaraldehyde-treated Bovine Pericardium.
Bovine pericardium (BP) is widely used as a biomaterial for tissue engineering. Glutaraldehyde and formaldehyde are commonly employed in the reticulation processes to enhance the material's resistance and preservation. In this study, we assessed the impact of long-term storage in 4% formaldehyde on the quantitative expression of immunophenotypic markers in glutaraldehyde-treated BP. Histologic and immunohistochemical analyses were performed on 2 BP patches, manufactured in 2009 and 2020, respectively. Braile Biomédica provided the BP patches. Sections of BP were stained with H&E, Weigert, and picrosirius red, followed by immunolabeling for vimentin, laminin 5, collagen I, and collagen IV using a standardized protocol. Microscopic images were captured using light and polarized microscopy, and the area of the antibody signal was quantified using Image J Software. Histologic analysis showed no autolysis or significant changes in the patches. Immunohistochemical analysis revealed a diffuse distribution of collagen I and collagen IV throughout the connective tissue of the patches. The 2020 specimen exhibited higher expression levels of collagen I (21.36%) and collagen IV (24.67%) compared with the 2009 specimen (collagen I: 15.87%; collagen IV: 12.02%). Laminin did not show reactivity in either specimen. Notably, vimentin immunopositivity differed significantly between the patches, with a larger area of expression observed in the 2020 patch (54%) compared with the 2009 patch (13%). In summary, there were no substantial differences in immunophenotypic expression between the 2009 and 2020 BP patches, except for the higher vimentin expression in the 2020 BP patch.
期刊介绍:
Applied Immunohistochemistry & Molecular Morphology covers newly developed identification and detection technologies, and their applications in research and diagnosis for the applied immunohistochemist & molecular Morphologist.
Official Journal of the International Society for Immunohistochemisty and Molecular Morphology.