Z Niknam, H Zali, V Mansouri, M Rezaei Tavirani, M Omidi
{"title":"聚己内酯/氧化镁/氧化石墨烯支架中间质干细胞分化的成骨细胞的形态学和分子分析","authors":"Z Niknam, H Zali, V Mansouri, M Rezaei Tavirani, M Omidi","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The loss or dysfunction of bone tissue that observed after bone tumor resections and severe nonunion fractures afflicts 200 million people worldwide. Bone tissue engineering is a promising approach to repair osteoporotic fractures.</p><p><strong>Objective: </strong>In this paper, polycaprolactone (PCL)/magnesium oxide (MgO)/graphene oxide (GO) nanofibrous scaffold was fabricated by electrospining method, and its biocompatibility and osteogenic differentiation of adipose-derived mesenchymal stem cells (MSCs) on this scaffold were evaluated and compared with pure PCL nanofibrous scaffold.</p><p><strong>Methods: </strong>SEM analysis, DAPI staining and MTT assay were used to evaluation biocompatibility of PCL/MgO/GO composite scaffold. In addition by ALP assay and proteomic approach, osteostimulatory effect of electrospun composite scaffold was investigated and the expression level of osteogenic markers including Runt-related transcription factor cbfa1/runx2 (runx2), collagen type I (Col1a1) and osteopontin (OPN) in MSCs seeded on PCL/MgO/GO composite scaffold was determined and compared with pure PCL scaffold. Then, RT-PCR technique was used to validate the level expression of these genes.</p><p><strong>Results: </strong>The obtained results showed that adhesion, viability and ALP activity of MSCs on PCL/MgO/GO scaffold considerably enhanced compared with pure PCL. As well as proteomic and real-time analysis illustrated the expression of osteogenic markers including runx2, Col1a1 and OPN increased (>2-fold) in cells seeded on PCL/MgO/GO composite scaffold.</p><p><strong>Conclusion: </strong>It was concluded that MgO and GO nanoparticles could improve the biocompatibility of PCL scaffold and enhance the osteogenic differentiation of MSCs.</p>","PeriodicalId":14242,"journal":{"name":"International Journal of Organ Transplantation Medicine","volume":null,"pages":null},"PeriodicalIF":0.3000,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7722513/pdf/ijotm-10-171.pdf","citationCount":"0","resultStr":"{\"title\":\"Morphological and Molecular Analysis of Osteoblasts Differentiated from Mesenchymal Stem Cells in Polycaprolactone/Magnesium Oxide/Graphene Oxide Scaffold.\",\"authors\":\"Z Niknam, H Zali, V Mansouri, M Rezaei Tavirani, M Omidi\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The loss or dysfunction of bone tissue that observed after bone tumor resections and severe nonunion fractures afflicts 200 million people worldwide. Bone tissue engineering is a promising approach to repair osteoporotic fractures.</p><p><strong>Objective: </strong>In this paper, polycaprolactone (PCL)/magnesium oxide (MgO)/graphene oxide (GO) nanofibrous scaffold was fabricated by electrospining method, and its biocompatibility and osteogenic differentiation of adipose-derived mesenchymal stem cells (MSCs) on this scaffold were evaluated and compared with pure PCL nanofibrous scaffold.</p><p><strong>Methods: </strong>SEM analysis, DAPI staining and MTT assay were used to evaluation biocompatibility of PCL/MgO/GO composite scaffold. In addition by ALP assay and proteomic approach, osteostimulatory effect of electrospun composite scaffold was investigated and the expression level of osteogenic markers including Runt-related transcription factor cbfa1/runx2 (runx2), collagen type I (Col1a1) and osteopontin (OPN) in MSCs seeded on PCL/MgO/GO composite scaffold was determined and compared with pure PCL scaffold. Then, RT-PCR technique was used to validate the level expression of these genes.</p><p><strong>Results: </strong>The obtained results showed that adhesion, viability and ALP activity of MSCs on PCL/MgO/GO scaffold considerably enhanced compared with pure PCL. As well as proteomic and real-time analysis illustrated the expression of osteogenic markers including runx2, Col1a1 and OPN increased (>2-fold) in cells seeded on PCL/MgO/GO composite scaffold.</p><p><strong>Conclusion: </strong>It was concluded that MgO and GO nanoparticles could improve the biocompatibility of PCL scaffold and enhance the osteogenic differentiation of MSCs.</p>\",\"PeriodicalId\":14242,\"journal\":{\"name\":\"International Journal of Organ Transplantation Medicine\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.3000,\"publicationDate\":\"2019-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7722513/pdf/ijotm-10-171.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International Journal of Organ Transplantation Medicine\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"TRANSPLANTATION\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Organ Transplantation Medicine","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"TRANSPLANTATION","Score":null,"Total":0}
引用次数: 0
摘要
背景:全球有 2 亿人在骨肿瘤切除和严重骨折后出现骨组织缺失或功能障碍。骨组织工程是一种修复骨质疏松性骨折的有效方法:本文采用电纺丝法制备了聚己内酯(PCL)/氧化镁(MgO)/氧化石墨烯(GO)纳米纤维支架,评估了该支架的生物相容性以及脂肪间充质干细胞(MSCs)在该支架上的成骨分化情况,并与纯PCL纳米纤维支架进行了比较:方法:采用 SEM 分析、DAPI 染色和 MTT 试验评估 PCL/MgO/GO 复合支架的生物相容性。此外,通过 ALP 检测和蛋白质组学方法,研究了电纺复合支架的成骨刺激作用,并测定了 Runt 相关转录因子 cbfa1/runx2 (runx2)、I 型胶原蛋白 (Col1a1) 和骨生成素 (OPN) 等成骨标志物在 PCL/MgO/GO 复合支架上的表达水平,并与纯 PCL 支架进行了比较。然后,采用 RT-PCR 技术验证了这些基因的表达水平:结果:研究结果表明,与纯 PCL 相比,间充质干细胞在 PCL/MgO/GO 复合支架上的粘附性、存活率和 ALP 活性均有显著提高。蛋白质组和实时分析表明,在 PCL/MgO/GO 复合支架上播种的细胞中,成骨标志物(包括 runx2、Col1a1 和 OPN)的表达量增加(>2 倍):结论:MgO 和 GO 纳米颗粒可改善 PCL 支架的生物相容性,并增强间充质干细胞的成骨分化。
Morphological and Molecular Analysis of Osteoblasts Differentiated from Mesenchymal Stem Cells in Polycaprolactone/Magnesium Oxide/Graphene Oxide Scaffold.
Background: The loss or dysfunction of bone tissue that observed after bone tumor resections and severe nonunion fractures afflicts 200 million people worldwide. Bone tissue engineering is a promising approach to repair osteoporotic fractures.
Objective: In this paper, polycaprolactone (PCL)/magnesium oxide (MgO)/graphene oxide (GO) nanofibrous scaffold was fabricated by electrospining method, and its biocompatibility and osteogenic differentiation of adipose-derived mesenchymal stem cells (MSCs) on this scaffold were evaluated and compared with pure PCL nanofibrous scaffold.
Methods: SEM analysis, DAPI staining and MTT assay were used to evaluation biocompatibility of PCL/MgO/GO composite scaffold. In addition by ALP assay and proteomic approach, osteostimulatory effect of electrospun composite scaffold was investigated and the expression level of osteogenic markers including Runt-related transcription factor cbfa1/runx2 (runx2), collagen type I (Col1a1) and osteopontin (OPN) in MSCs seeded on PCL/MgO/GO composite scaffold was determined and compared with pure PCL scaffold. Then, RT-PCR technique was used to validate the level expression of these genes.
Results: The obtained results showed that adhesion, viability and ALP activity of MSCs on PCL/MgO/GO scaffold considerably enhanced compared with pure PCL. As well as proteomic and real-time analysis illustrated the expression of osteogenic markers including runx2, Col1a1 and OPN increased (>2-fold) in cells seeded on PCL/MgO/GO composite scaffold.
Conclusion: It was concluded that MgO and GO nanoparticles could improve the biocompatibility of PCL scaffold and enhance the osteogenic differentiation of MSCs.
期刊介绍:
The International Journal of Organ Transplantation Medicine (IJOTM) is a quarterly peer-reviewed English-language journal that publishes high-quality basic sciences and clinical research on transplantation. The scope of the journal includes organ and tissue donation, procurement and preservation; surgical techniques, innovations, and novelties in all aspects of transplantation; genomics and immunobiology; immunosuppressive drugs and pharmacology relevant to transplantation; graft survival and prevention of graft dysfunction and failure; clinical trials and population analyses in the field of transplantation; transplant complications; cell and tissue transplantation; infection; post-transplant malignancies; sociological and ethical issues and xenotransplantation.