Immobilized Lentivirus Vector on Chondroitin Sulfate-Hyaluronate Acid-Silk Fibroin Hybrid Scaffold for Tissue-Engineered Ligament-Bone Junction
The lack of a fibrocartilage layer between graft and bone remains the leading cause of graft failure after anterior cruciate ligament (ACL) reconstruction. The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid...
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| Veröffentlicht in: | BioMed research international 2014-01, Vol.2014 (2014), p.1-10 |
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| creator | Sun, Liguo Li, Hongguo Qu, Ling Zhu, Rui Fan, Xiangli Xue, Yingsen Xie, Zhenghong Fan, Hongbin |
| description | The lack of a fibrocartilage layer between graft and bone remains the leading cause of graft failure after anterior cruciate ligament (ACL) reconstruction. The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid scaffold for reconstructing the fibrocartilage layer. The scaffold was fabricated by lyophilizing the CHS mixture with braided silk cables. The scanning electronic microscopy (SEM) showed that microporous CHS sponges were formed around silk cables. Each end of scaffold was modified with lentiviral-mediated transforming growth factor-β3 (TGF-β3) gene. The cells on scaffold were transfected by bonded lentivirus. In vitro culture demonstrated that mesenchymal stem cells (MSCs) on scaffolds proliferated vigorously and produced abundant collagen. The transcription levels of cartilage-specific genes also increased with culture time. After 2 weeks, the MSCs were distributed uniformly throughout scaffold. Deposited collagen was also found to increase. The chondral differentiation of MSCs was verified by expressions of collagen II and TGF-β3 genes in mRNA and protein level. Histology also confirmed the production of cartilage extracellular matrix (ECM) components. The results demonstrated that gene-modified silk cable-reinforced CHS scaffold was capable of supporting cell proliferation and differentiation to reconstruct the cartilage layer of interface. |
| doi_str_mv | 10.1155/2014/816979 |
| format | Article |
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The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid scaffold for reconstructing the fibrocartilage layer. The scaffold was fabricated by lyophilizing the CHS mixture with braided silk cables. The scanning electronic microscopy (SEM) showed that microporous CHS sponges were formed around silk cables. Each end of scaffold was modified with lentiviral-mediated transforming growth factor-β3 (TGF-β3) gene. The cells on scaffold were transfected by bonded lentivirus. In vitro culture demonstrated that mesenchymal stem cells (MSCs) on scaffolds proliferated vigorously and produced abundant collagen. The transcription levels of cartilage-specific genes also increased with culture time. After 2 weeks, the MSCs were distributed uniformly throughout scaffold. Deposited collagen was also found to increase. The chondral differentiation of MSCs was verified by expressions of collagen II and TGF-β3 genes in mRNA and protein level. Histology also confirmed the production of cartilage extracellular matrix (ECM) components. The results demonstrated that gene-modified silk cable-reinforced CHS scaffold was capable of supporting cell proliferation and differentiation to reconstruct the cartilage layer of interface.</description><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2014/816979</identifier><identifier>PMID: 25019087</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Puplishing Corporation</publisher><subject>Adsorption ; Animals ; Biocompatible Materials - chemical synthesis ; Biomechanics ; Bone and Bones - cytology ; Bone and Bones - physiology ; Bone surgery ; Cables ; Cell Adhesion - physiology ; Cell Differentiation - physiology ; Cell growth ; Cell Proliferation - physiology ; Cells, Cultured ; Chondroitin sulfate ; Chondroitin Sulfates - chemistry ; Collagen ; Collagens ; Culture ; Differentiation ; Fibroins - chemistry ; Genes ; Genetic Vectors - genetics ; Grafting ; Health aspects ; Hospitals ; Hyaluronic Acid - chemistry ; Lentivirus ; Lentivirus - genetics ; Ligaments ; Ligaments - cytology ; Ligaments - physiology ; Materials Testing ; Mesenchymal Stromal Cells - cytology ; Mesenchymal Stromal Cells - physiology ; Orthopedic surgery ; Rabbits ; Scaffolds ; Silk ; Tensile strength ; Tissue engineering ; Tissue Engineering - instrumentation ; Tissue Engineering - methods ; Tissue Scaffolds ; Transforming Growth Factor beta3 - genetics</subject><ispartof>BioMed research international, 2014-01, Vol.2014 (2014), p.1-10</ispartof><rights>Copyright © 2014 Liguo Sun et al.</rights><rights>COPYRIGHT 2014 John Wiley & Sons, Inc.</rights><rights>Copyright © 2014 Liguo Sun et al. Liguo Sun et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2014 Liguo Sun et al. 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c560t-bde6a359f45f32a048b355da223d2cb623f4bd6d8492f7344a85e1db3d0e2ebe3</citedby><cites>FETCH-LOGICAL-c560t-bde6a359f45f32a048b355da223d2cb623f4bd6d8492f7344a85e1db3d0e2ebe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4075190/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4075190/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25019087$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Pei, Guo-Xian</contributor><creatorcontrib>Sun, Liguo</creatorcontrib><creatorcontrib>Li, Hongguo</creatorcontrib><creatorcontrib>Qu, Ling</creatorcontrib><creatorcontrib>Zhu, Rui</creatorcontrib><creatorcontrib>Fan, Xiangli</creatorcontrib><creatorcontrib>Xue, Yingsen</creatorcontrib><creatorcontrib>Xie, Zhenghong</creatorcontrib><creatorcontrib>Fan, Hongbin</creatorcontrib><title>Immobilized Lentivirus Vector on Chondroitin Sulfate-Hyaluronate Acid-Silk Fibroin Hybrid Scaffold for Tissue-Engineered Ligament-Bone Junction</title><title>BioMed research international</title><addtitle>Biomed Res Int</addtitle><description>The lack of a fibrocartilage layer between graft and bone remains the leading cause of graft failure after anterior cruciate ligament (ACL) reconstruction. The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid scaffold for reconstructing the fibrocartilage layer. The scaffold was fabricated by lyophilizing the CHS mixture with braided silk cables. The scanning electronic microscopy (SEM) showed that microporous CHS sponges were formed around silk cables. Each end of scaffold was modified with lentiviral-mediated transforming growth factor-β3 (TGF-β3) gene. The cells on scaffold were transfected by bonded lentivirus. In vitro culture demonstrated that mesenchymal stem cells (MSCs) on scaffolds proliferated vigorously and produced abundant collagen. The transcription levels of cartilage-specific genes also increased with culture time. After 2 weeks, the MSCs were distributed uniformly throughout scaffold. Deposited collagen was also found to increase. The chondral differentiation of MSCs was verified by expressions of collagen II and TGF-β3 genes in mRNA and protein level. Histology also confirmed the production of cartilage extracellular matrix (ECM) components. The results demonstrated that gene-modified silk cable-reinforced CHS scaffold was capable of supporting cell proliferation and differentiation to reconstruct the cartilage layer of interface.</description><subject>Adsorption</subject><subject>Animals</subject><subject>Biocompatible Materials - chemical synthesis</subject><subject>Biomechanics</subject><subject>Bone and Bones - cytology</subject><subject>Bone and Bones - physiology</subject><subject>Bone surgery</subject><subject>Cables</subject><subject>Cell Adhesion - physiology</subject><subject>Cell Differentiation - physiology</subject><subject>Cell growth</subject><subject>Cell Proliferation - physiology</subject><subject>Cells, Cultured</subject><subject>Chondroitin sulfate</subject><subject>Chondroitin Sulfates - chemistry</subject><subject>Collagen</subject><subject>Collagens</subject><subject>Culture</subject><subject>Differentiation</subject><subject>Fibroins - chemistry</subject><subject>Genes</subject><subject>Genetic Vectors - genetics</subject><subject>Grafting</subject><subject>Health aspects</subject><subject>Hospitals</subject><subject>Hyaluronic Acid - chemistry</subject><subject>Lentivirus</subject><subject>Lentivirus - genetics</subject><subject>Ligaments</subject><subject>Ligaments - cytology</subject><subject>Ligaments - physiology</subject><subject>Materials Testing</subject><subject>Mesenchymal Stromal Cells - cytology</subject><subject>Mesenchymal Stromal Cells - physiology</subject><subject>Orthopedic surgery</subject><subject>Rabbits</subject><subject>Scaffolds</subject><subject>Silk</subject><subject>Tensile strength</subject><subject>Tissue engineering</subject><subject>Tissue Engineering - instrumentation</subject><subject>Tissue Engineering - methods</subject><subject>Tissue Scaffolds</subject><subject>Transforming Growth Factor beta3 - genetics</subject><issn>2314-6133</issn><issn>2314-6141</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>RHX</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqN0k1v0zAYB_AIgdg0duIMssQFgcL8nuSCVKqNDlXi0MHVcuLHrUdiDzsZKl-Cr4yrjvJyYb7Ykn_6P_ajpyieEvyGECHOKCb8rCayqZoHxTFlhJeScPLwcGbsqDhN6RrnlR1u5OPiiApMGlxXx8WPy2EIrevddzBoCX50ty5OCX2GbgwRBY_mm-BNDG50Hq2m3uoRysVW91MMPp_RrHOmXLn-C7pwbXYeLbZtdAatOm1t6A2yOejKpTRBee7XzgPEXTG31kMuWL4LHtCHyXejC_5J8cjqPsHp3X5SfLo4v5ovyuXH95fz2bLshMRj2RqQmonGcmEZ1ZjXLRPCaEqZoV0rKbO8NdLUvKG2YpzrWgAxLTMYKLTAToq3-9ybqR3AdPkhUffqJrpBx60K2qm_b7zbqHW4VRxXIvcuB7y8C4jh6wRpVINLHfS99hCmpIjklDaiquQ9KCGSiVrcgwouSN0wKTJ98Q-9DlP0uWlZCcplw6vqt1rrHpTzNuTfdLtQNeO0wpSLimX1eq-6GFKKYA-NIFjt5kzt5kzt5yzr53_27mB_TVUGr_Zg47zR39x_0p7tMWQCVh-wwJgLxn4CxUjkQA</recordid><startdate>20140101</startdate><enddate>20140101</enddate><creator>Sun, Liguo</creator><creator>Li, Hongguo</creator><creator>Qu, Ling</creator><creator>Zhu, Rui</creator><creator>Fan, Xiangli</creator><creator>Xue, Yingsen</creator><creator>Xie, Zhenghong</creator><creator>Fan, Hongbin</creator><general>Hindawi Puplishing Corporation</general><general>Hindawi Publishing Corporation</general><general>John Wiley & Sons, Inc</general><general>Hindawi Limited</general><scope>ADJCN</scope><scope>AHFXO</scope><scope>RHU</scope><scope>RHW</scope><scope>RHX</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>CWDGH</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>7TB</scope><scope>7U5</scope><scope>F28</scope><scope>L7M</scope><scope>5PM</scope></search><sort><creationdate>20140101</creationdate><title>Immobilized Lentivirus Vector on Chondroitin Sulfate-Hyaluronate Acid-Silk Fibroin Hybrid Scaffold for Tissue-Engineered Ligament-Bone Junction</title><author>Sun, Liguo ; Li, Hongguo ; Qu, Ling ; Zhu, Rui ; Fan, Xiangli ; Xue, Yingsen ; Xie, Zhenghong ; Fan, Hongbin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c560t-bde6a359f45f32a048b355da223d2cb623f4bd6d8492f7344a85e1db3d0e2ebe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Adsorption</topic><topic>Animals</topic><topic>Biocompatible Materials - 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The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid scaffold for reconstructing the fibrocartilage layer. The scaffold was fabricated by lyophilizing the CHS mixture with braided silk cables. The scanning electronic microscopy (SEM) showed that microporous CHS sponges were formed around silk cables. Each end of scaffold was modified with lentiviral-mediated transforming growth factor-β3 (TGF-β3) gene. The cells on scaffold were transfected by bonded lentivirus. In vitro culture demonstrated that mesenchymal stem cells (MSCs) on scaffolds proliferated vigorously and produced abundant collagen. The transcription levels of cartilage-specific genes also increased with culture time. After 2 weeks, the MSCs were distributed uniformly throughout scaffold. Deposited collagen was also found to increase. The chondral differentiation of MSCs was verified by expressions of collagen II and TGF-β3 genes in mRNA and protein level. Histology also confirmed the production of cartilage extracellular matrix (ECM) components. The results demonstrated that gene-modified silk cable-reinforced CHS scaffold was capable of supporting cell proliferation and differentiation to reconstruct the cartilage layer of interface.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Puplishing Corporation</pub><pmid>25019087</pmid><doi>10.1155/2014/816979</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | BioMed research international, 2014-01, Vol.2014 (2014), p.1-10 |
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| subjects | Adsorption Animals Biocompatible Materials - chemical synthesis Biomechanics Bone and Bones - cytology Bone and Bones - physiology Bone surgery Cables Cell Adhesion - physiology Cell Differentiation - physiology Cell growth Cell Proliferation - physiology Cells, Cultured Chondroitin sulfate Chondroitin Sulfates - chemistry Collagen Collagens Culture Differentiation Fibroins - chemistry Genes Genetic Vectors - genetics Grafting Health aspects Hospitals Hyaluronic Acid - chemistry Lentivirus Lentivirus - genetics Ligaments Ligaments - cytology Ligaments - physiology Materials Testing Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - physiology Orthopedic surgery Rabbits Scaffolds Silk Tensile strength Tissue engineering Tissue Engineering - instrumentation Tissue Engineering - methods Tissue Scaffolds Transforming Growth Factor beta3 - genetics |
| title | Immobilized Lentivirus Vector on Chondroitin Sulfate-Hyaluronate Acid-Silk Fibroin Hybrid Scaffold for Tissue-Engineered Ligament-Bone Junction |
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