Differential expression of GAP‐43 and neurofilament during peripheral nerve regeneration through bio‐artificial conduits
Nerve conduits are promising alternatives for repairing nerve gaps; they provide a close microenvironment that supports nerve regeneration. In this sense, histological analysis of axonal growth is a determinant to achieve successful nerve regeneration. To evaluate this process, the most‐used immunoh...
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description | Nerve conduits are promising alternatives for repairing nerve gaps; they provide a close microenvironment that supports nerve regeneration. In this sense, histological analysis of axonal growth is a determinant to achieve successful nerve regeneration. To evaluate this process, the most‐used immunohistochemical markers are neurofilament (NF), β‐III tubulin and, infrequently, GAP‐43. However, GAP‐43 expression in long‐term nerve regeneration models is still poorly understood. In this study we analysed GAP‐43 expression and its correlation with NF and S‐100, using three tissue‐engineering approaches with different regeneration profiles. A 10 mm gap was created in the sciatic nerve of 12 rats and repaired using collagen conduits or collagen conduits filled with fibrin–agarose hydrogels or with hydrogels containing autologous adipose‐derived mesenchymal stem cells (ADMSCs). After 12 weeks the conduits were harvested for histological analysis. Our results confirm the long‐term expression of GAP‐43 in all groups. The expression of GAP‐43 and NF was significantly higher in the group with ADMSCs. Interestingly, GAP‐43 was observed in immature, newly formed axons and NF in thicker and mature axons. These proteins were not co‐expressed, demonstrating their differential expression in newly formed nerve fascicles. Our descriptive and quantitative histological analysis of GAP‐43 and NFL allowed us to determine, with high accuracy, the heterogenic population of axons at different stages of maturation in three tissue‐engineering approaches. Finally, to perform a complete assessment of axonal regeneration, the quantitative immunohistochemical evaluation of both GAP‐43 and NF could be a useful quality control in tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd. |
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In this sense, histological analysis of axonal growth is a determinant to achieve successful nerve regeneration. To evaluate this process, the most‐used immunohistochemical markers are neurofilament (NF), β‐III tubulin and, infrequently, GAP‐43. However, GAP‐43 expression in long‐term nerve regeneration models is still poorly understood. In this study we analysed GAP‐43 expression and its correlation with NF and S‐100, using three tissue‐engineering approaches with different regeneration profiles. A 10 mm gap was created in the sciatic nerve of 12 rats and repaired using collagen conduits or collagen conduits filled with fibrin–agarose hydrogels or with hydrogels containing autologous adipose‐derived mesenchymal stem cells (ADMSCs). After 12 weeks the conduits were harvested for histological analysis. Our results confirm the long‐term expression of GAP‐43 in all groups. The expression of GAP‐43 and NF was significantly higher in the group with ADMSCs. Interestingly, GAP‐43 was observed in immature, newly formed axons and NF in thicker and mature axons. These proteins were not co‐expressed, demonstrating their differential expression in newly formed nerve fascicles. Our descriptive and quantitative histological analysis of GAP‐43 and NFL allowed us to determine, with high accuracy, the heterogenic population of axons at different stages of maturation in three tissue‐engineering approaches. Finally, to perform a complete assessment of axonal regeneration, the quantitative immunohistochemical evaluation of both GAP‐43 and NF could be a useful quality control in tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd.</description><identifier>ISSN: 1932-6254</identifier><identifier>EISSN: 1932-7005</identifier><identifier>DOI: 10.1002/term.1949</identifier><identifier>PMID: 25080900</identifier><language>eng</language><publisher>England: Hindawi Limited</publisher><subject>adipose‐derived mesenchymal stem cells ; Animals ; Axons - metabolism ; Biocompatible Materials - metabolism ; Collagen - metabolism ; Fibrin - chemistry ; GAP-43 Protein - metabolism ; growth‐associated protein 43 ; Hydrogels - chemistry ; immunohistochemistry ; Intermediate Filaments - metabolism ; Male ; Mesenchymal Stromal Cells - cytology ; nerve regeneration ; Nerve Regeneration - physiology ; neurofilament ; Rats ; Rats, Wistar ; Regenerative medicine ; Schwann Cells - cytology ; Sciatic Nerve - pathology ; Sepharose - chemistry ; Tissue engineering ; Tissue Engineering - methods</subject><ispartof>Journal of tissue engineering and regenerative medicine, 2017-02, Vol.11 (2), p.553-563</ispartof><rights>Copyright © 2014 John Wiley & Sons, Ltd.</rights><rights>Copyright © 2017 John Wiley & Sons, Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3029-151962bdf7cf84ef7d7c18eb393a0f5b74525243595f60473c8ade2d0d4203193</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,1416,27915,27916</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25080900$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Carriel, Víctor</creatorcontrib><creatorcontrib>Garzón, Ingrid</creatorcontrib><creatorcontrib>Campos, Antonio</creatorcontrib><creatorcontrib>Cornelissen, Maria</creatorcontrib><creatorcontrib>Alaminos, Miguel</creatorcontrib><title>Differential expression of GAP‐43 and neurofilament during peripheral nerve regeneration through bio‐artificial conduits</title><title>Journal of tissue engineering and regenerative medicine</title><addtitle>J Tissue Eng Regen Med</addtitle><description>Nerve conduits are promising alternatives for repairing nerve gaps; they provide a close microenvironment that supports nerve regeneration. In this sense, histological analysis of axonal growth is a determinant to achieve successful nerve regeneration. To evaluate this process, the most‐used immunohistochemical markers are neurofilament (NF), β‐III tubulin and, infrequently, GAP‐43. However, GAP‐43 expression in long‐term nerve regeneration models is still poorly understood. In this study we analysed GAP‐43 expression and its correlation with NF and S‐100, using three tissue‐engineering approaches with different regeneration profiles. A 10 mm gap was created in the sciatic nerve of 12 rats and repaired using collagen conduits or collagen conduits filled with fibrin–agarose hydrogels or with hydrogels containing autologous adipose‐derived mesenchymal stem cells (ADMSCs). After 12 weeks the conduits were harvested for histological analysis. Our results confirm the long‐term expression of GAP‐43 in all groups. The expression of GAP‐43 and NF was significantly higher in the group with ADMSCs. Interestingly, GAP‐43 was observed in immature, newly formed axons and NF in thicker and mature axons. These proteins were not co‐expressed, demonstrating their differential expression in newly formed nerve fascicles. Our descriptive and quantitative histological analysis of GAP‐43 and NFL allowed us to determine, with high accuracy, the heterogenic population of axons at different stages of maturation in three tissue‐engineering approaches. Finally, to perform a complete assessment of axonal regeneration, the quantitative immunohistochemical evaluation of both GAP‐43 and NF could be a useful quality control in tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd.</description><subject>adipose‐derived mesenchymal stem cells</subject><subject>Animals</subject><subject>Axons - metabolism</subject><subject>Biocompatible Materials - metabolism</subject><subject>Collagen - metabolism</subject><subject>Fibrin - chemistry</subject><subject>GAP-43 Protein - metabolism</subject><subject>growth‐associated protein 43</subject><subject>Hydrogels - chemistry</subject><subject>immunohistochemistry</subject><subject>Intermediate Filaments - metabolism</subject><subject>Male</subject><subject>Mesenchymal Stromal Cells - cytology</subject><subject>nerve regeneration</subject><subject>Nerve Regeneration - physiology</subject><subject>neurofilament</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Regenerative medicine</subject><subject>Schwann Cells - cytology</subject><subject>Sciatic Nerve - pathology</subject><subject>Sepharose - chemistry</subject><subject>Tissue engineering</subject><subject>Tissue Engineering - methods</subject><issn>1932-6254</issn><issn>1932-7005</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkctu1TAURS0EoqUw4AeQJSZMbnv8TDKs-gKpCITKOHLi43tdJXawE6BSB3wC38iX4KgXBowYecteXkf2JuQlg2MGwE9mTOMxa2TziByyRvBNBaAe77PmSh6QZznflk2llXhKDriCGhqAQ3J_7p3DhGH2ZqD4fUqYs4-BRkevTj_--vFTCmqCpQGXFJ0fzFhYapfkw5ZOmPy0w1SuBkxfkSbcYklmXhXzLsVlu6Odj8Vj0uyd79cxfQx28XN-Tp44M2R8sV-PyOfLi5uzt5vrD1fvzk6vN70A3myYYo3mnXVV72qJrrJVz2rsRCMMONVVUnHFpVCNchpkJfraWOQWrOQgyicckTcP3inFLwvmuR197nEYTMC45JbVuhaM17r6D5RrDQLUan39D3oblxTKQ1ahkiCE1IV6taeWbkTbTsmPJt21fyoowMkD8M0PePf3nEG7dtuu3bZrt-3Nxaf3axC_AZxHmNI</recordid><startdate>201702</startdate><enddate>201702</enddate><creator>Carriel, Víctor</creator><creator>Garzón, Ingrid</creator><creator>Campos, Antonio</creator><creator>Cornelissen, Maria</creator><creator>Alaminos, Miguel</creator><general>Hindawi Limited</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>M7Z</scope><scope>P64</scope><scope>7X8</scope><scope>7QO</scope></search><sort><creationdate>201702</creationdate><title>Differential expression of GAP‐43 and neurofilament during peripheral nerve regeneration through bio‐artificial conduits</title><author>Carriel, Víctor ; Garzón, Ingrid ; Campos, Antonio ; Cornelissen, Maria ; Alaminos, Miguel</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3029-151962bdf7cf84ef7d7c18eb393a0f5b74525243595f60473c8ade2d0d4203193</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>adipose‐derived mesenchymal stem cells</topic><topic>Animals</topic><topic>Axons - metabolism</topic><topic>Biocompatible Materials - metabolism</topic><topic>Collagen - metabolism</topic><topic>Fibrin - chemistry</topic><topic>GAP-43 Protein - metabolism</topic><topic>growth‐associated protein 43</topic><topic>Hydrogels - chemistry</topic><topic>immunohistochemistry</topic><topic>Intermediate Filaments - metabolism</topic><topic>Male</topic><topic>Mesenchymal Stromal Cells - cytology</topic><topic>nerve regeneration</topic><topic>Nerve Regeneration - physiology</topic><topic>neurofilament</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>Regenerative medicine</topic><topic>Schwann Cells - cytology</topic><topic>Sciatic Nerve - pathology</topic><topic>Sepharose - chemistry</topic><topic>Tissue engineering</topic><topic>Tissue Engineering - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Carriel, Víctor</creatorcontrib><creatorcontrib>Garzón, Ingrid</creatorcontrib><creatorcontrib>Campos, Antonio</creatorcontrib><creatorcontrib>Cornelissen, Maria</creatorcontrib><creatorcontrib>Alaminos, Miguel</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biochemistry Abstracts 1</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Journal of tissue engineering and regenerative medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Carriel, Víctor</au><au>Garzón, Ingrid</au><au>Campos, Antonio</au><au>Cornelissen, Maria</au><au>Alaminos, Miguel</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Differential expression of GAP‐43 and neurofilament during peripheral nerve regeneration through bio‐artificial conduits</atitle><jtitle>Journal of tissue engineering and regenerative medicine</jtitle><addtitle>J Tissue Eng Regen Med</addtitle><date>2017-02</date><risdate>2017</risdate><volume>11</volume><issue>2</issue><spage>553</spage><epage>563</epage><pages>553-563</pages><issn>1932-6254</issn><eissn>1932-7005</eissn><abstract>Nerve conduits are promising alternatives for repairing nerve gaps; they provide a close microenvironment that supports nerve regeneration. In this sense, histological analysis of axonal growth is a determinant to achieve successful nerve regeneration. To evaluate this process, the most‐used immunohistochemical markers are neurofilament (NF), β‐III tubulin and, infrequently, GAP‐43. However, GAP‐43 expression in long‐term nerve regeneration models is still poorly understood. In this study we analysed GAP‐43 expression and its correlation with NF and S‐100, using three tissue‐engineering approaches with different regeneration profiles. A 10 mm gap was created in the sciatic nerve of 12 rats and repaired using collagen conduits or collagen conduits filled with fibrin–agarose hydrogels or with hydrogels containing autologous adipose‐derived mesenchymal stem cells (ADMSCs). After 12 weeks the conduits were harvested for histological analysis. Our results confirm the long‐term expression of GAP‐43 in all groups. The expression of GAP‐43 and NF was significantly higher in the group with ADMSCs. Interestingly, GAP‐43 was observed in immature, newly formed axons and NF in thicker and mature axons. These proteins were not co‐expressed, demonstrating their differential expression in newly formed nerve fascicles. Our descriptive and quantitative histological analysis of GAP‐43 and NFL allowed us to determine, with high accuracy, the heterogenic population of axons at different stages of maturation in three tissue‐engineering approaches. Finally, to perform a complete assessment of axonal regeneration, the quantitative immunohistochemical evaluation of both GAP‐43 and NF could be a useful quality control in tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd.</abstract><cop>England</cop><pub>Hindawi Limited</pub><pmid>25080900</pmid><doi>10.1002/term.1949</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | adipose‐derived mesenchymal stem cells Animals Axons - metabolism Biocompatible Materials - metabolism Collagen - metabolism Fibrin - chemistry GAP-43 Protein - metabolism growth‐associated protein 43 Hydrogels - chemistry immunohistochemistry Intermediate Filaments - metabolism Male Mesenchymal Stromal Cells - cytology nerve regeneration Nerve Regeneration - physiology neurofilament Rats Rats, Wistar Regenerative medicine Schwann Cells - cytology Sciatic Nerve - pathology Sepharose - chemistry Tissue engineering Tissue Engineering - methods |
title | Differential expression of GAP‐43 and neurofilament during peripheral nerve regeneration through bio‐artificial conduits |
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