Scaffold‐free cartilage tissue engineering with a small population of human nasoseptal chondrocytes
Objective Cartilage tissue engineering is a promising approach to provide suitable materials for nasal reconstruction; however, it typically requires large numbers of cells. We have previously shown that a small number of chondrocytes cultivated within a continuous flow bioreactor can elicit substan...
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Veröffentlicht in: | The Laryngoscope 2017-03, Vol.127 (3), p.E91-E99 |
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creator | Chiu, Loraine L.Y. To, William T.H. Lee, John M. Waldman, Stephen D. |
description | Objective
Cartilage tissue engineering is a promising approach to provide suitable materials for nasal reconstruction; however, it typically requires large numbers of cells. We have previously shown that a small number of chondrocytes cultivated within a continuous flow bioreactor can elicit substantial tissue growth, but translation to human chondrocytes is not trivial. Here, we aimed to demonstrate the application of the bioreactor to generate large‐sized tissues from a small population of primary human nasoseptal chondrocytes.
Study Design
Experimental study.
Methods
Chondrocytes were cultured in the bioreactor using different medium compositions, with varying amounts of serum and with or without growth factors. Resulting engineered tissues were analyzed for physical properties, biochemical composition, tissue microstructure, and protein localization.
Results
Bioreactor‐cultivated constructs grown with serum and growth factors (basic fibroblast growth factor and transforming growth factor beta 2) had greater thickness, as well as DNA and glycosaminoglycan (GAG) contents, compared to low serum and no growth factor controls. These constructs also showed the most intense proteoglycan and collagen II staining.
Conclusion
The combination of bioreactor conditions, serum, and growth factors allowed the generation of large, thick scaffold‐free human cartilaginous tissues that resembled the native nasoseptal cartilage. There also may be implications for patient selection in future clinical applications of these engineered tissues because their GAG content decreased with donor age.
Level of Evidence
NA. Laryngoscope, 127:E91–E99, 2017 |
doi_str_mv | 10.1002/lary.26396 |
format | Article |
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Cartilage tissue engineering is a promising approach to provide suitable materials for nasal reconstruction; however, it typically requires large numbers of cells. We have previously shown that a small number of chondrocytes cultivated within a continuous flow bioreactor can elicit substantial tissue growth, but translation to human chondrocytes is not trivial. Here, we aimed to demonstrate the application of the bioreactor to generate large‐sized tissues from a small population of primary human nasoseptal chondrocytes.
Study Design
Experimental study.
Methods
Chondrocytes were cultured in the bioreactor using different medium compositions, with varying amounts of serum and with or without growth factors. Resulting engineered tissues were analyzed for physical properties, biochemical composition, tissue microstructure, and protein localization.
Results
Bioreactor‐cultivated constructs grown with serum and growth factors (basic fibroblast growth factor and transforming growth factor beta 2) had greater thickness, as well as DNA and glycosaminoglycan (GAG) contents, compared to low serum and no growth factor controls. These constructs also showed the most intense proteoglycan and collagen II staining.
Conclusion
The combination of bioreactor conditions, serum, and growth factors allowed the generation of large, thick scaffold‐free human cartilaginous tissues that resembled the native nasoseptal cartilage. There also may be implications for patient selection in future clinical applications of these engineered tissues because their GAG content decreased with donor age.
Level of Evidence
NA. Laryngoscope, 127:E91–E99, 2017</description><identifier>ISSN: 0023-852X</identifier><identifier>EISSN: 1531-4995</identifier><identifier>DOI: 10.1002/lary.26396</identifier><identifier>PMID: 27861930</identifier><language>eng</language><publisher>United States: Wiley Subscription Services, Inc</publisher><subject>Biomechanical Phenomena ; Cartilage tissue engineering ; Cell Culture Techniques ; Cells, Cultured ; Chondrocytes - cytology ; Chondrocytes - pathology ; continuous flow bioreactor ; Fibroblast Growth Factor 2 - administration & dosage ; Growth factors ; human nasoseptal cartilage ; Humans ; Immunohistochemistry ; Nasal Septum - cytology ; Nasal Surgical Procedures - methods ; primary cells ; Receptors, Transforming Growth Factor beta - administration & dosage ; Reconstructive Surgical Procedures - methods ; scaffold‐free ; Tensile Strength ; Tissue and Organ Harvesting ; Tissue engineering ; Tissue Engineering - methods ; Tissue Scaffolds</subject><ispartof>The Laryngoscope, 2017-03, Vol.127 (3), p.E91-E99</ispartof><rights>2016 The American Laryngological, Rhinological and Otological Society, Inc.</rights><rights>2017 The American Laryngological, Rhinological and Otological Society, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3576-41d8e2ef4b8b4e2861db2d5c53b25aea3f347960cce9640caf3c1faa245fad503</citedby><cites>FETCH-LOGICAL-c3576-41d8e2ef4b8b4e2861db2d5c53b25aea3f347960cce9640caf3c1faa245fad503</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Flary.26396$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Flary.26396$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27861930$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chiu, Loraine L.Y.</creatorcontrib><creatorcontrib>To, William T.H.</creatorcontrib><creatorcontrib>Lee, John M.</creatorcontrib><creatorcontrib>Waldman, Stephen D.</creatorcontrib><title>Scaffold‐free cartilage tissue engineering with a small population of human nasoseptal chondrocytes</title><title>The Laryngoscope</title><addtitle>Laryngoscope</addtitle><description>Objective
Cartilage tissue engineering is a promising approach to provide suitable materials for nasal reconstruction; however, it typically requires large numbers of cells. We have previously shown that a small number of chondrocytes cultivated within a continuous flow bioreactor can elicit substantial tissue growth, but translation to human chondrocytes is not trivial. Here, we aimed to demonstrate the application of the bioreactor to generate large‐sized tissues from a small population of primary human nasoseptal chondrocytes.
Study Design
Experimental study.
Methods
Chondrocytes were cultured in the bioreactor using different medium compositions, with varying amounts of serum and with or without growth factors. Resulting engineered tissues were analyzed for physical properties, biochemical composition, tissue microstructure, and protein localization.
Results
Bioreactor‐cultivated constructs grown with serum and growth factors (basic fibroblast growth factor and transforming growth factor beta 2) had greater thickness, as well as DNA and glycosaminoglycan (GAG) contents, compared to low serum and no growth factor controls. These constructs also showed the most intense proteoglycan and collagen II staining.
Conclusion
The combination of bioreactor conditions, serum, and growth factors allowed the generation of large, thick scaffold‐free human cartilaginous tissues that resembled the native nasoseptal cartilage. There also may be implications for patient selection in future clinical applications of these engineered tissues because their GAG content decreased with donor age.
Level of Evidence
NA. Laryngoscope, 127:E91–E99, 2017</description><subject>Biomechanical Phenomena</subject><subject>Cartilage tissue engineering</subject><subject>Cell Culture Techniques</subject><subject>Cells, Cultured</subject><subject>Chondrocytes - cytology</subject><subject>Chondrocytes - pathology</subject><subject>continuous flow bioreactor</subject><subject>Fibroblast Growth Factor 2 - administration & dosage</subject><subject>Growth factors</subject><subject>human nasoseptal cartilage</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Nasal Septum - cytology</subject><subject>Nasal Surgical Procedures - methods</subject><subject>primary cells</subject><subject>Receptors, Transforming Growth Factor beta - administration & dosage</subject><subject>Reconstructive Surgical Procedures - methods</subject><subject>scaffold‐free</subject><subject>Tensile Strength</subject><subject>Tissue and Organ Harvesting</subject><subject>Tissue engineering</subject><subject>Tissue Engineering - methods</subject><subject>Tissue Scaffolds</subject><issn>0023-852X</issn><issn>1531-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMuKFDEUhoMoTs_oxgeQgBsZqDHXuiyHwRs0CF5AV8Wp1El3hlRSJlUMvfMRfEafxAzdunDh6izOx8fPR8gzzq44Y-KVh3S4ErXs6gdkw7Xkleo6_ZBsylNWrRZfz8h5zreM8UZq9piciaateSfZhuAnA9ZGP_768dMmRGogLc7DDunicl6RYti5gJhc2NE7t-wp0DyB93SO8-phcTHQaOl-nSDQADlmnBfw1OxjGFM0hwXzE_LIgs_49HQvyJc3rz_fvKu2H96-v7neVkbqpq4UH1sUaNXQDgpF2TgOYtRGy0FoQJBWqqarmTHY1YqV5dJwCyCUtjBqJi_Iy6N3TvH7innpJ5cNeg8B45p73irelgpaFfTFP-htXFMo6wrVsDKn9CzU5ZEyKeac0PZzclPp3XPW38fv7-P34gQ_PynXYcLxL_qndgH4EbhzHg__UfXb64_fjtLfd4SSPg</recordid><startdate>201703</startdate><enddate>201703</enddate><creator>Chiu, Loraine L.Y.</creator><creator>To, William T.H.</creator><creator>Lee, John M.</creator><creator>Waldman, Stephen D.</creator><general>Wiley Subscription Services, Inc</general><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>K9.</scope><scope>7X8</scope></search><sort><creationdate>201703</creationdate><title>Scaffold‐free cartilage tissue engineering with a small population of human nasoseptal chondrocytes</title><author>Chiu, Loraine L.Y. ; To, William T.H. ; Lee, John M. ; Waldman, Stephen D.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3576-41d8e2ef4b8b4e2861db2d5c53b25aea3f347960cce9640caf3c1faa245fad503</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Biomechanical Phenomena</topic><topic>Cartilage tissue engineering</topic><topic>Cell Culture Techniques</topic><topic>Cells, Cultured</topic><topic>Chondrocytes - cytology</topic><topic>Chondrocytes - pathology</topic><topic>continuous flow bioreactor</topic><topic>Fibroblast Growth Factor 2 - administration & dosage</topic><topic>Growth factors</topic><topic>human nasoseptal cartilage</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Nasal Septum - cytology</topic><topic>Nasal Surgical Procedures - methods</topic><topic>primary cells</topic><topic>Receptors, Transforming Growth Factor beta - administration & dosage</topic><topic>Reconstructive Surgical Procedures - methods</topic><topic>scaffold‐free</topic><topic>Tensile Strength</topic><topic>Tissue and Organ Harvesting</topic><topic>Tissue engineering</topic><topic>Tissue Engineering - methods</topic><topic>Tissue Scaffolds</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chiu, Loraine L.Y.</creatorcontrib><creatorcontrib>To, William T.H.</creatorcontrib><creatorcontrib>Lee, John M.</creatorcontrib><creatorcontrib>Waldman, Stephen D.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>The Laryngoscope</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chiu, Loraine L.Y.</au><au>To, William T.H.</au><au>Lee, John M.</au><au>Waldman, Stephen D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Scaffold‐free cartilage tissue engineering with a small population of human nasoseptal chondrocytes</atitle><jtitle>The Laryngoscope</jtitle><addtitle>Laryngoscope</addtitle><date>2017-03</date><risdate>2017</risdate><volume>127</volume><issue>3</issue><spage>E91</spage><epage>E99</epage><pages>E91-E99</pages><issn>0023-852X</issn><eissn>1531-4995</eissn><abstract>Objective
Cartilage tissue engineering is a promising approach to provide suitable materials for nasal reconstruction; however, it typically requires large numbers of cells. We have previously shown that a small number of chondrocytes cultivated within a continuous flow bioreactor can elicit substantial tissue growth, but translation to human chondrocytes is not trivial. Here, we aimed to demonstrate the application of the bioreactor to generate large‐sized tissues from a small population of primary human nasoseptal chondrocytes.
Study Design
Experimental study.
Methods
Chondrocytes were cultured in the bioreactor using different medium compositions, with varying amounts of serum and with or without growth factors. Resulting engineered tissues were analyzed for physical properties, biochemical composition, tissue microstructure, and protein localization.
Results
Bioreactor‐cultivated constructs grown with serum and growth factors (basic fibroblast growth factor and transforming growth factor beta 2) had greater thickness, as well as DNA and glycosaminoglycan (GAG) contents, compared to low serum and no growth factor controls. These constructs also showed the most intense proteoglycan and collagen II staining.
Conclusion
The combination of bioreactor conditions, serum, and growth factors allowed the generation of large, thick scaffold‐free human cartilaginous tissues that resembled the native nasoseptal cartilage. There also may be implications for patient selection in future clinical applications of these engineered tissues because their GAG content decreased with donor age.
Level of Evidence
NA. Laryngoscope, 127:E91–E99, 2017</abstract><cop>United States</cop><pub>Wiley Subscription Services, Inc</pub><pmid>27861930</pmid><doi>10.1002/lary.26396</doi><tpages>9</tpages></addata></record> |
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subjects | Biomechanical Phenomena Cartilage tissue engineering Cell Culture Techniques Cells, Cultured Chondrocytes - cytology Chondrocytes - pathology continuous flow bioreactor Fibroblast Growth Factor 2 - administration & dosage Growth factors human nasoseptal cartilage Humans Immunohistochemistry Nasal Septum - cytology Nasal Surgical Procedures - methods primary cells Receptors, Transforming Growth Factor beta - administration & dosage Reconstructive Surgical Procedures - methods scaffold‐free Tensile Strength Tissue and Organ Harvesting Tissue engineering Tissue Engineering - methods Tissue Scaffolds |
title | Scaffold‐free cartilage tissue engineering with a small population of human nasoseptal chondrocytes |
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