Cargando…
Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo
OBJECTIVE: Lubricin expression in the superficial cartilage will be a crucial factor in the success of cartilage regeneration. Mesenchymal stem cells (MSCs) are an attractive cell source and the use of aggregates of MSCs has some advantages in terms of chondrogenic potential and efficiency of cell a...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750963/ https://www.ncbi.nlm.nih.gov/pubmed/26867127 http://dx.doi.org/10.1371/journal.pone.0148777 |
_version_ | 1782415511215144960 |
---|---|
author | Nakagawa, Yusuke Muneta, Takeshi Otabe, Koji Ozeki, Nobutake Mizuno, Mitsuru Udo, Mio Saito, Ryusuke Yanagisawa, Katsuaki Ichinose, Shizuko Koga, Hideyuki Tsuji, Kunikazu Sekiya, Ichiro |
author_facet | Nakagawa, Yusuke Muneta, Takeshi Otabe, Koji Ozeki, Nobutake Mizuno, Mitsuru Udo, Mio Saito, Ryusuke Yanagisawa, Katsuaki Ichinose, Shizuko Koga, Hideyuki Tsuji, Kunikazu Sekiya, Ichiro |
author_sort | Nakagawa, Yusuke |
collection | PubMed |
description | OBJECTIVE: Lubricin expression in the superficial cartilage will be a crucial factor in the success of cartilage regeneration. Mesenchymal stem cells (MSCs) are an attractive cell source and the use of aggregates of MSCs has some advantages in terms of chondrogenic potential and efficiency of cell adhesion. Lubricin expression in transplanted MSCs has not been fully elucidated so far. Our goals were to determine (1) whether cartilage pellets of human MSCs expressed lubricin in vitro chondrogenesis, (2) whether aggregates of human MSCs promoted lubricin expression, and (3) whether aggregates of MSCs expressed lubricin in the superficial cartilage after transplantation into osteochondral defects in rats. METHODS: For in vitro analysis, human bone marrow (BM) MSCs were differentiated into cartilage by pellet culture, and also aggregated using the hanging drop technique. For an animal study, aggregates of BM MSCs derived from GFP transgenic rats were transplanted to the osteochondral defect in the trochlear groove of wild type rat knee joints. Lubricin expression was mainly evaluated in differentiated and regenerated cartilages. RESULTS: In in vitro analysis, lubricin was detected in the superficial zone of the pellets and conditioned medium. mRNA expression of Proteoglycan4 (Prg4), which encodes lubricin, in pellets was significantly higher than that of undifferentiated MSCs. Aggregates showed different morphological features between the superficial and deep zone, and the Prg4 mRNA expression increased after aggregate formation. Lubricin was also found in the aggregate. In a rat study, articular cartilage regeneration was significantly better in the MSC group than in the control group as shown by macroscopical and histological analysis. The transmission electron microscope showed that morphology of the superficial cartilage in the MSC group was closer to that of the intact cartilage than in the control group. GFP positive cells remained in the repaired tissue and expressed lubricin in the superficial cartilage. CONCLUSION: Cartilage derived from MSCs expressed lubricin protein both in vitro and in vivo. Aggregation promoted lubricin expression of MSCs in vitro and transplantation of aggregates of MSCs regenerated cartilage including the superficial zone in a rat osteochondral defect model. Our results indicate that aggregated MSCs could be clinically relevant for therapeutic approaches to articular cartilage regeneration with an appropriate superficial zone in the future. |
format | Online Article Text |
id | pubmed-4750963 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-47509632016-02-26 Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo Nakagawa, Yusuke Muneta, Takeshi Otabe, Koji Ozeki, Nobutake Mizuno, Mitsuru Udo, Mio Saito, Ryusuke Yanagisawa, Katsuaki Ichinose, Shizuko Koga, Hideyuki Tsuji, Kunikazu Sekiya, Ichiro PLoS One Research Article OBJECTIVE: Lubricin expression in the superficial cartilage will be a crucial factor in the success of cartilage regeneration. Mesenchymal stem cells (MSCs) are an attractive cell source and the use of aggregates of MSCs has some advantages in terms of chondrogenic potential and efficiency of cell adhesion. Lubricin expression in transplanted MSCs has not been fully elucidated so far. Our goals were to determine (1) whether cartilage pellets of human MSCs expressed lubricin in vitro chondrogenesis, (2) whether aggregates of human MSCs promoted lubricin expression, and (3) whether aggregates of MSCs expressed lubricin in the superficial cartilage after transplantation into osteochondral defects in rats. METHODS: For in vitro analysis, human bone marrow (BM) MSCs were differentiated into cartilage by pellet culture, and also aggregated using the hanging drop technique. For an animal study, aggregates of BM MSCs derived from GFP transgenic rats were transplanted to the osteochondral defect in the trochlear groove of wild type rat knee joints. Lubricin expression was mainly evaluated in differentiated and regenerated cartilages. RESULTS: In in vitro analysis, lubricin was detected in the superficial zone of the pellets and conditioned medium. mRNA expression of Proteoglycan4 (Prg4), which encodes lubricin, in pellets was significantly higher than that of undifferentiated MSCs. Aggregates showed different morphological features between the superficial and deep zone, and the Prg4 mRNA expression increased after aggregate formation. Lubricin was also found in the aggregate. In a rat study, articular cartilage regeneration was significantly better in the MSC group than in the control group as shown by macroscopical and histological analysis. The transmission electron microscope showed that morphology of the superficial cartilage in the MSC group was closer to that of the intact cartilage than in the control group. GFP positive cells remained in the repaired tissue and expressed lubricin in the superficial cartilage. CONCLUSION: Cartilage derived from MSCs expressed lubricin protein both in vitro and in vivo. Aggregation promoted lubricin expression of MSCs in vitro and transplantation of aggregates of MSCs regenerated cartilage including the superficial zone in a rat osteochondral defect model. Our results indicate that aggregated MSCs could be clinically relevant for therapeutic approaches to articular cartilage regeneration with an appropriate superficial zone in the future. Public Library of Science 2016-02-11 /pmc/articles/PMC4750963/ /pubmed/26867127 http://dx.doi.org/10.1371/journal.pone.0148777 Text en © 2016 Nakagawa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Nakagawa, Yusuke Muneta, Takeshi Otabe, Koji Ozeki, Nobutake Mizuno, Mitsuru Udo, Mio Saito, Ryusuke Yanagisawa, Katsuaki Ichinose, Shizuko Koga, Hideyuki Tsuji, Kunikazu Sekiya, Ichiro Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo |
title | Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo |
title_full | Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo |
title_fullStr | Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo |
title_full_unstemmed | Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo |
title_short | Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo |
title_sort | cartilage derived from bone marrow mesenchymal stem cells expresses lubricin in vitro and in vivo |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750963/ https://www.ncbi.nlm.nih.gov/pubmed/26867127 http://dx.doi.org/10.1371/journal.pone.0148777 |
work_keys_str_mv | AT nakagawayusuke cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT munetatakeshi cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT otabekoji cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT ozekinobutake cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT mizunomitsuru cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT udomio cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT saitoryusuke cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT yanagisawakatsuaki cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT ichinoseshizuko cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT kogahideyuki cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT tsujikunikazu cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo AT sekiyaichiro cartilagederivedfrombonemarrowmesenchymalstemcellsexpresseslubricininvitroandinvivo |