Rose Ann G Franco, Eamonn McKenna, Pamela G Robey, Ross W Crawford, Michael R Doran, Kathryn Futrega
{"title":"SP7基因沉默会抑制骨髓基质细胞肥大,但同时也会抑制软骨生成。","authors":"Rose Ann G Franco, Eamonn McKenna, Pamela G Robey, Ross W Crawford, Michael R Doran, Kathryn Futrega","doi":"10.1177/20417314231177136","DOIUrl":null,"url":null,"abstract":"<p><p>For bone marrow stromal cells (BMSC) to be useful in cartilage repair their propensity for hypertrophic differentiation must be overcome. A single day of TGF-β1 stimulation activates intrinsic signaling cascades in BMSCs which subsequently drives both chondrogenic and hypertrophic differentiation. TGF-β1 stimulation upregulates <i>SP7</i>, a transcription factor known to contribute to hypertrophic differentiation, and <i>SP7</i> remains upregulated even if TGF-β1 is subsequently withdrawn from the chondrogenic induction medium. Herein, we stably transduced BMSCs to express an shRNA designed to silence <i>SP7</i>, and assess the capacity of <i>SP7</i> silencing to mitigate hypertrophy. <i>SP7</i> silencing dampened both hypertrophic and chondrogenic differentiation processes, resulting in diminished microtissue size, impaired glycosaminoglycan production and reduced chondrogenic and hypertrophic gene expression. Thus, while hypertrophic features were dampened by <i>SP7</i> silencing, chondrogenic differentation was also compromised. We further investigated the role of <i>SP7</i> in monolayer osteogenic and adipogenic cultures, finding that <i>SP7</i> silencing dampened characteristic mineralization and lipid vacuole formation, respectively. Overall, <i>SP7</i> silencing affects the trilineage differentiation of BMSCs, but is insufficient to decouple BMSC hypertrophy from chondrogenesis. These data highlight the challenge of promoting BMSC chondrogenesis whilst simultaneously reducing hypertrophy in cartilage tissue engineering strategies.</p>","PeriodicalId":17384,"journal":{"name":"Journal of Tissue Engineering","volume":null,"pages":null},"PeriodicalIF":6.7000,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288420/pdf/","citationCount":"0","resultStr":"{\"title\":\"<i>SP7</i> gene silencing dampens bone marrow stromal cell hypertrophy, but it also dampens chondrogenesis.\",\"authors\":\"Rose Ann G Franco, Eamonn McKenna, Pamela G Robey, Ross W Crawford, Michael R Doran, Kathryn Futrega\",\"doi\":\"10.1177/20417314231177136\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>For bone marrow stromal cells (BMSC) to be useful in cartilage repair their propensity for hypertrophic differentiation must be overcome. A single day of TGF-β1 stimulation activates intrinsic signaling cascades in BMSCs which subsequently drives both chondrogenic and hypertrophic differentiation. TGF-β1 stimulation upregulates <i>SP7</i>, a transcription factor known to contribute to hypertrophic differentiation, and <i>SP7</i> remains upregulated even if TGF-β1 is subsequently withdrawn from the chondrogenic induction medium. Herein, we stably transduced BMSCs to express an shRNA designed to silence <i>SP7</i>, and assess the capacity of <i>SP7</i> silencing to mitigate hypertrophy. <i>SP7</i> silencing dampened both hypertrophic and chondrogenic differentiation processes, resulting in diminished microtissue size, impaired glycosaminoglycan production and reduced chondrogenic and hypertrophic gene expression. Thus, while hypertrophic features were dampened by <i>SP7</i> silencing, chondrogenic differentation was also compromised. We further investigated the role of <i>SP7</i> in monolayer osteogenic and adipogenic cultures, finding that <i>SP7</i> silencing dampened characteristic mineralization and lipid vacuole formation, respectively. Overall, <i>SP7</i> silencing affects the trilineage differentiation of BMSCs, but is insufficient to decouple BMSC hypertrophy from chondrogenesis. These data highlight the challenge of promoting BMSC chondrogenesis whilst simultaneously reducing hypertrophy in cartilage tissue engineering strategies.</p>\",\"PeriodicalId\":17384,\"journal\":{\"name\":\"Journal of Tissue Engineering\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":6.7000,\"publicationDate\":\"2023-06-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288420/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Tissue Engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1177/20417314231177136\",\"RegionNum\":1,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q1\",\"JCRName\":\"CELL & TISSUE ENGINEERING\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Tissue Engineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1177/20417314231177136","RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/1/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"CELL & TISSUE ENGINEERING","Score":null,"Total":0}
SP7 gene silencing dampens bone marrow stromal cell hypertrophy, but it also dampens chondrogenesis.
For bone marrow stromal cells (BMSC) to be useful in cartilage repair their propensity for hypertrophic differentiation must be overcome. A single day of TGF-β1 stimulation activates intrinsic signaling cascades in BMSCs which subsequently drives both chondrogenic and hypertrophic differentiation. TGF-β1 stimulation upregulates SP7, a transcription factor known to contribute to hypertrophic differentiation, and SP7 remains upregulated even if TGF-β1 is subsequently withdrawn from the chondrogenic induction medium. Herein, we stably transduced BMSCs to express an shRNA designed to silence SP7, and assess the capacity of SP7 silencing to mitigate hypertrophy. SP7 silencing dampened both hypertrophic and chondrogenic differentiation processes, resulting in diminished microtissue size, impaired glycosaminoglycan production and reduced chondrogenic and hypertrophic gene expression. Thus, while hypertrophic features were dampened by SP7 silencing, chondrogenic differentation was also compromised. We further investigated the role of SP7 in monolayer osteogenic and adipogenic cultures, finding that SP7 silencing dampened characteristic mineralization and lipid vacuole formation, respectively. Overall, SP7 silencing affects the trilineage differentiation of BMSCs, but is insufficient to decouple BMSC hypertrophy from chondrogenesis. These data highlight the challenge of promoting BMSC chondrogenesis whilst simultaneously reducing hypertrophy in cartilage tissue engineering strategies.
期刊介绍:
The Journal of Tissue Engineering (JTE) is a peer-reviewed, open-access journal dedicated to scientific research in the field of tissue engineering and its clinical applications. Our journal encompasses a wide range of interests, from the fundamental aspects of stem cells and progenitor cells, including their expansion to viable numbers, to an in-depth understanding of their differentiation processes. Join us in exploring the latest advancements in tissue engineering and its clinical translation.