{"title":"低温保护剂在提高生物结构的低温打印所需的生物墨水性能方面的作用","authors":"Harshavardhan Budharaju, Dhakshinamoorthy Sundaramurthi, Swaminathan Sethuraman","doi":"10.1007/s10856-024-06855-2","DOIUrl":null,"url":null,"abstract":"<div><p>Preservation and long-term storage of readily available cell-laden tissue-engineered products are major challenges in expanding their applications in healthcare. In recent years, there has been increasing interest in the development of off-the-shelf tissue-engineered products using the cryobioprinting approach. Here, bioinks are incorporated with cryoprotective agents (CPAs) to allow the fabrication of cryopreservable tissue constructs. Although this method has shown potential in the fabrication of cryopreservable tissue-engineered products, the impact of the CPAs on the viscoelastic behavior and printability of the bioinks at cryo conditions remains unexplored. In this study, we have evaluated the influence of CPAs such as glycerol and dimethyl sulfoxide (DMSO) on the rheological properties of pre-crosslinked alginate bioinks for cryoprinting applications. DMSO-incorporated bioinks showed a reduction in viscosity and yield stress, while the addition of glycerol improved both the properties due to interactions with the calcium chloride used for pre-crosslinking. Further, tube inversion and printability experiments were performed to identify suitable concentrations and cryobioprinting conditions for bioinks containing CPAs & pre-crosslinked with CaCl<sub>2</sub>. Finally, based on the printability analysis & cell recovery results, 10% glycerol was used for cryobioprinting and preservation of cell-laden constructs at −80 °C and the viability of cells within the printed structures were evaluated after recovery. Cell viability results indicate that the addition of 10% glycerol to the pre-crosslinked bioink significantly improved cell viability compared to bioinks without CPAs, confirming the suitability of the developed bioink combination to fabricate tissue constructs for on-demand applications.</p><h3>Graphical abstract</h3><div><figure><div><div><picture><source><img></source></picture></div><div><p>Effect of cryoprotectants on the viscoelastic behavior of bioinks and cell recovery in cryobioprinted tissue constructs.</p></div></div></figure></div></div>","PeriodicalId":647,"journal":{"name":"Journal of Materials Science: Materials in Medicine","volume":"36 1","pages":""},"PeriodicalIF":4.2000,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s10856-024-06855-2.pdf","citationCount":"0","resultStr":"{\"title\":\"Insights on the role of cryoprotectants in enhancing the properties of bioinks required for cryobioprinting of biological constructs\",\"authors\":\"Harshavardhan Budharaju, Dhakshinamoorthy Sundaramurthi, Swaminathan Sethuraman\",\"doi\":\"10.1007/s10856-024-06855-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Preservation and long-term storage of readily available cell-laden tissue-engineered products are major challenges in expanding their applications in healthcare. In recent years, there has been increasing interest in the development of off-the-shelf tissue-engineered products using the cryobioprinting approach. Here, bioinks are incorporated with cryoprotective agents (CPAs) to allow the fabrication of cryopreservable tissue constructs. Although this method has shown potential in the fabrication of cryopreservable tissue-engineered products, the impact of the CPAs on the viscoelastic behavior and printability of the bioinks at cryo conditions remains unexplored. In this study, we have evaluated the influence of CPAs such as glycerol and dimethyl sulfoxide (DMSO) on the rheological properties of pre-crosslinked alginate bioinks for cryoprinting applications. DMSO-incorporated bioinks showed a reduction in viscosity and yield stress, while the addition of glycerol improved both the properties due to interactions with the calcium chloride used for pre-crosslinking. Further, tube inversion and printability experiments were performed to identify suitable concentrations and cryobioprinting conditions for bioinks containing CPAs & pre-crosslinked with CaCl<sub>2</sub>. Finally, based on the printability analysis & cell recovery results, 10% glycerol was used for cryobioprinting and preservation of cell-laden constructs at −80 °C and the viability of cells within the printed structures were evaluated after recovery. Cell viability results indicate that the addition of 10% glycerol to the pre-crosslinked bioink significantly improved cell viability compared to bioinks without CPAs, confirming the suitability of the developed bioink combination to fabricate tissue constructs for on-demand applications.</p><h3>Graphical abstract</h3><div><figure><div><div><picture><source><img></source></picture></div><div><p>Effect of cryoprotectants on the viscoelastic behavior of bioinks and cell recovery in cryobioprinted tissue constructs.</p></div></div></figure></div></div>\",\"PeriodicalId\":647,\"journal\":{\"name\":\"Journal of Materials Science: Materials in Medicine\",\"volume\":\"36 1\",\"pages\":\"\"},\"PeriodicalIF\":4.2000,\"publicationDate\":\"2025-01-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://link.springer.com/content/pdf/10.1007/s10856-024-06855-2.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Materials Science: Materials in Medicine\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s10856-024-06855-2\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ENGINEERING, BIOMEDICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Materials Science: Materials in Medicine","FirstCategoryId":"5","ListUrlMain":"https://link.springer.com/article/10.1007/s10856-024-06855-2","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ENGINEERING, BIOMEDICAL","Score":null,"Total":0}
Insights on the role of cryoprotectants in enhancing the properties of bioinks required for cryobioprinting of biological constructs
Preservation and long-term storage of readily available cell-laden tissue-engineered products are major challenges in expanding their applications in healthcare. In recent years, there has been increasing interest in the development of off-the-shelf tissue-engineered products using the cryobioprinting approach. Here, bioinks are incorporated with cryoprotective agents (CPAs) to allow the fabrication of cryopreservable tissue constructs. Although this method has shown potential in the fabrication of cryopreservable tissue-engineered products, the impact of the CPAs on the viscoelastic behavior and printability of the bioinks at cryo conditions remains unexplored. In this study, we have evaluated the influence of CPAs such as glycerol and dimethyl sulfoxide (DMSO) on the rheological properties of pre-crosslinked alginate bioinks for cryoprinting applications. DMSO-incorporated bioinks showed a reduction in viscosity and yield stress, while the addition of glycerol improved both the properties due to interactions with the calcium chloride used for pre-crosslinking. Further, tube inversion and printability experiments were performed to identify suitable concentrations and cryobioprinting conditions for bioinks containing CPAs & pre-crosslinked with CaCl2. Finally, based on the printability analysis & cell recovery results, 10% glycerol was used for cryobioprinting and preservation of cell-laden constructs at −80 °C and the viability of cells within the printed structures were evaluated after recovery. Cell viability results indicate that the addition of 10% glycerol to the pre-crosslinked bioink significantly improved cell viability compared to bioinks without CPAs, confirming the suitability of the developed bioink combination to fabricate tissue constructs for on-demand applications.
Graphical abstract
Effect of cryoprotectants on the viscoelastic behavior of bioinks and cell recovery in cryobioprinted tissue constructs.
期刊介绍：
The Journal of Materials Science: Materials in Medicine publishes refereed papers providing significant progress in the application of biomaterials and tissue engineering constructs as medical or dental implants, prostheses and devices. Coverage spans a wide range of topics from basic science to clinical applications, around the theme of materials in medicine and dentistry. The central element is the development of synthetic and natural materials used in orthopaedic, maxillofacial, cardiovascular, neurological, ophthalmic and dental applications. Special biomedical topics include biomaterial synthesis and characterisation, biocompatibility studies, nanomedicine, tissue engineering constructs and cell substrates, regenerative medicine, computer modelling and other advanced experimental methodologies.
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