Bioprinting最新文献

FK506 binding protein like, FKBPL, as a novel therapeutic target in 2D and 3D bioprinted, models of cardiac fibrosis

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-04 DOI: 10.1016/j.bprint.2025.e00397

Michael Chhor , Shreya Barman , Fatemeh Heidari , Amy L. Bottomley , Tracy Robson , Kristine McGrath , Lana McClements

Background

Cardiac fibrosis characterised by increased collagen deposition and extracellular matrix (ECM) remodeling is one of the main causes of heart failure. Inflammation and hypoxia are key processes leading to cardiac fibrosis although the mechanisms are poorly understood. In this study, we developed an innovative 3D bioprinted model of cardiac fibrosis using tunable matrices. The role of an anti-angiogenic protein, FK506 binding protein like (FKBPL) was then elucidated, for the first time, using both 2D and 3D bioprinted, models of cardiac fibrosis.

Methods

3D bioprinted model of cardiac fibrosis was developed using fetal fibroblast cells (HFF08), customised ECM cardiac components and pro-fibrotic/hypoxic factors (TGF-β, 10 ng/ml, DMOG, 1 mM) ± FKBPL mimetic (AD-01, 100 mM). In parallel, 2D in vitro models were also employed.

Results

In the 3D bioprinted model, fibroblasts formed networks spontaneously, which were stimulated by all treatments (p < 0.05–0.0001). This was in conjunction with a trend towards reduced FKBPL expression, particularly in the presence of DMOG/AD-01 treatment. In 2D cell culture, AD-01 potentiated TGF-β-induced col1a1 (p < 0.0001) and mmp2 mRNA (p < 0.05) expression whereas DMOG or reduced FKBPL expression with AD-01 abrogated this (p < 0.05–0.001). Following siRNA FKBPL transfection, α-SMA was reduced (p < 0.05).

Conclusion

This 3D bioprinted model of cardiac fibrosis in conjunction with 2D cell models could be used for biomarker and drug therapy screening towards accelerating the development of treatments for this hard-to-treat condition. Low FKBPL expression could be protective in cardiac fibrosis through the reduction in collagen production and α-SMA expression, or TGF-β/HIF-1α-mediated effects. Therapeutic strategies that inhibit FKBPL should be explored to abrogate cardiac fibrosis.

{"title":"FK506 binding protein like, FKBPL, as a novel therapeutic target in 2D and 3D bioprinted, models of cardiac fibrosis","authors":"Michael Chhor , Shreya Barman , Fatemeh Heidari , Amy L. Bottomley , Tracy Robson , Kristine McGrath , Lana McClements","doi":"10.1016/j.bprint.2025.e00397","DOIUrl":"10.1016/j.bprint.2025.e00397","url":null,"abstract":"<div><h3>Background</h3><div>Cardiac fibrosis characterised by increased collagen deposition and extracellular matrix (ECM) remodeling is one of the main causes of heart failure. Inflammation and hypoxia are key processes leading to cardiac fibrosis although the mechanisms are poorly understood. In this study, we developed an innovative 3D bioprinted model of cardiac fibrosis using tunable matrices. The role of an anti-angiogenic protein, FK506 binding protein like (FKBPL) was then elucidated, for the first time, using both 2D and 3D bioprinted, models of cardiac fibrosis.</div></div><div><h3>Methods</h3><div>3D bioprinted model of cardiac fibrosis was developed using fetal fibroblast cells (HFF08), customised ECM cardiac components and pro-fibrotic/hypoxic factors (TGF-β, 10 ng/ml, DMOG, 1 mM) ± FKBPL mimetic (AD-01, 100 mM). In parallel, 2D <em>in vitro</em> models were also employed.</div></div><div><h3>Results</h3><div>In the 3D bioprinted model, fibroblasts formed networks spontaneously, which were stimulated by all treatments (p < 0.05–0.0001). This was in conjunction with a trend towards reduced FKBPL expression, particularly in the presence of DMOG/AD-01 treatment. In 2D cell culture, AD-01 potentiated TGF-β-induced <em>col1a1</em> (p < 0.0001) and <em>mmp2</em> mRNA (p < 0.05) expression whereas DMOG or reduced FKBPL expression with AD-01 abrogated this (p < 0.05–0.001). Following siRNA FKBPL transfection, α-SMA was reduced (p < 0.05).</div></div><div><h3>Conclusion</h3><div>This 3D bioprinted model of cardiac fibrosis in conjunction with 2D cell models could be used for biomarker and drug therapy screening towards accelerating the development of treatments for this hard-to-treat condition. Low FKBPL expression could be protective in cardiac fibrosis through the reduction in collagen production and α-SMA expression, or TGF-β/HIF-1α-mediated effects. Therapeutic strategies that inhibit FKBPL should be explored to abrogate cardiac fibrosis.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"47 ","pages":"Article e00397"},"PeriodicalIF":0.0,"publicationDate":"2025-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143428692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Modeling of oral squamous cell carcinoma microenvironment- A 3D bioprinting approach

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00381

Akhilanand Chaurasia , Gowri Sivaramakrishnan , Farah Asa’ad , Lena Larsson , Arwa Daghrery , Joana Marques , Francesca Spirito , Vitória Batista Clemente , Ana Carolina Morais Apolônio , Mahdieh Alipour , Rini Tiwari

Background

Oral squamous cell carcinoma (OSCC) presents significant challenges due to its complex microenvironment and invasive characteristics. Traditional two-dimensional (2D) culture systems are inadequate for modelling the intricate features of OSCC, necessitating advanced techniques for better in vitro modelling.

Objective

This review aims to explore the applications of 3D bioprinting in modelling the OSCC microenvironment, highlighting the advantages over conventional methods and discussing recent advancements in the field.

Methods

The review synthesizes recent literature on 3D bioprinting technologies, focusing on their application in replicating OSCC's microenvironment. Key areas include the integration of various cell types within a biomimetic extracellular matrix, the use of microfluidic systems to study tumor-stromal interactions, and the incorporation of advanced imaging modalities.

Results

3D bioprinting allows for the precise fabrication of complex OSCC tumor architectures, incorporating cancer cells, stromal cells, and immune cells. The integration of microfluidic systems facilitates the study of tumor invasion, metastasis, and drug response. Recent advancements in bioink development, particularly the use of patient-derived cells and biomolecules, enhance the physiological relevance of these models. Emerging imaging technologies provide unprecedented insights into the dynamics of OSCC progression within these constructs.

Conclusion

3D bioprinting shows immense potential for advancing the understanding of OSCC pathobiology and developing personalized therapeutic strategies. However, challenges such as standardizing bioink formulations and scaling fabrication techniques must be addressed to effectively translate these innovations into clinical practice.

{"title":"Modeling of oral squamous cell carcinoma microenvironment- A 3D bioprinting approach","authors":"Akhilanand Chaurasia , Gowri Sivaramakrishnan , Farah Asa’ad , Lena Larsson , Arwa Daghrery , Joana Marques , Francesca Spirito , Vitória Batista Clemente , Ana Carolina Morais Apolônio , Mahdieh Alipour , Rini Tiwari","doi":"10.1016/j.bprint.2024.e00381","DOIUrl":"10.1016/j.bprint.2024.e00381","url":null,"abstract":"<div><h3>Background</h3><div>Oral squamous cell carcinoma (OSCC) presents significant challenges due to its complex microenvironment and invasive characteristics. Traditional two-dimensional (2D) culture systems are inadequate for modelling the intricate features of OSCC, necessitating advanced techniques for better <em>in vitro</em> modelling.</div></div><div><h3>Objective</h3><div>This review aims to explore the applications of 3D bioprinting in modelling the OSCC microenvironment, highlighting the advantages over conventional methods and discussing recent advancements in the field.</div></div><div><h3>Methods</h3><div>The review synthesizes recent literature on 3D bioprinting technologies, focusing on their application in replicating OSCC's microenvironment. Key areas include the integration of various cell types within a biomimetic extracellular matrix, the use of microfluidic systems to study tumor-stromal interactions, and the incorporation of advanced imaging modalities.</div></div><div><h3>Results</h3><div>3D bioprinting allows for the precise fabrication of complex OSCC tumor architectures, incorporating cancer cells, stromal cells, and immune cells. The integration of microfluidic systems facilitates the study of tumor invasion, metastasis, and drug response. Recent advancements in bioink development, particularly the use of patient-derived cells and biomolecules, enhance the physiological relevance of these models. Emerging imaging technologies provide unprecedented insights into the dynamics of OSCC progression within these constructs.</div></div><div><h3>Conclusion</h3><div>3D bioprinting shows immense potential for advancing the understanding of OSCC pathobiology and developing personalized therapeutic strategies. However, challenges such as standardizing bioink formulations and scaling fabrication techniques must be addressed to effectively translate these innovations into clinical practice.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00381"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143101938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Zirconia-calcium silicate bioactive composites for dental applications using DLP additive manufacturing

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00377

Ahmed Binobaid , Michele De Lisi , Josette Camilleri , Hany Hassanin , Khamis Essa

Zirconia has outstanding mechanical strength which made it a favourable material dental implants material. However, its use is limited by challenges in bone bonding and elasticity. This paper introduces a novel bioprinting ceramic material by mixing calcium silicate with zirconia to enhance bioactivity. Using the high precision and speed of Digital Light Processing (DLP), this study develops a novel zirconia-calcium silicate slurry for dental applications. The study reports the preparation of zirconia-calcium silicate, formulation of resin compositions, and optimization of the bioprinting, debinding and sintering. Employing a full factorial Design of Experiments (DOE), a systematic approach was implemented to identify optimal printing conditions such as the layer thickness, exposure time, and power. The results show that slurries formulated with BYK-111 as the dispersant and ACMO/PEGDA/TPO resin, coupled with 80 wt% solid loading, achieved the most favourable rheological properties, cure depth, and printing accuracy. The optimal printing conditions were 0.75 s exposure time, 300 % exposure power, and 30 μm layer thickness, ensured a relative density of the sintered implants exceeding 95 %. This study advances dental implant materials by introducing a novel DLP biomaterial with a slurry formulation, presenting significant implications for clinical applications and future research in developing advanced dental and medical implants.

{"title":"Zirconia-calcium silicate bioactive composites for dental applications using DLP additive manufacturing","authors":"Ahmed Binobaid , Michele De Lisi , Josette Camilleri , Hany Hassanin , Khamis Essa","doi":"10.1016/j.bprint.2024.e00377","DOIUrl":"10.1016/j.bprint.2024.e00377","url":null,"abstract":"<div><div>Zirconia has outstanding mechanical strength which made it a favourable material dental implants material. However, its use is limited by challenges in bone bonding and elasticity. This paper introduces a novel bioprinting ceramic material by mixing calcium silicate with zirconia to enhance bioactivity. Using the high precision and speed of Digital Light Processing (DLP), this study develops a novel zirconia-calcium silicate slurry for dental applications. The study reports the preparation of zirconia-calcium silicate, formulation of resin compositions, and optimization of the bioprinting, debinding and sintering. Employing a full factorial Design of Experiments (DOE), a systematic approach was implemented to identify optimal printing conditions such as the layer thickness, exposure time, and power. The results show that slurries formulated with BYK-111 as the dispersant and ACMO/PEGDA/TPO resin, coupled with 80 wt% solid loading, achieved the most favourable rheological properties, cure depth, and printing accuracy. The optimal printing conditions were 0.75 s exposure time, 300 % exposure power, and 30 μm layer thickness, ensured a relative density of the sintered implants exceeding 95 %. This study advances dental implant materials by introducing a novel DLP biomaterial with a slurry formulation, presenting significant implications for clinical applications and future research in developing advanced dental and medical implants.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00377"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143101937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

4D printing in skin tissue engineering: A revolutionary approach to enhance wound healing and combat infections

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2025.e00386

Laila A. Damiati , Samar A. Alsudir , Rean Y. Mohammed , Majed A. Majrashi , Shahad H. Albrahim , Aliyah algethami , Fatimah O. Alghamdi , Hala A. Alamari , Mai M. Alzaydi

Skin infection poses significant challenges in healthcare, demanding innovative solutions to enhance the efficacy of wound-repair interventions. 4D printing represents a revolutionary approach in addition to traditional wound-management strategies. 4D-printing materials, which are dynamic and responsive, can change their shape or properties over time in response to internal or external stimuli, creating a paradigm shift in how wounds are treated. This review explores the potential of 4D printing technology as a transformative solution addressing critical challenges in skin tissue engineering. It highlights the journey from 2D fabrication of skin implants to the current state of 4D printing focusing on skin tissue structures that allow for precise and sustained release of therapeutic agents while exhibiting self-healing properties. Also, the ability to integrate antimicrobials to the printed skin constructs that respond to specific stimuli, such as pH, light, temperature, humidity, or enzymes enables the on demand and controlled release of antimicrobial agents. Additionally, integrating artificial intelligence (AI) into the fabrication process of skin tissues represents a synergistic approach that combines advanced computational methodologies with biological principles to identify the optimal conditions for enhancing tissue regeneration. Indeed, 4D bioprinting and AI-driven precision in the customization of scaffolds based on patient-specific needs promise a new era of personalized medicine in skin tissue engineering.

{"title":"4D printing in skin tissue engineering: A revolutionary approach to enhance wound healing and combat infections","authors":"Laila A. Damiati , Samar A. Alsudir , Rean Y. Mohammed , Majed A. Majrashi , Shahad H. Albrahim , Aliyah algethami , Fatimah O. Alghamdi , Hala A. Alamari , Mai M. Alzaydi","doi":"10.1016/j.bprint.2025.e00386","DOIUrl":"10.1016/j.bprint.2025.e00386","url":null,"abstract":"<div><div>Skin infection poses significant challenges in healthcare, demanding innovative solutions to enhance the efficacy of wound-repair interventions. 4D printing represents a revolutionary approach in addition to traditional wound-management strategies. 4D-printing materials, which are dynamic and responsive, can change their shape or properties over time in response to internal or external stimuli, creating a paradigm shift in how wounds are treated. This review explores the potential of 4D printing technology as a transformative solution addressing critical challenges in skin tissue engineering. It highlights the journey from 2D fabrication of skin implants to the current state of 4D printing focusing on skin tissue structures that allow for precise and sustained release of therapeutic agents while exhibiting self-healing properties. Also, the ability to integrate antimicrobials to the printed skin constructs that respond to specific stimuli, such as pH, light, temperature, humidity, or enzymes enables the on demand and controlled release of antimicrobial agents. Additionally, integrating artificial intelligence (AI) into the fabrication process of skin tissues represents a synergistic approach that combines advanced computational methodologies with biological principles to identify the optimal conditions for enhancing tissue regeneration. Indeed, 4D bioprinting and AI-driven precision in the customization of scaffolds based on patient-specific needs promise a new era of personalized medicine in skin tissue engineering.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00386"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143101936","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advancing the in vitro drug screening models: Microbiome as a component of tissue-engineered skin

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00379

Vsevolod V. Shishkov , Polina Yu Bikmulina , Anna V. Kardosh , Sergey V. Tsibulnikov , Ekaterina V. Grekova , Yulia V. Kolesova , Polina A. Zakharova , Anastasiia M. Nesterova , Frederico David Alencar de Sena Pereira , Svetlana L. Kotova , Olga Yu Olisova , Massoud Vosough , Anastasia I. Shpichka , Peter S. Timashev

Currently, in vitro skin models are among the most advanced and frequently utilized tools in clinical practice and drug screening. The development of these models often involves the use of skin organoids and biofabrication techniques, such as 3D bioprinting. Despite this significant progress, the skin models employed in drug screening typically lack a microbiome component. Since the microbiome is recognized as a crucial element of healthy human skin, it is essential to integrate this aspect into existing skin models. This review outlines a pathway for the development of in vitro skin models that can be widely used as platforms for testing drugs and cosmetics. First, we discuss the diversity of the normal human microbiome and its interactions with human cells. Next, we examine current skin models, including those that incorporate microbiome components through various co-culturing methods. Finally, we discuss how biofabrication approaches can be combined with microbiome elements to create relevant and stable in vitro skin models.

{"title":"Advancing the in vitro drug screening models: Microbiome as a component of tissue-engineered skin","authors":"Vsevolod V. Shishkov , Polina Yu Bikmulina , Anna V. Kardosh , Sergey V. Tsibulnikov , Ekaterina V. Grekova , Yulia V. Kolesova , Polina A. Zakharova , Anastasiia M. Nesterova , Frederico David Alencar de Sena Pereira , Svetlana L. Kotova , Olga Yu Olisova , Massoud Vosough , Anastasia I. Shpichka , Peter S. Timashev","doi":"10.1016/j.bprint.2024.e00379","DOIUrl":"10.1016/j.bprint.2024.e00379","url":null,"abstract":"<div><div>Currently, in vitro skin models are among the most advanced and frequently utilized tools in clinical practice and drug screening. The development of these models often involves the use of skin organoids and biofabrication techniques, such as 3D bioprinting. Despite this significant progress, the skin models employed in drug screening typically lack a microbiome component. Since the microbiome is recognized as a crucial element of healthy human skin, it is essential to integrate this aspect into existing skin models. This review outlines a pathway for the development of in vitro skin models that can be widely used as platforms for testing drugs and cosmetics. First, we discuss the diversity of the normal human microbiome and its interactions with human cells. Next, we examine current skin models, including those that incorporate microbiome components through various co-culturing methods. Finally, we discuss how biofabrication approaches can be combined with microbiome elements to create relevant and stable in vitro skin models.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00379"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143101941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Nanocomposite hydrogel-based bioinks composed of a fucose-rich polysaccharide and nanocellulose fibers for 3D-bioprinting applications

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00382

Nicole S. Lameirinhas , João P.F. Carvalho , Maria C. Teixeira , Jorge L. Luís , Asiyah Esmail , Ricardo J.B. Pinto , Helena Oliveira , Filomena Freitas , José M. Oliveira , Carla Vilela , Armando J.D. Silvestre , Carmen S.R. Freire

Hydrogels are the most common type of bioinks, yet, finding adequate biomaterials to develop suitable bioinks for 3D bioprinting remains challenging. Herein, innovative hydrogel bioinks were developed by combining nanofibrillated cellulose (NFC) with a fucose-rich polysaccharide, FucoPol (FP), still unexplored for 3D bioprinting. NFC/FP bioinks with different mass proportions, namely 1:1, 2:1, 3:1 and 4:1, were prepared and denominated as NFC1FP, NFC2FP, NFC3FP and NFC4FP. A formulation without NFC was also prepared for comparison purposes (NFC0FP). The rheological properties of the bioinks were enhanced by the addition of NFC, as evidenced by the increase in shear viscosity from 1.39 ± 0.03 Pa s (NFC0FP) to 2933.7 ± 137.9 Pa s (ink NFC4FP) and by the 3D printing of complex structures with high shape fidelity (Pr ≈ 0.9). The stability and mechanical properties of the crosslinked hydrogels were also improved, with Young’s modulus increasing from 0.12 ± 0.04 MPa (NFC0FP) to 2.45 ± 0.06 MPa (NFC4FP). The successful 3D bioprinting of both A375 (melanoma) and HaCaT (keratinocyte) cell-laden bioinks translated into elevated cell viabilities (above 88 %) up to 21 days post-bioprinting. These results highlight the potential and versatility of NFC/FP bioinks for the bioprinting of 3D skin tissue analogues for biomedical applications.

{"title":"Nanocomposite hydrogel-based bioinks composed of a fucose-rich polysaccharide and nanocellulose fibers for 3D-bioprinting applications","authors":"Nicole S. Lameirinhas , João P.F. Carvalho , Maria C. Teixeira , Jorge L. Luís , Asiyah Esmail , Ricardo J.B. Pinto , Helena Oliveira , Filomena Freitas , José M. Oliveira , Carla Vilela , Armando J.D. Silvestre , Carmen S.R. Freire","doi":"10.1016/j.bprint.2024.e00382","DOIUrl":"10.1016/j.bprint.2024.e00382","url":null,"abstract":"<div><div>Hydrogels are the most common type of bioinks, yet, finding adequate biomaterials to develop suitable bioinks for 3D bioprinting remains challenging. Herein, innovative hydrogel bioinks were developed by combining nanofibrillated cellulose (NFC) with a fucose-rich polysaccharide, FucoPol (FP), still unexplored for 3D bioprinting. NFC/FP bioinks with different mass proportions, namely 1:1, 2:1, 3:1 and 4:1, were prepared and denominated as NFC1FP, NFC2FP, NFC3FP and NFC4FP. A formulation without NFC was also prepared for comparison purposes (NFC0FP). The rheological properties of the bioinks were enhanced by the addition of NFC, as evidenced by the increase in shear viscosity from 1.39 ± 0.03 Pa s (NFC0FP) to 2933.7 ± 137.9 Pa s (ink NFC4FP) and by the 3D printing of complex structures with high shape fidelity (<em>Pr</em> ≈ 0.9). The stability and mechanical properties of the crosslinked hydrogels were also improved, with Young’s modulus increasing from 0.12 ± 0.04 MPa (NFC0FP) to 2.45 ± 0.06 MPa (NFC4FP). The successful 3D bioprinting of both A375 (melanoma) and HaCaT (keratinocyte) cell-laden bioinks translated into elevated cell viabilities (above 88 %) up to 21 days post-bioprinting. These results highlight the potential and versatility of NFC/FP bioinks for the bioprinting of 3D skin tissue analogues for biomedical applications.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00382"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143101933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

3D printing of polysaccharide-based formulations: Opportunities for innovation

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00383

Fabian Hernandez-Tenorio , Edier Múnera-Gutiérrez , Alejandra M. Miranda , Alex A. Sáez , Luz Deisy Marín-Palacio , Catalina Giraldo-Estrada

3D printing is a technology that has gained significant interest due to its versatility in terms of design, as well as the wide variety of materials that can be used for the production of inks. Among the compounds with the greatest importance in the last decade for 3D printing are polysaccharides. These have been positioned as favorable compounds for the formulation of inks due to their properties such as flexibility, non-immunogenicity, pseudoplastic behavior, printability, biocompatibility, and biodegradability. Therefore, the implementation of polysaccharides in 3D printing promotes innovation in the development of materials and products for medical, food, pharmaceutical, and other applications. The objective of this review was to provide a comprehensive and exhaustive study of the technological advances in 3D printing of polysaccharide-based formulations. To this end, a bibliometric analysis was presented to establish trends using scientometric indicators that allowed us to delve deeper and identify the most relevant developments in the subject. Through this review, we sought to highlight the importance of polysaccharides and their wide range of applications in 3D printing and hope that it will provide a meaningful basis for the exploration of printable compounds from renewable sources.

{"title":"3D printing of polysaccharide-based formulations: Opportunities for innovation","authors":"Fabian Hernandez-Tenorio , Edier Múnera-Gutiérrez , Alejandra M. Miranda , Alex A. Sáez , Luz Deisy Marín-Palacio , Catalina Giraldo-Estrada","doi":"10.1016/j.bprint.2024.e00383","DOIUrl":"10.1016/j.bprint.2024.e00383","url":null,"abstract":"<div><div>3D printing is a technology that has gained significant interest due to its versatility in terms of design, as well as the wide variety of materials that can be used for the production of inks. Among the compounds with the greatest importance in the last decade for 3D printing are polysaccharides. These have been positioned as favorable compounds for the formulation of inks due to their properties such as flexibility, non-immunogenicity, pseudoplastic behavior, printability, biocompatibility, and biodegradability. Therefore, the implementation of polysaccharides in 3D printing promotes innovation in the development of materials and products for medical, food, pharmaceutical, and other applications. The objective of this review was to provide a comprehensive and exhaustive study of the technological advances in 3D printing of polysaccharide-based formulations. To this end, a bibliometric analysis was presented to establish trends using scientometric indicators that allowed us to delve deeper and identify the most relevant developments in the subject. Through this review, we sought to highlight the importance of polysaccharides and their wide range of applications in 3D printing and hope that it will provide a meaningful basis for the exploration of printable compounds from renewable sources.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00383"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143101940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Pneumatic extrusion-based bioprinting and flow cytometry: A method for analysing chemotherapy efficacy in 3D bioprinted A375 melanoma cell cultures

Q1 Computer Science

Bioprinting

Pub Date : 2025-02-01 DOI: 10.1016/j.bprint.2024.e00380

Maryke de Villiers, Awie F. Kotzé, Lissinda H. du Plessis

Melanoma, a highly aggressive form of skin cancer, continues to be a significant challenge due to its resistance to conventional chemotherapy treatments and the tendency for metastasis. Advancements in cell culture techniques, especially the transition from 2D cell cultures to more physiologically relevant 3D cell cultures, have provided valuable new insights into cancer biology and chemotherapy drug responses. Although various novel 3D cell culture techniques have been used in melanoma research, standardised and scalable 3D cell culture models suitable for high-throughput pre-clinical drug screening applications are still lacking. Therefore, the purpose of this study was to establish a 3D bioprinted melanoma cell culture model that allows the assessment of drug-induced apoptosis through a flow-cytometric analysis method in 96-well plates. To achieve this, the proposed method integrates the BIOX™ pneumatic extrusion-based 3D bioprinter to extrude reproducible cell-laden droplets in a 96-well plate, and an Annexin V/PI flow cytometric analysis technique optimised for 96-well plate format, to enable cell viability and apoptosis quantification in more physiologically relevant 3D bioprinted cell cultures. The proposed method was evaluated on A375 melanoma 2D and 3D bioprinted cell cultures assayed for drug-induced apoptosis through a flow cytometric method. In addition, a resazurin-based analysis method was also used and compared to determine the efficacy of the proposed flow cytometric analysis method. Compared to the 2D cell cultures, the 3D bioprinted cell cultures demonstrated higher levels of resistance to all chemotherapy drugs evaluated. Furthermore, the comparative analysis of the two methods concluded that the flow cytometric evaluation platform is more sensitive in detecting drug dose responses in 3D bioprinted cell culture models. This method is a proposed alternative to quantify drug-induced apoptosis in 3D melanoma research, thereby advancing the pre-clinical application of 3D bioprinting.

{"title":"Pneumatic extrusion-based bioprinting and flow cytometry: A method for analysing chemotherapy efficacy in 3D bioprinted A375 melanoma cell cultures","authors":"Maryke de Villiers, Awie F. Kotzé, Lissinda H. du Plessis","doi":"10.1016/j.bprint.2024.e00380","DOIUrl":"10.1016/j.bprint.2024.e00380","url":null,"abstract":"<div><div>Melanoma, a highly aggressive form of skin cancer, continues to be a significant challenge due to its resistance to conventional chemotherapy treatments and the tendency for metastasis. Advancements in cell culture techniques, especially the transition from 2D cell cultures to more physiologically relevant 3D cell cultures, have provided valuable new insights into cancer biology and chemotherapy drug responses. Although various novel 3D cell culture techniques have been used in melanoma research, standardised and scalable 3D cell culture models suitable for high-throughput pre-clinical drug screening applications are still lacking. Therefore, the purpose of this study was to establish a 3D bioprinted melanoma cell culture model that allows the assessment of drug-induced apoptosis through a flow-cytometric analysis method in 96-well plates. To achieve this, the proposed method integrates the BIOX™ pneumatic extrusion-based 3D bioprinter to extrude reproducible cell-laden droplets in a 96-well plate, and an Annexin V/PI flow cytometric analysis technique optimised for 96-well plate format, to enable cell viability and apoptosis quantification in more physiologically relevant 3D bioprinted cell cultures. The proposed method was evaluated on A375 melanoma 2D and 3D bioprinted cell cultures assayed for drug-induced apoptosis through a flow cytometric method. In addition, a resazurin-based analysis method was also used and compared to determine the efficacy of the proposed flow cytometric analysis method. Compared to the 2D cell cultures, the 3D bioprinted cell cultures demonstrated higher levels of resistance to all chemotherapy drugs evaluated. Furthermore, the comparative analysis of the two methods concluded that the flow cytometric evaluation platform is more sensitive in detecting drug dose responses in 3D bioprinted cell culture models. This method is a proposed alternative to quantify drug-induced apoptosis in 3D melanoma research, thereby advancing the pre-clinical application of 3D bioprinting.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"45 ","pages":"Article e00380"},"PeriodicalIF":0.0,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143164229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development and characterization of bioinks for 3D bioprinting of in vitro skeletal muscle constructs

Q1 Computer Science

Bioprinting

Pub Date : 2025-01-31 DOI: 10.1016/j.bprint.2025.e00396

Rodi Kado Abdalkader , Kosei Yamauchi , Satoshi Konishi , Takuya Fujita

The use of 3D bioprinting to construct in vitro skeletal muscle models presents a promising approach; however, selecting an optimal bioink remains a common challenge. This study focuses on the development and characterization of bioinks for extrusion-based 3D bioprinting, specifically targeting the creation of accurate skeletal muscle models. By exploring various compositions of alginate, gelatin, fibrinogen, and nanofiber cellulose, we evaluate these formulations based on printability and their support for the growth and differentiation of C2C12 myoblast cells.

While alginate provided a strong, stable matrix for printing scaffolds embedded with C2C12 cells, it did not effectively promote cell growth and differentiation. The addition of fibrinogen to alginate enhanced cell growth and differentiation but was limited mainly to the scaffold surfaces, even with the inclusion of gelatin as a sacrificial ink. Notably, replacing alginate with nanofiber cellulose (NFC) alongside fibrinogen significantly improved cell growth and differentiation, leading to the formation of mature myotubes. Cell distribution was observed both inside and on the surfaces of the scaffolds, indicating effective spatial cell distribution. Furthermore, the scaffolds were tailored to form skeletal muscle bundles anchored between PDMS pillars for contractility testing. Upon exposure to electrical stimulation, the cells displayed measurable displacement, demonstrating contractile function.

These findings offer valuable insights into optimizing bioink formulations that promote myoblast growth and differentiation into skeletal muscle in vitro, with potential applications in future neuromuscular disease modeling.

{"title":"Development and characterization of bioinks for 3D bioprinting of in vitro skeletal muscle constructs","authors":"Rodi Kado Abdalkader , Kosei Yamauchi , Satoshi Konishi , Takuya Fujita","doi":"10.1016/j.bprint.2025.e00396","DOIUrl":"10.1016/j.bprint.2025.e00396","url":null,"abstract":"<div><div>The use of 3D bioprinting to construct <em>in vitro</em> skeletal muscle models presents a promising approach; however, selecting an optimal bioink remains a common challenge. This study focuses on the development and characterization of bioinks for extrusion-based 3D bioprinting, specifically targeting the creation of accurate skeletal muscle models. By exploring various compositions of alginate, gelatin, fibrinogen, and nanofiber cellulose, we evaluate these formulations based on printability and their support for the growth and differentiation of C2C12 myoblast cells.</div><div>While alginate provided a strong, stable matrix for printing scaffolds embedded with C2C12 cells, it did not effectively promote cell growth and differentiation. The addition of fibrinogen to alginate enhanced cell growth and differentiation but was limited mainly to the scaffold surfaces, even with the inclusion of gelatin as a sacrificial ink. Notably, replacing alginate with nanofiber cellulose (NFC) alongside fibrinogen significantly improved cell growth and differentiation, leading to the formation of mature myotubes. Cell distribution was observed both inside and on the surfaces of the scaffolds, indicating effective spatial cell distribution. Furthermore, the scaffolds were tailored to form skeletal muscle bundles anchored between PDMS pillars for contractility testing. Upon exposure to electrical stimulation, the cells displayed measurable displacement, demonstrating contractile function.</div><div>These findings offer valuable insights into optimizing bioink formulations that promote myoblast growth and differentiation into skeletal muscle <em>in vitro</em>, with potential applications in future neuromuscular disease modeling.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"46 ","pages":"Article e00396"},"PeriodicalIF":0.0,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143212070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

3D printed cellulose nanofiber-reinforced and iron-crosslinked double network hydrogel composites for tissue engineering applications: Mechanical properties and cellular viability

Q1 Computer Science

Bioprinting

Pub Date : 2025-01-27 DOI: 10.1016/j.bprint.2025.e00392

Rohit Goyal , Soumyasri Nikhilesh Mahapatra , Rashmi Yadav , Santanu Mitra , Animesh Samanta , Anuj Kumar , Bimlesh Lochab

Additive manufacturing (i.e. 3D printing) is a promising technology for creating three-dimensional (3D) complex tissue-engineered hydrogel structures based on computer digital models resulting from patient-specific anatomical data of the organs. However, besides the printing process, it is worth studying the variation of individual components of the developed hydrogel composites to enable their suitability for tissue engineering. In this work, we shaped 3D printed multi-layered dual (UV- and Fe³⁺ ions)-crosslinked structures using hydrogel-inks composed of polyacrylamide (PAM), alginate (ALG), and cellulose nanofibres (CNFs). For extrusion, ALG in hydrogel precursor ink acted as a viscosity modifier owing to rapid gelation in the presence of low Ca²⁺ ions and CNF provided shear-thinning behavior. With the addition of optimal content of CNF (3 wt%), the mechanical properties of 3D printed composite hydrogel were enhanced and tuned using different fiber orientations. The maximum tensile stress of PAM/ALG_1.5/3CNF composite hydrogel is measured as ∼162 kPa, and maximum tensile toughness as ∼54 kJ/m³ supporting a good fracture resistance. Moreover, CNF-Fe³⁺ loaded 3D printed dual-networked composite hydrogels could disperse energy more efficiently and displayed maximum tensile stress as ∼285 kPa and maximum toughness as ∼200 kJ/m³. Further, In the current study, developed composite structures exhibited enhanced swelling ratio and thermal stability. In addition, finite element (FE) modelling was also exploited to analyze the novel anisotropic composite structures using efficient computational techniques. It is established that varying nanofiber content and fibrils orientation can be utilized to modulate the physicochemical, mechanical, and biological characteristics of printed structures. Overall, PAM/ALG_1.5/3CNF-Fe³⁺ printed composite structures present substantial stretchability, enhanced anisotropic mechanical and physicochemical properties with excellent cytocompatibility.

{"title":"3D printed cellulose nanofiber-reinforced and iron-crosslinked double network hydrogel composites for tissue engineering applications: Mechanical properties and cellular viability","authors":"Rohit Goyal , Soumyasri Nikhilesh Mahapatra , Rashmi Yadav , Santanu Mitra , Animesh Samanta , Anuj Kumar , Bimlesh Lochab","doi":"10.1016/j.bprint.2025.e00392","DOIUrl":"10.1016/j.bprint.2025.e00392","url":null,"abstract":"<div><div>Additive manufacturing (i.e. 3D printing) is a promising technology for creating three-dimensional (3D) complex tissue-engineered hydrogel structures based on computer digital models resulting from patient-specific anatomical data of the organs. However, besides the printing process, it is worth studying the variation of individual components of the developed hydrogel composites to enable their suitability for tissue engineering. In this work, we shaped 3D printed multi-layered dual (UV- and Fe<sup>3+</sup> ions)-crosslinked structures using hydrogel-inks composed of polyacrylamide (PAM), alginate (ALG), and cellulose nanofibres (CNFs). For extrusion, ALG in hydrogel precursor ink acted as a viscosity modifier owing to rapid gelation in the presence of low Ca<sup>2+</sup> ions and CNF provided shear-thinning behavior. With the addition of optimal content of CNF (3 wt%), the mechanical properties of 3D printed composite hydrogel were enhanced and tuned using different fiber orientations. The maximum tensile stress of PAM/ALG<sub>1.5</sub>/3CNF composite hydrogel is measured as ∼162 kPa, and maximum tensile toughness as ∼54 kJ/m<sup>3</sup> supporting a good fracture resistance. Moreover, CNF-Fe<sup>3+</sup> loaded 3D printed dual-networked composite hydrogels could disperse energy more efficiently and displayed maximum tensile stress as ∼285 kPa and maximum toughness as ∼200 kJ/m<sup>3</sup>. Further, In the current study, developed composite structures exhibited enhanced swelling ratio and thermal stability. In addition, finite element (FE) modelling was also exploited to analyze the novel anisotropic composite structures using efficient computational techniques. It is established that varying nanofiber content and fibrils orientation can be utilized to modulate the physicochemical, mechanical, and biological characteristics of printed structures. Overall, PAM/ALG<sub>1.5</sub>/3CNF-Fe<sup>3+</sup> printed composite structures present substantial stretchability, enhanced anisotropic mechanical and physicochemical properties with excellent cytocompatibility.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"46 ","pages":"Article e00392"},"PeriodicalIF":0.0,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143093895","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0