The Impact of Temperature and the Duration of Freezing on a Hydrogel Used for a 3D-Bioprinted In Vitro Skin Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Preparation of the Hydrogel for 3D Printing
2.3. Three-Dimensional Bioprinting and Postprocessing
2.3.1. Three-Dimensional Bioprinting
2.3.2. Lyophilization
2.4. Scaffold Characterization Methods
2.4.1. Rheological Measurements
2.4.2. Attenuated Total Reflectance Infrared Spectroscopy (AFT-IR)
2.4.3. Swelling Test
2.4.4. In Vitro Degradation Test
2.4.5. Internal Structure Analysis Using Nano-Computed Tomography
2.4.6. Morphology and Internal Structure Analysis Using Scanning Electron Microscopy
2.4.7. Mechanical Properties Analysis
2.4.8. Statistical Analysis
3. Results and Discussion
3.1. Printability of the Prepared Hydrogel Formulation
3.2. Chemical Composition of 3D-Bioprinted Scaffolds
3.3. Three-Dimensional-Bioprinted Scaffolds Water Uptake Capacity (Swelling Ratio)
3.4. In Vitro Degradation of the 3D-Bioprinted Scaffolds
3.5. Characterization of 3D-Printed Scaffolds
3.6. Mechanical Properties of 3D-Printed Scaffolds
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Choudhury, S.; Das, A. Advances in generation of three-dimensional skin equivalents: Pre-clinical studies to clinical therapies. Cytotherapy 2021, 23, 1–9. [Google Scholar] [CrossRef]
- Tabatabaei Hosseini, B.S.; Meadows, K.; Gabriel, V.; Hu, J.; Kim, K. Biofabrication of Cellulose-based Hydrogels for Advanced Wound Healing: A Special Emphasis on 3D Bioprinting. Macromol. Biosci. 2024, 24, 2300376. [Google Scholar] [CrossRef] [PubMed]
- Sadeghianmaryan, A.; Sadeghianmaryan, A.; Ahmadian, N.; Wheatley, S.; Sardroud, H.A.; Nasrollah, S.A.S.; Naseri, E.; Ahmadi, A. Advancements in 3D-printable polysaccharides, proteins, and synthetic polymers for wound dressing and skin scaffolding—A review. Int. J. Biol. Macromol. 2024, 266 Pt 1, 131207. [Google Scholar] [CrossRef]
- Liu, H.; Xing, F.; Yu, P.; Zhe, M.; Duan, X.; Liu, M.; Xiang, Z.; Ritz, U. A review of biomacromolecule-based 3D bioprinting strategies for structure-function integrated repair of skin tissues. Int. J. Biol. Macromol. 2024, 268 Pt 2, 131623. [Google Scholar] [CrossRef] [PubMed]
- Zidaric, T.; Milojević, M.; Gradišnik, L.; Kleinschek, K.S.; Maver, U.; Maver, T. Polysaccharide-Based Bioink Formulation for 3D Bioprinting of an In Vitro Model of the Human Dermis. Nanomaterials 2020, 10, 733. [Google Scholar] [CrossRef] [PubMed]
- Savoji, H.; Godau, B.; Hassani, M.S.; Akbari, M. Skin Tissue Substitutes and Biomaterial Risk Assessment and Testing. Front. Bioeng. Biotechnol. 2018, 6, 86. [Google Scholar] [CrossRef]
- Sanabria-de la Torre, R.; Fernández-González, A.; Quiñones-Vico, M.I.; Montero-Vilchez, T.; Arias-Santiago, S. Bioengineered Skin Intended as In Vitro Model for Pharmacosmetics, Skin Disease Study and Environmental Skin Impact Analysis. Biomedicines 2020, 8, 464. [Google Scholar] [CrossRef]
- Vig, K.; Chaudhari, A.; Tripathi, S.; Dixit, S.; Sahu, R.; Pillai, S.; Dennis, V.A.; Singh, S.R. Advances in Skin Regeneration Using Tissue Engineering. Int. J. Mol. Sci. 2017, 18, 789. [Google Scholar] [CrossRef]
- Tan, S.H.; Chua, D.A.C.; Tang, J.R.J.; Bonnard, C.; Leavesley, D.; Liang, K. Design of hydrogel-based scaffolds for in vitro three-dimensional human skin model reconstruction. Acta Biomater. 2022, 153, 13–37. [Google Scholar] [CrossRef]
- Kim, B.S.; Kwon, Y.W.; Kong, J.-S.; Park, G.T.; Gao, G.; Han, W.; Kim, M.-B.; Lee, H.; Kim, J.H.; Cho, D.-W. 3D cell printing of in vitro stabilized skin model and in vivo pre-vascularized skin patch using tissue-specific extracellular matrix bioink: A step towards advanced skin tissue engineering. Biomaterials 2018, 168, 38–53. [Google Scholar] [CrossRef]
- Taneja, H.; Salodkar, S.M.; Parmar, A.S.; Chaudhary, S. Hydrogel based 3D printing: Bio ink for tissue engineering. J. Mol. Liq. 2022, 367, 120390. [Google Scholar] [CrossRef]
- Muller, M.; Öztürk, E.; Arlov, Ø.; Gatenholm, P.; Zenobi-Wong, M. Alginate Sulfate-Nanocellulose Bioinks for Cartilage Bioprinting Applications. Ann. Biomed. Eng. 2017, 45, 210–223. [Google Scholar] [CrossRef]
- Zidaric, T.; Gradišnik, L.; Frangež, T.; Šoštarič, M.; Korunič, E.; Maver, T.; Maver, U. Novel 3D printed polysaccharide-based materials with prebiotic activity for potential treatment of diaper rash. Int. J. Biol. Macromol. 2024, 269 Pt 1, 131958. [Google Scholar] [CrossRef]
- Varaprasad, K.; Karthikeyan, C.; Yallapu, M.M.; Sadiku, R. The significance of biomacromolecule alginate for the 3D printing of hydrogels for biomedical applications. Int. J. Biol. Macromol. 2022, 212, 561–578. [Google Scholar] [CrossRef] [PubMed]
- Khan, S.A.; Ahmad, H.; Zhu, G.; Pang, H.; Zhang, Y. Three-Dimensional Printing of Hydrogels for Flexible Sensors: A Review. Gels 2024, 10, 187. [Google Scholar] [CrossRef] [PubMed]
- Footner, E.; Firipis, K.; Liu, E.; Baker, C.; Foley, P.; Kapsa, R.M.I.; Pirogova, E.; O’connell, C.; Quigley, A. Layer-by-Layer Analysis of In Vitro Skin Models. ACS Biomater. Sci. Eng. 2023, 9, 5933–5952. [Google Scholar] [CrossRef]
- Hou, L.X.; Ju, H.; Hao, X.P.; Zhang, H.; Zhang, L.; He, Z.; Wang, J.; Zheng, Q.; Wu, Z.L. Intrinsic Anti-Freezing and Unique Phosphorescence of Glassy Hydrogels with Ultrahigh Stiffness and Toughness at Low Temperatures. Adv. Mater. 2023, 35, 2300244. [Google Scholar] [CrossRef]
- Zeng, L.; He, J.; Cao, Y.; Wang, J.; Qiao, Z.; Jiang, X.; Hou, L.; Zhang, J. Tissue-adhesive and highly mechanical double-network hydrogel for cryopreservation and sustained release of anti-cancer drugs. Smart Mater. Med. 2021, 2, 229–236. [Google Scholar] [CrossRef]
- Jian, Y.; Handschuh-Wang, S.; Zhang, J.; Lu, W.; Zhou, X.; Chen, T. Biomimetic anti-freezing polymeric hydrogels: Keeping soft-wet materials active in cold environments. Mater. Horiz. 2021, 8, 351–369. [Google Scholar] [CrossRef]
- Liu, T.; Zhang, Y.; Sun, M.; Jin, M.; Xia, W.; Yang, H.; Wang, T. Effect of Freezing Process on the Microstructure of Gelatin Methacryloyl Hydrogels. Front. Bioeng. Biotechnol. 2021, 9, 810155. [Google Scholar] [CrossRef]
- Curic, L.C.; Šuligoj, M.; Ibic, M.; Marovič, N.; Vihar, B.; Vesenjak, M.; Dubrovski, P.D.; Novak, N.; Stergar, J.; Ban, I.; et al. Development of a novel NiCu nanoparticle-loaded polysaccharide-based hydrogel for 3D printing of customizable dressings with promising cytotoxicity against melanoma cells. Mater. Today Bio 2023, 22, 100770. [Google Scholar]
- Škrinjar, D.; Sever, M.; Maver, T.; Gradišnik, L.; Zidarič, T. Razvoj in vitro modela kože. Anali PAZU 2024, 14, 29–47. [Google Scholar] [CrossRef]
- Bidarra, S.J.; Barrias, C.C.; Granja, P.L. Injectable alginate hydrogels for cell delivery in tissue engineering. Acta Biomater. 2014, 10, 1646–1662. [Google Scholar] [CrossRef] [PubMed]
- Teoh, J.H.; Abdul Shakoor, F.T.; Wang, C.H. 3D Printing Methyl Cellulose Hydrogel Wound Dressings with Parameter Exploration Via Computational Fluid Dynamics Simulation. Pharm. Res. 2022, 39, 281–294. [Google Scholar] [CrossRef]
- Dumanli, A.G. Nanocellulose and its Composites for Biomedical Applications. Curr. Med. Chem. 2017, 24, 512–528. [Google Scholar] [CrossRef]
- ISO 3219:1993(en); Plastics—Polymers/Resins in the Liquid State or as Emulsions or Dispersions—Determination of Viscosity Using a Rotational Viscometer with Defined Shear Rate. International Standard Standardization: Geneva, Switzerland, 1993.
- Giesekus, H.; Langer, G. Determination of the true flow curves of non-Newtonian liquids and plastics using the representative viscosity method. Rheol. Acta 1977, 16, 1–22. [Google Scholar] [CrossRef]
- Garcia, M.C.; Alfaro, M.C.; Calero, N.; Muñoz, J. Influence of polysaccharides on the rheology and stabilization of alpha-pinene emulsions. Carbohydr. Polym. 2014, 105, 177–183. [Google Scholar] [CrossRef]
- Maver, U.; Xhanari, K.; Žižek, M.; Gradišnik, L.; Repnik, K.; Potočnik, U.; Finšgar, M. Carboxymethyl cellulose/diclofenac bioactive coatings on AISI 316LVM for controlled drug delivery, and improved osteogenic potential. Carbohydr. Polym. 2020, 230, 115612. [Google Scholar] [CrossRef]
- Maver, U.; Xhanari, K.; Žižek, M.; Korte, D.; Gradišnik, L.; Franko, M.; Finšgar, M. A combination of interdisciplinary analytical tools for evaluation of multi-layered coatings on medical grade stainless steel for biomedical applications. Eur. J. Pharm. Biopharm. 2018, 128, 230–246. [Google Scholar] [CrossRef]
- Zhang, Y.; Ye, L.; Cui, J.; Yang, B.; Sun, H.; Li, J.; Yao, F. A Biomimetic Poly(vinyl alcohol)-Carrageenan Composite Scaffold with Oriented Microarchitecture. ACS Biomater. Sci. Eng. 2016, 2, 544–557. [Google Scholar] [CrossRef]
- Lee, V.; Singh, G.; Trasatti, J.P.; Bjornsson, C.; Xu, X.; Tran, T.N.; Yoo, S.-S.; Dai, G.; Karande, P. Design and fabrication of human skin by three-dimensional bioprinting. Tissue Eng. Part C Methods 2014, 20, 473–484. [Google Scholar] [CrossRef] [PubMed]
- Olejnik, A.; Semba, J.A.; Kulpa, A.; Dańczak-Pazdrowska, A.; Rybka, J.D.; Gornowicz-Porowska, J. 3D Bioprinting in Skin Related Research: Recent Achievements and Application Perspectives. ACS Synth. Biol. 2022, 11, 26–38. [Google Scholar] [CrossRef]
- Choi, K.Y.; Ajiteru, O.; Hong, H.; Suh, Y.J.; Sultan, T.; Lee, H.; Lee, J.S.; Lee, Y.J.; Lee, O.J.; Kim, S.H.; et al. A digital light processing 3D-printed artificial skin model and full-thickness wound models using silk fibroin bioink. Acta Biomater. 2023, 164, 159–174. [Google Scholar] [CrossRef]
- Tosini, M.; Tänzer, T.; Villata, S.; Baruffaldi, D.; Monica, V.; Peracino, B.; Primo, L.; Frascella, F.; Pirri, F.; Audenino, A.; et al. A Methodological Approach for Interpreting and Comparing the Viscoelastic Behaviors of Soft Biological Tissues and Hydrogels at the Cell-Length Scale. Appl. Sci. 2024, 14, 1093. [Google Scholar] [CrossRef]
- Plesser, H.E. Reproducibility vs. Replicability: A Brief History of a Confused Terminology. Front. Neuroinform. 2017, 11, 76. [Google Scholar] [CrossRef] [PubMed]
- Aljohani, W.; Ullah, M.W.; Zhang, X.; Yang, G. Bioprinting and its applications in tissue engineering and regenerative medicine. Int. J. Biol. Macromol. 2018, 107 Pt A, 261–275. [Google Scholar] [CrossRef]
- Markstedt, K.; Mantas, A.; Tournier, I.; Martínez Ávila, H.; Hägg, D.; Gatenholm, P. 3D Bioprinting Human Chondrocytes with Nanocellulose-Alginate Bioink for Cartilage Tissue Engineering Applications. Biomacromolecules 2015, 16, 1489–1496. [Google Scholar] [CrossRef] [PubMed]
- Schwab, A.; Levato, R.; D’este, M.; Piluso, S.; Eglin, D.; Malda, J. Printability and Shape Fidelity of Bioinks in 3D Bioprinting. Chem. Rev. 2020, 120, 11028–11055. [Google Scholar] [CrossRef]
- Rutz, A.L.; Hyland, K.E.; Jakus, A.E.; Burghardt, W.R.; Shah, R.N. A multimaterial bioink method for 3D printing tunable, cell-compatible hydrogels. Adv. Mater. 2015, 27, 1607–1614. [Google Scholar] [CrossRef]
- Lee, S.C.; Gillispie, G.; Prim, P.; Lee, S.J. Physical and Chemical Factors Influencing the Printability of Hydrogel-based Extrusion Bioinks. Chem. Rev. 2020, 120, 10834–10886. [Google Scholar] [CrossRef]
- Amorim, P.A.; D’ávila, M.; Anand, R.; Moldenaers, P.; Van Puyvelde, P.; Bloemen, V. Insights on shear rheology of inks for extrusion-based 3D bioprinting. Bioprinting 2021, 22, e00129. [Google Scholar] [CrossRef]
- Zhong, Q.; Chen, Y.; Zhang, X.; Yang, G.; Jin, W.; Peng, D.; Huang, Q. Correlation between 3D printability and rheological properties of biopolymer fluid: A case study of alginate-based hydrogels. J. Food Eng. 2024, 370, 111970. [Google Scholar] [CrossRef]
- Picout, D.R.; Ross-Murphy, S.B. Rheology of biopolymer solutions and gels. Sci. World J. 2003, 3, 105–121. [Google Scholar] [CrossRef]
- Rau, D.A.; Bortner, M.J.; Williams, C.B. A rheology roadmap for evaluating the printability of material extrusion inks. Addit. Manuf. 2023, 75, 103745. [Google Scholar] [CrossRef]
- Bom, S.; Ribeiro, R.; Ribeiro, H.M.; Santos, C.; Marto, J. On the progress of hydrogel-based 3D printing: Correlating rheological properties with printing behaviour. Int. J. Pharm. 2022, 615, 121506. [Google Scholar] [CrossRef]
- Ning, L.; Sun, H.; Lelong, T.; Guilloteau, R.; Zhu, N.; Schreyer, D.J.; Chen, X. 3D bioprinting of scaffolds with living Schwann cells for potential nerve tissue engineering applications. Biofabrication 2018, 10, 035014. [Google Scholar] [CrossRef]
- Guvendiren, M.; Lu, H.D.; Burdick, J.A. Shear-thinning hydrogels for biomedical applications. Soft Matter 2012, 8, 260–272. [Google Scholar] [CrossRef]
- Enoch, K.; Rakavi, C.S.; Somasundaram, A.A. Tuning the rheological properties of chitosan/alginate hydrogels for tissue engineering application. Colloids Surf. A Physicochem. Eng. Asp. 2024, 697, 134434. [Google Scholar] [CrossRef]
- Waresindo, W.X.; Rahmi Luthfianti, H.; Priyanto, A.; Hapidin, D.A.; Edikresnha, D.; Aimon, A.H.; Suciati, T.; Khairurrijal, K. Freeze–thaw hydrogel fabrication method: Basic principles, synthesis parameters, properties, and biomedical applications. Mater. Res. Express 2023, 10, 024003. [Google Scholar] [CrossRef]
- Tang, Y.; Wang, X.; Li, Y.; Lei, M.; Du, Y.; Kennedy, J.F.; Knill, C.J. Production and characterisation of novel injectable chitosan/methylcellulose/salt blend hydrogels with potential application as tissue engineering scaffolds. Carbohydr. Polym. 2010, 82, 833–841. [Google Scholar] [CrossRef]
- Mohan, T.; Štiglic, A.D.; Beaumont, M.; Konnerth, J.; Gürer, F.; Makuc, D.; Maver, U.; Gradišnik, L.; Plavec, J.; Kargl, R.; et al. Generic Method for Designing Self-Standing and Dual Porous 3D Bioscaffolds from Cellulosic Nanomaterials for Tissue Engineering Applications. ACS Appl. Bio Mater. 2020, 3, 1197–1209. [Google Scholar] [CrossRef] [PubMed]
- dos Santos Carvalho, J.D.; Rabelo, R.S.; Hubinger, M.D. Thermo-rheological properties of chitosan hydrogels with hydroxypropyl methylcellulose and methylcellulose. Int. J. Biol. Macromol. 2022, 209, 367–375. [Google Scholar] [CrossRef] [PubMed]
- Nunes, M.R.; de Souza Maguerroski Castilho, M.; de Lima Veeck, A.P.; da Rosa, G.C.; Montanheiro Noronha, C.; Maciel, M.V.O.B.; Manique Barreto, P. Antioxidant and antimicrobial methylcellulose films containing Lippia alba extract and silver nanoparticles. Carbohydr. Polym. 2018, 192, 37–43. [Google Scholar] [CrossRef] [PubMed]
- Derkach, S.R.; Voron’ko, N.G.; Sokolan, N.I.; Kolotova, D.S.; Kuchina, Y.A. Interactions between gelatin and sodium alginate: UV and FTIR studies. J. Dispers. Sci. Technol. 2020, 41, 690–698. [Google Scholar] [CrossRef]
- Wang, M.; Zang, Y.; Hong, K.; Zhao, X.; Yu, C.; Liu, D.; An, Z.; Wang, L.; Yue, W.; Nie, G. Preparation of pH-sensitive carboxymethyl cellulose/chitosan/alginate hydrogel beads with reticulated shell structure to deliver Bacillus subtilis natto. Int. J. Biol. Macromol. 2021, 192, 684–691. [Google Scholar] [CrossRef]
- Serafin, A.; Culebras, M.; Collins, M.N. Synthesis and evaluation of alginate, gelatin, and hyaluronic acid hybrid hydrogels for tissue engineering applications. Int. J. Biol. Macromol. 2023, 233, 123438. [Google Scholar] [CrossRef]
- Caliari, S.R.; Burdick, J.A. A practical guide to hydrogels for cell culture. Nat. Methods 2016, 13, 405–414. [Google Scholar] [CrossRef]
- Zhang, X.; Morits, M.; Jonkergouw, C.; Ora, A.; Valle-Delgado, J.J.; Farooq, M.; Ajdary, R.; Huan, S.; Linder, M.B.; Rojas, O.J.; et al. Three-Dimensional Printed Cell Culture Model Based on Spherical Colloidal Lignin Particles and Cellulose Nanofibril-Alginate Hydrogel. Biomacromolecules 2020, 21, 1875–1885. [Google Scholar] [CrossRef]
- Bittner, S.M.; Pearce, H.A.; Hogan, K.J.; Smoak, M.M.; Guo, J.L.; Melchiorri, A.J.; Scott, D.W.; Mikos, A.G. Swelling Behaviors of 3D Printed Hydrogel and Hydrogel-Microcarrier Composite Scaffolds. Tissue Eng. Part A 2021, 27, 665–678. [Google Scholar] [CrossRef]
- Kopač, T.; Abrami, M.; Grassi, M.; Ručigaj, A.; Krajnc, M. Polysaccharide-based hydrogels crosslink density equation: A rheological and LF-NMR study of polymer-polymer interactions. Carbohydr. Polym. 2022, 277, 118895. [Google Scholar] [CrossRef]
- Ashammakhi, N.; Ahadian, S.; Xu, C.; Montazerian, H.; Ko, H.; Nasiri, R.; Barros, N.; Khademhosseini, A. Bioinks and bioprinting technologies to make heterogeneous and biomimetic tissue constructs. Mater. Today Bio 2019, 1, 100008. [Google Scholar] [CrossRef]
- Nair, L.S.; Laurencin, C.T. Biodegradable polymers as biomaterials. Prog. Polym. Sci. 2007, 32, 762–798. [Google Scholar] [CrossRef]
- Mao, H.; Yang, L.; Zhu, H.; Wu, L.; Ji, P.; Yang, J.; Gu, Z. Recent advances and challenges in materials for 3D bioprinting. Prog. Nat. Sci. Mater. Int. 2020, 30, 618–634. [Google Scholar] [CrossRef]
- Mohabatpour, F.; Duan, X.; Yazdanpanah, Z.; Tabil, X.L.; Lobanova, L.; Zhu, N.; Papagerakis, S.; Chen, X.; Papagerakis, P. Bioprinting of alginate-carboxymethyl chitosan scaffolds for enamel tissue engineering in vitro. Biofabrication 2022, 15, 015022. [Google Scholar] [CrossRef] [PubMed]
- Zhang, H.; Cheng, J.; Ao, Q. Preparation of Alginate-Based Biomaterials and Their Applications in Biomedicine. Mar. Drugs 2021, 19, 264. [Google Scholar] [CrossRef]
- Salvatore, L.; Calò, E.; Bonfrate, V.; Pedone, D.; Gallo, N.; Natali, M.L.; Sannino, A.; Madaghiele, M. Exploring the effects of the crosslink density on the physicochemical properties of collagen-based scaffolds. Polym. Test. 2021, 93, 106966. [Google Scholar] [CrossRef]
- Li, L.; Guo, J.; Xiong, R. Synthesis and swelling behavior of a fully degradable physical cross-linked high strength hydrogel. Polym. Test. 2021, 94, 106982. [Google Scholar] [CrossRef]
- Aston, R.; Sewell, K.; Klein, T.; Lawrie, G.; Grøndahl, L. Evaluation of the impact of freezing preparation techniques on the characterisation of alginate hydrogels by cryo-SEM. Eur. Polym. J. 2016, 82, 1–15. [Google Scholar] [CrossRef]
- Zhang, D.; Liu, Y.; Gross, W.; Tang, Y.; Zheng, J. Designing Antifreezing Hydrogels with Enhanced Mechanical Properties Using a Simple Crosslinker. Giant 2023, 16, 100203. [Google Scholar] [CrossRef]
- Xu, D.; Miranda, M.L.H.; Evans, N.D.; Sengers, B.G.; Browne, M.; Cook, R.B. Depth profiling via nanoindentation for characterisation of the elastic modulus and hydraulic properties of thin hydrogel layers. J. Mech. Behav. Biomed. Mater. 2023, 148, 106195. [Google Scholar] [CrossRef]
- Wekwejt, M.; Michalska-Sionkowska, M.; Bartmański, M.; Nadolska, M.; Łukowicz, K.; Pałubicka, A.; Osyczka, A.; Zieliński, A. Influence of several biodegradable components added to pure and nanosilver-doped PMMA bone cements on its biological and mechanical properties. Mater. Sci. Eng. C 2020, 117, 111286. [Google Scholar] [CrossRef]
- Stanisławska, A.; Szkodo, M.; Staroszczyk, H.; Dawidowska, K.; Kołaczkowska, M.; Siondalski, P. Effect of the ex situ physical and in situ chemical modification of bacterial nanocellulose on mechanical properties in the context of its potential applications in heart valve design. Int. J. Biol. Macromol. 2024, 269, 131951. [Google Scholar] [CrossRef]
Nanoindentation Mechanical Properties | ||||
---|---|---|---|---|
Hardness [MPa] | Indentation Modulus of Elasticity [MPa] | Plastic Work [nJ] | Elastic Work [nJ] | |
CTRL | 26 ± 19 | 585 ± 239 | 54.4 ± 19.6 | 27.1 ± 5.3 |
−20 °C (7 days) | 229 ± 109 | 2971 ± 983 | 12.8 ± 3.6 | 10.6 ± 1.7 |
−20 °C (29 days) | 59 ± 14 | 1062 ± 230 | 22.3 ± 4.4 | 20.8 ± 3.5 |
−80 °C (7 days) | 20 ± 7 | 249 ± 45 | 43 ± 11 | 41.5 ± 3.3 |
−80 °C (29 days) | 28 ± 17 | 644 ± 240 | 49.3 ± 18 | 28.1 ± 6.4 |
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Sever, M.; Škrinjar, D.; Maver, T.; Belak, M.; Zupanič, F.; Anžel, I.; Zidarič, T. The Impact of Temperature and the Duration of Freezing on a Hydrogel Used for a 3D-Bioprinted In Vitro Skin Model. Biomedicines 2024, 12, 2028. https://doi.org/10.3390/biomedicines12092028
Sever M, Škrinjar D, Maver T, Belak M, Zupanič F, Anžel I, Zidarič T. The Impact of Temperature and the Duration of Freezing on a Hydrogel Used for a 3D-Bioprinted In Vitro Skin Model. Biomedicines. 2024; 12(9):2028. https://doi.org/10.3390/biomedicines12092028
Chicago/Turabian StyleSever, Maja, Dominik Škrinjar, Tina Maver, Monika Belak, Franc Zupanič, Ivan Anžel, and Tanja Zidarič. 2024. "The Impact of Temperature and the Duration of Freezing on a Hydrogel Used for a 3D-Bioprinted In Vitro Skin Model" Biomedicines 12, no. 9: 2028. https://doi.org/10.3390/biomedicines12092028