Experimental and analytical study of novel rapid freeze casting technique to fabricate 3D-shaped gelatin nanofibers

Document Type: Original Article


1 Biomaterials Research Group, Nanotechnology and Advanced Materials Department, Materials and Energy Research Center, Karaj, Iran.

2 Department of Biomedical Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran.


Nanofibrous scaffolds are often used to reconstruct damaged tissues/organs. Unfortunately the lack of producing three-dimensional (3D) nanofiber results in their restricted applications. Therefore, bulky-shaped gelatin nanofibers were fabricated through novel rapid freeze casting (RFC) technique to simulate extracellular matrix (ECM) and accelerate the regeneration. Formation of 3D-shaped fibers in the range of 200-1000 nm with approximately 98% porosity and significantly improved mechanical stability compared with conventional freeze casting (CFC) technique is one of the strengths of this study even though both RFC and CFC macrostructures are similar. Outcomes proved this novel technique reduced hydrophilicity and controlled biodegradation rate owing to applying a high freezing gradient in order to the production of thin pores. The viability of more than 90% cells compared with control group confirmed the biocompatibility of constructs and supporting cellular proliferation. In brief, novel RFC gelatin nanofibers represented original physicochemical and mechanical features for further in-vitro and in-vivo studies.

Graphical Abstract

Experimental and analytical study of novel rapid freeze casting technique to fabricate 3D-shaped gelatin nanofibers


1.        Sangkert S, Kamonmattayakul S, Chai WL, Meesane J. A biofunctional-modified silk fibroin scaffold with mimic reconstructed extracellular matrix of decellularized pulp/collagen/fibronectin for bone tissue engineering in alveolar bone resorption. Mater Lett [Internet]. 2016 Mar;166:30–4. Available from: http://linkinghub.elsevier.com/retrieve/pii/S0167577X15309824

2.        Boccaccini AR, Ma PX. Tissue engineering using ceramics and polymers. 2014.

3.        Unnithan AR, Arathyram RS, Kim CS. Electrospinning of polymers for tissue engineering [Internet]. Nanotechnology Applications for Tissue Engineering. Elsevier Inc.; 2015. 45-55 p. Available from: http://linkinghub.elsevier.com/retrieve/pii/B9780323328890000030

4.        Ghorbani F, Nojehdehyan H, Zamanian A, Gholipourmalekabadi M, Mozafari M. Synthesis, physico-chemical characteristics and cellular behavior of poly (lactic-co-glycolic acid)/gelatin nanofibrous scaffolds for engineering soft connective tissues. Adv Mater Lett. 2016;7(2):163–9.

5.        Mi HY, Jing X, Yu E, McNulty J, Peng XF, Turng LS. Fabrication of triple-layered vascular scaffolds by combining electrospinning, braiding, and thermally induced phase separation. Mater Lett [Internet]. 2015;161:305–8. Available from: http://dx.doi.org/10.1016/j.matlet.2015.08.119

6.        Ariga K, Hill JP, Lee M V, Vinu A, Charvet R, Acharya S. Challenges and breakthroughs in recent research on self-assembly. Sci Technol Adv Mater. 2008;9(1):14109.

7.        Vinogradov G V., Yarlykov B V., Tsebrenko M V., Yudin A V., Ablazova TI. Fibrillation in the flow of polyoxymethylene melts. Polymer (Guildf). 1975;16(8):609–14.

8.        Ndaro MS, Xiangyu J, Ting C, Yu C. Effect of impact force on tensile properties and fiber splitting of splittable bicomponent hydroentangled fabrics. Fibers Polym. 2007;8(4):421–6.

9.        Benavides RE, Jana SC, Reneker DH. Nanofibers from Scalable Gas Jet Process. ACS Macro Lett [Internet]. 2012 Aug 21;1(8):1032–6. Available from: http://pubs.acs.org/doi/abs/10.1021/mz300297g

10.      Ghorbani F, Zamanian A, Nojehdehian H. Effects of pore orientation on in-vitro properties of retinoic acid-loaded PLGA/gelatin scaffolds for artificial peripheral nerve application. Mater Sci Eng C [Internet]. 2017 Mar;77:159–72. Available from: http://linkinghub.elsevier.com/retrieve/pii/S0928493116321634

11.      Ghorbani F, Nojehdehian H, Zamanian A. Physicochemical and mechanical properties of freeze cast hydroxyapatite-gelatin scaffolds with dexamethasone loaded PLGA microspheres for hard tissue engineering applications. Mater Sci Eng C [Internet]. 2016 Dec;69:208–20. Available from: http://linkinghub.elsevier.com/retrieve/pii/S092849311630649X

12.      Salgado AJ, Gomes ME, Chou A, Coutinho OP, Reis RL, Hutmacher DW. Preliminary study on the adhesion and proliferation of human osteoblasts on starch-based scaffolds. Mater Sci Eng C [Internet]. 2002 May;20(1–2):27–33. Available from: http://www.sciencedirect.com/science/article/pii/S0928493102000097

13.      Wu X, Liu Y, Li X, Wen P, Zhang Y, Long Y, et al. Preparation of aligned porous gelatin scaffolds by unidirectional freeze-drying method. Acta Biomater [Internet]. 2010 Mar [cited 2013 Nov 24];6(3):1167–77. Available from: http://www.ncbi.nlm.nih.gov/pubmed/19733699

14.      Cai J, Ziemba KS, Smith GM, Jin Y. Evaluation of cellular organization and axonal regeneration through linear PLA foam implants in acute and chronic spinal cord injury. J Biomed Mater Res Part A [Internet]. 2007 Nov;83A(2):512–20. Available from: http://doi.wiley.com/10.1002/jbm.a.31296

15.      Zamanian A, Farhangdoust S, Yasaei M, Khorami M, Hafezi M. The effect of particle size on the mechanical and microstructural properties of freeze-casted macroporous hydroxyapatite scaffolds. Int J Appl Ceram Technol [Internet]. 2014 Jan 4;11(1):12–21. Available from: http://doi.wiley.com/10.1111/ijac.12031

16.      Farhangdoust S, Zamanian A, Yasaei M, Khorami M. The effect of processing parameters and solid concentration on the mechanical and microstructural properties of freeze-casted macroporous hydroxyapatite scaffolds. Mater Sci Eng C Mater Biol Appl [Internet]. 2013 Jan 1 [cited 2015 Apr 25];33(1):453–60. Available from: http://www.ncbi.nlm.nih.gov/pubmed/25428095

17.      Deville S. Freezing Colloids: Observations, Principles, Control, and Use [Internet]. Cham: Springer International Publishing; 2017. (Engineering Materials and Processes). Available from: http://link.springer.com/10.1007/978-3-319-50515-2

18.      Li X-K, Cai S-X, Liu B, Xu Z-L, Dai X-Z, Ma K-W, et al. Characteristics of PLGA-gelatin complex as potential artificial nerve scaffold. Colloids Surf B Biointerfaces [Internet]. 2007 Jun 15 [cited 2015 Oct 11];57(2):198–203. Available from: http://www.ncbi.nlm.nih.gov/pubmed/17368867

19.      Zhang S, Huang Y, Yang X, Mei F, Ma Q, Chen G, et al. Gelatin nanofibrous membrane fabricated by electrospinning of aqueous gelatin solution for guided tissue regeneration. J Biomed Mater Res A. 2009;90:671–9.