Experimental Investigation of Governing Parameters in the Electrospinning of Poly(3-hydroxybutyrate)-Starch Scaffolds: Structural Characterization en en In this study, for the first time, electrospinning of polyhydroxybutyrate/starch was done and optimized to produce uniform-size and bead-free nanofibers following Taguchi methodology. It was found that main parameters such as type of solvent, applied voltage, solution flow rate and distance from nozzle tip to collector surface play an important role in the characteristics of the obtained nanofibrous morphologies. It was also found out from the main effects that the solvent type (chloroform/dimethylformamide, trifluoroacetic acid) is the most effective factor in the diameter and quality of the fibers. The structure and the average diameter of the fibers were assessed by a scanning electron microscope. The results indicated that all electrospun scaffolds have three-dimensional structures with high interconnected porosity. The optimum levels of factors were determined as follows: 18kV of voltage, 0.75 µL/h of flow rate, 15 cm of distance, and TFA as solvent in order to obtain the thinnest bead-free nanofibers. 4 12 Maryam Abdollahi Asl Tissue Engineering and Regenerative Medicine Institute Central Tehran Branch, Islamic Azad University Iran info@jourcc.com Mohammad Mohammadalipour Department of Chemical Engineering Isfahan University of Technology Iran info@jourcc.com Saeed Karbasi Department of Biomaterials and Tissue Engineering Isfahan University of Medical Sciences Iran karbasi@med.mui.ac.ir Electrospinning Polyhydroxybutyrate Scaffold Starch Taguchi method Article2.pdf [1] P.K. Chandra, S. Soker, A. Atala, Chapter 1 - Tissue engineering: current status and future perspectives, in: R. Lanza, R. Langer, J.P. Vacanti, A. Atala (Eds.), Principles of Tissue Engineering (Fifth Edition), Academic Press2020, pp. 1-35. ## [2] M.A. asl, H. Ghomi, Fabrication of highly porous merwinite scaffold using the space holder method, International Journal of Materials Research 111(9) (2020) 711-718. ## [3] S.S. Patil, R.D.K. Misra, The significance of macromolecular architecture in governing structure-property relationship for biomaterial applications: an overview, Materials Technology 33(5) (2018) 364-386. ## [4] M.R. Foroughi, S. Karbasi, M. Khoroushi, A.A. Khademi, Polyhydroxybutyrate/chitosan/bioglass nanocomposite as a novel electrospun scaffold: fabrication and characterization, Journal of Porous Materials 24(6) (2017) 1447-1460. ## [5] Z. Mohammadalizadeh, S. Karbasi, S. Arasteh, Physical, mechanical and biological evaluation of poly (3-hydroxybutyrate)-chitosan/MWNTs as a novel electrospun scaffold for cartilage tissue engineering applications, Polymer-Plastics Technology and Materials 59(4) (2020) 417-429. ## [6] D. Sadeghi, S. Karbasi, S. Razavi, S. Mohammadi, M.A. Shokrgozar, S. Bonakdar, Electrospun poly(hydroxybutyrate)/chitosan blend fibrous scaffolds for cartilage tissue engineering, Journal of Applied Polymer Science 133(47) (2016). ## [7] P. Naderi, M. Zarei, S. Karbasi, H. Salehi, Evaluation of the effects of keratin on physical, mechanical and biological properties of poly (3-hydroxybutyrate) electrospun scaffold: Potential application in bone tissue engineering, European Polymer Journal 124 (2020) 109502. ## [8] J. Su, L. Chen, L. Li, Characterization of polycaprolactone and starch blends for potential application within the biomaterials field, African Journal of Biotechnology 11(3) (2012) 694-701. ## [9] A. Shafqat, A. Tahir, A. Mahmood, A.B. Tabinda, A. Yasar, A. Pugazhendhi, A review on environmental significance carbon foot prints of starch based bio-plastic: A substitute of conventional plastics, Biocatalysis and Agricultural Biotechnology 27 (2020) 101540. ## [10] M.E. Gomes, H.S. Azevedo, A.R. Moreira, V. Ellä, M. Kellomäki, R.L. Reis, Starch–poly(ε-caprolactone) and starch–poly(lactic acid) fibre-mesh scaffolds for bone tissue engineering applications: structure, mechanical properties and degradation behaviour, Journal of Tissue Engineering and Regenerative Medicine 2(5) (2008) 243-252. ## [11] J. Sukyte, E. Adomaviciute, R. Milasius, J. Bendoraitiene, P.P. Danilovas, Formation of poly (vinyl alcohol)/cationic starch blend nanofibres via the electrospinning technique: The influence of different factors, Fibres and Textiles in Eastern Europe (2012). ## [12] H.B. Zhang, M. Zhu, R.Q. You, Modified Biopolymer Scaffolds by Co-Axial Electrospinning, Advanced Materials Research 160-162 (2011) 1062-1066. ## [13] T. Wu, M. Ding, C. Shi, Y. Qiao, P. Wang, R. Qiao, X. Wang, J. Zhong, Resorbable polymer electrospun nanofibers: History, shapes and application for tissue engineering, Chinese Chemical Letters 31(3) (2020) 617-625. ## [14] V. Sgarminato, C. Tonda-Turo, G. Ciardelli, Reviewing recently developed technologies to direct cell activity through the control of pore size: From the macro- to the nanoscale, Journal of Biomedical Materials Research Part B: Applied Biomaterials 108(4) (2020) 1176-1185. ## [15] L. Kong, G.R. Ziegler, Quantitative relationship between electrospinning parameters and starch fiber diameter, Carbohydrate Polymers 92(2) (2013) 1416-1422. ## [16] A.H. Tehrani, A. Zadhoush, S. Karbasi, S.N. Khorasani, Experimental investigation of the governing parameters in the electrospinning of poly(3-hydroxybutyrate) scaffolds: Structural characteristics of the pores, Journal of Applied Polymer Science 118(5) (2010) 2682-2689. ## [17] V. Varkey, E. Tomlal Jose, U.S. Sajeev, Electrospinning technique for the fabrication of poly(styrene-co-methyl methacrylate) nanofibers and the effect of fiber diameter on UV–Visible absorption and thermal properties, Materials Today: Proceedings 33 (2020) 2077-2081. ## [18] L. Wannatong, A. Sirivat, P. Supaphol, Effects of solvents on electrospun polymeric fibers: preliminary study on polystyrene, Polymer International 53(11) (2004) 1851-1859. ## [19] G. Barouti, C.G. Jaffredo, S.M. Guillaume, Advances in drug delivery systems based on synthetic poly(hydroxybutyrate) (co)polymers, Progress in Polymer Science 73 (2017) 1-31. ## [20] E.B. Toloue, S. Karbasi, H. Salehi, M. Rafienia, Potential of an electrospun composite scaffold of poly (3-hydroxybutyrate)-chitosan/alumina nanowires in bone tissue engineering applications, Materials Science and Engineering: C 99 (2019) 1075-1091. ## [21] F. Amini, D. Semnani, S. Karbasi, S.N. Banitaba, A novel bilayer drug-loaded wound dressing of PVDF and PHB/Chitosan nanofibers applicable for post-surgical ulcers, International Journal of Polymeric Materials and Polymeric Biomaterials 68(13) (2019) 772-777. ## [22] T. Yao, H. Chen, M.B. Baker, L. Moroni, Effects of Fiber Alignment and Coculture with Endothelial Cells on Osteogenic Differentiation of Mesenchymal Stromal Cells, Tissue Engineering Part C: Methods 26(1) (2019) 11-22. ## [23] S. Tungprapa, T. Puangparn, M. Weerasombut, I. Jangchud, P. Fakum, S. Semongkhol, C. Meechaisue, P. Supaphol, Electrospun cellulose acetate fibers: effect of solvent system on morphology and fiber diameter, Cellulose 14(6) (2007) 563-575. ## [24] A.P. Kishan, E.M. Cosgriff-Hernandez, Recent advancements in electrospinning design for tissue engineering applications: A review, Journal of Biomedical Materials Research Part A 105(10) (2017) 2892-2905. ## [25] A. Timnak, J.A. Gerstenhaber, K. Dong, Y.-e. Har-el, P.I. Lelkes, Gradient porous fibrous scaffolds: a novel approach to improving cell penetration in electrospun scaffolds, Biomedical Materials 13(6) (2018) 065010. ## [26] A.R. Jabur, M.A. Najim, S.A.A. Al- Rahman, Study the effect of flow rate on some physical properties of different polymeric solutions, Journal of Physics: Conference Series 1003 (2018) 012069. ## [27] S. Mahalingam, B.T. Raimi-Abraham, D.Q.M. Craig, M. Edirisinghe, Solubility–spinnability map and model for the preparation of fibres of polyethylene (terephthalate) using gyration and pressure, Chemical Engineering Journal 280 (2015) 344-353. ## [28] A. Haider, S. Haider, I.-K. Kang, A comprehensive review summarizing the effect of electrospinning parameters and potential applications of nanofibers in biomedical and biotechnology, Arabian Journal of Chemistry 11(8) (2018) 1165-1188. ## [29] N. Bhardwaj, S.C. Kundu, Electrospinning: A fascinating fiber fabrication technique, Biotechnology Advances 28(3) (2010) 325-347. ## [30] H.H. Kim, M.J. Kim, S.J. Ryu, C.S. Ki, Y.H. Park, Effect of fiber diameter on surface morphology, mechanical property, and cell behavior of electrospun poly(ε-caprolactone) mat, Fibers and Polymers 17(7) (2016) 1033-1042. ## [31] C. Viezzer, M.M.d.C. Forte, F.A. Berutti, A.K. Alves, C.P. Bergmann, Effect of electrospun phb and hap-phb composite scaffolds characteristics on mesenchymal stem cell growth viability, MOJ Applied bionics and biomechanics [recurso eletrônico]. Edmond. vol. 1, no. 6 (2017), art. 00035, 8 p. (2017). ## [32] Z. Li, C. Wang, Effects of Working Parameters on Electrospinning, in: Z. Li, C. Wang (Eds.), One-Dimensional nanostructures: Electrospinning Technique and Unique Nanofibers, Springer Berlin Heidelberg, Berlin, Heidelberg, 2013, pp. 15-28. ## [33] H.-W. Chen, M.-F. Lin, Characterization, Biocompatibility, and Optimization of Electrospun SF/PCL/CS Composite Nanofibers, Polymers 12(7) (2020) 1439. ## [34] C. Zandén, Functional Fiber Based Materials for Microsystem Applications, Chalmers Tekniska Hogskola (Sweden)2014. ## [35] H.S. SalehHudin, E.N. Mohamad, W.N.L. Mahadi, A. Muhammad Afifi, Multiple-jet electrospinning methods for nanofiber processing: A review, Materials and Manufacturing Processes 33(5) (2018) 479-498.