Comparison of the Biological Behaviour of Human Dermal Fibroblasts seeded on 3D Printed Polylactic acid, Polycaprolactone and Polyethylene Terephthalate Scaffolds in vitro

Year 2021, , 7 - 13, 07.07.2021

Ufkay Karabay , R. Bugra Husemoglu , Mehtap Yuksel Egrılmez , Selma Aydemir , Başak Baykara , Serhat Cagiral , Hasan Havıtçıoğlu

https://doi.org/10.51934/jomit.957164

Cited By: 1

Abstract

Regenerative medicine is a scientific field that improves and repairs diseased and injured tissues. Three-dimensional (3D) printing is an innovative technology that provides a new application field for regenerative medicine. 3D printed scaffolds by programming pore sizes and shapes serve as a temporary basis for cells until the natural extracellular matrix (ECM) is reconstructed. Dermal fibroblasts are mesenchymal cells located in the dermal skin layer that produce and organize ECM components. They play an essential role in skin wound healing and fibrosis. The aim of this study is to analyze the viability, adhesion, distribution, and collagen IV expression of human dermal fibroblasts (HDFs) seeded on 3D printed polylactic acid (PLA), polyethylene terephthalate (PET), and poly-ε-caprolactone (PCL) scaffolds in vitro. HDFs were seeded on scaffolds or tissue culture plastic plates as control and were cultured for 1 and 3 days. 3D PLA, PCL, and PET scaffolds were prepared using a custom made fused deposition modeling printer. The cell viability was measured by WST-1 assay on days 1 and 3. The cell adhesion was evaluated by scanning electron microscopy (SEM). The distribution was analyzed by hematoxylin and eosin (H&E) staining. Collagen IV expression was analyzed by immunohistochemical (IHC) staining. On day 1, the viability of HDFs on the 3D PLA scaffolds was significantly higher than PCL scaffolds. On day 3, the viability of HDFs on 3D PLA and PET scaffolds was significantly higher than PCL scaffolds. SEM images showed that HDFs on 3D PLA scaffolds attached the surfaces, filled the interfiber gaps and maintained their tissue specific morphology on day 3 compared to PCL and PET scaffolds. Histological images stained with H&E demonstrated that the distribution of HDFs on 3D PLA scaffolds was uniform on days 1 and 3. Collagen IV staining was more intense in HDFs on 3D PLA scaffolds on days 1 and 3. This study shows that 3D PLA scaffolds create a appropriate environment for cell viability, adhesion, distribution and may provide a high advantage in skin tissue regeneration.

Keywords

Human dermal fibroblasts, 3D printed scaffolds, Skin tissue regeneration

References

• Watt FM, Fujiwara H. Cell-Extracellular matrix interactions in normal and diseased skin. Cold Spring Harb Perspect Biol 2011;3(4):1-14.
• Driskell RR, Watt FM. Understanding fibroblast heterogeneity in the skin. Trends Cell Biol 2015;25(2):92-9.
• Witherel CE, Abebayehu D, Barker TH, Kara LS. Macrophage and fibroblast interactions in biomaterial-mediated fibrosis. Adv Healthc Mater 2019;8(4):1-16.
• Dorati R, Colonna C, Tomasi C, Genta I, Bruni G, Conti B. Design of 3D scaffolds for tissue engineering testing a tough polylactide-based graft copolymer. Mater Sci Eng C 2014;34(1):130-9.
• Aladdad AM, Amer MH, Sidney L, et al. A thermoresponsive three-dimensional fibrous cell culture platform for enzyme-free expansion of mammalian cells. Acta Biomaterialia 2019;95:427-38.
• Zhang G, Varkey M, Wang Z, Xie B, Hou R, Atala A. ECM concentration and cell-mediated traction forces play a role in vascular network assembly in 3D bioprinted tissue. Biotechnolog Bioeng 2020;117(4):1148-58.
• Tyler B, Gullotti D, Mangraviti A, Utsuki T, Brem H. Polylactic acid (pla) controlled delivery carriers for biomedical applications. Adv Drug Deliv Rev 2016;107: 163-75.
• Bridge JC, Aylott JW, Brightling CE, et al. Adapting the electrospinning process to provide three unique environments for a tri-layered in vitro model of the airway wall. J Vis Exp 2015;101:e52986.
• Pinzon-Garcia AD, Cassini-Vieira P, Ribeiro CC, et al. Efficient cutaneous wound healing using bixin-loaded PCL nanofibers in diabetic mice. J Biomed Mater Res Part B Appl. Biomater 2017;105:1938-49.
• Farrugia BL, Brown TD, Upton Z, Hutmacher DW, Dalton PD, Dargaville TR. Dermal fibroblast infiltration of poly(ε-caprolactone) scaffolds fabricated by melt electrospinning in a direct writing mode. Biofabrication. 2013;5(2): 025001.
• Huerta RR, Silva E, Ekaette I, El-Bialy T, Saldaña MDA. High-Intensity ultrasound-assisted formation of cellulose nanofiber scaffold with low and high lignin content and their cytocompatibility with gingival fibroblast cells. Ultrason Sonochem 2020;64:104759.
• Chen WC, Wei YH, Chu IM, Yao CL. Effect of chondroitin sulphate C on the in vitro and in vivo chondrogenesis of mesenchymal stem cells in crosslinked type II collagen scaffolds. J Tissue Eng Regen Med 2013;7(8):665-72.
• Griffin MF, Naderi N, Kalaskar DM, Seifalian AM, Butler PE. Argon plasma surface modification promotes the therapeutic angiogenesis and tissue formation of tissue-engineered scaffolds in vivo by adipose-derived stem cells. Stem Cell Res Ther 2019;10(1):1-14.
• Nagarajan S, Reddy BSR. Bio-absorbable polymers in implantation - an overview. J Sci Ind Res 2009;8:993-1009.
• Theryo G, Jing F, Pitet LM, Hillmyer MA. Tough polylactide graft copolymers. Macromolecules 2010; 43:7394-7.
• Karabay U, Husemoglu RB, Egrilmez MY, Havitcioglu H. 3D printed polylactic acid scaffold for dermal tissue engineering application : the fibroblast proliferation in vitro. Journal of Medical Innovation and Technology 2019;1(2):51-6.
• Wang J, Huang N, Yang P, Leng YX, Sun H, Liu ZY, Chu PK. The effects of amorphous carbon films deposited on polyethylene terephthalate on bacterial adhesion. Biomaterials 2004;25:3163-70.
• Zhang X (Edited by). Science and Principles of Biodegradable and Bioresorbable Medical Polymers. In: Zhang X, Peng X, Zhang S. Synthetic biodegradable medical polymers: polymer blends. Woodhead Publishing, 2017;217-54.
• Petretta M, Gambardella A, Boi M, et al. Composite scaffolds for bone tissue regeneration based on PCL and Mg-containing bioactive glasses. Biology (Basel) 2021;10(5):398.
• Patrício T, Glória A, Bártolo P. Mechanical and biological behaviour of PCL and PCL/PLA scaffolds for tissue engineering applications. Chem Eng Trans 2013;32:1645-50.
• Hewitt E, Mros S, McConnell M, Cabral JD and Ali A. Melt-Electrowriting with novel milk protein/PCL biomaterials for skin regeneration. Biomedical Materials 2019;4(5): 055013.
• ÖG Geyik, Nalbant B, Hüsemoğlu RB, Yüce Z, Ünek T, Havıtçıoğlu H. Investigation of Surface Adhesion of MCF-7 Cells in 3D Printed PET and PLA Tissue Scaffold Models. Journal of Medical Innovation and Technology 2019;1(2):45-50.
• Hasegawa H, Naito I, Nakano K, Momota R, Nishida K, Taguchi T, et al. The distributions of type IV collagen alpha chains in basement membranes of human epidermis and skin appendages. Arch Histol Cytol 2007;70(4):255-65.
• Olsen DR, Peltonen J, Jaakkola S, Chu ML, Uitto J. Collagen gene expression by cultured human skin fibroblasts. Abundant steady-state levels of type VI procollagen messenger RNAs. J Clin Invest 1989;83(3): 791-5.
• Betz P, Nerlich A, Wilske J, Tübel J, Wiest I, Penning R, et al. The time-dependent rearrangement of the epithelial basement membrane in human skin wounds-immunohistochemical localization of Collagen IV and VII. Int J Legal Med 1992;105:93-7.