Bioengineering of fibroblast-conditioned polycaprolactone/gelatin electrospun scaffold for skin tissue engineering.

Publisher: Wiley-Blackwell Country of Publication: United States NLM ID: 7802778 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1525-1594 (Electronic) Linking ISSN: 0160564X NLM ISO Abbreviation: Artif Organs Subsets: MEDLINE

Publication: Cambridge, MA : Wiley-Blackwell
Original Publication: Cleveland, International Society for Artificial Organs.

Gelatin*/chemistry ; Tissue Engineering*/methods; Animals ; Fibroblasts ; Mice ; Polyesters/chemistry ; Tissue Scaffolds/chemistry

Background: Synthetic tissue engineering scaffolds has poor biocompatiblity with very low angiogenic properties. Conditioning the scaffolds with functional groups, coating with biological components, especially extracellular matrix (ECM), is an excellent strategy for improving their biomechanical and biological properties.
Methods: In the current study, a composite of polycaprolactone and gelatin (PCL/Gel) was electrospun in the ratio of 70/30 and surface modified with 1% gelatin-coating (G-PCL/Gel) or plasma treatment (P-PCL/Gel). The surface modification was determined by SEM and ATR-FTIR spectroscopy, respectively. The scaffolds were cultured with fibroblast 3T3, then decellularized during freeze-thawing process to fabricate a fibroblast ECM-conditioned PCL/Gel scaffold (FC-PCL/Gel). The swelling and degaradtion as well as in vitro and in vivo biocompatibility and angiogenic properties of the scaffolds were evaluated.
Results: The structure of the surface-modified G-PCL/Gel and P-PCL/Gel were unique and not changed compared with the PCL/Gel scaffolds. ATR-FTIR analysis admitted the formation of oxygen-containing groups, hydroxyl and carboxyl, on the surface of the P-PCL/Gel scaffold. The SEM micrographs and DAPI staining confirmed the cell attachment and the ECM deposition on the platform and successful removal of the cells after decellularization. P-PCL/Gel showed better cell attachment, ECM secretion and deposition after decellularization compared with G-PCL/Gel. The FC-PCL/Gel was considered as an optimized scaffold for further assays in this study. The FC-PCL/Gel showed increased hydrophilic behavior and cytobiocompatibility compared with P-PCL/Gel. The ECM on the FC-PCL/Gel scaffold showed a gradual degradation during 30 days of degradation time, as a small amount of ECM remained over the FC-PCL/Gel scaffold at day 30. The FC-PCL/Gel showed significant biocompatibility and improved angiogenic property compared with P-PCL/Gel when subcutaneously implanted in a mouse animal model for 7 and 28 days.
Conclusions: Our findings suggest FC-PCL/Gel as an excellent biomimetic construct with high angiogenic properties. This bioengineered construct can serve as a possible application in our future pre-clinical and clinical studies for skin regeneration.
(© 2022 International Center for Artificial Organs and Transplantation and Wiley Periodicals LLC.)

Seal B, Otero T, Panitch A. Polymeric biomaterials for tissue and organ regeneration. Mater Sci Eng R Rep. 2001;34(4-5):147-230.
Marler JJ, Upton J, Langer R, Vacanti JP. Transplantation of cells in matrices for tissue regeneration. Adv Drug Deliv Rev. 1998;33(1-2):165-82.
Gholipourmalekabadi M, Samadikuchaksaraei A, Seifalian AM, Urbanska AM, Ghanbarian H, Hardy JG, et al. Silk fibroin/amniotic membrane 3D bi-layered artificial skin. Biomed Mater. 2018;13(3):035003.
Gholipourmalekabadi M, Seifalian AM, Urbanska AM, Omrani MD, Hardy JG, Madjd Z, et al. 3D protein-based bilayer artificial skin for the guided scarless healing of third-degree burn wounds in vivo. Biomacromol. 2018;19(7):2409-22.
Mogoşanu GD, Grumezescu AM. Natural and synthetic polymers for wounds and burns dressing. Int J Pharm. 2014;463(2):127-36.
Assunção M, Dehghan-Baniani D, Yiu CHK, Später T, Beyer S, Blocki A. Cell-Derived extracellular matrix for tissue engineering and regenerative medicine. Front Bioeng Biotechnol. 2020;8:1378.
Wang TW, Wu HC, Huang YC, Sun JS, Lin FH. Biomimetic bilayered gelatin-chondroitin 6 sulfate-hyaluronic acid biopolymer as a scaffold for skin equivalent tissue engineering. Artif Organs. 2006;30(3):141-9.
Dhivya S, Padma VV, Santhini E. Wound dressings-a review. BioMedicine. 2015;5(4):24-28.
Bhardwaj N, Kundu SC. Electrospinning: a fascinating fiber fabrication technique. Biotechnol Adv. 2010;28(3):325-47.
Keshvardoostchokami M, Majidi SS, Huo P, Ramachandran R, Chen M, Liu B. Electrospun nanofibers of natural and synthetic polymers as artificial extracellular matrix for tissue engineering. Nanomaterials. 2021;11(1):21.
Nejaddehbashi F, Hashemitabar M, Bayati V, Abbaspour M, Moghimipour E, Orazizadeh M. Application of polycaprolactone, chitosan, and collagen composite as a nanofibrous mat loaded with silver sulfadiazine and growth factors for wound dressing. Artif Organs. 2019;43(4):413-23.
Norouzi M, Boroujeni SM, Omidvarkordshouli N, Soleimani M. Advances in skin regeneration: application of electrospun scaffolds. Adv Healthc Mater. 2015;4(8):1114-33.
Yoshimoto H, Shin Y, Terai H, Vacanti J. A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering. Biomaterials. 2003;24(12):2077-82.
Cipitria A, Skelton A, Dargaville T, Dalton P, Hutmacher D. Design, fabrication and characterization of PCL electrospun scaffolds-a review. J Mater Chem. 2011;21(26):9419-53.
Molina MIE, Malollari KG, Komvopoulos K. Design challenges in polymeric scaffolds for tissue engineering. Front Bioeng Biotechnol. 2021;9:617141.
Capellato P, Silva G, Popat K, Simon-Walker R, Alves Claro AP, Zavaglia C. Cell investigation into the biocompatibility of adult human dermal fibroblasts with PCL nanofibers/TiO2 nanotubes on the surface of Ti-30Ta alloy for biomedical applications. Artif Organs. 2020;44(8):877-82.
Siddiqui N, Kishori B, Rao S, Anjum M, Hemanth V, Das S, et al. Electropsun polycaprolactone fibres in bone tissue engineering: a review. Mol Biotechnol. 2021;63(5):1-26.
Siddiqui N, Asawa S, Birru B, Baadhe R, Rao S. PCL-based composite scaffold matrices for tissue engineering applications. Mol Biotechnol. 2018;60(7):506-32.
Mistura DV, Messias AD, Duek EA, Duarte MA. Development, characterization, and cellular adhesion of poly (L-lactic acid)/Poly (caprolactone triol) membranes for potential application in bone tissue regeneration. Artif Organs. 2013;37(11):978-84.
Shekaran A, Lam A, Sim E, Jialing L, Jian LI, Wen JTP, et al. Biodegradable ECM-coated PCL microcarriers support scalable human early MSC expansion and in vivo bone formation. Cytotherapy. 2016;18(10):1332-44.
Feng B, Ji T, Wang X, Fu W, Ye L, Zhang H, et al. Engineering cartilage tissue based on cartilage-derived extracellular matrix cECM/PCL hybrid nanofibrous scaffold. Mater Des. 2020;193:108773.
Venugopal JR, Zhang Y, Ramakrishna S. In vitro culture of human dermal fibroblasts on electrospun polycaprolactone collagen nanofibrous membrane. Artif Organs. 2006;30(6):440-6.
Turner NJ, Badylak SF. The use of biologic scaffolds in the treatment of chronic nonhealing wounds. Adv Wound Care. 2015;4(8):490-500.
Blackstone BN, Gallentine SC, Powell HM. Collagen-based electrospun materials for tissue engineering: a systematic review. Bioengineering. 2021;8(3):39.
Mirzadeh H, Bagheri-Khoulenjani S. Recent achievements in bone and skin tissue engineering in Iran. Artif Organs. 2018;42(6):585-588.
Samadikuchaksaraei A, Gholipourmalekabadi M, Erfani Ezadyar E, Azami M, Mozafari M, Johari B, et al. Fabrication and in vivo evaluation of an osteoblast-conditioned nano-hydroxyapatite/gelatin composite scaffold for bone tissue regeneration. J Biomed Mater Res Part A. 2016;104(8):2001-10.
Franco-Barraza J, Beacham DA, Amatangelo MD, Cukierman E. Preparation of extracellular matrices produced by cultured and primary fibroblasts. Curr Protoc Cell Biol. 2016;71(1):10.9.1-10.9.34.
Safaeijavan R, Soleimani M, Divsalar A, Eidi A, Ardeshirylajimi A. Biological behavior study of gelatin coated PCL nanofiberous electrospun scaffolds using fibroblasts. Arch Adv Biosci. 2014;5(1):67-73.
Meghdadi M, Atyabi S-M, Pezeshki-Modaress M, Irani S, Noormohammadi Z, Zandi M. Cold atmospheric plasma as a promising approach for gelatin immobilization on poly (ε-caprolactone) electrospun scaffolds. Progr Biomater. 2019;8(2):65-75.
Gholipourmalekabadi M, Mozafari M, Salehi M, Seifalian A, Bandehpour M, Ghanbarian H, et al. Development of a cost-effective and simple protocol for decellularization and preservation of human amniotic membrane as a soft tissue replacement and delivery system for bone marrow stromal cells. Adv Healthc Mater. 2015;4(6):918-26.
Mohammed Z, Jeelani S, Rangari V. Effect of low-temperature plasma treatment on surface modification of polycaprolactone pellets and thermal properties of extruded filaments. JOM. 2020;72(4):1523-32.
Darain F, Chan WY, Chian KS. Performance of surface-modified polycaprolactone on growth factor binding, release, and proliferation of smooth muscle cells. Soft Mater. 2010;9(1):64-78.
Afsharian YP, Rahimnejad M. Bioactive electrospun scaffolds for wound healing applications: a comprehensive review. Polym Testing. 2020;93:106952.
Behtouei E, Zandi M, Askari F, Daemi H, Zamanlui S, Arabsorkhi-Mishabi A, et al. Bead-free and tough electrospun PCL/gelatin/PGS ternary nanofibrous scaffolds for tissue engineering application. J Appl Polym Sci. 2021;139(2):51471.
Chong E, Phan T, Lim I, Zhang Y, Bay B, Ramakrishna S, et al. Evaluation of electrospun PCL/gelatin nanofibrous scaffold for wound healing and layered dermal reconstitution. Acta Biomater. 2007;3(3):321-30.
Zhang Y, Ouyang H, Lim CT, Ramakrishna S, Huang ZM. Electrospinning of gelatin fibers and gelatin/PCL composite fibrous scaffolds. J Biomed Mater Res B Appl Biomater. 2005;72(1):156-65.
Li WJ, Laurencin CT, Caterson EJ, Tuan RS, Ko FK. Electrospun nanofibrous structure: a novel scaffold for tissue engineering. J Biomed Mater Res. 2002;60(4):613-21.
Samadian H, Farzamfar S, Vaez A, Ehterami A, Bit A, Alam M, et al. A tailored polylactic acid/polycaprolactone biodegradable and bioactive 3D porous scaffold containing gelatin nanofibers and Taurine for bone regeneration. Sci Rep. 2020;10(1):1-12.
Bhattacharjee P, Kundu B, Naskar D, Kim H-W, Bhattacharya D, Maiti TK, et al. Potential of inherent RGD containing silk fibroin-poly (Є-caprolactone) nanofibrous matrix for bone tissue engineering. Cell Tissue Res. 2016;363(2):525-40.
Harris GM, Raitman I, Schwarzbauer JE. Cell-derived decellularized extracellular matrices. Methods Cell Biol. 2018;143:97-114.
Beachley V, Wen X. Polymer nanofibrous structures: fabrication, biofunctionalization, and cell interactions. Prog Polym Sci. 2010;35(7):868-92.
Sharif S, Ai J, Azami M, Verdi J, Atlasi MA, Shirian S, et al. Collagen-coated nano-electrospun PCL seeded with human endometrial stem cells for skin tissue engineering applications. J Biomed Mater Res B Appl Biomater. 2018;106(4):1578-86.
Li M, Zhang T, Jiang J, Mao Y, Zhang A, Zhao J. ECM coating modification generated by optimized decellularization process improves functional behavior of BMSCs. Mater Sci Eng C. 2019;105:110039.
Kim J, Ma T. Autocrine fibroblast growth factor 2-mediated interactions between human mesenchymal stem cells and the extracellular matrix under varying oxygen tension. J Cell Biochem. 2013;114(3):716-27.
Xing Q, Yates K, Tahtinen M, Shearier E, Qian Z, Zhao F. Decellularization of fibroblast cell sheets for natural extracellular matrix scaffold preparation. Tissue Eng Part C Methods. 2015;21(1):77-87.
Noh YK, Du P, Kim IG, Ko J, Kim SW, Park K. Polymer mesh scaffold combined with cell-derived ECM for osteogenesis of human mesenchymal stem cells. Biomater Res. 2016;20(1):1-7.
Theocharis AD, Skandalis SS, Gialeli C, Karamanos NK. Extracellular matrix structure. Adv Drug Deliv Rev. 2016;97:4-27.
Li F, Li W, Johnson S, Ingram D, Yoder M, Badylak S. Low-molecular-weight peptides derived from extracellular matrix as chemoattractants for primary endothelial cells. Endothelium. 2004;11(3-4):199-206.
Vorotnikova E, McIntosh D, Dewilde A, Zhang J, Reing JE, Zhang LI, et al. Extracellular matrix-derived products modulate endothelial and progenitor cell migration and proliferation in vitro and stimulate regenerative healing in vivo. Matrix Biol. 2010;29(8):690-700.
Hodde J, Record R, Liang H, Badylak S. Vascular endothelial growth factor in porcine-derived extracellular matrix. Endothelium. 2001;8(1):11-24.
Daley WP, Peters SB, Larsen M. Extracellular matrix dynamics in development and regenerative medicine. J Cell Sci. 2008;121(3):255-64.
Guilak F, Cohen DM, Estes BT, Gimble JM, Liedtke W, Chen CS. Control of stem cell fate by physical interactions with the extracellular matrix. Cell Stem Cell. 2009;5(1):17-26.
Westhauser F, Senger AS, Obert D, Ciraldo FE, Schuhladen K, Schmidmaier G, et al. Gelatin coating increases in vivo bone formation capacity of three-dimensional 45S5 bioactive glass-based crystalline scaffolds. J Tissue Eng Regener Med. 2019;13(2):179-90.
Merk M, Chirikian O, Adlhart C. 3D PCL/gelatin/genipin nanofiber sponge as scaffold for regenerative medicine. Materials. 2021;14(8):2006.
Alev C, Ii M, Asahara T. Endothelial progenitor cells: a novel tool for the therapy of ischemic diseases. Antioxid Redox Signal. 2011;15(4):949-65.
Otero-Viñas M, Falanga V. Mesenchymal stem cells in chronic wounds: the spectrum from basic to advanced therapy. Advances in Wound Care. 2016;5(4):149-63.
Terrovitis JV, Smith RR, Marbán E. Assessment and optimization of cell engraftment after transplantation into the heart. Circ Res. 2010;106(3):479-94.
Christman KL, Lee RJ. Biomaterials for the treatment of myocardial infarction. J Am Coll Cardiol. 2006;48(5):907-13.
Riis S, Hansen AC, Johansen L, Lund K, Pedersen C, Pitsa A, et al. Fabrication and characterization of extracellular matrix scaffolds obtained from adipose-derived stem cells. Methods. 2020;171:68-76.
Sun M, He Y, Zhou T, Zhang P, Gao J, Lu F. Adipose extracellular matrix/stromal vascular fraction gel secretes angiogenic factors and enhances skin wound healing in a murine model. Biomed Res Int. 2017;2017:1-11.
Sapru S, Das S, Mandal M, Ghosh AK, Kundu SC. Sericin-chitosan-glycosaminoglycans hydrogels incorporated with growth factors for in vitro and in vivo skin repair. Carbohyd Polym. 2021;258:117717.

33225 Iran University of Medical Sciences

Keywords: 3T3 fibroblast; cell adhesion; cell attachment; electrospun; extracellular matrix; plasma treatment; polycaprolactone

0 (Polyesters)
24980-41-4 (polycaprolactone)
9000-70-8 (Gelatin)

Date Created: 20220110 Date Completed: 20220520 Latest Revision: 20220520

20231215

10.1111/aor.14169

35006608