AccScience Publishing / IJB / Online First / DOI: 10.36922/IJB025090069
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RESEARCH ARTICLE

Three-dimensional bioprinting of gelatin methacryloyl hydrogel with a tri-layered vascularized architecture for full-thickness skin regeneration

Yichen Luo1 ,  Dan Li2 ,  Cai Lin3 ,  Xue Zhou1 ,  Jien Ma4* ,  Bin Zhang1*
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1 State Key Laboratory of Fluid Power & Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, Hangzhou, Zhejiang, China
2 Department of Electro-Hydraulic Technology and Equipment Research Center, Binhai Industrial Technology Research Institute of Zhejiang University, Tianjin, China
3 Department of Burn, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China
4 Department of Electrical Engineering, College of Electrical Engineering, Zhejiang University, Hangzhou, Zhejiang, China
IJB, 025090069 https://doi.org/10.36922/IJB025090069
Received: 26 February 2025 | Accepted: 15 April 2025 | Published online: 16 April 2025
(This article belongs to the Special Issue Advances in 3D Bioprinting)
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

The skin is the largest organ of the human body and is the primary barrier against external stressors. However, in cases of severe skin damage or pathological conditions, the body’s natural physiological repair mechanisms are often insufficient to support effective skin tissue repair and regeneration. Bioprinting, a form of three-dimensional (3D) printing technology, utilizes various biomaterials and cells to construct complex 3D structures, offering the potential to overcome the limitations of conventional tissue-engineered skin and to develop functional skin substitutes. In this study, we developed a 3D bioprinter with excellent printing performance to fabricate vascularized skin substitutes. Through methacrylic anhydride-mediated modification of gelatin, we synthesized gelatin methacryloyl (GelMA) with varying degrees of substitution. The resulting GelMA hydrogel exhibited excellent mechanical properties, swelling ratio, porosity, and rheological properties. To create a hydrogel-multicellular composite bio-ink, we adjusted the concentration of the GelMA solution and co-cultured human immortalized epidermal cells, human foreskin fibroblasts, and human umbilical vein endothelial cells to optimize biological function. Importantly, by fine-tuning the printing parameters, the 3D extrusion-printed lines successfully fused into a continuous membrane, enhancing interlayer bonding and mechanical integrity. This process enabled the construction of a vascularized skin substitute with distinct reticular and papillary layers. In addition, the 3D-printed vascularized skin was implanted into skin defect models established in BALB/c nude mice and New Zealand rabbits to investigate its regenerative capabilities. These findings hold significant implications for the utilization of 3D-printed vascularized skin for improving skin injury repair, thereby advancing the field of skin tissue engineering.  

Graphical abstract
Keywords
Three-dimensional bioprinting
Gelatin methacryloyl
Skin injury
Vascularized skin
Funding
This work was supported by the National Key Research and Development Program of China (Grant No. 2018YFA0703000) and the Key Science and Technology Program of Zhejiang Province (Grant No. 2023C03170 and 2023C03071).
Conflict of interest
The authors declare no conflict of interest.
References

 

  1. Lee SG, Lee S, Bae HK, et al. Evaluation of the therapeutic efficacy of human skin equivalents manufactured through droplet-based bioprinting/nebulization technology. Mol cell Toxicol. 2024;20(1):129-138. doi: 10.1007/s13273-023-00330-9
  2. Michael S, Sorg H, Peck CT, et al. Tissue engineered skin substitutes created by laser-assisted bioprinting form skin-like structures in the dorsal skin fold chamber in mice. PLoS ONE. 2013;8(3):e57741. doi: 10.1371/journal.pone.0057741
  3. Kafrashian Z, Brück S, Rogin P, et al. Segmented, side‐emitting hydrogel optical fibers for multimaterial extrusion printing. Adv Mater. 2025;37(4): e2309166. doi: 10.1002/adma.202309166
  4. Lin Z, Qiu X, Cai Z, et al. High internal phase emulsions gel ink for direct-ink-writing 3D printing of liquid metal. Nat Commun. 2024;15(1):4806. doi: 10.1038/s41467-024-48906-w
  5. Zhang C, Hua W, Mitchell K, Jin Y. Multiscale embedded printing of engineered human tissue and organ equivalents. Proc Natl Acad Sci USA. 2024;121(9): e2313464121. doi: 10.1073/pnas.2313464121
  6. Sobreiro‐Almeida R, Santos SC, Decarli MC, et al. Leveraging blood components for 3D printing applications through programmable ink engineering approaches. Adv Sci. 2024;11(47):2406569. doi: 10.1002/advs.202406569
  7. Dong T, Hu J, Dong Y, et al. Advanced biomedical and electronic dual-function skin patch created through microfluidic-regulated 3D bioprinting. Bioact Mater. 2024;40: 261-274. doi: 10.1016/j.bioactmat.2024.06.015
  8. Bebiano LB, Presa R, Vieira F, Lourenco BN, Pereira RF. Bioinspired and photo-clickable thiol-ene bioinks for the extrusion bioprinting of mechanically tunable 3D skin models. Biomimetics (Basel). 2024;9(4):228. doi: 10.3390/biomimetics9040228
  9. Gu Z, Fu J, Lin H, He Y. Development of 3D bioprinting: from printing methods to biomedical applications. Asian J Pharm Sci. 2020;15(5): 529-557. doi: 10.1016/j.ajps.2019.11.003
  10. Selvam SP, Ayyappan S, Jamir SI, Sellappan LK, Manoharan S. Recent advancements of hydroxyapatite and polyethylene glycol (PEG) composites for tissue engineering applications–a comprehensive review. Eur Polym J. 2024;215: 113226. doi: 10.1016/j.eurpolymj.2024.113226
  11. Taite LJ, Rowland ML, Ruffino KA, Smith BRE, Lawrence MB, West J L. Bioactive hydrogel substrates: probing leukocyte receptor–ligand interactions in parallel plate flow chamber studies. Ann Biomed Eng. 2006;34: 1705-1711. doi: 10.1007/s10439-006-9173-x
  12. Coudane J, Nottelet B, Mouton J, Garric X, Berghe HVD. Poly (ε-caprolactone)-based graft copolymers: synthesis methods and applications in the biomedical field: a review. Molecules. 2022;27(21):7339. doi: 10.3390/molecules27217339
  13. Milosevic M, Stojanovic DB, Simic V, et al. Preparation and modeling of three‐layered PCL/PLGA/PCL fibrous scaffolds for prolonged drug release. Sci Rep. 2020;10(1):11126. doi: 10.1038/s41598-020-68117-9
  14. Chen Y, Li J, Lu J, Ding M, Chen Y. Synthesis and properties of Poly (vinyl alcohol) hydrogels with high strength and toughness. Polym Test. 2022;108:107516. doi: 10.1016/j.polymertesting.2022.107516
  15. Zhao X, Lang Q, Yildirimer L, et al. Photocrosslinkable gelatin hydrogel for epidermal tissue engineering. Adv Health Mater. 2016;5(1):108-118. doi: 10.1002/adhm.201500005
  16. Xu W, Molino BZ, Cheng F, et al. On low-concentration inks formulated by nanocellulose assisted with gelatin methacrylate (GelMA) for 3D printing toward wound healing application. ACS Appl Mater Interfaces. 2019;11(9):8838-8848. doi: 10.1021/acsami.8b21268
  17. Mazio C, Casale C, Imparato G, et al. Pre-vascularized dermis model for fast and functional anastomosis with host vasculature. Biomaterials. 2019;192:159-170. doi: 10.1016/j.biomaterials.2018.11.018
  18. Xu H, Liu Z, Wei Y, et al. Complexation‐induced resolution enhancement pleiotropic small diameter vascular constructs with superior antibacterial and angiogenesis properties. Adv Healthc Mater. 2023;12(29):2301809. doi: 10.1002/adhm.202301809
  19. Wan H, Cao Y, Lo LW, Zhao J, Sepúlveda N, Wang C. Flexible carbon nanotube synaptic transistor for neurological electronic skin applications. ACS Nano. 2020;14(8):10402-10412. doi: 10.1021/acsnano.0c04259
  20. Vidal SEL, Tamamoto KA, Nguyen H, Abbott RD, Cairns DM, Kaplan DL. 3D biomaterial matrix to support long term, full thickness, immuno-competent human skin equivalents with nervous system components. Biomaterials. 2019;198:194-203. doi: 10.1016/j.biomaterials.2018.04.044
  21. Xiong M, Yang X, Shi Z, et al. Programmable artificial skins accomplish antiscar healing with multiple appendage regeneration. Adv Mater. 2024;36(50): 2407322. doi: 10.1002/adma.202407322
  22. Xia Y, Yan S, Wei H, et al. Multifunctional porous bilayer artificial skin for enhanced wound healing. ACS Appl Mater Interfaces. 2024;16(27):34578-34590. doi: 10.1021/acsami.4c05074
  23. Ma J, Qin C, Wu J, et al. 3D multicellular micropatterning biomaterials for hair regeneration and vascularization. Mater Horiz. 2023;10(9):3773-3784. doi: 10.1039/d3mh00528c
  24. Motter Catarino C, Cigaran Schuck D, Dechiario L, Karande P. Incorporation of hair follicles in 3D bioprinted models of human skin. Sci Adv. 2023;9(41):eadg0297. doi: 10.1126/sciadv.adg0297
  25. Chen H, Ma X, Gao T, et al. Robot-assisted in situ bioprinting of gelatin methacrylate hydrogels with stem cells induces hair follicle-inclusive skin regeneration. Biomed Pharmacother. 2023;158:114140. doi: 10.1016/j.biopha.2022.114140
  26. Zhao W, Chen H, Zhang Y, et al. Adaptive multi‐degree‐of‐freedom in situ bioprinting robot for hair‐follicle‐inclusive skin repair: a preliminary study conducted in mice. Bioeng Transl Med. 2022;7(3):e10303. doi: 10.1002/btm2.10303
  27. Dai LG, Dai NT, Chen TY, Kang LY, Hsu SH. A bioprinted vascularized skin substitute with fibroblasts, keratinocytes, and endothelial progenitor cells for skin wound healing. Bioprinting. 2022;28:e00237. doi: 10.1016/j.bprint.2022.e00237
  28. Ma J, Qin C, Wu J, et al. 3D printing of strontium silicate microcylinder‐containing multicellular biomaterial inks for vascularized skin regeneration. Adv Healthc Mater. 2021;10(16):2100523. doi: 10.1002/adhm.202100523
  29. Barros NR, Kim HJ, Gouidie MJ, et al. Biofabrication of endothelial cell, dermal fibroblast, and multilayered keratinocyte layers for skin tissue engineering. Biofabrication. 2021;13(3):035030. doi: 10.1088/1758-5090/aba503
  30. Xiang P, Yan L, Ge L, He X, Du N, Liu X. Development of a radial-flux machine with multi-shaped magnet rotor and non-ferromagnetic yoke for low torque ripple and rotor mass. IEEE Trans Ind Appl. 61;2025:2897-2910. doi: 10.1109/TIA.2025.3532558
  31. Xiang P, Yan L, Guo Y, He X, Gerada C, Chen IM. A concentrated-flux-type pm machine with irregular magnets and iron poles. IEEE/ASME Trans Mech. 2023;29(1): 691-702. doi: 10.1109/TMECH.2023.3293505
  32. Guo J, Gu H, Yin S, et al. Hepatocyte-derived Igκ promotes HCC progression by stabilizing electron transfer flavoprotein subunit α to facilitate fatty acid β-oxidation. J Exp Clin Cancer Res. 2024;43(1):280. doi: 10.1186/s13046-024-03203-8
  33. Liu C, Qin W, Wang Y, et al. 3D printed gelatin/sodium alginate hydrogel scaffolds doped with nano-attapulgite for bone tissue repair. Int J Nanomed. 2021;16:8417-8432. doi: 10.2147/IJN.S339500
  34. Pierce MC, Strasswimmer J, Hyle Park B, Cense B, De Boer JF. Birefringence measurements in human skin using polarization-sensitive optical coherence tomography. J Biomed Opt. 2004;9(2):287-291. doi: 10.1117/1.1645797
  35. Wang Y, Liu Y, Chen S, et al. Enhancing bone regeneration through 3D printed biphasic calcium phosphate scaffolds featuring graded pore sizes. Bioact Mater. 2024;46:21–36. doi: 10.1016/j.bioactmat.2024.11.024
  36. Kim BS, Yang WK, Jeong SK, et al. 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. doi: 10.1016/j.biomaterials.2018.03.040
  37. Won-woo C, Minjun A, Byoung SK, Dong-Woo C. Blood‐lymphatic integrated system with heterogeneous melanoma spheroids via in‐bath three‐dimensional bioprinting for modelling of combinational targeted therapy. Adv Sci. 2022;9(29):2202093. doi: 10.1002/advs.202202093
  38. Song J, Liu T, Liao Z, et al. Digital light processing bioprinting neural systems with porous hydrogel in structure and function for disease models. Cell Rep Phys Sci. 2024;5(12):102311. doi: 10.1016/j.xcrp.2024.102311
  39. Zandi N, Daniele M, Brown A. Advances in fibrin-based bioprinting for skin tissue regeneration: exploring design, and innovative approaches. Biomed Mater Devices. 2025;3:330-348. doi: 10.1007/s44174-024-00198-w
  40. Fauzi MB, Rashidbenam Z, Bin Saim A, Binti Hj Idrus R. Preliminary study of in vitro three-dimensional skin model using an ovine collagen type i sponge seeded with co-culture skin cells: submerged versus air-liquid interface conditions. Polymers. 2020;12(12): 2784. doi: 10.3390/polym12122784
  41. Monsuur HN, Boink MA, Weijers EM, et al. Methods to study differences in cell mobility during skin wound healing in vitro. J Biomech. 2016;49(8):1381-1387. doi: 10.1016/j.jbiomech.2016.01.040
  42. Petry L, Kippenberger S, Meissner M, et al. Directing adipose‐derived stem cells into keratinocyte‐like cells: impact of medium composition and culture condition. J Eur Acad Dermatol Venereol. 2018;32(11):2010-2019. doi: 10.1111/jdv.15010
  43. Colin E, Plyer A, Golzio M, Meyer N, Faver G, Orlik X. Imaging of the skin microvascularization using spatially depolarized dynamic speckle. J Biomed Opt. 2022;27(4):046003. doi: 10.1117/1.Jbo.27.4.046003
  44. Hu X, Wang L, Deng J, et al. Dietary nitrate accelerates the healing of infected skin wounds in mice by increasing microvascular density. Biochem Biophys Res Commun. 2023;686:149176. doi: 10.1016/j.bbrc.2023.149176
  45. Fu T, Sullivan DP, Gonzalez AM, et al. Mechanotransduction via endothelial adhesion molecule CD31 initiates transmigration and reveals a role for VEGFR2 in diapedesis. Immunity. 2023;56(10):2311-2324.e6. doi: 10.1016/j.immuni.2023.08.001
  46. Park H, Collignon AM, Lepry WC, et al. Acellular dense collagen-S53P4 bioactive glass hybrid gel scaffolds form more bone than stem cell delivered constructs. Mater Sci Eng C Mater Biol Appl. 2021;120:111743. doi: 10.1016/j.msec.2020.111743
  47. Trappmann B, Gautrot JE, Connelly JT, et al. Extracellular-matrix tethering regulates stem-cell fate. Nat Mater. 2012;11(7): 642-649. doi: 10.1038/nmat3339
  48. Singh A, Dalal N, Tayalia P. An interplay of matrix stiffness, dimensionality and adhesivity on cellular behavior. Biomed Mater. 2023;18(2): 025010. doi: 10.1088/1748-605X/acb7c0
  49. Trujillo S, Gonzalez-Garcia C, Rico P, et al. Engineered 3D hydrogels with full-length fibronectin that sequester and present growth factors. Biomaterials. 2020; 252:120104. doi: 10.1016/j.biomaterials.2020.120104
  50. Ito M, Hiramatsu H, Kobayashi K, et al. NOD/SCID/γ c null mouse: an excellent recipient mouse model for engraftment of human cells. Blood. 2002;100(9):3175-3182. doi: 10.1182/blood-2001-12-0207
  51. Albanna M, Binder KW, Murphy SV, et al. In situ bioprinting of autologous skin cells accelerates wound healing of extensive excisional full-thickness wounds. Sci Rep. 2019;9(1):1856. doi: 10.1038/s41598-018-38366-w
  52. Wei Q, Su J, Meng S, et al. MiR‐17‐5p‐engineered sEVs encapsulated in GelMA hydrogel facilitated diabetic wound healing by targeting PTEN and p21. Adv Sci (Weinh). 2024;11(13):2307761. doi: 10.1002/advs.202307761
  53. Zhang G, Zhang Z, Cao G, et al. Engineered dermis loaded with confining forces promotes full-thickness wound healing by enhancing vascularisation and epithelialisation. Acta Biomater. 2023; 170: 464-478. doi: 10.1016/j.actbio.2023.08.049
  54. Jin T, Fu Z, Zhou L, et al. GelMA loaded with platelet lysate promotes skin regeneration and angiogenesis in pressure ulcers by activating STAT3. Sci Rep. 2024; 14(1): 18345. doi: 10.1038/s41598-024-67304-2
  55. Chen L, Ye JL, Gao C, Deng F, Liu W, Zhang Q. Design and fabrication of gelatin-based hydrogel loaded with modified amniotic extracellular matrix for enhanced wound healing. Heliyon. 2023;9(10):e20521. doi: 10.1016/j.heliyon.2023.e20521
  56. Hao X, Luo J, Huang Y, et al. Injectable antiswelling and high-strength bioactive hydrogels with a wet adhesion and rapid gelling process to promote sutureless wound closure and scar-free repair of infectious wounds. ACS Nano. 2023;17(21): 22015-22034. doi: 10.1021/acsnano.3c08625

 

 

 

 

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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
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