Gelatin-based micro-hydrogel carrying genetically engineered human endothelial cells for neovascularization

Young Hwan Choi, Su Hwan Kim, In Seon Kim, Kyung Min Kim, Seong Keun Kwon, Nathaniel S. Hwang

Research output: Contribution to journalArticleResearchpeer-review

1 Citation (Scopus)

Abstract

Cell delivery systems based on micro-hydrogels may facilitate the long-term survival of cells upon transplantation. Micro-hydrogels may effectively support cell proliferation, attachment, and migration in ischemic environments. In this study, we report the fabrication of a gelatin methacrylate (GelMA)-based micro-hydrogel for efficient in vivo delivery of genetically engineered endothelial cells. Micro-hydrogels were initially processed via electrospraying of GelMA and alginate (ALG) mixtures (at different ratios) on to calcium chloride (CaCl2) solution. Electrospraying of the GelMA/ALG mixture resulted in the formation of a micro-hydrogel, owing to ALG crosslinking. Secondary crosslinking of GelMA with UV light and ALG hydrogel chelation using sodium citrate solution resulted in GelMA-based micro-hydrogel formation. We observed the angiogenic response of human umbilical vein endothelial cells (HUVECs) in GelMA concentration-dependent manner. The seeding of HUVECs engineered to express human vascular endothelial growth factor on to the GelMA micro-hydrogel and the subsequent transplantation of the micro-hydrogel into a hindlimb ischemia model effectively attenuated the ischemia condition. This facile and simple micro-hydrogel fabrication strategy may serve as a robust method to fabricate efficient cell carriers for various ischemic diseases. Statement of Significance: For the therapeutic angiogenesis, it is important to provide the therapeutic cells with a carrier that could stabilize therapeutic cells and facilitate long-term survival of cells. Furthermore, it is also important to administer as many therapeutic cells as possible in a fixed volume. From these cues, we fabricated ECM-based micro-hydrogel produced by the high through-put system. And we intended to facilitate activation of therapeutic cells by coating the therapeutic cells onto the micro-hydrogel. In this manuscript, we fabricated methacrylate gelatin (GelMA) based micro-hydrogels using the electro-spraying method and coated HUVECs engineered to express hVEGF onto the micro-hydrogels. Then, we identified that GelMA concentration-dependent angiogenic response of HUVECs. Furthermore, we demonstrated that the VEGF secreting HUVEC-GelMA micro-hydrogels induced the restoration of blood flow and neovascularization in a hind-limb ischemia mouse model. These findings demonstrate that the high-throughput fabrication of ECM micro-hydrogels could be a novel platform to apply in neovascularization and tissue engineering.

Original languageEnglish
Pages (from-to)285-296
Number of pages12
JournalActa Biomaterialia
Volume95
DOIs
StatePublished - 1 Sep 2019

Fingerprint

Methacrylates
Hydrogel
Endothelial cells
Gelatin
Hydrogels
Endothelial Cells
Human Umbilical Vein Endothelial Cells
Alginate
Military electronic countermeasures
Ischemia
Fabrication
Crosslinking
Therapeutics
Cells
Calcium Chloride
Cell Transplantation
Cell proliferation
Tissue Engineering
Ultraviolet Rays
Spraying

Keywords

  • Electrospray
  • Gelatin methacrylate (GelMA)
  • Hindlimb ischemia
  • Micro-hydrogel
  • hVEGF-secreting HUVEC

Cite this

Choi, Young Hwan ; Kim, Su Hwan ; Kim, In Seon ; Kim, Kyung Min ; Kwon, Seong Keun ; Hwang, Nathaniel S. / Gelatin-based micro-hydrogel carrying genetically engineered human endothelial cells for neovascularization. In: Acta Biomaterialia. 2019 ; Vol. 95. pp. 285-296.
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Gelatin-based micro-hydrogel carrying genetically engineered human endothelial cells for neovascularization. / Choi, Young Hwan; Kim, Su Hwan; Kim, In Seon; Kim, Kyung Min; Kwon, Seong Keun; Hwang, Nathaniel S.

In: Acta Biomaterialia, Vol. 95, 01.09.2019, p. 285-296.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Gelatin-based micro-hydrogel carrying genetically engineered human endothelial cells for neovascularization

AU - Choi, Young Hwan

AU - Kim, Su Hwan

AU - Kim, In Seon

AU - Kim, Kyung Min

AU - Kwon, Seong Keun

AU - Hwang, Nathaniel S.

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N2 - Cell delivery systems based on micro-hydrogels may facilitate the long-term survival of cells upon transplantation. Micro-hydrogels may effectively support cell proliferation, attachment, and migration in ischemic environments. In this study, we report the fabrication of a gelatin methacrylate (GelMA)-based micro-hydrogel for efficient in vivo delivery of genetically engineered endothelial cells. Micro-hydrogels were initially processed via electrospraying of GelMA and alginate (ALG) mixtures (at different ratios) on to calcium chloride (CaCl2) solution. Electrospraying of the GelMA/ALG mixture resulted in the formation of a micro-hydrogel, owing to ALG crosslinking. Secondary crosslinking of GelMA with UV light and ALG hydrogel chelation using sodium citrate solution resulted in GelMA-based micro-hydrogel formation. We observed the angiogenic response of human umbilical vein endothelial cells (HUVECs) in GelMA concentration-dependent manner. The seeding of HUVECs engineered to express human vascular endothelial growth factor on to the GelMA micro-hydrogel and the subsequent transplantation of the micro-hydrogel into a hindlimb ischemia model effectively attenuated the ischemia condition. This facile and simple micro-hydrogel fabrication strategy may serve as a robust method to fabricate efficient cell carriers for various ischemic diseases. Statement of Significance: For the therapeutic angiogenesis, it is important to provide the therapeutic cells with a carrier that could stabilize therapeutic cells and facilitate long-term survival of cells. Furthermore, it is also important to administer as many therapeutic cells as possible in a fixed volume. From these cues, we fabricated ECM-based micro-hydrogel produced by the high through-put system. And we intended to facilitate activation of therapeutic cells by coating the therapeutic cells onto the micro-hydrogel. In this manuscript, we fabricated methacrylate gelatin (GelMA) based micro-hydrogels using the electro-spraying method and coated HUVECs engineered to express hVEGF onto the micro-hydrogels. Then, we identified that GelMA concentration-dependent angiogenic response of HUVECs. Furthermore, we demonstrated that the VEGF secreting HUVEC-GelMA micro-hydrogels induced the restoration of blood flow and neovascularization in a hind-limb ischemia mouse model. These findings demonstrate that the high-throughput fabrication of ECM micro-hydrogels could be a novel platform to apply in neovascularization and tissue engineering.

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KW - Electrospray

KW - Gelatin methacrylate (GelMA)

KW - Hindlimb ischemia

KW - Micro-hydrogel

KW - hVEGF-secreting HUVEC

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DO - 10.1016/j.actbio.2019.01.057

M3 - Article

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SP - 285

EP - 296

JO - Acta Biomaterialia

JF - Acta Biomaterialia

SN - 1742-7061

ER -