TY - JOUR
T1 - Plant Recombinant Human Collagen Type I Hydrogels for Corneal Regeneration
AU - Haagdorens, Michel
AU - Edin, Elle
AU - Fagerholm, Per
AU - Groleau, Marc
AU - Shtein, Zvi
AU - Ulčinas, Artūras
AU - Yaari, Amit
AU - Samanta, Ayan
AU - Cepla, Vytautas
AU - Liszka, Aneta
AU - Tassignon, Marie José
AU - Simpson, Fiona
AU - Shoseyov, Oded
AU - Valiokas, Ramūnas
AU - Pintelon, Isabel
AU - Ljunggren, Monika Kozak
AU - Griffith, May
N1 - Funding Information: Open access funding provided by Linkoping University. This research was funded with research grants from The Research Foundation Flanders (#FWO - 11ZB315N) to MH; EuroNanoMed II (G0D5615N to MJT) for the REGENERATE project in partnership with the Swedish Research Council (529-2014-7490 to MKL) and the Research Council of Lithuania (EuroNanoMed2-01/2015 to RV), and the European Cooperation in Science and Technology (EU-COST BM1302). MG acknowledges salary and research support from her Canada Research Chair Tier 1 in Biomaterials and Stem Cell in Ophthalmology and a Caroline Durand Foundation Research Chair in Cellular Therapy in the Eye, as well as a Honorary Professorship at the School of Optometry and Vision Science, Cardiff University. FS is supported by an NSERC Canada doctoral studentship. Funding Information: We thank Dr. Christopher McTiernan, University of Ottawa Heart Institute, for the nerve counts in Fig. 4E. We acknowledge Science Advances (AAAS) for reproduction of results in Fig. 4 from McTiernan et al. We thank Prof. Dr. Sorcha Ní Dhubhghaill for her help with the pig surgeries, Prof. Dr. Nadia Zakaria for administration and logistics, and Ms. Nezahat Bostan and Ms. Eline Melsbach for procurement of the amniotic membranes and cadaveric donor eyes. Ms. Sofie Thys is acknowledged for performing the scanning electron microscopy and Ms. Bieke Vanhoutte for assisting in the microbial challenge testing. The tobacco plant in the graphical abstract is adapted from “Tobacco and its adulterations”, this rendering is in the public domain [42 ]. Publisher Copyright: © 2021, The Author(s).
PY - 2022/6
Y1 - 2022/6
N2 - Purpose: To determine feasibility of plant-derived recombinant human collagen type I (RHCI) for use in corneal regenerative implants Methods: RHCI was crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form hydrogels. Application of shear force to liquid crystalline RHCI aligned the collagen fibrils. Both aligned and random hydrogels were evaluated for mechanical and optical properties, as well as in vitro biocompatibility. Further evaluation was performed in vivo by subcutaneous implantation in rats and corneal implantation in Göttingen minipigs. Results: Spontaneous crosslinking of randomly aligned RHCI (rRHCI) formed robust, transparent hydrogels that were sufficient for implantation. Aligning the RHCI (aRHCI) resulted in thicker collagen fibrils forming an opaque hydrogel with insufficient transverse mechanical strength for surgical manipulation. rRHCI showed minimal inflammation when implanted subcutaneously in rats. The corneal implants in minipigs showed that rRHCI hydrogels promoted regeneration of corneal epithelium, stroma, and nerves; some myofibroblasts were seen in the regenerated neo-corneas. Conclusion: Plant-derived RHCI was used to fabricate a hydrogel that is transparent, mechanically stable, and biocompatible when grafted as corneal implants in minipigs. Plant-derived collagen is determined to be a safe alternative to allografts, animal collagens, or yeast-derived recombinant human collagen for tissue engineering applications. The main advantage is that unlike donor corneas or yeast-produced collagen, the RHCI supply is potentially unlimited due to the high yields of this production method. Lay Summary: A severe shortage of human-donor corneas for transplantation has led scientists to develop synthetic alternatives. Here, recombinant human collagen type I made of tobacco plants through genetic engineering was tested for use in making corneal implants. We made strong, transparent hydrogels that were tested by implanting subcutaneously in rats and in the corneas of minipigs. We showed that the plant collagen was biocompatible and was able to stably regenerate the corneas of minipigs comparable to yeast-produced recombinant collagen that we previously tested in clinical trials. The advantage of the plant collagen is that the supply is potentially limitless.
AB - Purpose: To determine feasibility of plant-derived recombinant human collagen type I (RHCI) for use in corneal regenerative implants Methods: RHCI was crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form hydrogels. Application of shear force to liquid crystalline RHCI aligned the collagen fibrils. Both aligned and random hydrogels were evaluated for mechanical and optical properties, as well as in vitro biocompatibility. Further evaluation was performed in vivo by subcutaneous implantation in rats and corneal implantation in Göttingen minipigs. Results: Spontaneous crosslinking of randomly aligned RHCI (rRHCI) formed robust, transparent hydrogels that were sufficient for implantation. Aligning the RHCI (aRHCI) resulted in thicker collagen fibrils forming an opaque hydrogel with insufficient transverse mechanical strength for surgical manipulation. rRHCI showed minimal inflammation when implanted subcutaneously in rats. The corneal implants in minipigs showed that rRHCI hydrogels promoted regeneration of corneal epithelium, stroma, and nerves; some myofibroblasts were seen in the regenerated neo-corneas. Conclusion: Plant-derived RHCI was used to fabricate a hydrogel that is transparent, mechanically stable, and biocompatible when grafted as corneal implants in minipigs. Plant-derived collagen is determined to be a safe alternative to allografts, animal collagens, or yeast-derived recombinant human collagen for tissue engineering applications. The main advantage is that unlike donor corneas or yeast-produced collagen, the RHCI supply is potentially unlimited due to the high yields of this production method. Lay Summary: A severe shortage of human-donor corneas for transplantation has led scientists to develop synthetic alternatives. Here, recombinant human collagen type I made of tobacco plants through genetic engineering was tested for use in making corneal implants. We made strong, transparent hydrogels that were tested by implanting subcutaneously in rats and in the corneas of minipigs. We showed that the plant collagen was biocompatible and was able to stably regenerate the corneas of minipigs comparable to yeast-produced recombinant collagen that we previously tested in clinical trials. The advantage of the plant collagen is that the supply is potentially limitless.
KW - Cornea regeneration
KW - Limbal stem cells
KW - Plant collagen
KW - Recombinant human collagen type 1
KW - Tissue engineering
UR - http://www.scopus.com/inward/record.url?scp=85112622343&partnerID=8YFLogxK
U2 - https://doi.org/10.1007/s40883-021-00220-3
DO - https://doi.org/10.1007/s40883-021-00220-3
M3 - Article
SN - 2364-4133
VL - 8
SP - 269
EP - 283
JO - Regenerative Engineering and Translational Medicine
JF - Regenerative Engineering and Translational Medicine
IS - 2
ER -
