scholarly journals Extemporaneous Preparation of Injectable and Enzymatically Degradable 3D Cell Culture Matrices from an Animal‐Component‐Free Recombinant Protein Based on Human Collagen Type I

2019 ◽  
pp. 1900127
Author(s):  
Hiroyuki Kamata ◽  
Satoko Ashikari‐Hada ◽  
Yusuke Mori ◽  
Akihiko Azuma ◽  
Ken‐ichiro Hata
Keyword(s):  
Cell Culture ◽  
Recombinant Protein ◽  
Collagen Type ◽  
3D Cell Culture ◽  
Collagen Type I ◽  
Type I ◽  
Human Collagen ◽  
Extemporaneous Preparation ◽  
Animal Component

scholarly journals Plant Recombinant Human Collagen Type I Hydrogels for Corneal Regeneration

2021 ◽  
Author(s):  
Michel Haagdorens ◽  
Elle Edin ◽  
Per Fagerholm ◽  
Marc Groleau ◽  
Zvi Shtein ◽  
...  
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Collagen Fibrils ◽  
Type I ◽  
Safe Alternative ◽  
Human Donor ◽  
Human Collagen ◽  
Donor Corneas

Abstract 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.

scholarly journals Hydroxylation of recombinant human collagen type I alpha 1 in transgenic maize co-expressed with a recombinant human prolyl 4-hydroxylase

2011 ◽  
Vol 11 (1) ◽  
pp. 69 ◽  
Author(s):  
Xing Xu ◽  
Qinglei Gan ◽  
Richard C Clough ◽  
Kameshwari M Pappu ◽  
John A Howard ◽  
...  
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Transgenic Maize ◽  
Type I ◽  
Human Collagen

scholarly journals Cosmetic Potential of a Recombinant 50 kDa Protein

Cosmetics ◽  
2022 ◽  
Vol 9 (1) ◽  
pp. 8
Author(s):  
Nesma Aly ◽  
Emilie Benoit ◽  
Jean-Luc Chaubard ◽  
Kavyasree Chintalapudi ◽  
Soojin Choung ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Collagen Type ◽  
Collagen Type I ◽  
Protein Product ◽  
Dermal Fibroblasts ◽  
Type Iii Collagen ◽  
Type I ◽  
Type Iii ◽  
Extraction And Purification ◽  
Recombinant Type

Collagen and its derivative proteins have been widely used as a major component for cosmetic formulations as a natural ingredient and moisturizer. Most commercially available collagens are animal-derived collagen type I and other forms of collagen, such as type III collagen, are far less prevalent in animals, making extraction and purification extremely difficult and expensive. Here, we report the production of a 50 kDa protein produced in yeast that is 100% identical to the N-terminus of the human type III collagen. This recombinant protein has a larger molecular weight than most incumbent recombinant collagen proteins available for personal care applications. We report the industrialization of both the fermentation and purification processes to produce a final recombinant protein product. This final protein product was shown to be safe for general applications to human skin and compatible with common formulation protocols, including ethanol-based formulations. This recombinant collagen type III protein was also shown to uniquely stimulate both collagen type I and type III production and secretion by primary human dermal fibroblasts. The unique combination of biostimulation, compatibility with beauty product formulations and demonstrated commercial production, make this novel recombinant type III collagen a good candidate for broad application in the cosmetics industry.

scholarly journals Sources of Collagen for Biomaterials in Skin Wound Healing

2019 ◽  
Vol 6 (3) ◽  
pp. 56 ◽  
Author(s):  
Evan Davison-Kotler ◽  
William S. Marshall ◽  
Elena García-Gareta
Keyword(s):  
Collagen Type ◽  
Recombinant Expression ◽  
Collagen Type I ◽  
Skin Wound ◽  
Type I ◽  
Expression Systems ◽  
Skin Wound Healing ◽  
Skin Collagen ◽  
Human Collagen ◽  
Collagen Type Vii

Collagen is the most frequently used protein in the fields of biomaterials and regenerative medicine. Within the skin, collagen type I and III are the most abundant, while collagen type VII is associated with pathologies of the dermal–epidermal junction. The focus of this review is mainly collagens I and III, with a brief overview of collagen VII. Currently, the majority of collagen is extracted from animal sources; however, animal-derived collagen has a number of shortcomings, including immunogenicity, batch-to-batch variation, and pathogenic contamination. Recombinant collagen is a potential solution to the aforementioned issues, although production of correctly post-translationally modified recombinant human collagen has not yet been performed at industrial scale. This review provides an overview of current collagen sources, associated shortcomings, and potential resolutions. Recombinant expression systems are discussed, as well as the issues associated with each method of expression.

Sequence Alignment between vWF and Peptides Inhibiting the vWF-collagen Interaction Does not Result in the Identification of a Collagen-binding Site in vWF

2000 ◽  
Vol 84 (10) ◽  
pp. 621-625 ◽  
Author(s):  
R. M. van der Plas ◽  
G. Vandecasteele ◽  
S. Vauterin ◽  
E. G. Huizinga ◽  
J. J. Sixma ◽  
...  
Keyword(s):  
Binding Site ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Binding Studies ◽  
Collagen Types ◽  
Collagen Binding ◽  
Human Collagen ◽  
Rat Tail ◽  
Calf Skin

SummaryWe previously found that two peptides (N- and Q-peptide) selected by phage display for binding to an anti-vWF antibody, were able to inhibit vWF-binding to collagen (1). The sequence of those peptides could be aligned with the sequence in vWF at position 1129-1136 just outside the A3-domain. As the peptides represent an epitope or mimotope of vWF for binding to collagen we next wanted to study whether the alignment resulted in the identification of a new collagen binding site in vWF. We mutated the 1129-1136 VWTLPDQC sequence in vWF to VATAPAAC. Expressing this mutant vWF (7.8-vWF) in a fur-BHK cell line resulted in well processed 7.8-vWF containing a normal distribution of molecular weight multimers. However, binding studies of this mutant vWF to rat tail, human and calf skin collagens type I, to human collagen types III and VI, revealed no decrease in vWF-binding to any of these collagens. Thus, although the N-and Q-peptides did inhibit the vWF-collagen interaction, the resulting alignment with the vWF sequence did not identify a collagen binding site, pointing out that alignments (although with a high percentage of identity) do not always result in identification of binding epitopes. However, suprisingly removal of the A3-domain or changing the vWF sequence at position 1129-1136 resulted in an increase of vWF-binding to human collagen type VI and to rat tail collagen type I, implying that these changes result in a different conformation of vWF with an increased binding to these collagens as a consequence.

scholarly journals Potent Costimulation of Effector T Lymphocytes by Human Collagen Type I

2000 ◽  
Vol 165 (9) ◽  
pp. 4935-4940 ◽  
Author(s):  
Wei Hong Rao ◽  
Jonathan M. Hales ◽  
Richard D. R. Camp
Keyword(s):  
T Lymphocytes ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Human Collagen

The Collagen-binding Leech Products rLAPP and Calin Prevent both von Willebrand Factor and α2β1 (GPIa/IIa)-I-domain Binding to Collagen in a Different Manner

1999 ◽  
Vol 82 (09) ◽  
pp. 1160-1163 ◽  
Author(s):  
H. Deckmyn ◽  
H. Depraetere ◽  
A. Kerekes
Keyword(s):  
Binding Sites ◽  
Collagen Type ◽  
Collagen Type I ◽  
The Other ◽  
Type I ◽  
Flow Conditions ◽  
Human Collagen ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Natural Inhibitors

SummaryCalin and rLAPP are two natural inhibitors that are able to inhibit the vWF-binding and platelet adhesion to collagen both under static and flow conditions. In this study we demonstrate that both rLAPP and Calin prevent α2I-domain binding to human collagen type I with an IC50 of 5 μg/ml. However, although both vWF and α2I-domain binding to collagen is prevented by rLAPP and Calin, the latter two do not bind to the same collagen site since Calin only partially could compete with rLAPP for binding to collagen. Also vWF and the α2I-domain were unable to compete completely with each other for the binding to collagen. So the following hypothesis can be made: the binding sites of vWF and of the α2I-domain on human collagen type I are different but close to each other since rLAPP could inhibit both interactions, and thus should bind to an overlapping epitope. The Calin preparation on the other hand may still contain two active principles, one interfering with vWF-binding, the other with the α2I-domain-binding to collagen.

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