Oil body bound oleosin-rhFGF9 fusion protein expressed in safflower (Carthamus tinctorius L.) stimulates hair growth and wound healing in mice

Abstract

BackgroundFibroblast growth factor 9 (FGF9) is a heparin-binding growth factor, secreted by both mesothelial and epithelial cells, which participates in hair follicle regeneration, wound healing, and bone development. A suitable source of recombinant human FGF9 (rhFGF9) is needed for research into potential clinical applications. We present that expression of oleosin-rhFGF9 fusion protein in safflower (Carthamus tinctorius L.) seeds stimulates hair growth and wound healing.ResultsThe oleosin-rhFGF9 expressed in safflower seeds, in which it localizes to the surface of oil bodies. The expression of oleosin-rhFGF9 was confirmed by polyacrylamide gel electrophoresis and western blotting. According to BCA and Enzyme-linked immunosorbent assay (ELISA) assay, the results show that the expression level of oleosin-rhFGF9 was 0.14% of oil body protein. The oil body bound oleosin-rhFGF9 showed mitogenic activity towards NIH3T3 cells in a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The efficacy of oil body bound oleosin-rhFGF9 in promoting hair growth and wound healing was investigated in C57BL/6 mice. In a hair regeneration experiment, 50 μg/μl oil body bound oleosin-rhFGF9 was applied to the dorsal skin of mice in the resting phase of the hair growth cycle. After 15 days, thicker hair and increased number of new hairs were seen compared with controls. Furthermore, the number of new hairs was greater compared with rhFGF9-treated mice. The hair follicles of mice treated with oil body bound oleosin-rhFGF9 expressed β-catenin more abundantly. In a wound healing experiment, dorsal skin wounds were topically treated with 50 μg/μl oil body bound oleosin-rhFGF9. Wound healing was quicker compared with mice treated with rhFGF9 and controls, especially in the earlier stages of healing.ConclusionsThe oil body bound oleosin-rhFGF9 promotes both hair growth and wound healing. It appears to promote hair growth, at least in part, by up-regulating β-catenin expression. The potential of oil body bound oleosin-rhFGF9 as an external drug can treat the alopecia and wounds or use in further clinical application.Electronic supplementary materialThe online version of this article (10.1186/s12896-018-0433-2) contains supplementary material, which is available to authorized users.

Phenol Content and Antioxidant and Antiaging Activity of Safflower Seed Oil (Carthamus Tinctorius L.)

“…Moreover, a natural skin toning cream has been developed from safflower oil [55]. The oil body bound oleosin-rhFGF9 expressed in safflower seed stimulates hair growth and wound healing in mice [56]. Dakhil et al [57] have reported that safflower oil characteristics can make it a main ingredient in the preparation of topical agents for the treatment of various skin problems.…”

Fabricating a novel HLC-hBMP2 fusion protein for the treatment of bone defects

“…The idea of using fusion protein as a delivery vehicle to effectively control the release of growth factors have been reported but mainly on the skin defects repairing [44][45][46][47][48][49].…”

Microparticles and Nanoparticles from Plants—The Benefits of Bioencapsulation

“…The lipophilic nature of oil bodies makes them easy to separate from the aqueous extraction medium by floating centrifugation, which simplifies downstream processing [ 95 ]. This technology has enabled the accumulation of proteins such as β-glucuronidase (the first to be reported), xylanase and hirudin in Brassica napus seeds [ 95 , 96 , 97 ], insulin, human fibroblast growth factor 9 (hFGF-9) and hEGF in Arabidopsis thaliana seeds [ 98 , 99 , 100 ], and hFGF-9, hEGF and antimicrobial peptides in safflower seeds [ 66 , 67 , 101 , 102 ]. All of these proteins remained functional in vitro.…”

“…All of these proteins remained functional in vitro. The emulsifying properties of oil bodies in safflower seeds were shown to promote absorption when topically applied to the skin of wounded rodents [ 66 ]. The transdermal drug delivery of oil body-linked hEGF significantly accelerated wound healing and tissue regeneration, and the mechanism may be similar to that seen with drugs delivered via liposomes [ 66 ].…”

“…For example, xylanase expressed as an oleosin fusion lacked the N-linked glycans found on the native protein, but these are not required for its activity [97]. Similarly, the N-glycans normally found on human growth factors such as hFGF-9 were also missing from the oleosin fusion protein [66]. Oleosin fusion technology is therefore unsuitable for proteins that require complex PTMs for activity.…”