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Wound healing assay. Left column: normal saline control group; center column: plant KGF1 group; right column: KGF1 standard protein group.
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Keratinocyte growth factor 1 (KGF1) is a growth factor that promotes epidermal cell proliferation, migration, differentiation, and wound repair. It is expressed at low levels in a form of inclusion body in E. coli. In order to increase its expression and activity, we produced tobacco plants expressing KGF1 via Agrobacterium-mediated transformation...
Citations
... FGF7 is especially involved in insulin secretion of pancreatic b-cells [11] . Recombinant FGF7 derived from tobacco plants was found to promote wound healing in diabetic rats [12] . FGF7 is also produced in osteogenically differentiated cells and improves bone marrow engraftment through a regulatory-T-cell-dependent mechanism [13] . ...
Fibroblast growth factor 7 (FGF7) is involved in lipid metabolism, which is considered as a candidate gene with close relation with muscle development by eGWAs and RNA-Seq analyses. To date, limited research has been conducted on the relationship between FGF7 gene and growth traits. The main objective of this work was to further investigate the association between novel InDel within FGF7 gene and growth traits in goat. Herein, FGF7 mRNA expression levels were investigated in various Fuqing goat tissues. We found that FGF7 gene was expressed in six adult goat tissues with the highest mRNA levels in adipose tissue. This result suggested that FGF7 gene might play a critical role in fat deposition. We also detected potential polymorphisms in Fuqing, Nubian and Jianyang Daer breeds. A 22-bp InDel polymorphism in FGF7 gene was detected in 396 goats and the three genotypes were designated as II, ID, and DD. Correlation analysis revealed that InDel polymorphism was significantly associated with growth traits (P < 0.05). Goats with genotypes ID and/or II had superior growth traits compared to those with genotype DD. In summary, our findings suggested that the 22-bp InDel within FGF7 gene could act as a molecular marker to improve the growth traits of goats in breeding programs.
... Cells derived from the mesenchyme produce and secrete KGF1, which promotes epidermal cell proliferation, differentiation, migration, and wound repair (Harrell et al. 2021). KGF1 is produced in Escherichia coli at low expression levels and formed in the inclusion bodies, which makes its purification difficult and limits its use in basic and applied pharmacological and pathological research (Feng et al. 2014). ...
Recombinant human KGF1 (rhKGF) is a fibroblast growth factor that promotes epidermal cell proliferation, differentiation, migration, and wound repair. In this study, we used the cell culture of a hazelnut expression system to produce rhKGF. These cells have a high growth rate. A cassette containing the rhKGF gene fused to hydrophobin tag was modified based on the hazelnut optimized codon usage and subsequently introduced into a hazelnut suspension culture under the control of a RAmy3D promoter, and secreted into the plant cell culture medium. The results showed that the highest transcription rate of the rhKGF gene was obtained under the RAmy3D promoter on the third day of transgenic cell placement in the promoter induction medium. To optimize the production of rhKGF secreted into the culture medium, the pH of the medium was adjusted during the production phase. After exposing the hazelnut cells to a pH of 7 for 12 d in the production phase, it was observed that approximately 95.35% of the cells remained viable. Western blotting analysis revealed the presence of two specific bands at approximately 30 and 40 kDa, corresponding to the fusion protein in the purified hydrophobin-rhKGF. The higher molecular weight of the fusion protein (~ 40 kDa) compared to the calculated weight is likely due to glycosylation. These results indicate that the transient expression system is effective in producing a significant amount of rhKGF within a few weeks using hazelnut cells. The amount of biomass produced could increase up to 7 times within a 24-d period, indicating that cell suspension could be a viable option for producing foreign proteins.
... Polymeric nanoparticles are biocompatible colloidal systems that have risen in importance for both biomedical and bioengineering applications [99]. They are generally integrated by charged polymers and connected by interactivity of cationic and anionic chains of groups [100]. When drugs are incorporated into polymeric systems, this process prevents the deterioration caused by proteases found in the injury and delivered in stages to lower the frequency of administration [101]. ...
Medicinal plants have been used since ancient times for their various therapeutic activities and are safer compared to modern medicines, especially when properly identifying and preparing them and choosing an adequate dose administration. The phytochemical compounds present in plants are progressively yielding evidence in modern drug delivery systems by treating various diseases like cancers, coronary heart disease, diabetes, high blood pressure, inflammation, microbial, viral and parasitic infections, psychotic diseases, spasmodic conditions, ulcers, etc. The phytochemical requires a rational approach to deliver the compounds to enhance the efficacy and to improve patients’ compatibility. Nanotechnology is emerging as one of the most promising strategies in disease control. Nano-formulations could target certain parts of the body and control drug release. Different studies report that phytochemical-loaded nano-formulations have been tested successfully both in vitro and in vivo for healing of skin wounds. The use of nano systems as drug carriers may reduce the toxicity and enhance the bioavailability of the incorporated drug. In this review, we focus on various nano-phytomedicines that have been used in treating skin burn wounds, and how both nanotechnology and phytochemicals are effective for treating skin burns.
... The generation of recombinant growth factors becomes an interest of the biopharmaceutical industry due to its potential application in regenerative medicine [26]. A range of growth factors including acid fibroblast growth factor (aFGF) [27,28], keratinocyte growth factor 1 (KGF1) [29], human epidermal growth factor (hEGF) [21,30] where produced in plants. hEGF was expressed in different plant hosts such as tomato [31], soybean [32], and also in Nicotiana species [21,22]. ...
Human epidermal growth factor (hEGF) has gained clinical importance due to its ability to promote wound healing. Due to its commercial applications and high market demand, recombinant EGF has been produced in several forms. Currently, plant expression system is considered as potential alternative for low-cost recombinant protein production. Hence, this study focused on improving the production of hEGF in plants by effective gene construct design and optimizing the Agrobacterium culture conditions for high protein production. In this context, hEGF gene was cloned into plant geminiviral expression vector pBYR2e and transformed in to N. benthamiana leaves via., agroinfiltration. The recombinant hEGF was purified from the plant crude extracts by single-step affinity chromatography. Furthermore, the plant-produced hEGF has shown to promote cell migration comparable to commercial hEGF in HaCaT cells in vitro. These results indicated the potential of plant expression system for the production of recombinant hEGF for tissue engineering applications.
... To the best of our knowledge, this is the first report about the transient expression of HSA gene in plant system. The Agrobacterium-mediated transient expression system in intact tobacco leaves have been successfully used [23], lumbrokinase [24], Camelid nanobodies [25] and keratinocyte growth factor 1 [26]. ...
Today, recombinant human proteins make up a considerable part of FDA-approved biotechnological drugs. The selection of proper expression platform for manufacturing recombinant protein is a vital factor in achieving the optimal yield and quality of a biopharmaceutical in a timely fashion. This experiment was aimed to compare the transient expression level of human serum albumin gene in different tobacco genotype. For this, the Agrobacterium tumefaciens strains LB4404 and GV3101 harboring pBI121-HSA binary vector were infiltered in leaves of three tobacco genotypes, including Nicotiana benthamiana and N. tabacum cv Xanthi and Samsun. The qRT-PCR, SDS-PAGE, western blotting and ELISA analysis were performed to evaluate the expression of HSA gene in transgenic plantlets. Our results illustrated that the expression level of rHSA in tobacco leaves was highly dependent on Agrobacterium strains, plant genotypes and harvesting time. The highest production of recombinant HSA protein was obtained in Samsun leaves infected with A. tumefaciens strain GV3101 after 3 days of infiltration.
... Các nghiên cứu cho thấy rằng KGF có nhiều chức năng khác nhau bao gồm kích thích phân chia, tăng sinh và biệt hóa các dòng tế bào biểu mô và tế bào sừng [1,3,4]. Bên cạnh đó, KGF còn có vai trò trong việc tái tạo, sửa chữa các mô và cơ quan tổn thương thông kích thích sự di chuyển của các tế bào sừng và biểu mô ở miệng vết thương, từ đó thúc đẩy tiến trình làm lành vết thương [5,6]. Hiện nay, KGF đã được nghiên cứu rộng rãi và ứng dụng trong nhiều lĩnh vực như mỹ phẩm, dược phẩm và hỗ trợ trong điều trị ung thư [7][8][9]. ...
... Theo theo thống kê, trung bình một bệnh nhân có trọng lượng 70 kg sẽ tốn khoảng 5000 euro cho một lần điều trị bằng Palifermin [11], tương đương khoảng hơn 100 triệu VNĐ. Với mục tiêu thu nhận KGF với lượng lớn hơn, có nhiều chiến lược biểu hiện tái tổ hợp đã và đang được tiến hành trên nhiều hệ thống như vi khuẩn E.coli [12,13], tế bào cây thuốc lá [6], tế bào con tằm [14], tế bào chuột lang Trung Quốc -CHO [15], tuy nhiên vẫn còn tồn tại nhiều nhược điểm như lượng KGF thu nhận thấp, bản thân KGF gây độc cho tế bào chủ biểu hiện như ở hệ thống E. coli [12]; đồng thời, hiệu quả sản xuất thấp do chi phí cao ở hệ thống biểu hiện tế bào động vật có vú,… Năm 2018, Bahadori và cộng sự đã biểu hiện thành công KGF dạng tiết ở hệ thống tế bào nấm men P. pastoris với hoạt tính kích thích tăng sinh trên dòng tế bào A549 và NIH3T3. Nhận thấy P. pastoris là hệ thống tế bào chủ biểu hiện với nhiều ưu điểm như khả năng biến đổi sau dịch mã và gấp cuộn protein tương đồng với tế bào động vật, đặc biệt promoter mạnh và khả năng tiết protein ngoại bào giúp protein tái tổ hợp ở hệ thống này biểu hiện ở mức độ cao, có thể chiếm tới hơn 80% tổng protein tiết [16][17][18], chúng tôi lựa chọn P. pastoris nhằm biểu hiện KGF tái tổ hợp. ...
Keratinocyte Growth Factor (KGF) is a paracrine-acting and epithelium-specific growth factor produced by cells of mesenchymal origin, play an important role in promoting proliferation, differentiation, motility of epithelial cells and stimulating regeneration of damaged epithelial tissues. Recent studies indicated that recombinant KGF is produced in many different expression systems such as bacteria, insect cells, plant and mammalian cells. However, KGF’s yields obtained from these systems is low and production’s cost is high especially in mammalian cells. In this study, the yeast Pichia pastoris was chosen as a host for KGF expression through induction of methanol by promoter AOX on pPICzαA vector system. The results demonstrated that the Pichia pastoris X33:kgf transformants secreted KGF directly into BMMY medium after inducing by 0.5% methanol. The recombinant protein was purified by heparin affinity chromatography with the yield of 1.35 mg/l and the purity of 99.89% showed by SDS-PAGE. In addition, MTT assay showed the purified recombinant KGF had a proliferation effect on A549 cell line since A549 known as a cell has KGF’s receptor.
... These include therapies-mainly antibodies-for influenza, Ebola virus, dengue, norovirus, human immunodeficiency virus and acquired immune deficiency syndrome (HIV/AIDS), hepatitis A, hepatitis B, hepatitis C, severe acute respiratory syndrome (SARS) coronavirus, plague, malaria, West Nile virus, anthrax, human papillomavirus, bluetongue virus, Streptococcus pneumoniae, rabies, and cholera (107). They even include the production, in N. benthamiana, of vitronectin (30), human growth factors (33), cancer treatments (56), and cocaine addiction treatments (70). The most publicly visible of these therapies has been ZMapp, an anti-Ebola antibody cocktail expressed in N. benthamiana and used to treat two infected American health care workers during the 2014 Ebola outbreak (5,79). ...
A decade ago, the value of Nicotiana benthamiana as a tool for plant molecular biologists was beginning to be appreciated. Scientists were using it to study plant-microbe and protein-protein interactions, and it was the species of choice with which to activate plasmid-encoded viruses, screen for gene functions with virus-induced gene silencing (VIGS), and transiently express genes by leaf agroinfiltration. However, little information about the species' origin, diversity, genetics, and genomics was available, and biologists were asking the question of whether N. benthamiana is a second fiddle or virtuoso. In this review, we look at the increased knowledge about the species and its applications over the past decade. Although N. benthamiana may still be the sidekick to Arabidopsis, it shines ever more brightly with realized and yet-to-be-exploited potential.
... KGF is a potent growth factor involved in re-epithelization of skin wounds [158]. In two recent investigations it was shown that that KGF contained in self-assembled NPs could stimulate healing of skin wounds, giving increased reepithelization, better tissue remodeling and skin regeneration [159,160]. PLGA is a FDA approved material used in the production of polymeric NPs (100-200 nm) due to its advantages including biodegradability, biocompatibility and non-toxicity [161]. PLGA NPs are normally fabricated by emulsification of hydrophobic components, using an organic solvent and various surfactants [162]. ...
... Feng et al. 9 carried out a study in diabetic rats; to do so, they made wounds on the animals' backs and injected keratinocyte growth factors to assess healing. Two wounds measuring 2 cm in diameter were made on each side of the spinal column; growth factor was injected in one side, whereas saline solution was injected in the other side. ...
INTRODUCTION:
Healing is a process that restores the physical integrity of body structures. It is a dynamic, complex, multicellular process that involves the extracellular matrix, cytokines, blood cells, and growth factors. Growth factors are proteins that activate and stimulate cell proliferation through the activation of angiogenesis, mitogenesis, and gene transcription, accelerating the healing process.
OBJECTIVE:
To assess the influence of growth factors on the healing process of wounds made on the backs of female rats compared to the control wound, through macro and microscopy.
METHODS:
This study used 45 female Wistar rats, in which three wounds were made on the back. The first was the control wound, the second received epithelial growth factor injection, and the third received a combination of factors. Macroscopic and microscopic assessments were performed on the third, seventh, and 15th days of the experiment. For microscopic analysis, hematoxylin-eosin staining was utilized to assess the inflammatory process; vimentin, for assessment of blood vessels and fibroblasts, and Sirius Red for collagen assessment.
RESULTS:
In the macroscopic assessment, the use of growth factors resulted in faster healing and decrease of granulation tissue on days seven and 15; (80.31% reduction in the control wound vs. 83.24% in the epithelial wound vs. 100% in the mixed wound). Utilizing microscopy, at the three stages of the experiment, there were no significant differences between the three wounds; however, when comparing the day of euthanization for each type of wound, there was a favorable outcome for epithelial and mixed wounds (between the third vs. 15th day, p < 0.001, and in the comparison of the seventh vs. 15th day; p = 0.002 and p = 0.001 for epithelial and mixed wounds, respectively) with a higher number of fibroblasts, angiogenesis, and collagen type I.
CONCLUSION:
The use of growth factors accelerates healing, stimulates greater angiogenic activity, and accelerates fibroplasia and collagen maturation.
