scholarly journals Important metabolites in maintaining folic acid, homocysteine and polyamine metabolism associated with ranibizumab treatment in cultured Human Tenon’s Fibroblast

Author(s):  
Siti Munirah Md Noh ◽  
Siti Hamimah Sheikh Abdul Kadir ◽  
Sushil Kumar Vasudevan

Anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to Vascular Endothelial Growth Factor (VEGF), ranibizumab works by binding and neutralizes all active VEGF, thus limits the progressive cell growth and proliferation. Its application in ocular diseases has shown remarkable desired effects, however to date its anti-fibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated Human Tenos’s fibroblast (HTF). Cultured HTFs were treated for48 hours with 0.5 mg/ml of ranibizumab and 0.5 mg/ml control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF Liquid Chromatography/ Mass Spectrometer (LC/MS) System to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analysed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At statistical analysis of p-value < 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly up-regulated in ranibizumab-treated group, with 6 of the mostly up-regulated have insignificant role in fibroblast’s cell cycle and wound healing regulations Meanwhile, 121 identified metabolites were down-regulated with seven of the mostly down-regulated are significantly involved in cell cycle and proliferation. Our findings demonstrated that ranibizumab abrogates the tissue scarring process and wound healing formation by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folic acid cycle, nucleotide synthesis, homocysteine and spermidine metabolism. This study provides an insight of ranibizumab mechanism of actions on HTFs from the perspective of metabolomics.

2019 ◽  
Author(s):  
Siti Munirah Md Noh ◽  
Siti Hamimah Sheikh Abdul Kadir ◽  
Sushil Kumar Vasudevan

Anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to Vascular Endothelial Growth Factor (VEGF), ranibizumab works by binding and neutralizes all active VEGF, thus limits the progressive cell growth and proliferation. Its application in ocular diseases has shown remarkable desired effects, however to date its anti-fibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated Human Tenos’s fibroblast (HTF). Cultured HTFs were treated for48 hours with 0.5 mg/ml of ranibizumab and 0.5 mg/ml control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF Liquid Chromatography/ Mass Spectrometer (LC/MS) System to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analysed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At statistical analysis of p-value < 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly up-regulated in ranibizumab-treated group, with 6 of the mostly up-regulated have insignificant role in fibroblast’s cell cycle and wound healing regulations Meanwhile, 121 identified metabolites were down-regulated with seven of the mostly down-regulated are significantly involved in cell cycle and proliferation. Our findings demonstrated that ranibizumab abrogates the tissue scarring process and wound healing formation by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folic acid cycle, nucleotide synthesis, homocysteine and spermidine metabolism. This study provides an insight of ranibizumab mechanism of actions on HTFs from the perspective of metabolomics.


Biomolecules ◽  
2019 ◽  
Vol 9 (6) ◽  
pp. 243
Author(s):  
Siti Munirah Md Noh ◽  
Siti Hamimah Sheikh Abdul Kadir ◽  
Sushil Vasudevan

The anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to vascular endothelial growth factor (VEGF), ranibizumab works by binding and neutralizing all active VEGF-A, thus limiting progressive cell growth and proliferation. Ranibizumab application in ocular diseases has shown remarkable desired effects; however, to date, its antifibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated human Tenon’s fibroblasts (HTFs). Cultured HTFs were treated for 48 h with 0.5 mg/mL of ranibizumab and 0.5 mg/mL control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF liquid chromatography/mass spectrometer (LC/MS) system to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analyzed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At p-value < 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly upregulated in ranibizumab-treated group, with six of the mostly upregulated having insignificant role in fibroblast cell cycle and wound healing regulations. Meanwhile, 121 identified metabolites that were downregulated, and seven of the mostly downregulated are significantly involved in cell cycle and proliferation. Our findings suggest that ranibizumab abrogates the tissue scarring and wound healing process by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folate cycle, nucleotide synthesis, and homocysteine and spermidine metabolism. This study provides an insight into ranibizumab’s mechanism of action in HTFs from the perspective of metabolomics.


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Wubante Demilew ◽  
Getnet Mequanint Adinew ◽  
Seyfe Asrade

Background. Medicinal plants play indispensable roles to treat various ailments. Acanthus polystachyus is one of the medicinal plants used traditionally for treatment of wounds. However, there were no scientific reports documented so far on the wound healing activities of this plant. Thus, the present study provides a scientific evaluation for the wound healing potential of the crude extract of A. polystachyus leaves. Methods. The crude extraction was carried out using 80% methanol. The crude extract was prepared in 5% (w/w) and 10% (w/w) ointment and evaluated for wound healing activity using excision, infected, and incision wound models in Swiss albino mice. Results. Both 5%  and 10% (w/w) ointments significantly reduced period of epithelialization and increased wound contraction rate and tensile strength compared to the negative control group (P < 0.05). The wound healing activity of 10% (w/w) ointment treated group was greater than 5% (w/w) and nitrofurazone ointment treated groups in S. aureus infected wound model. Conclusion. These results demonstrate that the crude extract of A. polystachyus leaves possesses wound healing activities. This justifies the traditional claimed use of the plant for treating uninfected and infected wounds caused by S. aureus.


Author(s):  
Hana M. Hammad ◽  
Amer Imraish ◽  
Maysa Al-Hussaini ◽  
Malek Zihlif ◽  
Amani A. Harb ◽  
...  

Objective: Achillea fragrantissima L. (Asteraceae) is a traditionally used medicinal herb in the rural communities of Jordan. Methods: The present study evaluated the efficacy of the ethanol extract of this species on angiogenesis in both, ex vivo using rat aortic ring assay and in vivo using rat excision wound model. Results: In concentrations of 50 and 100 µg/ml, the ethanol extract showed angiogenic stimulatory effect and significantly increased length of capillary protrusions around aorta rings of about 60% in comparison to those of untreated aorta rings. In MCF-7 cells, the ethanol extract of A. fragrantissima stimulates the production of VEGF in a dose-dependent manner. 1% and 5% of ethanol extract of A. fragrantissima containing vaseline based ointment was applied on rat excision wounds for six days and was found to be effective in wound healing and maturation of the scar. Both preparations resulted in better wound healing when compared to the untreated control group and vaseline-treated group. This effect was comparable to that induced by MEBO, the positive control. Conclusion: The results indicate that A. fragrantissima has a pro-angiogenic effect, which may act through the VEGF signaling pathway.


Author(s):  
Joon M. Jung ◽  
Hae K. Yoon ◽  
Chang J. Jung ◽  
Soo Y. Jo ◽  
Sang G. Hwang ◽  
...  

Cold plasma can be beneficial for promoting skin wound healing and has a high potential of being effectively used in treating various wounds. Our aim was to verify the effect of cold plasma in accelerating wound healing and investigate its underlying mechanism in vitro and in vivo. For the in vivo experiments, 2 full-thickness dermal wounds were created in each mouse (n = 30). While one wound was exposed to 2 daily plasma treatments for 3 min, the other wound served as a control. The wounds were evaluated by imaging and histological analyses at 4, 7, and 11 days post the wound infliction process. Immunohistochemical studies were also performed at the same time points. In vitro proliferation and scratch assay using HaCaT keratinocytes and fibroblasts were performed. The expression levels of wound healing–related genes were analyzed by real-time polymerase chain reaction and western blot analysis. On day 7, the wound healing rates were 53.94% and 63.58% for the control group and the plasma-treated group, respectively. On day 11, these rates were 76.05% and 93.44% for the control and plasma-treated groups, respectively, and the difference between them was significant ( P = .039). Histological analysis demonstrated that plasma treatment promotes the formation of epidermal keratin and granular layers. Immunohistochemical studies also revealed that collagen 1, collagen 3, and alpha-smooth muscle actin appeared more abundantly in the plasma-treated group than in the control group. In vitro, the proliferation of keratinocytes was promoted by plasma exposure. Scratch assay showed that fibroblast exposure to plasma increased their migration. The expression levels of collagen 1, collagen 3, and alpha-smooth muscle actin were elevated upon plasma treatment. In conclusion, cold plasma can accelerate skin wound healing and is well tolerated.


Author(s):  
Endang Sri Purwanti Ningsih ◽  
Noorlaila Noorlaila ◽  
Ikhwan Rizki Muhammad ◽  
Windy Yuliana Budianto

Background: The process of wound healing is influenced by various factors such as age, hormones, and wound care. Wound care is done to accelerate wound healing which can be done by various methods, one of them is traditional care. Traditional wound care can use medicinal plants. Rhodomyrtus tomentosa is a medicinal plant that has an antioxidant, anti-inflammatory, antitumor and antibacterial content. Thus this study aims to evaluate the effectiveness of the antiseptic solution of the Rodhomyrtus tomentosa leaf extract on wound healing in male Wistar rats. Method: this research is pure experimental research with post test only control group design. Thirty male white rats were divided into five groups, namely negative control, positive control, Rhodomyrtus tomentosa leaf extract 15%, 30%, and 60%. Rhodomyrtus tomentosa leaf extraction was carried out by maceration method with 70% ethano solvent. The extraction results are divided into 3 concentrations (15%, 30% and 60%). The wound healing process was evaluated by measuring the length of the wound manually from 0 to 10 days in each group. Meanwhile, the number of fibroblast cells was calculated through hematoxylin eosin (HE) staining and observed using an Olympus CX41 microscope with a 10x magnification and objective lens magnification in 3 fields. Result: There was a significant difference in the reduction in wound length (p =< 0,000) between the five experimental groups (Rhodomyrtus tomentosa leaf extract solution 15%, 30% and 60%, negative control and positive control. Solution of rhodomyrtus tomentosa leaf extract accelerated the increase in the number of fibroblasts compared to the negative control group (p = 0.003), but did not make a difference (p = 0.403) with the positive control group. Rhodomyrtus tomentosa leaf extraction solution had the same microscopic effect on the number of fibroblasts with a positive control group given 0.9% NaCl solution. Conclusion: There was a significant difference in the number of fibroblasts between all groups, but no difference in wound healing length.


Revista CERES ◽  
2011 ◽  
Vol 58 (2) ◽  
pp. 149-154
Author(s):  
Alexandre Couto Tsiomis ◽  
Andréa Pacheco Batista Borges ◽  
Ana Paula Falci Daibert ◽  
Tatiana Schmitz Duarte ◽  
Emily Correna Carlo Reis ◽  
...  

Bone loss, either by trauma or other diseases, generates an increasing need for substitutes of this tissue. This study evaluated Bioglass as a bone substitute in the regeneration of the alveolar bone in mandibles of dogs by clinical, surgical and radiological analysis. Twenty-eight adult dogs were randomly separated into two equal groups. In each animal, a bone defect was created on the vestibular surface of the alveolar bone between the roots of the fourth right premolar tooth. In the treated group, the defect was immediately filled with bioglass, while in the control, it remained unfilled. Clinical evaluations were performed daily for a week, as well as x-rays immediately after surgery and at 8, 14, 21, 42, 60, 90 and 120 days post-operative. Most animals in both groups showed no signs of inflammation and wound healing was similar. Radiographic examination revealed a gradual increase of radiopacity in the region of the defect in the control group. In the treated group, initial radiopacity was higher than that of adjacent bone, decreasing until 21 days after surgery. Then it gradually increased until 120 days after surgery, when the defect became undetectable. The results showed that Bioglass integrates into bone tissue, is biocompatible and reduced the period for complete bone regeneration.


2019 ◽  
Vol 7 (2) ◽  
pp. 66
Author(s):  
Richard Fritzgerald ◽  
Cecilia Lunardhi ◽  
Ruslan Effendy ◽  
Tamara Yuanita

Background. Root canal treatment is a main role in decreasing infection from root canal and pulp. The main cause of periapical damage mostly are bacteries. E.faecalis is a bactery that is found as an etiology of endodontic treatment failure. Cell wall of this bacteria is containing Lipoteichoic acid (LTA). LTA can penetrate into the periradicular tissue, act as endotoxin in host and cause periradicular inflammation then lead to bone destruction. LTA stimulates immunology reaction that produce Tumor Necrosis Factor alpha (TNF-α) and Transforming growth factors beta (TGF-ß). TNF-α is a main mediator and also have an important role in inflamation response otherwise TGF-ß is working as a multifunction  regulator of cell growth and differentiation during reforming and remodelling.  Purpose. The aim of this study is to know about the expression of TNF-α and TGF-ß during the periapical tissue damage due to induction of E.faecalis. Method. This study used laboratory experimental with the post test only control group design. A total of 30 male rats were randomly divided into 3 main groups, Group A (control negative) : normal tooth. Group B (control positive) : every tooth was induced only by sterile BHI-b. Group C (treated group) : every tooth  was induced by 10 μl BHI-b E.faecalis ATCC212(106 CFU). The animals were sacrificed 21 days later and prepared for histological examination of tissue damage, then we did the immunohistochemistry  followed by calculation on the light microscope. Result. The analysis revealed that the expression of TNF-α at treated group are higher than negative control and positive control but the expression of  TGF-ß at treated group are higher than the negative control group but lower than positive control. Conclusion. From this study we know that the expression of TNF-α and TGF-ß are changing during the periapical tissue damage that induced by E.faecalis.


2021 ◽  
Vol 24 (1) ◽  
pp. 1-5
Author(s):  
Olivia Avriyanti Hanafiah ◽  
Denny Satria ◽  
Avi Syafitri

Tooth extraction is a process of removing teeth from the alveolar bone. In wound healing, fibroblast are very important cells. The main purpose of this study was to determine the effect of mobe leaf 1% and 3% extract gel (Artocarpus lakoocha) on fibroblast proliferation in post extraction tooth socket wound healing. This research used 16 samples of wistar rats, divided into 4 groups, a positive control group, a negative control group and a 1% and 3% mobe leaf extract gel group. The left mandibular incisors were extracted, then 1% and 3% gels of mobe leaf extract were applied on day 1 to day 7. Data analysis was calculated using the Kruskal-Wallis test on clinical data and one way ANOVA test for microscopic. The result of the socket wound healing activity test for a good concentration of mobe leaf extract gel was 3%. This research shows significant resultith p-value of 0.018 (< 0.05) on the closure of the socket wound clinically which means the closure of the wound accelerates because of the mobe leaf 3% extract gel treatment. The distance of fibroblast on microscopically shows significant resultith a p-value of 0.002 (< 0.05), which means that there was an enlargement of the distance fibroblast at the socket wound closure with application of mobe leaf 3% extract gel. From the results of the study it can be concluded that mobe leaf 3% extract gel has the best ability to show acceleration the closure of the socket wound either clinically or microscopically.


Parasite ◽  
2019 ◽  
Vol 26 ◽  
pp. 1 ◽  
Author(s):  
Wilfried Lebon ◽  
Jacques Guillot ◽  
Maria-Jesús Álvarez ◽  
José Antonio Bazaga ◽  
Marie-Laure Cortes-Dubly ◽  
...  

In the past decade, canine thelaziosis due to Thelazia callipaeda has been diagnosed in an increasing number of European countries, with endemic areas being identified. A multi-center field trial was conducted in endemic areas in France and Spain to evaluate the efficacy of monthly administrations of the oral milbemycin oxime/afoxolaner combination (NexGard Spectra®) for the prevention of T. callipaeda infection in at-risk dogs. A total of 79 dogs negative for T. callipaeda and with a clinical history of eyeworm infection in the past two years completed the study. Dogs were randomly allocated either to a negative control group (42 dogs) or to the NexGard Spectra® treated group (37 dogs). All dogs were followed up for a 6-month period and assessed monthly for the presence of nematodes on the eyes and for the signs of ocular thelaziosis (e.g., conjunctivitis, keratitis, and ocular discharge). When the presence of nematodes was confirmed, the conjunctival fornix was flushed with a saline solution for parasite recovery and counting, and the dogs were treated appropriately. Recovered parasites were stored in 70% alcohol for subsequent morphological identification. During the course of the study, 57.1% (24/42) of the control dogs were diagnosed positive for Thelazia infection, which illustrates a high incidence rate of parasite infection. Conversely, no eyeworm was recovered from any of the 37 dogs that received NexGard Spectra®. All parasites sampled were confirmed to be T. callipaeda. This clinical field study demonstrated that monthly administrations of NexGard Spectra® provided 100% preventive efficacy against canine thelaziosis.


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