In this paper, we aimed at investigating antioxidant activity of waste extracts from different medicinal plants, endemic of the Mediterranean area, on fibroblast proliferation and wound healing. (I) In vitro wound-healing assay of mouse keratinocytes (at 16 h of wound healing) in the treatments in F. (J) The y axis shows the percentage of area covered (at 16 h of wound healing) by keratinocytes transfected with mock, scramble siRNA (si Scra), or Slug siRNA (si Slug) oligos for 2 d and incubated in 6-d WT or VIM−/− MDF-conditioned medium. Vimentin reconstitution in VIM−/− fibroblasts restored both their proliferation and TGF-β1 production. Botanical.com. J. Phytomedicine. Maioli M. et al. 2013;12:189, 11. The column was equilibrated for 15min before the next sample injections. Bookshelf Fibroblast Proliferation and Migration in Wound Healing by Phytochemicals: Evidence for a Novel Synergic Outcome. Mouse skin keratinocytes were isolated from WT newborn mice and cultured in FAD medium (DMEM/Ham’s F-12 Nutrient Mixture; 3.5:1.1) supplemented with 10% (vol/vol) Chelex-treated FCS (Invitrogen), 0.18 mM adenine, 0.5 μg/mL hydrocortisone, 5 μg/mL insulin, 100 pM cholera toxin (all from Sigma), 10 ng/mL EGF, 100 U/mL sodium pyruvate, 100 μg/mL penicillin/streptomycin, and 2 mM Glutamax (all from Invitrogen) in 5% CO2 and 32 °C. The skin is the ideal organ to study cellular heterogeneity … Sirius Red, Fast Green, and picric acid (all from Sigma) were used in Picro-Sirius Red staining for histological visualization of collagen fibers in tissue sections under polarized light microscopy (33) at 20× magnification, and pictures were taken at a fixed exposure. 2013;7:1063-1073, 42. TGF-β has been shown to modulate integrin expression of keratinocytes toward a more migratory phenotype (34), suggesting its supportive role in reepithelialization. Burns or excisional skin wounds in mice are common experimental approaches to assess molecular, cellular, and tissue movements associated with repair. Bars indicate the mean fold changes ± SEM relative to day 0 WT; **P < 0.01; n = 3. The contents of the active ingredients were expressed as milligrams of active ingredient per L of extract. As reported in Table 1, total phenolic amounts, expressed as GAE were higher in extracts from L and H as compared to C. If we considered the differences between linear regression, the comparison of slopes, in C (14.60 ± 0.06) (light blue line), in H (24.74 ± 3.11) (pink line) and in L (26.55 ± 8.49) (purple line) were not significantly different. Reactive oxygen species (ROS) at high concentration counteract wound healing processes, due to cellular membranes damages. Slug siRNAs (ON-TARGET plus Mouse Snai2 siRNA-SMARTpool; Dharmacon) and scrambled siRNAs (AllStars Negative Control; Qiagen) were used in this study. S4 B and C), there was no significant change in the myeloperoxidase activity of neutrophils (MPO+) in WT and VIM−/− wounds at day 9 postinjury (Fig. Actually, after volatile oils removal, waste plant materials are still abundant in phenolic compounds, oligomers and flavonoid glycosides with various biological activities [28], therefore these compounds could still have an application [29], [30]. However, in cells cultured with C waste extract cell viability was significantly induced after 96-120 hours of culturing, as compared to control untreated cells, being more effective at a lower concentration (1 µL/mL) (Figure 3, panel d-e). 2019;24(8):1515, 49. The discoloration percentage at 734 nm was calculated for each point; the antioxidant capacity of the test compound was expressed in percent inhibition (%), and the IC50 value, the amount of an extract that neutralize 50% of the radical, was calculated from regression analysis and expressed as mean ± SD. Food Chem. (D) Quantification of average epidermis thickness on day 15 after burn injury. Pharm. L and also H extracts exhibited a strong antioxidant activity, significantly higher (P<0.01) as compared to the effect elicited by C, along all the experimental time points of the ABTS assay (0-50 minutes, Figure 2 panel a). Der Pharm. Reepithelialization was defined as the area within the pan-keratin marks that was covered with epithelium. Miraj S. Lavandula stoechas l: A systematic review of medicinal and molecular perspectives. Found insideListing 294 products in alphabetical order, this handy product guide for wound care practitioners across all practice settings offers detailed information—sizes, action, indications, contraindications, application, and removal ... Nym. Fibroblasts play an important role in this process and participate in transformation into myofibroblasts, the synthesis of extracellular matrix (ECM) and fibers, and the secretion of a variety of growth factors. A circular reference was placed alongside to permit correction for the distance between the camera and the animals. S2 A–D). Cells cultured with 1 μL/mL CH or CLH exhibited a complete wound closure within 48h, earlier than in control untreated cells. var thismailbody2 = ""; (A and B) Representative pictures of confocal images of keratin 6 (green) and DAPI (red) (A) and keratin 14 (green) and DAPI (red) (B) in VIM−/− and WT wounds on day 9 and 15 after skin burn injury. (B) Quantitation of pan-keratin+ cell colony numbers in dermis–epidermis interface regions on day 9 postinjury. Loss of vimentin disrupts this coordination, leading to slow, poor, and incomplete wound healing. Traditional and folk medicinal uses include treatment of skin problems, fevers, chronic infections, wounds, bites and stings [14], [15]. Božin BN, Lakić NS, Mišan A. et al. Media was removed and cells were washed in PBS before adding the medium conditioned with different concentrations (1, 5, or 10 µL/mL) of each waste extracts of C, or L, or H, or with the combination of 2 or 3 plants waste extracts (CLH, CL, CH, LH). Int. Phenolic profile, antioxidant and antimicrobial activity of oregano (Origanum vulgare L, Lamiaceae) postdistillation waste extracts. Ind. a dynamic skin reparative process that results in a sequence of events, including inflammation, proliferation, and migration of different cell types as fibroblasts. Melatonin and vitamin D interfere with the adipogenic fate of adipose-derived stem cells. Although on day 9 postinjury, 1.5-fold fewer neutrophil cells had infiltrated to the edges of WT wounds than to the edges of VIM−/− wounds (Fig. The ABTS·+ solution was diluted with bi-distillated water to an absorbance of 0.7 (±0.02) at 734 nm. 2020 Dec 17;2020:8878370. doi: 10.1155/2020/8878370. Wound-healing is a dynamic process, which, following skin injury, arises through a sequence of events, including inflammation, proliferation, and migration of different cell types as fibroblasts [1], [2]. Chevallier A. Encyclopedia of Medicinal Plants. Within this context, identifying tools able to implement cell proliferation and migration could improve tissue regeneration. b) Migration of fibroblasts after scratch and treatment with 1 μL/mL CLH extracts. 2E). DISCUSSION AND CONCLUSIONS. Food Chem. Maddaluno L, Urwyler C, Werner S. Fibroblast growth factors: Key players in regeneration and tissue repair. We also show (Table 5) a positive Spearman correlation between antioxidant activity measured by ABTS and total phenolic contents (P<0.05). In B and D data are shown as means ± SEM; n = 3; *P < 0.05; **P < 0.01. The final concentration of combined extracts was 38.90 µg/µL for CLH, 45.32 µg/µL for CL, 33.25 µg/µL for CH and 38.13 µg/µL for LH. Thank you for your interest in spreading the word on PNAS. A Review of the Contribution of Mast Cells in Wound Healing: Involved Molecular and Cellular Mechanisms. 2014;808:25-39, 44. performed research; J.I. However, there is very limited information on the wound-healing effect of proteins in human cord blood plasma exosomes (CBPexo). 6B). thismailbody3 = thismailbody1; The normal wound healing involves three successive but overlapping phases, including hemostasis/inflammatory phase, proliferative phase, and remodeling phase. Vimentin heterozygous mice (129/Sv × C57BL/6) were used to generate vimentin-deficient homozygotes (VIM−/−) and WT offspring (20). The experiments were performed a few days after transfection as indicated in Results and in the figure legends. Sasidharan S, Chen Y, Saravanan D. et al. D, dermis region; S, scab region. 6 C and D). It has been hypothesized that fibroblasts play an active role. The active fibroblast TGF-β induces Slug–EMT signaling in keratinocytes, promoting EMT-like transdifferentiation and keratinocyte migration. A new approach to discriminate Rosmarinus officinalis L. plants with antioxidant activity, based on HPTLC fingerprint and targeted phenolic analysis combined with PCA. This hypothesis is supported by our preliminary observations showing that, although complete wound closure seemed to be achieved eventually (>20–25 d after wounding) in VIM−/− mice, the reepithelialization was still incomplete, as already reflected in the weak reepithelization of Vim−/− skin at day 15 postinjury. Variability of chemical composition and antioxidant activity of essential oils between Myrtus communis var. Recent Advances: The amniotic fluids … Agyare C, Amuah E, Adarkwa-Yiadom M. et al. The figures are representative of different independent experiments. favour and accelerate wound closure. Data are shown as means ± SEM; n = 3. Vimentin-deficient wounds were conspicuously devoid of collagen accumulation, apparently as a direct consequence of the severe inhibition of fibroblast proliferation and expansion. Fibroblasts are known to play an essential role in wound healing, and cell migration is one of the most essential steps, during the proliferative phase, responsible for wound closure [1], [21], [64], [65]. Chemistry and pharmacology of saponins: special focus on cytotoxic properties. Quantification of collagen was performed as previously described (39) using Image Pro Plus version 4.5.0.29 imaging software (Media Cybernetics Inc.). Data are shown as means ± SEM; n = 12. To diminish the variance within samples, at least 10 consecutive sections from each wound sample and three to six samples per time point were counted, as indicated in the figure legends. PMC 2016;94:665-672, 33. (B) Representative confocal pictures of immunofluorescent labeling of DAPI (blue), keratin 5 (green), and vimentin (red) (Left) or keratin 14 (green) and N-cadherin (red) (Right) from isolated mouse keratinocytes. (B) Representative pictures of wounds in WT and VIM−/− mice 15 d after excisional injury. European Surgical Research. Mass Spectrom. 4C). (Scale bars, 20 μm.) (Scale bar, 100 μm.) In order to measure the antioxidant activity of waste extracts and their synergic effects, 100 µl of aqueous extracts solution at various concentrations (from 0.1 to 100 µg/mL) were added to 900 µL of diluted ABTS·+ and the absorbance recorded at time zero and after 50 min at which point the absorbance was stable. Plast Reconstr Surg Glob Open. thismailbody4 = thismailbody4 + ""; 2005;41:35-40, 13. bFGF is a well-known member of the FGF family protein that modulates the growth, differentiation, migration and survival of a wide variety of cell types . 2020 Sep 26;25(19):4422. doi: 10.3390/molecules25194422. The central concept in this study is that a major cytoskeletal component, vimentin, acts as a signal integrator during wound healing, operating in both signal-triggering and signal-receiving cells. Mitomycin C (10 μg/mL) was included in the medium to prevent cell proliferation. During the formation of granulation tissue in a dermal wound, platelets, monocytes and other cellular blood constituents release various peptide growth factors to stimulate fibroblasts to migrate into the wound site and proliferate, in order to reconstitute the various connective tissue components. Interestingly, after 15 d of recovery, Slug expression in VIM−/− skin was restored to the same levels as in WT skin. 4B). By day 15 postinjury, keratinocyte colonies had fused successfully with the newly formed stratified mature epidermis in WT wounds. 4B), reinforcing the notion that TGF-β signaling is a major driver in keratinocyte transdifferentiation and migration induced by fibroblast vimentin. A study estimates that there are 50 billion birds in the world, with the majority in palearctic and nearctic realms. original wound. Crops Prod. S2). 2019(6):1-12, 50. This revised second edition contains several new chapters on lymphoedema, nutrition, skin care, continence, and scarring. Than cells were cultured with a medium conditioned with different concentrations (1, 5, 10 µL/mL) of each waste extracts of C, or L or H, or with the combination of 2 or 3 plants waste extracts (CLH, CL, CH, LH), for 24h, 48h, 72h, 96h or 120 hours. The right lower corner of each panel shows an enlarged image of the area in the white box. The pipeline detects the nuclei from the Hoechst channel and counts them. The growth medium of mouse keratinocytes was replaced with the MDF-conditioned medium for 5 d. Cell lysates were collected and blotted with antibodies against desmoplakin, E-cadherin, N-cadherin, vimentin, Slug, and loading control GAPDH. Plants. Cells were washed three times with TBS and were lysed in SDS/PAGE loading buffer. (A) Representative pictures showing immunohistochemical labeling of pan-keratin in WT and VIM−/− wounds on days 9 (D9) and 15 (D15) postinjury. Correspondingly, the levels of active TGF-β1 in the supernatants of VIM−/− MDFs were significantly lower than in the supernatants of WT MDFs (Fig. It was destroyed by acid hydrolysis. Data are shown as means ± SEM; n = 3. Table 4 (DA FARE) shows quantitative results of pool of CLH, CL, CH and LH. 2008;119(1):141-144, 23. Immunohistochemical analysis of the expression of Collagen type I and type III was assessed in fibroblasts after scratch and treatment with different concentrations of single waste extract (C, L, H) or pooled waste extracts (CLH, CL, CH, LH) for 72 hours. Phenolic compounds quantified in CLH, CL, CH and LH decoction water. Image credit: Luca Bindi and Paul J. Steinhardt. (£) Caffeoylquinic acid derivatives were expressed as chlorogenic acid. Adv. Ihn H, Kikuchi K, Soma Y, Sato S, Fujimoto M, Tamaki T, Igarashi A, Takehara K. Acta Derm Venereol. Fibroblasts play a crucial role in tissue repair, from the late inflammatory phase until the fully final epithelization of the injured tissue, by secreting growth factors, cytokines, collagens and others extracellular matrix (ECM) components [3]-[6]. Vladić J, Zeković Z, Jokić S. et al. The results are expressed as the mean ± SEM. Results are expressed as mean ± standard deviation of 3 determinations. 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