Modulation of scarring in a liquid environment in the Yorkshire pig
Richard G. ReishBaraa ZuhailiJuri BergmannPejman AflakiTaro KoyamaFlorian HacklEmily WaisbrenJosé A. CansecoKapil VermaElof ErikssonFeng Yao
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ABSTRACT Decreased inflammatory response seen in wet wound healing may be correlated with diminished scarring. This study seeks to test this hypothesis and to validate a model of scarring in the Yorkshire pig. Four Yorkshire pigs were used to create 36 dorsal wounds per pig (144 wounds total) in the following groups: full-thickness excisional, partial thickness, meshed split-thickness skin grafts, sheet split-thickness skin grafts, minced skin, and incisional wounds. Wounds were randomized into wet and dry groups. Wet wounds were enclosed in polyurethane chambers with 2 mL of normal saline. Dry wounds were covered with regular gauze. Terminal biopsies were performed at 72 hours and day 28. Histology demonstrated significantly less inflammatory infiltrate, thicker neoepidermis, more pronounced rete ridge formation, and decreased scar tissue thickness in wet wounds. The mean macroscopic scar surface area was significantly decreased in full-thickness excisional wet wounds compared with dry wounds (61.2 mm(2) vs. 150.8 mm(2), p<0.01). Hydroxyproline content was decreased in full-thickness wet compared with dry groups (44.81 vs. 62.21 mg/g, p<0.01). Tensile strength was 90% greater in full-thickness wet compared with dry groups (p<0.01). Healing in the liquid environment significantly reduced scar formation. This model will allow for future investigation of high-concentration topical scar-modulating agents in the liquid environment.Keywords:
Hydroxyproline
Histology
Diabetes-impaired wound healing and other tissue abnormalities are considered to be a major concern.The aim of this study is to assess the wound healing activity of the methanolic extracts of Berula angustifolia leaves.Seven- week-old male Wistar rats with diabetes induced by streptozotocin injection were randomized into 5 groups of 6 rats based on allocated treatment. Wounds were created by an excision-based or incision-based wound model. For wound healing activity, the extracts were applied topically in the form of ointment and compared with the control groups. The healing of the wound was assessed based on excision, incision, hydroxyproline estimation, biomechanical, and biochemical studies.The healing rate of the extract-treated groups was significantly different compared with the control group (P < .05). Hydroxyproline contents increased significantly in the extract-treated groups (P < .05). There were significant differences in the extract-treated versus nonextract-treated groups, particularly in terms of cellular infiltration, acute hemorrhage, congestion, edema, collagen production and density, reepithelialization, and neovascularization.The methanolic extract of B angustifolia enhances wound healing activity significantly in both studied wound models. From this animal study, enhanced wound contraction, decreased epithelialization time, increased hydroxyproline content, improved mechanical indices, histological characteristics, and biochemical studies suggest the extract of B angustifolia leaves may have therapeutic benefits in diabetes-impaired wound healing.
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The methanolic extract of dried pomegranate (Punica granatum) peels showed the presence of a high content of phenolic compounds (44.0%) along with other constituents. This extract was formulated as a 10% (wt/wt) water-soluble gel and was studied for its wound healing property against an excision wound on the skin of Wistar rats. The activity was compared with that of a commercial topical antibacterial applicant. The wound healing activity was assessed by measuring the percent contraction in skin and estimation of collagen content in terms of hydroxyproline content. Healed skin was also subjected to histopathological studies to examine the microscopic changes. The animals treated with 2.5% gel showed moderate healing (55.8% and 40.8% healing compared with negative and positive controls, respectively), whereas the group treated with 5.0% gel showed good healing (59.5% and 44.5% healing compared with negative and positive controls, respectively). The amount of hydroxyproline increased by twofold in the group treated with 5.0% gel. Histopathological studies also supported the wound healing on application of the gels. The group of rats that received 5.0% gel showed complete healing after 10 days, whereas in rats treated with 2.5% gel, healing was observed on day 12, in contrast to the positive control animals receiving the blank gel, which took 16-18 days for complete healing. The results of this study may be extended to different types of wounds so that the formulation could be exploited to develop it as a topical dermatological formulation. High-performance liquid chromatography analysis of the extract showed the presence of gallic acid and catechin as major components.
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The effects of fibronectin (Fn) on burn healing were observed in mice. A split thickness burn covered 7% approximately 8% TBSA was created on the back of mouse. Experimentally 0.05ml of Fn (0.5mg/ml) was topically applied daily. Wound healing time and healing rate were determined. On 5 and 10 postburn days, biopsy sample of wound tissue was obtained. Total protein, DNA, and hydroxyproline contents were measured according to Bradford method, Hoechest 33258 dye fluorometry, and routine measure respectively. The mean wound healing time in mice treated with Fn was 18.67 +/- 4.50 days compared to 22.89 +/- 2.85 days in the controls. The wound healing rate was significantly higher in the Fn-treated mice. The contents of wound tissue DNA and hydroxyproline in Fn group were significantly increased on day 5, and total protein was significantly increased on day 10. This study indicated that topical application of Fn is beneficial to wound healing.
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The present study was conducted to investigate the histological changes and wound healing effect of aqueous extract of Elaeagnus angustifolia. After creating full-thickness skin wounds on the back of 45 male Sprague-Dawley rats they were randomly divided into three groups. Treated group received the extract, positive control group were treated with mupirocin ointment 2% and control group did not receive any treatment. Wound healing rates were calculated on days 3, 5, 8, 10, 12 and 15 post-wounding and the wound tissues were harvested at 5, 10, and 15 days for histological analysis and hydroxyproline content measurement. The results indicated a significant increase in the percentage of wound contraction and hydroxyproline content in the treated group comparing to the control and positive control groups. A significant increase in the assigned histological scores was observed at 10 and 15 days in the treated and positive control groups compared to the control group. The results demonstrate that aqueous extract of Elaeagnus angustifolia accelerates cutaneous wound healing, and its effect may be due to the increased re-epithelialization and collagen deposition in wound and so it can be considered as a therapeutic agent for wound healing.
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Background: Wound healing is a complex process, dependent on available nutrition substrates. When used together with β‐hydroxy β‐methylbutyrate, arginine and glutamine have been shown to increase collagen deposition in human subjects. However, there are no experimental investigations on the influence of this amino acid mixture with regard to secondary wound healing. The aim of this study is to investigate the effects of the supplementation of these 3 amino acids on the healing of open wounds in otherwise healthy animals. Materials and Methods : Twelve rats were divided into control and treatment groups. Two 2‐cm × 1‐cm full‐thickness skin defects were prepared on each subject. The rats in both groups received a diet containing 1.2 g of protein per 100 g of body weight per day. The treatment group, in addition, received 200 mg/kg L‐arginine, 200 mg/kg L‐glutamine, and 40 mg/kg β‐hydroxy β‐methylbutyrate every day. Wound sizes were measured every 2 days. On the 10th day, tissue samples were taken for histopathologic evaluation and also for the measurement of hydroxyproline concentrations. Results : There was no statistically significant difference between mean wound sizes for the 2 groups ( P > .05). There was also no statistically significant difference between the groups with regard to histological healing parameters (reepithelialization [ P = 1.00], granulation tissue [ P = 1.00], collagen accumulation [ P = .455], inflammatory cell accumulation [ P = .455], angiogenesis [ P = .242]) or tissue hydroxyproline concentrations ( P = .240). Conclusion : Diet supplemented with arginine, glutamine, and β‐hydroxy β‐methylbutyrate is not beneficial in enhancing secondary healing of open wounds in rats. Further research regarding this topic is warranted.
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In an experimental study the local effect of i.v. applied xantinol-nicotinate (Complamin) on healing skin incisions in rats was studied. In conclusion, there was a higher ultimate tensile strength of wounds treated by the agent, based on the increased synthesis of collagen fibres. This statement was proved by the alteration of free hydroxyproline in serum. Histologically, there was an intensive vascularization of the wound healing area. Concerning the elastic behaviour we found a more rigid regenerative tissue in the early phases of healing on account of the enhanced thickness of the wound area.
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Ethanolic extract of leaves of O. sanctum was investigated for normal wound healing and dexamethasone depressed healing using incision, excision and dead space wound models in albino rats. The extract of O. sanctum significantly increased the wound breaking strength in incision wound model. The extract treated wounds were found to epithelialize faster and the rate of wound contraction was significantly increased as compared to control wounds. Significant increase in wet and dry granulation tissue weight, granulation tissue breaking strength and hydroxyproline content in dead space wound model was observed. The extract significantly decreased the antihealing activities of dexamethasone in all the wound models. The results indicated that the leaf extract promotes wound healing significantly and able to overcome the wound healing suppressing action of dexamethasone. Histological examination of granulation tissue to determine the pattern of lay-down for collagen confirmed the results.
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Objective By detecting the influence of hydroxyproline on collagen metabolism of wound healing of rabbit ear to investigate the inhibition effect of hyaluronic acid on wound healing.Methods Wound healing model were made on the ears of 18 rabbits.They were randomly divided into 2%HA treated-group(groupA),1%HA treated-group(groupB) and normal saline treated-group(groupC).The wound healing time and scare formation were observed and recorded.The wound area skin was obtained at different days after operation.Tissue hydroxyproline was detected.Data was analyzed statisticaly.Results The hydroxyproline content in group A and B were lower than group C(P0.01).The difference between group A and B was also prominent(P0.05).The wound healing time of group A and B were later than group C,the scars also smaller than group C.Conclusion Hyaluronic acid can inhibit the collagen synthesis,delay the wound healing,reduce the scar formation.The dependence relationship between effect and dosage is notable.
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The objective of this study was to evaluate the wound healing potential of L-glutamine in laboratory rats using excision and incision wound models. Excision wounds of size 500 mm(2) and depth 2 mm were made on the dorsal portion of male Wistar rats (230-250 g) and were used for the study of oral L-glutamine (1 g/kg) treatment on the rate of contraction of wound and epithelisation. Histological evaluation of wound tissue was also performed. Six-centimetre-long two linear-paravertebral incisions in male Wistar rats (230-250 g) were used to study the effect of L-glutamine (1 g/kg, p.o.) treatment on tensile strength, total protein and hydroxyproline content in the incision model. Oral administration of L-glutamine (1 g/kg) significantly decreased wound area, epithelisation period and wound index, whereas the rate of wound contraction significantly increased (P < 0·001) when compared with vehicle control rats in the excision wound model. Tensile strength, hydroxyproline content and protein level were significantly increased (P < 0·001) in L-glutamine (1 g/kg, p.o.)-treated rats when compared with vehicle control rats in the incision wound model. Histological evaluation of wound tissue from L-glutamine (1 g/kg, p.o.)-treated rats showed complete epithelialisation with new blood vessel formation and high fibrous tissues in the excision wound model. In conclusion, oral administration of l-glutamine (1 g/kg) promotes wound healing by acting on various stages of wound healing such as collagen synthesis, wound contraction and epithelialisation.
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