Figure 3 - uploaded by Jordan R. Yaron
Content may be subject to copyright.
Assessment of peri-wound angiogenesis in wounds treated with M-T7. (A) ELISA quantification of TNFα and VEGF in wound tissues treated with saline or M-T7 collected on days 1, 4 and 7 post-wounding normalized to total protein. Bars are mean and standard error. Statistics were calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. (B) Quantification of CD31+ cells and vessels per 20× field in the peri-wound area of wounds treated with saline or M-T7 collected on days 4 and 7 post-wounding. Bars are mean and standard error. Two non-overlapping fields were quantified per mouse and statistics were performed on the average per mouse with the N = 4 per group. Statistics were calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. (C) Representative peri-wound CD31 IHC fields (10×) collected on days 4 and 7 post-wounding. Scale bars are 50 μm. Zoom areas indicated by boxes. N = 3-4 in each group and time point.

Assessment of peri-wound angiogenesis in wounds treated with M-T7. (A) ELISA quantification of TNFα and VEGF in wound tissues treated with saline or M-T7 collected on days 1, 4 and 7 post-wounding normalized to total protein. Bars are mean and standard error. Statistics were calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. (B) Quantification of CD31+ cells and vessels per 20× field in the peri-wound area of wounds treated with saline or M-T7 collected on days 4 and 7 post-wounding. Bars are mean and standard error. Two non-overlapping fields were quantified per mouse and statistics were performed on the average per mouse with the N = 4 per group. Statistics were calculated by two-way ANOVA with Fisher's LSD post-hoc analysis. (C) Representative peri-wound CD31 IHC fields (10×) collected on days 4 and 7 post-wounding. Scale bars are 50 μm. Zoom areas indicated by boxes. N = 3-4 in each group and time point.

Source publication
Article
Full-text available
Complex dermal wounds represent major medical and financial burdens, especially in the context of comorbidities such as diabetes, infection and advanced age. New approaches to accelerate and improve, or "fine tune" the healing process, so as to improve the quality of cutaneous wound healing and management, are the focus of intense investigation. He...

Contexts in source publication

Context 1
... the context of tissue injury, TNFα is critical for the downstream production of vascular endothelial growth factor (VEGF), an essential growth factor in regulating angiogenesis [46]. We performed ELISA analyses of the healing wound's bed tissue on days 1, 4 and 7 post-wounding to quantitatively measure levels of TNFα and VEGF ( Figure 3A). Recombinant M-T7 induced a significant increase in wound bed levels of TNFα on days 1 and 4 (p < 0.05), and of VEGF by day 7 (p < 0.05), versus saline treatment alone. ...
Context 2
... M-T7 induced a significant increase in wound bed levels of TNFα on days 1 and 4 (p < 0.05), and of VEGF by day 7 (p < 0.05), versus saline treatment alone. We performed immunohistochemistry of wound tissues on days 4 and 7 post-wounding to determine the degree of angiogenesis by staining for CD31 (also called PECAM-1), a canonical marker for endothelial cells in the vasculature ( Figure 3B, C). A quantification of the number of CD31+ cells and vessels per 20× field in the peri-wound area indicated a significant increase on day 4 post-wounding (p < 0.05) versus saline treatment alone. ...
Context 3
... observed no significant difference on day 7 post-wounding. Qualitatively, we noted that the CD31+ cells formed more robust vessels in the wounds treated with M-T7, with increased length and thickness versus saline treatment alone ( Figure 3C). Taken together, these results suggest that M-T7 stimulates an early TNFα response, which stimulates a more robust VEGF response, ultimately leading to accelerated angiogenesis in the peri-wound area associated with accelerated wound closure. ...
Context 4
... found that M-T7 treatment significantly increased the accumulation of CD4+ cells in the epithelial tongue of healing wounds versus saline treatment alone ( Figure 4G,H, Figure S2). We did not observe an effect on neutrophil infiltration ( Figure S3). Thus, decoupling the chemokine-glycosaminoglycan gradient with M-T7 modulates the immune response in the wound environment to accelerate healing. ...
Context 5
... example, topical simvastatin and asperosaponin VI both act to accelerate wound healing by enlisting the VEGF signaling cascade, and VEGF-C applied directly to wounds also accelerates healing [60][61][62]. Here, we show that a recombinant M-T7 treatment resulted in a significant increase in local TNFα during the earliest stages of wound healing, which temporally transitions into a significant increase in VEGF in the healing bed ( Figure 3A). This coincides with the canonical, early inflammatory phase of healing, and the transition to the proliferation phase of healing [63]. ...
Context 6
... coincides with the canonical, early inflammatory phase of healing, and the transition to the proliferation phase of healing [63]. Accordingly, increased angiogenesis was directly observed by immunohistochemistry for CD31 in the peri-wound area ( Figure 3B,C). Thus, the data suggest that topical M-T7 modulates the chemokine environment, resulting in the augmentation of pro-healing molecules in the wound environment, engaging the pro-angiogenesis signaling cascade at the level of both cytokines and growth factors, and resulting in a significant induction of angiogenesis at the boundaries of healing wounds associated with increased wound closure. ...

Citations

... The splinted full thickness model was used study the effects of recombinant myxoma virus-derived immune modulator M-T7 [49], of Babassu oil [50], of three-dimensional printed scaffolds and electrospun mats [51], of adult gingival multipotent mensenchymal stem cells [52], of induced pluripotent stem cells clay [53], of poly(lactic-co-glycolic acid) and vascular endothelial growth factor [54], and of renal dysfunction [44] on cutaneous wound healing. ...
Article
Full-text available
A large number of models are now available for the investigation of skin wound healing. These can be used to study the processes that take place in a phase-specific manner under both physiological and pathological conditions. Most models focus on wound closure, which is a crucial parameter for wound healing. However, vascular supply plays an equally important role and corresponding models for selective or parallel investigation of microcirculation regeneration and angiogenesis are also described. In this review article, we therefore focus on the different levels of investigation of skin wound healing (in vivo to in virtuo) and the investigation of angiogenesis and its parameters.
... This finding is entirely consistent with prior research both in pre-clinical or clinical studies wherein Serp-1 demonstrated no significant toxicity and no neutralizing antibodies were detected in the Phase Further studies should also aim to investigate the molecular mechanism of action of Serp-1 in its anti-inflammatory role. Finally, a recent publication from our group demonstrated that administration of recombinant M-T7 protein (another Myxoma virus protein) could also accelerate dermal wound healing (24). This provides an additional example of a Myxoma virus derived protein in facilitating tissue repair. ...
Article
Full-text available
Purpose: Chemical corneal injuries carry a high morbidity and commonly lead to visual impairment. Here, we investigate the role of Serp-1, a serine protease inhibitor, in corneal wound healing. Methods: An alkaline-induced corneal injury was induced in 14 mice. Following injury, five mice received daily topical saline application while nine mice received Serp-1 100 μL topically combined with a daily subcutaneous injection of 100 ng/gram body weight of Serp-1. Corneal damage was monitored daily through fluorescein staining and imaging. Cross sectional corneal H&E staining were obtained. CD31 was used as marker for neovascularization. Results: Serp-1 facilitates corneal wound healing by reducing fibrosis and neovascularization while mitigating inflammatory cell infiltration with no noticeable harm related to its application. Conclusions: Serp-1 effectively mitigates inflammation, decreases fibrosis, and reduce neovascularization in a murine model of corneal injury without affecting other organs. Translational Relavence: Our study provides preclinical data for topical application of Serp-1 to treat corneal wounds.
Article
Bacterial infection is a major obstacle to the wound healing process. The hydrogel dressings with a simpler structure and good antibacterial and wound healing performance are appealing for clinical application. Herein, a robust hydrogel was synthesized from acrylamide (AM), acrylic acid (AA) and N,N′-methylene diacrylamide (MBA) via a redox initiating polymerization. The polymerization conditions were optimized to obtain the hydrogel with minimum unreacted monomers, which were 0.25% and 0.12% for AM and AA, respectively. The hydrogel had good mechanical strength, and could effectively resist damage by external forces and maintain a good macroscopic shape. It showed large water uptake capacity, and could post load a wide range of molecules via hydrogen bonding and electrostatic interaction. Loading of antibiotic doxycycline (DOX) enabled the hydrogel with good antibacterial activity against both Gram-positive bacteria and Gram-negative bacteria in vitro and in vivo. In a rat model of methicillin-resistant Staphylococcus aureus (MRSA)-infected full-thickness skin defect wound, the DOX-loaded hydrogel showed good therapeutic effect. It could significantly promote the wound closure, increased the collagen coverage area, down-regulate the expressions of pro-inflammatory TNF-α and IL-1β factors, and up-regulate the expressions of anti-inflammatory IL-4 factor and CD31 neovascularization factor.