Adenoviral Gene Delivery to Primary Human Cutaneous Cells and Burn Wounds

Hirsch, Tobias; Peter, Sebastian von; Dubin, Grzegorz; Mittler, D.; Jacobsen, Frank; Lehnhardt, M.; Eriksson, Elof; Steinau, Hans-Ulrich; Steinstraesser, Lars

doi:10.2119/2006-00031.hirsch

Cited by 30 publications

(24 citation statements)

References 42 publications

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“…To address this issue, we have utilized a murine model in which uniform, graded severity of thermal injury could reliably be produced. The branding iron type burn (9, 20) was chosen over scald or flame burn (21–23). We prefer this model because it allowed us to produce burns of graded severity.…”

Section: Introductionmentioning

confidence: 99%

Association of Increasing Burn Severity in Mice With Delayed Mobilization of Circulating Angiogenic Cells

et al. 2010

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Angiogenesis is an important component of wound healing. Mobilization of circulating angiogenic cells (CACs) has been observed in patients with cutaneous burn wounds, but a systematic exploration of the phenomenon in an animal model has not been carried out. Using a murine model, in which burn depth can be varied precisely, and a validated culture method for quantifying circulating CACs, we found that increasing burn depth resulted in a progressive delay in the time to mobilization of circulating CACs. This delay in CAC mobilization was associated with a delay in perfusion and vascularization of the burn wound tissue. Analysis of CACs in the peripheral blood of human burn patients, using the same culture assay, confirmed results previously obtained by flow cytometry, that CAC levels peak early after the burn wound, and point to the clinical relevance of findings from the murine model.

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Section: Introductionmentioning

confidence: 99%

Association of Increasing Burn Severity in Mice With Delayed Mobilization of Circulating Angiogenic Cells

et al. 2010

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“…To date however, the deployment of gene-based medicines in conjunction with these grafts has not been fully exploited in the post-genomic era. Accordingly, while the benefit of specific cellular and acellular graft matrices is clinically accepted [1–3], the delivery of genes to the wound bed has largely focused on strategies using incisional and punch wounds [4–9] that are distinct from accepted clinical protocols for severe burn injury and wound healing. With the widespread use of biosynthetic grafts, advances in noninvasive imaging, and a better understanding of the cell biology of synthetic graft models [2,10], we are now able to better examine gene delivery to the wound bed through quantitative and noninvasive techniques.…”

Section: Introductionmentioning

confidence: 99%

The noninvasive, quantitative, in vivo assessment of adenoviral-mediated gene delivery in skin wound biomaterials

et al. 2009

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Because there are few reports using gene delivery in clinically-approved synthetic matrices, we examined the feasibility of using a noninvasive imaging system to study the kinetics of luciferase gene expression when delivered in an adenoviral vector. Using a mouse model of full thickness injury, we quantified the kinetics of gene expression, determined the optimal dose of particle delivery, and established the temporal importance of drug delivery in obtaining optimal gene expression. Specifically, we found that the ideal time to deliver adenovirus to a graft is during the early phase of graft wound closure (days 0-3 post-operatively) for a peak of gene expression to occur 7 days after delivery. Under these conditions, there is a saturating dose of 6 × 10 8 adenoviral particles per graft. In light of these findings, we examined whether the efficacy of delivery could be increased by modulating the composition of the grafts. When a collagen gene-activated matrix (GAM) containing basic fibroblast growth factor (FGF2) was compared to matrix alone, a significant increase in gene expression is observed when identical amounts of vector are delivered (p < 0.05). Taken together, these results show how a noninvasive and quantitative assessment of gene expression can be used to optimize gene delivery and that the composition of matrices can dramatically influence gene expression in the wound bed.

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“…Our previous data supports that adenoviral-mediated gene therapy efficiently delivers the transgene and significantly improves neovascularization and wound healing outcomes 2,56 . An advantage of the adenoviral mediated gene transfer is that the vector begins to work within 48 hours and has transgene expression decreasing from 2 to 7 days with no significant expression after 14 days 57 . In support, several studies have shown that growth factor and/or transgene delivery during the initial critical phase of wound healing may be sufficient to correct the neovascularization and wound healing deficit and facilitate favorable wound healing progression.…”

Section: Discussionmentioning

confidence: 99%

Adenoviral-mediated gene transfer of insulin-like growth factor 1 enhances wound healing and induces angiogenesis

Balaji

LeSaint

Bhattacharya

et al. 2014

Journal of Surgical Research

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Background Chronic wounds are characterized by a wound healing and neovascularization deficit. Strategies to increase neovascularization can significantly improve chronic wound healing. Insulin like growth factor (IGF-1) is reported to be a keratinocyte mitogen and is believed to induce angiogenesis via a vascular endothelial growth factor (VEGF) dependent pathway. Using a novel ex vivo human dermal wound model and a diabetic impaired wound healing murine model, we hypothesized that adenoviral over expression of IGF-1 (Ad-IGF-1) will enhance wound healing and induce angiogenesis through a VEGF dependent pathway. Methods Ex vivo: 6 mm full thickness punch biopsies were obtained from normal human skin, and 3 mm full thickness wounds were created at the center. Skin explants were maintained at air liquid interface. Db/db murine model: 8 mm full thickness dorsal wounds in diabetic (db/db) mice were created. Treatment groups in both human ex vivo and in vivo db/db wound models include 1×108 PFU of Ad-IGF-1 or Ad-LacZ, and PBS (n=4–5/group). Cytotoxicity (LDH) was quantified at days 3, 5 and 7 for the human ex vivo wound model. Epithelial gap closure (H&E; Trichrome), VEGF expression (ELISA) and capillary density (CD 31+ CAPS/HPF) were analyzed at day 7. Results In the human ex vivo organ culture, the adenoviral vectors did not demonstrate any significant difference in cytotoxicity compared to PBS. Ad-IGF-1 over expression significantly increases basal keratinocyte migration, with no significant effect on epithelial gap closure. There was a significant increase in capillary density in the Ad-IGF-1 wounds. However, there was no effect on VEGF levels in Ad-IGF-1 samples compared to controls. In db/db wounds, Ad-IGF-1 over expression significantly improves epithelial gap closure and granulation tissue with a dense cellular infiltrate compared to controls. Ad-IGF-1 also increases capillary density, again with no significant difference in VEGF levels in the wounds compared to control treatments. Conclusions In two different models, our data demonstrates that adenoviral mediated gene transfer of IGF-1 results in enhanced wound healing and induces angiogenesis via a VEGF-independent pathway. Understanding the underlying mechanisms of IGF-1 effects on angiogenesis may help produce novel therapeutics for chronic wounds or diseases characterized by a deficit in neovascularization.

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Adenoviral Gene Delivery to Primary Human Cutaneous Cells and Burn Wounds

Cited by 30 publications

References 42 publications

Association of Increasing Burn Severity in Mice With Delayed Mobilization of Circulating Angiogenic Cells

Association of Increasing Burn Severity in Mice With Delayed Mobilization of Circulating Angiogenic Cells

The noninvasive, quantitative, in vivo assessment of adenoviral-mediated gene delivery in skin wound biomaterials

Adenoviral-mediated gene transfer of insulin-like growth factor 1 enhances wound healing and induces angiogenesis

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