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Preconditioning of surgical pedicle flaps with DNA plasmid expressing hypoxia-inducible factor-1α (HIF-1α) promotes tissue viability

Abstract

Ischemic necrosis of surgical flaps after reconstruction is a major clinical problem. Hypoxia-inducible factor-1α (HIF-1α) is considered the master regulator of the adaptive response to hypoxia. Among its many properties, it regulates the expression of genes encoding angiogenic growth factors, which have a short half-life in vivo. To achieve a continuous application of the therapeutic, we utilized DNA plasmid delivery. Transcription of the DNA plasmid confirmed by qRT-PCR showed significantly increased mRNA for HIF-1α in the transfected tissue compared to saline control tissue. Rats were preconditioned by injecting with either HIF-1α DNA plasmid or saline intradermally in the designated flap region on each flank. Seven days after preconditioning, each rat had two isolated pedicle flaps raised with a sterile silicone sheet implanted between the skin flap and muscle layer. The flaps preconditioned with HIF-1α DNA plasmid had significantly less necrotic area. Angiogenesis measured by CD31 staining showed a significant increase in the number of vessels per high powered field in the HIF-1α group (p < 0.05). Our findings offer a potential therapeutic strategy for significantly promoting the viability of surgical pedicle flaps by ischemic preconditioning with HIF-1α DNA plasmid.

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Fig. 1: The surgical flap model used for the evaluation.
Fig. 2: Efficacy of dermal DNA delivery.
Fig. 3: HIF-1α mRNA expression was increased by plasmid transfection.
Fig. 4: Photographic assessment of necrosis.
Fig. 5: Histology of pedicle skin flap tissue.
Fig. 6: Measurement of H&E score of skin flaps based on H&E scoring table.
Fig. 7: NTC9385 CA5-HIF-1α DNA plasmid transfection increased CD31+ vessel density in skin flaps on day 14.

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Acknowledgements

Funding for this work was provided by the Hendrix Burn/Wound Fund, Johns Hopkins Bayview Medical Center, Johns Hopkins University.

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Correspondence to Kai-Hua Chang or John W. Harmon.

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JWH, GPM, and FL are founders of Canton Biotechnologies Inc. (CBI), Baltimore, MD. CBI is a biotech company embedded in Johns Hopkins, founded to develop the plasmid DNA delivery of HIF for wound repair and regeneration.

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Chang, KH., Shoureshi, P., Lay, F. et al. Preconditioning of surgical pedicle flaps with DNA plasmid expressing hypoxia-inducible factor-1α (HIF-1α) promotes tissue viability. Gene Ther 28, 319–328 (2021). https://doi.org/10.1038/s41434-020-00199-6

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