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Matrix metalloproteinase-inspired suicidal treatments of diabetic ulcers with siRNA-decorated nanofibrous meshes

Gene Therapy volume 20, pages 378–385 (2013)Cite this article

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Abstract

Matrix metalloproteinase (MMP)-assisted siRNA treatment was accomplished with a nanofibrous matrix for suicidal gene therapy of diabetic ulcers. We fabricated a MMP-responsive nanofibrous matrix to control release of small interfering RNA (siRNA) in response to a high concentration of MMPs in diabetic ulcers. For MMP-responsive release of siRNA, linear polyethyleneimine (LPEI) was chemically conjugated on the surface of the nanofibrous matrix via a MMP-cleavable linker. To control the abnormally elevated MMP-2 expression in diabetic ulcers, MMP-2 siRNA was electrostatically incorporated into LPEI-immobilized nanofibrous meshes with various nitrogen/phosphate (N/P) ratios. The release profiles of siRNA and LPEI were monitored to confirm that MMP responsiveness of the matrix and MMP-2 significantly increased the release of both siRNA and LPEI for 72 h. The released fractions were transfected to dermal fibroblasts. Quantitative reverse transcription (qRT)-PCR for endogenous MMP-2 expression confirmed that the gene-silencing effects of siRNA were dependent on the charge ratio of LPEI to siRNA on the mesh. Diabetic animals with dorsal burns were treated with siRNA-incorporated nanofibrous mesh for 7days. siRNA-incorporated nanofibrous meshes dramatically increased the MMP-2 gene-silencing effects of the siRNA and neo-collagen accumulation at the wound sites. RT-PCR also confirmed the highest expression levels of the keratinocyte-specific markers and the lowest expression levels of MMP-2 in the nanofibrous mesh-treated groups, suggesting that wound recovery is restored to normal levels. The wound recovery rates of diabetic ulcers were significantly increased when siRNA-incorporated nanofibrous meshes were administered. Thus, the suicidal treatment with the MMP-2 siRNA-decorated nanofibrous mesh is expected to improve prognosis of diabetic ulcers with reduced side effects.

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Acknowledgements

This research was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (grant# 2012005857).

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  1. Department of Biomaterials Engineering, School of Bioscience and Bioengineering, Kangwon National University, Chuncheon, Republic of Korea

    H S Kim & H S Yoo

  2. Institute of Bioscience and Bioengineering, Kangwon National University, Chuncheon, Republic of Korea

    H S Yoo

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Correspondence to H S Yoo.

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Kim, H., Yoo, H. Matrix metalloproteinase-inspired suicidal treatments of diabetic ulcers with siRNA-decorated nanofibrous meshes. Gene Ther 20, 378–385 (2013). https://doi.org/10.1038/gt.2012.49

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