Matrix metalloproteinase-inspired suicidal treatments of diabetic ulcers with siRNA-decorated nanofibrous meshes.
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.
Department of Biomaterials Engineering, School of Bioscience and Bioengineering, Kangwon National University, Chuncheon, Republic of Korea.
H S
HS
Kim
H S
HS
Yoo
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002056:Burns / Q000628:therapy; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D017719:Diabetic Foot / Q000628:therapy*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D020868:Gene Silencing; D015316:Genetic Therapy; D020778:Matrix Metalloproteinase 2 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D057139:Nanofibers / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic; D014945:Wound Healing
D001672:Biocompatible Materials; D034741:RNA, Small Interfering; D003094:Collagen; D020778:Matrix Metalloproteinase 2