Tissue Eng Regen Med.  2018 Dec;15(6):711-719. 10.1007/s13770-018-0166-2.

Local Silencing of Connective Tissue Growth Factor by siRNA/Peptide Improves Dermal Collagen Arrangements

• Cho Lee AR
• Woo I

• KMID: 2441263
• DOI: http://doi.org/10.1007/s13770-018-0166-2

Abstract

BACKGROUND
Collagen organization within tissues has a critical role in wound regeneration. Collagen fibril diameter, arrangements and maturity between connective tissue growth factor (CTGF) small interfering RNA (siRNA) and mismatch scrambled siRNA-treated wound were compared to evaluate the efficacy of CTGF siRNA as a future implement for scar preventive medicine.
METHODS
Nanocomplexes of CTGF small interfering RNA (CTGF siRNA) with cell penetrating peptides (KALA and MPGΔNLS) were formulated and their effects on CTGF downregulation, collagen fibril diameter and arrangement were investigated. Various ratios of CTGF siRNA and peptide complexes were prepared and down-regulation were evaluated by immunoblot analysis. Control and CTGF siRNA modified cells-populated collagen lattices were prepared and rates of contraction measured. Collagen organization in rabbit ear 8 mm biopsy punch wound at 1 day to 8 wks post injury time were investigated by transmission electron microscopy and histology was investigated with Olympus System and TS-Auto software.
RESULTS
CTGF expression was down-regulated to 40% of control by CTGF siRNA/KALA (1:24) complexes (p < 0.01) and collagen lattice contraction was inhibited. However, down-regulated of CTGF by CTGF siRNA/MPGΔNLS complexes was not statistically significant. CTGF KALA-treated wound appeared with well formed-basket weave pattern of collagen fibrils with mean diameter of 128 ± 22 nm (n = 821). Mismatch siRNA/KALA-treated wound showed a high frequency of parallel small diameter fibrils (mean 90 ± 20 nm, n = 563).
CONCLUSION
Controlling over-expression of CTGF by peptide-mediated siRNA delivery could improve the collagen orientation and tissue remodeling in full thickness rabbit ear wound.

Keyword

Connective tissue growth factor siRNA; Scar; Collagen; Dermal matrix

MeSH Terms

Biopsy
Cell-Penetrating Peptides
Cicatrix
Collagen*
Connective Tissue Growth Factor*
Connective Tissue*
Down-Regulation
Ear
Microscopy, Electron, Transmission
Preventive Medicine
Regeneration
RNA, Small Interfering
Wounds and Injuries
Cell-Penetrating Peptides
Collagen
Connective Tissue Growth Factor
RNA, Small Interfering