Tissue Eng Regen Med.  2018 Aug;15(4):453-466. 10.1007/s13770-018-0123-0.

A Novel Dorsal Slit Approached Non-Ischemic Partial Nephrectomy Method for a Renal Tissue Regeneration in a Mouse Model

Affiliations
  • 1BioMedical Research Institute, Kyungpook National University Hospital, 130 Dongdeok-ro, Jung-gu, Daegu 41944, South Korea.
  • 2Department of Laboratory Animal Research Support Team, Yeungnam University Medical Center, 170 Hyunchung-ro, Nam-gu, Daegu 42415, South Korea.
  • 3Department of Pathology, Central Hospital, 480 Munsu-ro, Nam-gu, Ulsan 44667, South Korea.
  • 4Department of Urology, School of Medicine, Kyungpook National University, Kyungpook National University Hospital, 130 Dongdeok-ro, Jung-gu, Daegu 41944, South Korea. urokbs@knu.ac.kr, tgkwon@knu.ac.kr
  • 5Department of Urology, College of Medicine, Yeungnam University, 170 Hyunchung-ro, Nam-gu, Daegu 42415, South Korea.
  • 6Center for Biomaterials, Korea Institute of Science and Technology, 5 Hwarangro, Seongbuk-gu, Seoul 02792, South Korea.
  • 7Center for Biomaterials, Biomedical Research Institute, Korea Institute of Science and Technology, 5 Hwarangro, Seongbuk-gu, Seoul 02792, South Korea.
  • 8Department of Urology, Kyungpook National University Chilgok Hospital, 807 Hoguk-ro, Buk-gu, Daegu 41404, South Korea.

Abstract

BACKGROUND
Kidney ischemia-reperfusion (IR) via laparotomy is a conventional method for kidney surgery in a mouse model. However, IR, an invasive procedure, can cause serious acute and chronic complications through apoptotic and inflammatory pathways. To avoid these adverse responses, a Non-IR and dorsal slit approach was designed for kidney surgery.
METHODS
Animals were divided into three groups, 1) sham-operated control; 2) IR, Kidney IR via laparotomy; and 3) Non-IR, Non-IR and dorsal slit. The effects of Non-IR method on renal surgery outcomes were verified with respect to animal viability, renal function, apoptosis, inflammation, fibrosis, renal regeneration, and systemic response using histology, immunohistochemistry, real-time polymerase chain reaction, serum chemistry, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, and Masson's trichrome staining.
RESULTS
The Non-IR group showed 100% viability with mild elevation of serum blood urea nitrogen and creatinine values at day 1 after surgery, whereas the IR group showed 20% viability and lethal functional abnormality. Histologically, renal tubule epithelial cell injury was evident on day 1 in the IR group, and cellular apoptosis enhanced TUNEL-positive cell number and Fas/caspase-3 and KIM-1/NGAL expression. Inflammation and fibrosis were high in the IR group, with enhanced CD4/CD8-positive T cell infiltration, inflammatory cytokine secretion, and Masson's trichrome stain-positive cell numbers. The Non-IR group showed a suitable microenvironment for renal regeneration with enhanced host cell migration, reduced immune cell influx, and increased expression of renal differentiation-related genes and anti-inflammatory cytokines. The local renal IR influenced distal organ apoptosis and inflammation by releasing circulating pro-inflammatory cytokines.
CONCLUSION
The Non-IR and dorsal slit method for kidney surgery in a mouse model can be an alternative surgical approach for researchers without adverse reactions such as apoptosis, inflammation, fibrosis, functional impairment, and systemic reactions.

Keyword

Ischemia-reperfusion; Renal regeneration; Inflammation; Apoptosis; Fibrosis

MeSH Terms

Animals
Apoptosis
Blood Urea Nitrogen
Cell Count
Cell Movement
Chemistry
Creatinine
Cytokines
DNA Nucleotidylexotransferase
Epithelial Cells
Fibrosis
Immunohistochemistry
Inflammation
Kidney
Laparotomy
Methods*
Mice*
Nephrectomy*
Real-Time Polymerase Chain Reaction
Regeneration*
Creatinine
Cytokines
DNA Nucleotidylexotransferase
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