3D Humanized Bioprinted Tubulointerstitium Model to Emulate Renal Fibrosis In Vitro.

Area of Science:

• Biotechnology
• Regenerative Medicine
• Nephrology

Background:

• Chronic kidney disease (CKD) involves progressive kidney function loss and fibrosis, traditionally studied using in vivo models.
• Ethical concerns and the 3Rs principles necessitate advanced humanized 3D in vitro models for studying kidney fibrosis.
• Current models lack the complexity to fully replicate the fibrotic microenvironment of the kidney.

Purpose of the Study:

• To develop and validate a novel 3D bioprinted in vitro model for studying kidney fibrosis.
• To create a humanized platform that mimics the renal tubulointerstitium and fibrosis progression.
• To establish a physiologically relevant model for drug screening and disease research.

Main Methods:

• A protocol for decellularizing pig kidney tissue to create decellularized and partially digested extracellular matrix (ddECM) was established.
• Primary human renal cells were encapsulated within the ddECM hydrogel to create a 3D bioprinted model.
• Fibrosis was induced using transforming growth factor beta 1 (TGF-β1), and hydrogel mechanics were modulated via vitamin B2 crosslinking.

Main Results:

• The bioprinted model successfully replicated the renal tubulointerstitium and showed increased stiffness (Young's modulus) over time.
• Treatment with TGF-β1 led to increased extracellular matrix deposition, cell dedifferentiation into myofibroblasts, and upregulation of fibrotic genes.
• The model demonstrated collagen deposition patterns similar to human fibrotic kidney tissue.

Conclusions:

• The developed 3D bioprinted model provides a more physiologically relevant platform for studying kidney fibrosis.
• This humanized in vitro model can advance research into kidney disease progression.
• The model holds potential for high-throughput drug screening and the development of novel therapeutic strategies for kidney fibrosis.