Optimization of the Renal Decellularization Process
obtaining Acellular Frames
Name: AFRÂNIO CÔGO DESTEFANI
Publication date: 31/10/2018
Advisor:
Name |
Role |
|---|---|
| BRENO VALENTIM NOGUEIRA | Advisor * |
Examining board:
| Name |
Role |
|---|---|
| ALEXANDRE MARTINS COSTA SANTOS | Internal Examiner * |
| BRENO VALENTIM NOGUEIRA | Advisor * |
| CAMILLE DE MOURA BALARINI | External Examiner * |
| FAUSTO EDMUNDO LIMA PEREIRA | Internal Examiner * |
| MARCO CESAR CUNEGUNDES GUIMARÃES | Internal Examiner * |
Summary: Chronic Kidney Disease (CKD) is characterized by progressive deterioration of renal function, which can compromise various tissues and organs. The main therapy indicated for
patients with CKD is renal transplantation. However, the absence of available organs, as
well as an organ rejection rate, is required for new therapies. Thus, the implementation of
bioengineering for organ regeneration has emerged as an alternative to traditional organ
transplantation. Currently, overtime has been expended on total renal decellularization by
perfusion with detergents. In these cases, there is a loss of essential components of the
extracellular matrix (ECM) and derangements in the scaffold architecture obtained which
can substantially influence subsequent cell repopulation. In this work, the total discoloration
of rat kidneys was measured in reduced time with the use of detergent solution (SDS, 1%)
by renal technique under pressure (100 mmHg) and flow (1.0 mL/min). At the end of this
stage, the scaffolds were incorporated as translucent physical characteristics and preserved
vascular conduits. An electron microscopic and histochemical analysis revealed glomeruli
with a preserved basement membrane, as well as a tubular and vascular network with no cell
and DNA residues. Proteomic analysis identified the preservation of majority proteins
related to ECM compounds. Thus, the use of low concentration of SDS for 6 hours promoted
a successful thawing, in addition to preserving a MEC with the minimum residue of SDS
(<0,01%). Also, we developed a retraction index as a correction calculation due to elastic
retraction of the kidney after processing. Electron microscopy, spectrophotometric
quantification, and mass spectrometry analyzes were performed to compare the results. We
demonstrated that the histological analysis, easy technique, and low cost, presented excellent
results when the correction index was applied, allowing the comparison of samples before
and after the process with reliable data. With these data, we demonstrate that it is possible
to reduce the time of decellularization with the preservation of essential components of the
ECM.
