Potential of human deciduous periodontal ligament stem cells for use in hepatic lineage and their exosome application on hepatic injuries / Punitha R. Vasanthan
Introduction: Human deciduous periodontal ligament stem cells represent as another source of stem cells from the dental origin. These cells were shown to have multi-potential attributes and also self renewing ability. These easily accessible stem cells candidate could also resolve issues conce...
Saved in:
Main Author: | |
---|---|
Format: | Thesis |
Published: |
2020
|
Subjects: | |
Online Access: | http://studentsrepo.um.edu.my/12908/4/punitha.pdf http://studentsrepo.um.edu.my/12908/ |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Summary: | Introduction: Human deciduous periodontal ligament stem cells represent as
another source of stem cells from the dental origin. These cells were shown to have
multi-potential attributes and also self renewing ability. These easily accessible
stem cells candidate could also resolve issues concerning ethical and compatability
for the use in regenerative purposes. However, their efficiency towards hepatic
lineage differentiation has not been addressed so far. Meanwhile, the main
contributor for regenerative properties were supposedly mediated by soluble
factors and extracellular vesicles such as exosomes secreted by cells. Objectives:
The aims of this study are to isolate, characterize and also to investigate the
propensity of stem cells derived from periodontal ligament towards hepatic lineage.
Besides that, the emergence of cell free based treatment have prompted the study
on exosomes derived from periodontal ligament stem cells and their relevance in
hepatic injury. Methods: Periodontal stem cells derived from human deciduous
tooth were isolated and characterized using the gold standard characterization
method. Differentiation of periodontal ligament stem cells into hepatocyte-like
cells were conducted by the exposure of two phases media induction. First phase
was performed in the presence of Hepatocyte growth factors and Nicotinamide to
induce a definitive endoderm formation. In the subsequent phase, the cells were
treated with Oncostatin M, Dexamethosone and Insulin Transferrin to generate
hepatocyte-like cells. Hepatic-related characters of the generated hepatocyte-like
cells were determined at both mRNA and protein level. Furthermore, functional
assay was conducted to examine the functionality of the hepatocyte-like cells. On
the other hand, the extracellular vesicles components were characterized prior to
the application into assisting in in vitro hepatic injury induced by hydrogen
peroxide on HepG2 cells. Polymer based precipitation were employed to isolate
iii
exosomes and subjected to assessments on surface markers and protein
quantification. Results: Foremost changes observed in the generation of
hepatocyte-like cells were the morphological features in which these cells were
transformed from fibroblastic shape to polygonal shape. Temporal expression of
hepatic markers ranging from early endodermal up to late markers were detected
in the hepatocyte-like cells. Differentiated periodontal ligament stem cells
exhibited functional features of a hepatocyte during and at the end of the
experiment duration. Crucial hepatic markers such as glycogen storage, albumin
and urea secretion were also shown. In addition, exosomes isolated from
periodontal ligament stem cells displayed positive exosome markers with
correlation between cell confluence and exosome release. As for the in vitro hepatic
injury, exosome from periodontal ligament stem cells conditioned media showed
reduction in terms of HepG2 cells injuries after being exposed to oxidative stress.
Conclusions: These findings exhibited the ability of periodontal ligament stem
cells to be directed into hepatic lineage fate contributing towards usage beyond
tooth regeneration. Along with that, the exosomes from periodontal ligament stem
cells displayed hepatoprotective attributes in terms of reduction of oxidative stress.
Ultimately, these cells can be regarded as an alternative autologous source potential
not only for stem cell based treatment but also as a compelling alternative for non
viable or cell free treatment in liver diseases.
Keywords: periodontal ligament, stem cells, hepatocytes, exosomes
|
---|