Effect of RGD peptide concentration on cell adhesion and viability in 2D and 3D PEG hydrogel environment

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Effect of RGD peptide concentration on cell adhesion and viability in 2D and 3D PEG hydrogel environment


Author: Qin, Xueshan
Title: Effect of RGD peptide concentration on cell adhesion and viability in 2D and 3D PEG hydrogel environment
Degree: M.Sc.
Year: 2009
Subject: Hong Kong Polytechnic University -- Dissertations.
Aspartic acid.
Polyethylene glycol.
Cell adhesion.
Department: Dept. of Health Technology and Informatics
Pages: xv, 62 leaves : ill. ; 30 cm.
Language: English
InnoPac Record: http://library.polyu.edu.hk/record=b2303067
URI: http://theses.lib.polyu.edu.hk/handle/200/4098
Abstract: Mesenchymal stem cells (MSC) are pluripotent cells that can differentiate into a wide range of specialized cell types. They have been used for tissue regeneration in three-dimensional (3D) scaffold through the control and guidance of MSC differentiation. However, there still exist some major obstacles to their applications for tissue regeneration. One of the major obstacles is the poor understanding of the mechanisms underlying MSC differentiation and their interaction with the biomaterial scaffold. This project is to study the interaction between Mesenchymal stem cells (MSC) and the biomaterials-RGD grafted poly (ethylene glycol) (PEG) hydrogel. PEG hydrogel is an ideal scaffold for tissue engineering because of its biocompatible, non-toxicity and highly hydrated nature which are highly resemble the native extracellular matrix (ECM). The PEG hydrogels were fabricated using a photo-polymerization technique and a biomolecular-RGD peptide was incorporated into the PEG hydrogel to investigate the relationship between MSC adhesion and RGD concentration. RGD (Arg-Gly-Asp) tripeptide has been proved that it is the active sequence of adhesive proteins, and different RGD concentration have been demonstrated to exhibit different effects on cell adhesion which plays a central role in development and disease as it can affect cytoskeletal organization, cell polarity, cell proliferation, and gene expression. In theory, higher RGD concentration would result more cell adhesion, but when RGD concentration reaches a certain value, its effect on cell adhesion will be saturated, it means the cell adhesion numbers will be no any change. This study is to investigate this phenomenon, find out the statistical relationship between the number of stem cell adhesion and the RGD concentrations, and find the optimal RGD concentration for cell viability, adhesion, proliferation and differentiation of MSC in gradient PEG hydrogel. In this study, MSC were also encapsulated in the PEG hydrogel in situ by photo-polymerization to investigate their growth and death conditions in 3 dimensions. Other, interaction of MSC with biomaterials, such as proliferation and differentiation were also studied and the statistical analysis was conducted. Results showed the number of cell adhesion was significant different (P <0.05) among different RGD concentrations in 2D cell culture, and higher RGD concentration indeed lead to more cell adhesion. However, the number of cell adhesion had no obvious change among days of culture. In 3D cell culture, cell viability was significant different among different RGD concentrations, and cell viability increased as RGD concentration increased. It proved that RGD had effect on cell viability. Also, cell viability was significant different among different culture lengths (P<0.05), but cell viability was no significant different between fifth day and seventh day of culture (P>0.05). In addition, 30% (w/v) PEGDA was more suitable for hydrogel fabrication in 3B cell culture than 10% (w/v) PEGDA.

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