Chapter 1

Introduction

1.1. INTRODUCTION Nanotechnology is an emerging field focusing on the development of biomaterial design using simple biological building blocks. Molecular self-assembly is recognised as the process in which small molecules organise spontaneously into well-ordered nanostructures with specific functionality via reversible, non-covalent interactions including hydrogen bonding, disulphide bonds, van der Waals forces, electrostatic interactions, hydrophobic interactions and - stacking [1-3]. Peptide, as building blocks have served to be excellent building blocks for designing nanostructures that are highly relevant to tissue engineering, drug delivery systems, protein therapeutics and molecular bio-sensing
[4]

. On a molecular level, self-assembled nanostructures form stable

[1-2,4-7]

secondary structures; -sheets

16]

, -hairpins

[8-11]

, -helices and coiled-coils

[6, 12-13]

respectively. Non-natural peptide self-assemblies such as peptide amphiphiles (PAs) [14have been reported to form fibrous networks designed to mimic the dimensions of

the extra-cellular matrix (ECM). Aromatic peptides have also shown to undergo molecular self-assembly via - interactions to give rise to organised supramolecular nanostructures
[17-18]

. More specifically, peptide self-assembly resulted in the formation

[17-18]

of nanotubes (hollow cylindrical structures formed from ordered nanofibres)

Recently, several researchers have reported the use of another class of aromatic peptide, Fmoc protected di-peptides [4,19-23] which self-assembled to form fibrous hydrogels with anti-inflammatory and tuneable characteristics. The three-dimensional gel networks could be beneficial for different biomedical applications where the hydrogel can act as a carrier of drugs or other therapeutic agents or as a medium for encapsulating and maintaining viable cells and accumulating extra cellular matrix (ECM)[1-3,24-25]. Researchers are now focusing on constructing de novo
[25]

designs for tissue repair and

regeneration. Zhang and co-workers described the use of ionic-complementary peptides with alternating charged (hydrophobic and hydrophilic) amino acids to be used as peptide scaffolds to culture nerve cells and chondrocytes for tissue repair
[25-27]

Progressively more efforts were made by researchers to study different scaffolds, for example, Stupp and colleagues described the use of PAs scaffolds to support stem cell growth and differentiation, bone and cartilage regeneration, and therapeutic cell delivery
[28-31]

. Jayawarna et al. have used aromatic di-peptides which organised into well[21-23]

defined nanofibrous scaffolds to support the culture of chondrocytes and human dermal fibroblasts for cell culture applications in vitro . In all of the above examples,

hydrogels have been used as support matrices. The successes demonstrate that gels have considerable promise as scaffolds for in vitro and in vivo applications. Although a number of ionic-complementary peptide designs have been found to promote cell culture applications, Saiani and colleagues developed new self-assembled octapeptide systems FEFEFKFK and FEFKFEFK containing alternating charged and non-charged amino acid residues
[5-7]

. It was reported; gelation occurred by simply

mixing the peptides in distilled water at room temperature. The resultant gels consisted of -sheet conformation with fibre dimensions capable of mimicking the dimensions of ECM [6]. The current research will focus on testing these novel octapeptide systems for cell culture applications to support 2D and 3D cell culture in vitro. The first part of the thesis focuses on the synthesis and characterise self-assembled peptide gels under neutral physiological conditions and to compare and contrast in relation to alternating charge distribution of the two peptides on fibrous morphologies and mechanical properties at pH 7. Second part of the thesis enfolds the development of highly homogenous octapeptide gels for 2D and 3D culture of bovine chondrocytes. The stability of the two scaffolds in terms of cell morphology, viability and proliferation were compared respectively. Moreover, the mechanical properties in terms of ECM formation under cell culture conditions will be assessed. A preliminary study on the potential use of octapeptides gels as injectable biomaterials has also been introduced. The thesis is divided as follows; Chapter 1 and 2 describe the literature review based on the molecular self-assembly of different peptide systems which have been employed to promote cell culture and tissue engineering applications. The following chapters (3, 4, 5 and 6) comprehensively describe the research aim, results, discussion and conclusion of the findings obtained. Chapter 7 summarises the overall findings of the current research and suggestions for setting out the future research agenda.

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