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Essay/Term paper: Dionysus

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Dionysus


Dionysus was the most widely worshipped and popular god in ancient
Greece. It's not difficult to see why; he was their god of wine, merriment,
ritual dance, warm moisture, and later, civilization. He was often depicted as
a handsome young man, dressed in fawnskin, and carrying a goblet and an ivy-
covered staff.
Some myths hold that Dionysus was the son of Zeus--the king of the god--
and Persephone--queen of the underworld--but most myths state that he is the son
of Zeus and a mortal woman named Semel. This woman Semele was not any mortal,
though. She was a princess, and a beautiful one at that. Zeus was notorious
for being rather prolific, and when his wife, the goddess Hera heard that he had
gone off and mated with a mortal, she became quite upset. Hera, in an attempt
to exact her revenge, appeared to Semele and told her to ask Zeus to appear to
her in his divine form. When Zeus obliged, Semele was immediately consumed in
flames, for no mortal can look upon a god in his natural state. However, Zeus
saved the unborn Dionysus by sewing him up in his thigh, thus incubating him.
What happened next is different in every story. Some myths say he lived
with a king and queen loyal to Zeus until Hera discovered him, and, in a jealous
rage, warped their brains. In this version of the story, Dionysus was turned
into a goat by his father in an attempt to hide him from Hera; from then on he
had small horns on his head.
After he was safe, he went to live with the nymphs, who taught him to
make wine. Hera eventually found him again, and this time she also warped his
brain. The nymphs rejected him, and he went to live with the satyrs, who were
men with goat legs and horns, and their leader Silenus. Dionysus traveled with
the satyrs, who disgusted everyone they encountered with their rude, drunken
behavior.
Silenus is usually portrayed as a fat drunken man who rides on an ass.
He was once captured by King Midas. When Dionysus intervened, Midas freed
Silenus in exchange for the power to turn all he touched into gold. Dionysus
and his band eventually encountered the maenads. The maenads were a group of
wild, warlike creatures. They were horribly vicious, and unfortunately, they
were also incredibly stupid. They started quite a few unsuccessful wars against
kingdoms in Africa.
When Zeus finally found Dionysus again, he returned his mind to normal.
However, Dionysus refused to give up his unruly traveling companions. The people
of Achaea loved Dionysus, but hated his satyr and maenad friends. One time,
when Dionysus was visiting a port city, he was captured by a group of pirates
who weren't aware of his divine powers. He destroyed the pirates and sailed
their ship to the Island of Naxos. It was there that he met his bride and only
love Ariadne. Dionysus then made a journey into the depths of Hades to bring
back his mother Semele. He took her to Mt. Olympus, and changed her name to
Thyone. This fooled Hera, and Semele managed to remain safe. It was at this
time that Dionysus was thought to have fully returned to his godly state.
The Greeks worshipped Dionysus with two festivals known as Dionysia.
The lesser Dionysia was held in December, and the greater was held in March.
These Dionysia were usually marked by drunken orgies. The highlight of the
festivities was the sparagmos. During this part of the ceremony, a live goat
would be disemboweled, and the partygoers would feast upon its raw flesh. In
Rome, Dionysus was known as Bacchus, and his festival, or Bacchanalia, was also
extremely immoral, even by Rome's standards. So immoral in fact, that the
Roman Senate in 186 B.C., made a law forbidding the celebration of the
Bacchanalia. The law apparently didn't stop those fun-loving Romans,


 

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1. Introduction

The design of regenerative devices relies heavily on the use of three-dimensional (3D) scaffolds to provide the appropriate environment, mechanical support and an initial cell anchorage site for the regeneration of tissues and organs [1]. As a biomaterial, hydrogels provide a 3D hydrated framework with tissue-like elasticity for culturing cells. They also possess the ability to encapsulate cells and molecules prior to gelation, thus affording a minimally invasive avenue to deliver cells and bioactive factors [2,3,4].

Collagen type I is a natural polymer and is the most abundant structural building block in connective tissues such as bone, tendon and cartilage [5,6].

The basic structure of collagen is a triple helix of three left-handed polypeptide chains, which can self-aggregate to form collagen fibrils, fibres and fascicles in vivo [6,7,8]. As a scaffold material, collagen is inherently biocompatible and bioactive, and has a macromolecular organisation similar to the natural extracellular matrix (ECM); however, its use as a biomaterial building block may be limited due to batch-to-batch variability, potential antigenicity and difficulty in tuning features such as stiffness, degradation and bioactivity [9].

Cross-linking methods are necessary to stabilise collagen in aqueous solutions ex vivo and can be divided into three groups: physical interactions (e.g., ionically cross-linking and hydrogen bonds), chemical reactions (e.g., with glutaraldehyde [10], diisocyanates [11], carbodiimide-activated diacids [12], or photochemical polymerisation [13] or enzyme (e.g., glutaminase) catalysed mechanisms [14,15].

Chemical cross-linking can often be performed via covalent modification of lysine and hydroxylysine residues and amino termini, whose cumulative occurrence counts for ~3 × 10−4 mol·g−1 of collagen [16,17,18,19,20]. An advantage of chemical cross-linking is that the mechanical performance of the resulting hydrogel can be systematically adjusted via variation in cross-linker type and cross-link density, so that controlled tissue-specific ECM analogues can be successfully obtained [9,21,22]. Limitations include reagent toxicity, side reactions and lack of preservation of collagen triple helical conformation in resulting cross-linked systems, the latter being key to ensuring high affinity for cells and mechanical stability in physiological conditions [12,23].

“Click” chemistry is a cross-linking method which invokes high reactivity and selectivity [24,25,26,27]. Other attributes of click chemistry include insensitivity to oxygen and water, high yields, mild reaction conditions and no generation of potentially harmful by-products, thus reducing the toxicity of the resulting product [27,28,29]. As such, click chemistry is an appealing approach for the formation of collagen hydrogels.

Among click reactions, thiol-click chemistry has been shown to be a powerful tool for the efficient formation of thioether linkages (i.e., C–S–C) [30]. Examples of two thiol-click reactions include: Michael-addition, which requires an electron-deficient carbon-carbon double bond (vinyl) and a base [31]; and the thiol-ene reaction, which proceeds via a radical mediated, step-growth mechanism between a thiol and a vinyl group that can be initiated using a light source and a photoinitiator [32,33]. Here, the vinyl group should be incapable of homo-polymerisation to ensure that the only reaction taking place is the thiol-ene click reaction, such being the case for vinyl ether or norbornene-based mono-/macromers [34].

As a result of the reaction selectivity and oxygen insensitivity, an interesting feature of thiol-ene mixtures is their potential to be delivered as injectable liquid promptly leading to the formation of a tissue cavity-conformable hydrogel following application of a suitable external stimulus. As a result of the quick gelation capability, thiol-ene mixtures can also be formulated with cells and molecules prior to gel formation and implantation in vivo [27,35]. However, to achieve collagen gelation in situ, a major hurdle for the thiol-ene photo-click reaction is the necessary use of a non-toxic and water-soluble photoinitiator [1,36]. Upon light exposure, the photoinitiator generates free radicals that may cause cellular damage to a varying extent, depending on the gelation kinetics of the reacting mixture [37]. Additionally, most of the commercially-available photoinitiators are intended for use in organic solvents or dental resins and are therefore mostly insoluble in aqueous solutions of collagen.

2-Hydroxy-1-[4-(2-hydroxyethoxy) phenyl]-2-methyl-1-propanone (Irgacure™ 2959 (I2959)) and Lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) are two water-soluble photoinitiators capable of producing radicals when exposed to ultraviolet (UV) light (365 nm). I2959 is a commonly used photoinitiator for encapsulation of cells within hydrogels due to its well-established cytocompatibility at low concentrations [38].

In this work, we explored the challenge associated with the formation of an injectable, regenerative device via a thiol-ene photo-click reaction in the presence of 2-iminothiolane (2IT)-functionalised collagen and 8-arm norbornene-terminated poly(ethylene glycol) (PEG-NB). 2IT was selected as a flexible thiol-forming compound and was covalently-coupled to type I rat tail collagen via ring-opening reaction using the amino group from (hydroxy-)lysine terminations. PEG-NB was employed as the synthetic hydrogel building block with the NB function providing the carbon-carbon double bond incapable of homo-polymerisation required for the thiol-ene reaction. The presence of NB-terminated multi-arms was expected to minimise reaction-hindering steric effects, thereby promoting faster gelation compared to linear PEG-NB variants.We hypothesised that the orthogonality offered by the thiol-ene reaction could result in the rapid formation of a defined collagen based hydrogel network (~seconds) and therefore minimal toxic response [39]. Although thiol-ene hydrogels have been reported, none have been prepared using collagen as the precursor material [27,34,35,40,41]. The introduction of a PEG phase in the collagen-based hydrogel provided improved user control of gel formation and mechanical properties [39,42]. Two water-soluble photoinitiators were employed to establish the photoinitiator-gelation-cytotoxicity relationship involved in the formation of thiol-ene photo-click collagen-PEG hydrogels.

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