Shalika Neelaveni & Abhinav Adhikari

Loudoun Academy of Science

Background Method Statistical

Statistical Analysis
Analysis
Current Cancer Treatment v Mann-Whitney (trials 1-7)
v Cancer drugs have negative side effects due to the inaccurate nature of current ATRP Synthesis of pHEMA Hydrogels o Purpose: Compare initial dye solution concentration with concentration of
dye solutions after hydrogels were placed
treatment.
v Much of the healthy, non-cancerous tissue can be harmed. HEMA monomer and To this degassed Ethyl-α- o Summary:
Hydrogels • P-values for all tests are not considered significant
EGDMA crosslinker solution, the bpy bromoisobutyrate • Results were inconclusive because of an insufficient amount of data
v Hydrogels are a major development as a targeted drug delivery system to cancer were dissolved in a ligand and Cu(I)Br initiator was added to
cells. v Unpaired t test (trials 1-7)
v Hydrogels consist of monomers as the framework of the gel, and cross linkers as
methanol/water catalyst were added complete o Purpose: Compare initial dye solution concentration with concentration of
the bonding for the network. mixture. to the stirred solution. polymerization. dye solutions after hydrogels were placed
v Hydrogels for controlled drug delivery can be created with more than one o Summary:
polymer network partially interlaced (without covalent bonding) caused by radical • P-values for all tests are considered significant
polymerization; this can allow for easy separation, which can result in the release Synthesis of Antigen Comonomer • This indicates that if data with similar distribution and difference was
of a loaded therapeutic. observed, extremely statistically significant data would likely yield as a
v Poly(hydroxyethyl methacrylate), otherwise known as pHEMA, hydrogels are N-succinimidyl acrylate was result Table 5: Statistical Analysis (α = 0.05)
added to a phosphate The resulting vinyl-Rabbit The vinyl-Rabbit IgG was
biocompatible and non-toxic making them successful in many drug delivery Statistical Test P-value
buffer solution containing IgG was purified by split into five units.
systems. Mann Whitney (trials 1-3) 0.1000
Rabbit IgG. The reaction removing the phosphate Methanol and water was
Antigens & Antibodies Antigens was carried out at 36°C for buffer solution through added to each unit to Unpaired t test (trials 1-3) 0.0002
v An antigen is a toxin or foreign substance which an hour to introduce vinyl centrifuge tube filtration reduce the concentration. Mann Whitney (trials 4-7) 0.1000
induces an immune response in the body, such groups into the Rabbit IgG. Unpaired t test (trials 4-7) 0.0002
as the creation of antibodies.
v Antibodies are a blood protein produced by the Cell
immune system in response to and Discussion
Discussion
counteracting a specific antigen. Antibodies ATRP Synthesis of pHEMA-EGDMA-Antigen-Antibody Hydrogels
ATRP The results of this research indicate that both the pHEMA hydrogels and vinyl-Rabbit
v Atom transfer radical
HEMA monomer, EGDMA To this degassed solution, IgG were synthesized correctly. FTIR graph 1, which compares the IR spectra of
polymerization (ATRP) is a
crosslinker, vinyl-Rabbit the bpy ligand and Ethyl-α- pHEMA hydrogels synthesized within this study with an IR spectra from a sample
polymerization method that bromoisobutyrate library displays a 76.26 match rating. Both spectra include peaks at 1725
requires a monomer, initiator, IgG, and GAR Anti-Rabbit Cu(I)Br catalyst were
IgG were dissolved in a added to the stirred initiator was added to wavenumbers and at 1150 wavenumbers suggesting carbon-oxygen stretch bonds.
catalyst, ligand, and solvent. complete polymerization. FTIR graph 2, which compares the IR spectra of the vinyl-Rabbit IgG, Rabbit IgG
Current Research https://pubs.acs.org/doi/10.1021/ma0359032 methanol/water mixture. solution.
antigen, and N-succinimidyl acrylate, displays that the graphs of NSA and the vinyl-
v In 1999, scientists from Kansai University in Rabbit IgG contain peaks at 1100 wavenumbers, which suggests a carbon-oxygen
Japan developed a hydrogel with an antibody- stretch, and 1650 wavenumbers, which suggests a vinylidene bond. Both of these
antigen complex which swelled in response to a
specific free floating antigen.
Dye Loading support the idea that a vinyl group was attached to the antigen. These peaks can
also be found on IR spectra graphs of methylmethacrylate and succinimide, both of
v The hydrogel was prepared by grafting the rabbit The absorbance of the dye
Hydrogels were submerged Once the hydrogels were solution was then which have similar chemical structures to N-succinimidyl acrylate. Furthermore, FTIR
immunoglobulin G (rabbit IgG) antigen and goat for 24 hours in phosphate removed, the absorbance graph 3, which compares the pHEMA-EGDMA-antigen-antibody hydrogels with other
anti-rabbit IgG (GAR IgG) antibody, while subtracted from the initial
buffer solution containing of the solution was components of the hydrogels, displays a peak formed at 1650 wavenumbers which
acrylamide was used as the monomer.
https://www.ncbi.nlm.nih.gov/pubmed/10391240 absorbance to find the
dye, and later centrifuged recorded using a expresses modification of original pHEMA hydrogel structure to now incorporate the
v The results from the studies indicated that the hydrogel can swell and become difference. A standard curve
to separate the hydrogel spectrophotometer, using a vinyl-antigen and antibody. Lastly, the dye absorbance data for pHEMA hydrogels
porous in the presence of a free floating antigen because the bonds between was used to determine the
from dye solution. wavelength of 630nm. change in concentration. was statistically significant, and suggested that the concentration of dye after the
the antibody and antigen within the polymer dissociate due to the antibody’s
hydrogels were placed in was significantly lower than the initial concentration of the
much higher binding affinity for the free floating antigens.
dye.

Purpose Table 1: Change in absorbance based on hydrogel dye absorption

Conclusion
Conclusion & Application
Data & Results Trial
Number
Initial Absorbance
Absorbance after
Hydrogel
Dye Absorbance
by Hydrogel
Cancer drugs and therapeutics often have adverse side effects due to their 1 0.690 0.607 0.083 Through this research it was concluded that the pHEMA hydrogel had
interaction with healthy tissues that are not the intended target, thus it is necessary 2 0.690 0.617 0.073
to develop a treatment which is able to specifically target cancerous tissue. The
been modified by the vinyl-Rabbit IgG and the Goat-anti-rabbit IgG.
3 0.690 0.618 0.072
purpose of this research is to synthesize a targeted, anti-cancer drug delivery system Figure #1: FTIR comparison between synthesized pHEMA hydrogels and sample Current problems in the targeted therapy field include the inability of
library Average 0.690 0.614 0.076 Table 5: Absorbance of dye solution as a function of amount of antibody in
using antigen responsive hydrogels, with HEMA as a biocompatible monomer. Table 2: Change in concentration based on hydrogel dye absorption pHEMA-EGDMA-Antigen-Antibody hydrogel applying the research to real life medical uses. For instance, some
Trial Initial Concentration after Dye Absorbance Amount of
Initial Absorbance
Absorbance after Dye Absorbance monomers, such as acrylamide, are toxic and carcinogenic and thus
Number Concentration (%) Hydrogel (%) by Hydrogel (%) antibody (mg) hydrogel by Hydrogel
0 0.372 0.387 -0.015
cannot be applied to the human body. This research provides an
1 0.693 0.601 0.092
2 0.693 0.618 0.075 0.054 0.372 0.386 -0.014
alternative to current treatments by creating non-toxic and
3 0.693 0.619 0.074 0.074 0.372 0.35 0.022 biocompatible hydrogels that display potential for drug delivery to
Average 0.693 0.613 0.080 0.094 0.372 0.338 0.034 cancerous sites. By using materials found within the body as
Table 3: Change in absorbance based on hydrogel dye absorption Average 0.372 0.316 0.056 biomarkers, the cancerous regions can be effectively targeted and
Figure #2 FTIR comparison between synthesized pHEMA-EGDMA-Antigen-Antibody hydrogels, Trial Number Initial Absorbance
Absorbance after Dye Absorbance Table 6: Concentration of dye solution as a function of amount of antibody treated with this novel hydrogel.
hydrogel by Hydrogel in pHEMA-EGDMA-Antigen-Antibody hydrogel
vinyl-rabbit IgG, and GAR IgG antibody
4 0.382 0.398 -0.016 Amount of Initial Concentration after Dye Absorbance
antibody (mg) Concentration (%) hydrogel (%) by Hydrogel (%)
5 0.382 0.456 -0.074
0 0.376 0.391 0.015
6
7
0.382
0.382
0.343
0.349
0.039
0.033
0.054 0.376 0.390 0.014 Future
Future Direction
Work
0.074 0.376 0.355 -0.021
Average 0.382 0.387 -0.005
Hypothesis
Hypotesis Table 4: Change in concentration based on hydrogel dye absorption
0.094 0.376 0.343 -0.033 In the future, this research will focus on optimizing model drug absorption and
Initial Concentration Concentration after Dye Absorbance Average 0.376 0.321 -0.055 testing release of model drug in a free floating antigen solution. Additionally, this
Trial Number
(%) hydrogel (%) by Hydrogel (%) research looks toward using human cancer antigens and antibodies instead of
It is hypothesized that if an antibody-antigen complex is used as a secondary monomer
4 0.378 0.394 -0.016 models to replicate the environment of the human body and cancerous tissue. More
in a pHEMA hydrogel, the hydrogel will be able to respond to antigens in a free floating
5 0.378 0.453 -0.075 thorough analysis will be conducted on modification of pHEMA hydrogels through
solution and release a model drug. It is also hypothesized that if the amount of the
6 0.378 0.337 0.041 the incorporation of antigens and antibodies to understand the change in chemical
antibody in the hydrogel increases, the amount of model drug absorbed by the hydrogel
7 0.378 0.345 0.033 structure.
will increase as the polymer network will become more secure.
Average 0.378 0.382 -0.004

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