The Preparation And Application Of Cross-linked Egg White In Ovarian Cancer Cell 3D Culture

Ovarian cancer creates heavy personal and societal burdens,primarily because it is difficult to prevent.The majority of patients who are diagnosed early can still be cured.The characteristics of its invasion and metastasis—that its molecular and protease synergies engage in a dynamic process that continually reconstructthe extracellular matrix(ECM)—lead to a mortality rate of 90%among women with ovarian cancer.Cancer is closely associated with aging.As the body ages,the extracellular matrix microenvironment can promote malignant changes in epithelial cells.According to the cross-linking theory of aging,proteins and large other molecules,such as nucleic acids,form macromolecules through covalent cross-linking,which interfere with the functioning of normal cells.The cross-linking reaction involves nucleic deoxyribonucleic acid(DNA)and extracellular collagen fibers.Egg white(EW)is inexpensive,easily obtained,and can be used for cell culture with good biocompatibility.In recent years,scientists have used native EW in cell cultures and achieved good results using matrigel and collagen.However,native EW has low strength,toughness,and resistance to water and is easily degraded.Cross-linking,however,not only simulates an aging ECM but also increases mechanical strength of EW and slows its degradation.Through cross-linking between EW and chemicals with different hardness gradients,surface roughness,and porosity,the aim of this study was to investigate the effects of the variation in the morphologies and mechanical properties of the ECM on the proliferation and aggregation of ovarian cancer SKOV-3 cells in vitro,in an attempt to clarify the mechanisms of ovarian cancer,thereby advancing its prevention and treatment and the screening of antitumor drugs.This doctoral study included the following:(1)A preparation of genipin(GP)cross-linked EW to simulate an aging ECM in a 3D culture of ovarian cancer SKOV-3 cells.As a natural product with low biological toxicity,GP is a good protein cross-linking agent.We used GP cross-linked EW as a cell culture scaffold to simulate aging ECM with different morphologies and mechanical properties.The results suggest that the stiffness of the ECM may increase with cross-linking,simultaneously increasing the speed of ovarian cancer SKOV-3 cell proliferation.The cross-linked EW scaffold was a suitable model for investigating age-induced changes in the ECM and ovarian cancer SKOV-3 cell proliferation and progression.Further studies aimed at preventing tumoral invasion and metastasis should focus on drug screening to identify candidates who do not have age-induced degenerative changes in the ECM.(2)A discussion of the possible molecular mechanisms of ovarian cancer SKOV-3 cell aggregates and drug resistancein GP cross-linked EW.To simulate the microenvironmentof tumor cell growth by building a 3D cross-linked EW cell culture model of ovarian cancer SKOV-3 cells,this study looked at the phenotype of ovarian cancer cell formation,cell aggregation distribution,the expression of the ABCG2/CD44 molecule,key adhesion molecules such as E-cadherin,and N-cadherin expression.After ovarian cancer SKOV-3 cell adhesion and aggregation,the molecular mechanisms of invasion and metastasis were further studied to fully simulate the complex extracellular microenvironment in vivo.(3)A discussion of the effects of porosity and stiffness in glutaraldehyde(GA)cross-linked EW in simulating an aging ECM upon the distribution and aggregation of ovarian cancer SKOV-3 cells.GA was the first cross-linking agent and is most commonly used as a cross-linking agent in gelatin.It is water-soluble,can be used in a double functional group,and is relatively inexpensive.In addition,it can be completely cross-linked with gelatin in low concent rations to improve its hydrophilicity and mechanical properties.The cross-linking of GA and EW was done with high-activity,quick-response stable products.Ovarian cancer SKOV-3 cells can be easily aggregated by 3D culture,which is why they were selected as seed cells.To determine the biological characteristics of tumors and to track cell growth and tumor formation and development,we simulated the effect of the ECM microenvironment on the proliferation,differentiation,invasion,and metastasis of ovarian cancer SKOV-3 cells in vitro through different cross-linked EW.The different GA cross-linked Ews were studied in terms of their elasticity,aperture size,and cytocompatibility.(4)A discussion of the effects of macroporosity and stiffness in P(MVE-alt-MA)cross-linked EW to simulate an aging ECM and on the proliferation and metastasis of ovarian cancer SKOV-3 cells.Poly[(methyl vinyl ether)-a/t-(maleic acid)](P(MVE-alt-MA))can be used for biomedical polymer cross-linking modification in the engineering of macroporous scaffolds due to its non-toxicity.The P(MVE-alt-MA)cross-linked EW prepared in this study had sufficient cytocompatibility,mechanical strength,plasticity,and capacity to support sustained cell growth.As a result,the cross-linked porous EW scaffolds provided a platform for maintaining the viability and function of ovarian SKOV-3 cancer cells for the reconstruction of cancer tissue.The modification of EW simulated the microenvironment of an aging ECM and revealed the relationship between aging and tumor progression.This study intended to simulate aging,including the formation of macroporosity and change in stiffness in the ECM.We then assessed the effects of aging on cell invasion and the metastasis of ovarian cancer.The consistency of anti-aging and cancer treatment were put forward at same time.This is of great significance for the prevention and treatment of cancer.