(M-497) Evaluation of the host responses to designer self-assembling growth factor mimics

Introduction:: Introduction: The objective of this study is to investigate how the chemical properties of Multidomain Peptide (MDP) hydrogels influences immune response in an in vivo environment. MDP hydrogels are a type of nanofibrous biomaterials with potential applications in regenerative medicine, drug delivery, and tissue engineering. Their unique sequence and the ability to form into self-assembled structures makes them adaptable in many potential areas of research in drug delivery. Despite their potential for biomedical use, the way in which these hydrogels interact with the immune system of the host needs to be studied substantially. Our study utilizes Western blotting and flow cytometry techniques to investigate the impact of domains on the immunogenicity of MDPs. It can also help us characterize the peptide hydrogel and better understand its ability to stimulate an immune response.


Materials and Methods:: Method: The study utilized MDPs which are prepared in neutral DI water and equimolar 10x PBS as a counterion to promote fibrillization. Western blotting and flow cytometry techniques were used to investigate the impact of charged domains on the immunogenicity of MDPs. To prepare cell suspensions for flow cytometry analysis, we will add 500,000 to 250,000 cells from a subcutaneous implantation of hydrogel in a murine model to each tube and diluted them 1:1 with FACS buffer. Before co-staining the samples with standard panels of immunophenotyping antibodies, we will incubate them with antibodies for 5 minutes at 4°C. After staining, cells will be washed and resuspended in PBS and then stained with Blue live/dead fixable dead cell stain for 30 minutes on ice. For Western blotting analysis, tissues were homogenized in a RIPA buffer containing protease inhibitors and then centrifuged before determining protein concentration using a BCA protein assay kit. The samples then were loaded onto SDS-PAGE gels for analysis. The membrane was probed with the primary and then secondary-HRP antibodies which was imaged in Azure gel imager.


Results, Conclusions, and Discussions:: Result: Based on the initial trials, the MDPs have shown promising results in invitro myoblast culture where the phosphorylation of the Serine threonine Kinase was observed. The biological and chemical functionality of these peptides has a substantial impact on the immune cell response based on the histological analysis, however via flow cytometry analysis the degree of inflammation and the host response can be quantified which would be crucial in designing the future peptide constructs via simulations to optimize their effects in an in vivo environment.



Conclusion: In the future, we hope to pursue in vivo work by administering the peptide hydrogels to rodent animal models through subcutaneous injection. We will complete qualitative analysis to better understand a potential immune response elicited from these animal models. In conclusion, by investigating the impact of bioactive domains in MDP hydrogels on their immunogenicity and exploring future in vivo studies, we hope to gain a deeper understanding of how to optimize these materials for various biomedical applications.




Acknowledgements (Optional): :

References (Optional): :