Photothermal Interface Mediated Intracellular Molecules Delivery And Its Application In Cell Therapy

Cell therapies with engineering cells have shown significant efficacy on the treatment of a variety of diseases,which represents an important development direction of medicine in the future.As an important technology of engineering cells,physical membrane rupture mediated intracellular molecular delivery is one of the key bottlenecks of cell therapy application transformation.For constructing an ideal intracellular molecular delivery system,researchers have designed and constructed a serious of photothermal interfaces to adjust local reversible membrane rupture and exogenous molecular transport.It provides theoretical basis and solution for the development of intracellular molecular delivery system.Based on the above background,this work focused on the topic of "photothermal interface mediated intramolecular delivery and its application in cell therapy".As the laser hotspots density,interface microstructure and exogenous molecular transmission are the three important factors of delivery and harvesting efficiency,we design the latent photothermal interface delivery system and transparent photothermal pump patch delivery system,respectively,to regulate the above three factors.Furthermore,we explored their potential application in cell therapy,such as neural differentiation of embryonic stem cells,engineering dendritic vaccine and transdermal engineering cells.The details are as follows:Firstly,focusing on the regulation of the hotspot density and interface microstructure,we constructed the latent photothermal interface mediated intracellular macromolecular delivery platform.Multiwall carbon nanotubes(MWNTs)were doped with polydimethylsiloxane(PDMS)to construct photothermal interface;The laser hotspot density and microstructure of interface were regulated by adjusting the content of MWNTs;The effects of MWNTs content on cell adhesion,cell membrane rupture and cell harvesting were systematically studied;The latent photothermal interface delivery system with high delivery efficiency,cell viability and cell harvesting rate was successfully constructed.The delivery efficiency,cell viability,cell release and cell harvesting rate of Hela cells were increased to 95%,80%,99%and 85%,respectively,which made it possible to obtain engineering cells by simple,efficient and safe method.Secondly,through the above mentioned latent photothermal delivery platform,we have engineered hard-to-transfect cells and then explored their application potential in cell therapy.We efficiently delivered exogenous macromolecules into cells,including mouse dendritic cells(mDC),embryonic stem cells(mESC),human embryonic fibroblasts(hEF),human umbilical vein endothelial cells(HUVEC),primary dendritic cells(BMDC)and primary T cells,and then harvested the engineering cells by latent photothermal delivery platform.The delivery efficiency,cell viability and harvesting rate of the above cells were increased to 85%,80%and 80%,respectively.On this basis,the delivery of RA into mESC can significantly enhance the differentiation of neural cells;and the delivery of pHTP into mDC can enhance the function of mDC,thus effectively activate the immune activity of T cells,which proved the application potential of latent photothermal delivery platform in the field of cell therapy.Finally,on the basis of the above,we further focused on the regulation of exogenous molecular transport pathway and rate,and build transparent photothermal pump delivery system,and then applied it in non-invasive transdermal engineering cells.The transparent photothermal pump patch with excellent transparency and photothermal conversion efficiency was constructed by gold nanorod@polydopamine doped PDMS;The exogenous molecule transport path was shortened by the design of the photothermal pump,and the exogenous molecule transport rate was improved by the micro thermal expansion performance of PDMS;The delivery efficiency and cell viability of photothermal interface delivery system were further improved to more than 90%.Then,we combined the epidermal delivery of photothermal pump patch with the dermal delivery of exosomes,and then efficiently transdermal engineered dermal cells by a non-invasive way.It brings new possibilities for subcutaneous cell therapy.In this work,through the design and construction of latent photothermal interface and photothermal pump patch,we systematically studied the effects of hotspot density,interface microstructure,exogenous molecular transport pathway and rate on intracellular molecular delivery.It provided theoretical guidance and application basis for further development of intracellular molecular delivery.Moreover,in our work,we have explored some solutions about the delivery problems of cell therapy,which contributed to promote the development and application of cell therapy.