Effect And Mechanism Of HiPSCs-KCs-derived Exosome In Deep Second Degree Burn Wound Healing In Mice

Skin is wrapped on the surface of people and other animals,directly in contact with the outside environment,with protection,excretion,regulating body temperature and feeling the effects of external stimulation,an organ,is the first barrier against external intrusion of human beings.Therefore,skin repair after injury is one of the more common clinical problems,such as:deep Ⅱ degree burn,Ⅲ degree burns,unable to heal itself,require a doctor’s intervention,wound healing time is very long,but the function recovery after the skin is bad,can cause mental and physical double injury for patients.At present,it is urgent for researchers to find new methods to promote rapid wound healing and improve skin healing function.Exosomes are small membranous vesicles containing complex RNA and proteins that are involved in intercellular communication.In recent years,with the research of researchers,people have a deeper understanding of exosomes,which play an important role in a variety of physiological activities of the human body.At the beginning of the 21st century,stem cells have become the most potential cells in the repair of intractable wounds.Induced pluripotent stem cells(iPSCs)are the reprogramming of terminally differentiated somatic cells into pluripotent stem cells by the introduction of specific transcription factors.The application of induced pluripotent stem cells does not have the ethical problems of ordinary stem cells,and promotes the innovation of treatment methods and strategies for a variety of diseases related to inflammation,tissue damage and regeneration and repair.At present,exploring the exosomes derived from hiPSCS-KCS provides a new direction and hope for wound repair.To deepen the research and development of iPSCs in skin wound healing.Study on the effect and mechanism of hiPSCs-KCs-derived exosomes in deep Ⅱ degree burn wound healing in mice.1.The effect of hiPSCS-KCS-derived exosome on the healing of deep Ⅱ degree burn skin in mice(1)PurposeObjective to investigate the effect of hiPSCS-KCS-exosome on the healing of deep Ⅱ degree skin burn in mice(2)Methods1.The hiPSCs were cultured and differentiated,and the conditional medium was collected to extract and identify hiPSCS-KCS-exosome.2.hiPSCS-KCS-exosome was applied to the treatment of deep Ⅱ degree burn in mice.Wound healing and epithelialization were observed by gross and HE staining.(3)Results1.Exosome extracted from the conditional medium had a double-film-forming structure with an average particle size of 70 nm,namely hiPSCS-KCs-exosome.2.Being compared with the PBS group,the effect of burn wound healing and epithelialization of the hiPSCs-KCS-exosome group were enhanced.(4)ConclusionhiPSCs-KCs-exosome promoted wound healing by promoting the re-epithelialization of deep Ⅱ burn in mice.2.The signaling mechanism of hiPSCS-KCS-exosome promoting deep degree Ⅱburn repair(1)PurposeTo investigate the mechanism of the involvement of hiPSCs-KCS-exosome in burn repair(2)Methods1.The effects of hiPSCS-KCS-exosome at different concentrations on cell proliferation and migration were observed by cell EDU-488 test,cell scratch test and Transwell test.2.Main miRNAs and their effects on cell proliferation and migration were screened by miRNA high-throughput sequencing,cell scratch assay,Transwell assay and cell EDU assay.3.Cell scratch assay and transwell assay were used to verify the effect of miR-762 on the migration of HaCaT,Human Dermal Fibroblasts(HDF)and Human Umbilical Venous Endothelial Cells(HUVEC).(3)Results1.hiPSCS-KCS-exosome can be absorbed and utilized by HaCaT and HUVEC,and can promote cell migration.2.miR-762 is rich in hiPSCS-KCS-exosome,miR-762 can promote the migration of HaCaT,HDF and HUVEC.(4)ConclusionmiR-762 reduces cell adhesion,thereby promoting cell migration.3.Repair effect of miR-762 on deep Ⅱ degree burn in mice(1)PurposeTo verify if miR-762 could promote the healing of deep Ⅱ degree burn in mice(2)MethodsmiR-762 was applied to the treatment of deep Ⅱ degree burn in mice.The wound healing and epithelialization were observed by gross observation,morphological observation and immunohistochemistry.(3)ResultsmiR-762 could promote the healing of deep Ⅱ degree burn and improve wound healing rate in mice.(4)ConclusionmiR-762 reduced the adhesion ability of keratinocytes,fibroblasts and human umbilical venous endothelial cells,promoted cell migration,and thus accelerated the healing of skin wounds.