| ObjectiveThe occurrence of bone-related diseases such as bone injury will seriously affect people’s health and work and study.The repair process is slow and the medical expenses are large.It is one of the difficult problems in the medical field to be solved.With the development of regeneration medicine,cell therapy has gradually become a hot spot for bone injury treatment.Among them,stem cells during the birth period have always been considered to be reliable cell sources in medical research and clinical trials.A variety of stem cell types have been identified in human placenta.Human amniotic epithelial cell(h AECs)has a new type of potential seed cells for cell therapy and clinical applications with its obvious advantages.Based on bibliometrics,this study will predict the future direction and research hotspot of human amniotic epithelial cells(h AECs),and on this basis,by constructing a co-culture system of h AECs and osteoblasts(OB)in vitro,explore the differentiation and potential mechanism of HAECS during bone injury repair.It is expected to provide the theoretical basis and research foundation for h AECs to repair bone injury to supplement its mechanism of action.Method1、Part IIncluded in all the data are from January 2012 to December 2022 Webof Science core literature database,with subject "Human amniotic epithelial cells,Human amniotic epithelial cell,h AECs" search.Cite Space is used to analyze the development status and research direction of h AECs related fields.2、Part IIh AECs were isolated from the full-term placenta by trypsin digestion,and the growth curve of h AECs was observed and recorded with x CELLigence RTCAS16.After that,the obtained h AECs were characterized and identified by immunofluorescence technology and flow cytometry,and induced by lipogenic differentiation medium,chondrogenic differentiation medium and osteogenic differentiation medium,and the differentiation ability of cells was identified.h AECs were inducted into osteogenesis using OB and h AECs coculture induction technology,and si RNA was used to interfere with the expression of YAP1 gene in the induction system.The experimental groups and treatment methods of induction process were blank group(no coculture induction,no si RNA inhibition),control group(no coculture induction,no si RNA inhibition),induction group(coculture induction,no si RNA inhibition),and inhibition group(coculture induction,no si RNA inhibition).The medium was changed every 48 hours.The results were processed and analyzed by RT-q PCR,westernblot and alkaline phosphatase staining.Result1、Part IA total of 1583 papers and reviews on h AECs published in the last 10 years were retrieved and included.According to the analysis results,the country with the most published papers in the field of h AECs is China,followed by the United States.The United States has the highest intermediate centrality.Monashuniversity is the most published research institution,and Harvard Medical School has the highest mediated centrality;Through keyword cooccurrence,topic clustering and keyword emergence analysis,it is found that stem cell transplantation,tissue engineering,regenerative medicine and other research directions may be the hot spots of h AECs research in the future.1、Part IIThe amniotic membrane separated by placenta was colorless and transparent after rinsing,and had certain toughness.The isolated human amniotic epithelial cells showed typical path-stone growth after culture.The newly inoculated h AECs will complete the wall adhesion and start to grow in about 18 h,and start to stabilize in 48 h.Immunofluorescence showed positive expression of human amniotic epithelial fine marker CK19 and negative expression of mesenchymal stem cell characteristic marker vimentin.Expression flow cytometry showed that CD73,CD90,CD105,CD29,EPCAM and CD324 were positive,while CD34,CD45 and HLA-DR were negative.The results of adipogenic differentiation,chondrogenic differentiation and osteogenic differentiation showed that h AECs showed specific staining results.In the process of coculture induction,the expression level of YAP1 in human amniotic epithelial cells was increased,the proliferation ability was relatively enhanced,and the osteoblast differentiation trend was displayed,and the osteoblast-related markers were increased to varying degrees.In the inhibitory group,YAP1 expression was inhibited,and the proliferation ability was relatively weakened,as well as the osteogenic differentiation ability,and the osteogenic markers were also reduced to varying degrees.ConclusionCite Space visualization shows that h AECs has great application potential in tissue engineering,stem cell transplantation and regenerative medicine.h AECs showed the ability of adipogenic differentiation,chondrogenic differentiation and osteogenic differentiation,and showed stem-cell related characteristics.In the presence of OB,h AECs gradually differentiate toward osteogenesis,and YAP1 in HIPPO signaling pathway can affect the proliferation and osteogenic differentiation of h AECs... |
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