Cell sheet formation enhances the therapeutic effects of human umbilical cord mesenchymal stem cells on myocardial infarction as a bioactive material

Stem cell-based therapy has been used to treat ischaemic heart diseases for two decades.However,optimal cell types and transplantation methods remain unclear.This study evaluated the therapeutic effects of human umbilical cord mesenchymal stem cell(hUCMSC)sheet on myocardial infarction(MI).Methods:hUCMSCs expressing luciferase were generated by lentiviral transduction for in vivo bio-luminescent imaging tracking of cells.We applied a temperature-responsive cell culture surface-based method to form the hUCMSC sheet.Cell retention was evaluated using an in vivo bio-luminescent imaging tracking system.Unbiased transcriptional profiling of infarcted hearts and further immunohistochemical assessment of monocyte and macrophage subtypes were used to determine the mechanisms underlying the therapeutic effects of the hUCMSC sheet.Echocardiography and pathological analyses of heart sections were performed to evaluate cardiac function,angiogenesis and left ventricular remodelling.Results:When transplanted to the infarcted mouse hearts,hUCMSC sheet significantly improved the retention and survival compared with cell suspension.At the early stage of MI,hUCMSC sheet modulated inflammation by decreasing Mcp1-positive monocytes and CD68-positive macrophages and increasing Cx3cr1-positive non-classical macrophages,preserving the cardiomyocytes from acute injury.Moreover,the extracellular matrix produced by hUCMSC sheet then served as bioactive scaffold for the host cells to graft and generate new epicardial tissue,providing mechanical support and routes for revascularsation.These effects of hUCMSC sheet treatment significantly improved the cardiac function at days 7 and 28 post-MI.Conclusions:hUCMSC sheet formation dramatically improved the biological functions of hUCMSCs,mitigating adverse post-MI remodelling by modulating the inflammatory response and providing bioactive scaffold upon transplantation into the heart.Translational perspective:Due to its excellent availability as well as superior local cellular retention and survival,allogenic transplantation of hUCMSC sheets can more effectively acquire the biological functions of hUCMSCs,such as modulating inflammation and enhancing angiogenesis.Moreover,the hUCMSC sheet method allows the transfer of an intact extracellular matrix without introducing exogenous or synthetic biomaterial,further improving its clinical applicability.

Resveratrol promotes the survival and neuronal differentiation of hypoxia-conditioned neuronal progenitor cells in rats with cerebral ischemia

Hypoxia conditioning could increase the survival of transplanted neuronal progenitor cells(NPCs)in rats with cerebral ischemia but could also hinder neuronal differentiation partly by suppressing mitochondrial metabolism.In this work,the mitochondrial metabolism of hypoxia-conditioned NPCs(hcNPCs)was upregulated via the additional administration of resveratrol,an herbal compound,to resolve the limitation of hypoxia conditioning on neuronal differentiation.Resveratrol was first applied during the in vitro neuronal differentiation of hcNPCs and concurrently promoted the differentiation,synaptogenesis,and functional development of neurons derived from hcNPCs and restored the mitochondrial metabolism.Furthermore,this herbal compound was used as an adjuvant during hcNPC transplantation in a photothrombotic stroke rat model.Resveratrol promoted neuronal differentiation and increased the long-term survival of transplanted hcNPCs.18-fluorine fluorodeoxyglucose positron emission tomography and rotarod test showed that resveratrol and hcNPC transplantation synergistically improved the neurological and metabolic recovery of stroke rats.In conclusion,resveratrol promoted the neuronal differentiation and therapeutic efficiency of hcNPCs in stroke rats via restoring mitochondrial metabolism.This work suggested a novel approach to promote the clinical translation of NPC transplantation therapy.

MiR-551b-5p Contributes to Pathogenesis of Vein Graft Failure via Upregulating Early Growth Response-1 Expression

Background： Vein graft failure （VGF） is a serious complication of coronary artery bypass graft, although the mechanism remains unclear. The study aimed to investigate the effects of microRNAs （miRNAs） on the endothelial dysfunction involved in VGF. Methods： Human umbilical vein endothelial cells （HUVECs） were subjected to mechanical stretch stimulation to induce endothelial dysfunction. Genome-wide transcriptome profiling was performed using the Human miRNA OneArray＂ V4 （PhalanxBio Inc., San Diego, USA）. The miRNA-messenger RNA （mRNA） network was investigated using gene ontology and Kyoto Encyclopedia of Genes and Genomes. The miR-55 1b-5p mimic and inhibitor were applied to regulate miR-55 lb-5p expression in the HUVECs. The 5-ethynyl-2＇-deoxyuridine assay, polymerase chain reaction （PCR）, and Western blotting （WB） were used to assess HUVECs proliferation, mRNA expression, and protein expression, respectively. The vein graft model was established in early growth response （Egr）-I knockout （KO） mice and wide-type （WT） C57BL/6J mice for pathological and immunohistochemical analysis. Endothelial cells isolated from the veins of WT and Egr-1 KO mice were subjected to mechanical stretch stimulation; PCR and WB were conducted to confirm the regulatory effect of Egr- 1 on Intercellular adhesion molecule （loam-1）. One-way analysis of variance and independent t-test were performed for data analysis. Results： Thirty-eight rniRNAs were differentially expressed in HUVECs after mechanical stretch stimulation. The bioinforrnatics analysis revealed that Egr-1 might be involved in VGF and was a potential target gene of miR-551b-5p. The mechanical stretch stimulation increased miR-55 1b-5p expression by 2.93 ± 0.08 told （t= 3.07, P 〈 0.05）, compared with the normal HUVECs. Transfection with the miR-551b-5p mimic or inhibitor increased expression of miR-551b-5p by 793.1 ± 171.6 fold （t = 13.84, P 〈 0.001） or decreased by 26.3% ± 2.4% （t= 26.39, P 〈 0.05） in the HUVECs, respectively. HUVECs proliferation and EGR-I mRNA expression were significantly suppressed by inhibiting miR-551b-5p expression （P 〈 0.05）. The lumens of the vein grafts in the Egr-1 KO mice were wider than that in the WT mice. lcam-I expression was suppressed significantly in the Egr-1 KO vein grafts （P 〈 0.05）. Conclusions： Increased miR-55 1b-5p expression leads to endothelial dysfunction by upregulating Egr-1 expression. EGR-1 KO can improve the function of a grafted vein through suppressing loam-1.

ASIC2 Synergizes with TRPV1 in the Mechano-Electrical Transduction of Arterial Baroreceptors

Mechanosensitive ion channels(MSCs)are key molecules in the mechano-electrical transduction of arterial baroreceptors.Among them,acid-sensing ion channel 2(ASIC2)and transient receptor potential vanilloid subfamily member 1(TRPV1)have been studied extensively and documented to play important roles.In this study,experiments using aortic arch-aortic nerve preparations isolated from rats revealed that both ASIC2 and TRPV1 are functionally necessary,as blocking either abrogated nearly all pressure-dependent neural discharge.However,whether ASIC2 and TRPV1 work in coordination remained unclear.So we carried out cell-attached patch-clamp recordings in HEK293T cells co-expressing ASIC2 and TRPV1 and found that inhibition of ASIC2 completely blocked stretch-activated currents while inhibition of TRPV 1 only partially blocked these currents.Immunofluorescence staining of aortic arch-aortic adventitia from rats showed that ASIC2 and TRPV1 are co-localized in the aortic nerve endings,and co-immunoprecipitation assays confirmed that the two proteins form a compact complex in HEK293T cells and in baroreceptors.Moreover,protein modeling analysis,exogenous co-immunoprecipitation assays,and biotin pull-down assays indicated that ASIC2 and TRPV1 interact directly.In summary,our research suggests that ASIC2 and TRPV1 form a compact complex and function synergisti-cally in the mechano-electrical transduction of arterial baroreceptors.The model of synergism between MSCs may have important biological significance beyond ASIC2 and TRPV 1.