BACKGROUND:Mesenchymal stem cell (MSC)-based therapy may be a future treatment for myocardial infarction (MI). However, few studies have assessed the therapeutic efficacy of adipose tissue-derived MSCs (ADSCs) obtained from elderly patients in comparison to that of bone marrow-derived MSCs (BMSCs) from the same elderly patients. The metabolomics results revealed a significantly higher L-arginine excretion from aged ADSCs vs BMSCs in hypoxic conditions. This was hypothesized as the possible mechanism that ADSCs showed an improved angiogenic capacity and enhanced the therapeutic effect on ischemic heart diseases. AIM:To investigate the role of L-arginine in enhancing angiogenesis and cardiac protection by comparing ADSCs and BMSCs in hypoxic conditions for MI therapy. METHODS:Metabolomic profiling of supernatants from ADSCs and BMSCs under hypoxic conditions were performed. Then, arginine succinate lyase (ASL) overexpression and short hairpin RNA plasmid were prepared and transfected into BMSCs. Subsequently, in vitro wound healing and Matrigel tube formation assays were used to verify the proangiogenetic effects of ADSC positive control, BMSCs, BMSCs ASL short hairpin RNA, BMSCs ASL overexpressed, and BMSC negative control on cocultured human umbilical vein endothelial cells. All sample sizes, which were determined to meet the statistical requirements and be greater than 3, were established on the basis of previously established literature standards. The protein levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor, etc. were detected. In vivo, the five types of cells were transplanted into the infarcted area of MI rat models, and the therapeutic effects of the transplanted cells were evaluated by echocardiography on cardiac function and by Masson's staining/terminal-deoxynucleotidyl transferase mediated nick end labeling assay/immunofluorescence detection on the infarcted area. RESULTS:Metabolomic analysis showed that L-arginine was increased. Using ASL gene transfection, we upregulated the production of L-arginine in aged patient-derived BMSCs in vitro, which in turn enhanced mitogen activated protein kinase and VEGF receptor 2 protein expression, VEGF and basic fibroblast growth factor secretion, and inductive angiogenesis to levels comparable to donor-matched ADSCs. After the cell transplantation in vivo, the modified BMSCs as well as ADSCs exhibited decreased apoptotic cells, enhanced vessel formation, reduced scar size, and improved cardiac function in the MI rat model. The therapeutic efficacy decreased by inhibiting L-arginine synthesis. CONCLUSION:L-arginine is important for inducing therapeutic angiogenesis for ADSCs and BMSCs in hypoxic conditions. ADSCs have higher L-arginine secretion, which leads to better angiogenesis induction and cardiac protection. ADSC transplantation is a promising autologous cell therapy strategy in the context of the present aging society.

Engineered heart tissues (EHTs) are regarded as being the most promising alternative to synthetic materials, and autologous mesenchymal stem cells (MSCs) are widely used as seeding cells. However, few studies have evaluated the feasibility of using MSCs from patients with cyanotic congenital heart disease (C-CHD) as seeding cells for EHTs, in comparison with cells from patients of acyanotic congenital heart disease (A-CHD). In the present study, we cultured MSCs from A-CHD and C-CHD patients in normoxia or hypoxia conditions, and compared their pro-angiogenic, anti-apoptotic and inflammation-modulatory potentials. In vivo, we seeded the cells into collagen patches conjugated with, or without, proangiogenic cytokines, which were used to repair the right ventricular outflow tract (RVOT) of rats. The in vitro results showed that C-CHD MSCs expressed higher levels of VEGFA and VEGFR2, and secreted more pro-angiogenic and anti-inflammatory cytokines under hypoxic conditions. On the other hand, apoptosis-related genes from C-CHD MSCs were modulated adaptably, converting these cells into an anti-apoptotic phenotype. In vivo studies demonstrated that in 4 weeks after RVOT reconstruction, cytokine-immobilized patches seeded with C-CHD MSCs exhibited preserved morphology, prolonged cell survival and enhanced angiogenesis compared to A-CHD MSCs. C-CHD MSCs that undergo “naturally hypoxic precondition” present a better cell source for EHTs, which would provide a promising individualized biomaterial for C-CHD patients.

Adipose‑derived stem cells (ADSCs) and bone marrow‑derived stem cells (BMSCs) are considered to be prospective sources of mesenchymal stromal cells (MSCs), that can be used in cell therapy for atherosclerosis. The present study investigated whether ADSCs co‑cultured with M1 foam macrophages via treatment with oxidized low‑density lipoprotein (ox‑LDL) would lead to similar or improved anti‑inflammatory effects compared with BMSCs. ADSCs, peripheral blood monocytes, BMSCs and ox‑LDL were isolated from ten coronary heart disease (CHD) patients. After three passages, the supernatants of the ADSCs and BMSCs were collected and systematically analysed by liquid chromatography‑quadrupole time‑of‑flight‑mass spectrometry (6530; Agilent Technologies, Inc., Santa Clara, CA, USA). Cis‑9, trans‑11 was deemed to be responsible for the potential differences in the metabolic characteristics of ADSCs and BMSCs. These peripheral blood monocytes were characterized using flow cytometry. Following peripheral blood monocytes differentiation into M1 macrophages, the formation of M1 foam macrophages was achieved through treatment with ox‑LDL. Overall, 2x106 ADSCs, BMSCs or BMSCs+cis‑9, trans‑11 were co‑cultured with M1 foam macrophages. Anti‑inflammatory capability, phagocytic activity, anti‑apoptotic capability and cell viability assays were compared among these groups. It was demonstrated that the accumulation of lipid droplets decreased following ADSCs, BMSCs or BMSCs+cis‑9, trans‑11 treatment in M1 macrophages derived from foam cells. Consistently, ADSCs exhibited great advantageous anti‑inflammatory capabilities, phagocytic activity, anti‑apoptotic capability activity and cell viability over BMSCs or BMSCs+cis‑9, trans‑11. Additionally, BMSCs+cis‑9, trans‑11 also demonstrated marked improvement in anti‑inflammatory capability, phagocytic activity, anti‑apoptotic capability activity and cell viability in comparison with BMSCs. The present results indicated that ADSCs would be more appropriate for transplantation to treat atherosclerosis than BMSCs alone or BMSCs+cis‑9, trans‑11. This may be an important mechanism to regulate macrophage immune function.

Surgical ventricular reconstruction (SVR) can restore cardiac function for left ventricular aneurysm to some extent. However, the patches used in this treatment have some limitations such as stiffness and calcification. Engineering heart tissues (EHTs) have emerged as a promising biomaterial to repair damaged heart. Nevertheless, selecting optimal candidate cells for EHTs has been controversial. Aging is a major consideration for seed cells derived from elderly patients. Hence, this study was aimed to assess the proliferation of, antiapoptosis potential of, and expression of senescence-associated factors (eg, SA-β-Gal, cyclin-dependent kinase inhibitor 2A (P16), cyclin-dependent kinase inhibitor 1 (P21) in adipose-derived stem cells (ADSCs), and bone marrow stem cells (BMSCs) in vitro. In addition, cardiac function, cell survival, and angiogenesis of ADSCs and BMSCs after SVR were assessed in vivo. The in vitro results showed that old ADSCs (OAs) grew faster; expressed lower levels of SA-β-Gal, P16, and P21; and possessed more pronounced antiapoptosis activity than old BMSCs (OBs). The in vivo results demonstrated that 28 days after patch implantation, animals that received OAs patches showed better restoration of cardiac function than animals that received OBs patches. Meanwhile, old ADSCs possessed more potential regarding cell survival and angiogenesis. These results suggest that ADSCs may be superior to BMSCs with regard to autologous cell transplantation in elderly patients.

Background. Engineered heart tissues (EHTs) present a promising alternative to current materials for surgical ventricular restoration (SVR); however, the clinical application remains limited by inadequate vascularization postimplantation. Moreover, a suitable and economic animal model for primary screening is another important issue. Methods. Recently, we used 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride chemistry (EDC) to initiate a strengthened, cytokine-conjugated collagenous platform with a controlled degradation speed. In vitro, the biomaterial exhibited an enhanced mechanical strength maintaining a porous ultrastructure, and the constant release of cytokines promoted the proliferation of seeded human mesenchymal stem cells (hMSCs). In vivo, with the hMSC-seeded, cytokine-immobilized patch (MSCs + GF patch), we performed modified SVR for rats with left ventricular aneurysm postmyocardial infarction (MI). Overall, the rats that underwent modified SVR lost less blood and had lower mortality. After 4 weeks, the rats repaired with this cell-seeded, cytokine-immobilized patch presented preserved cardiac function, beneficial morphology, enhanced cell infiltration, and functional vessel formation compared with the cytokine-free (MSC patch), cell-free (GF patch), or blank controls (EDC patch). Furthermore, the degradable period of the collagen patch in vivo extended up to 3 months after EDC treatment. Conclusions. EDC may substantially modify collagen scaffold and provide a promising and practical biomaterial for SVR.

Previous studies have demonstrated that extracellular matrix (ECM) can be used in tissue engineering due to its bioactivity. However, adipose-derived ECM (A-dECM) has never been applied in bone tissue engineering, and it is unknown whether it would be beneficial to the growth of bone marrow mesenchymal stem cells (BMSCs). In this study, we produced chitosan/gelatin/A-dECM (C/G/A-dECM) scaffolds via lyophilization and crosslinking; chitosan/gelatin (C/G) scaffolds were used as controls. For the C/G/A-dECM scaffolds, the average pore size was 285.93 ± 85.39 μm; the average porosity was 90.62 ± 3.65%; the average compressive modulus was 0.87 ± 0.05 kPa; and the average water uptake ratio was 13.73 ± 1.16. In vitro, A-dECM scaffolds could promote the attachment and proliferation of BMSCs. In the same osteogenic-inducing reagent, better osteogenic differentiation could be observed for the C/G/A-dECM scaffolds than for the C/G scaffolds. Thus, we conclude that A-dECM is a promising material and that C/G/A-dECM scaffolds are a candidate for bone tissue engineering.

Adipose-tissue derived mesenchymal stem cell (ADSC)-based therapy is a promising option for patients with atherosclerotic conditions, including coronary artery disease. However, the potential differences in the metabolic characteristics between bone marrow-derived mesenchymal stem cells (BMSCs) and ADSCs have remained to be fully elucidated. The present study aimed to compare the metabolic profiles of BMSCs and ADSCs via liquid chromatography quadrupole time-of-flight mass spectrometry. BMSCs and ADSCs obtained from elderly coronary heart disease patients were cultured, and after three passages, supernatants of each cell type were collected and systematically analysed. Substantial differences were detected between the metabolite signatures of ADSCs and BMSCs. In addition, further analysis using partial least-squares discriminant analysis score plots indicated significant differences between the supernatants of the two cell types. The following metabolites were deemed to be responsible for the potential differences in the metabolic characteristics of BMSCs and ADSCs: D-lactic acid, hydroxyindoleacetaldehyde, -D-glucose, bovinic acid, 9,10-epoxyoctadecenoic acid, glyceraldehyde, phenylpyruvic acid, L-octanoylcarnitine, retinyl ester, -ketoisovaleric acid, guanidoacetic acid, N-acetylneuraminic acid, imidazoleacetic acid riboside, sphingosine and pseudouridine 5-phosphate. Based on these findings, there may be significant differences in the following metabolic pathways: The linoleic acid metabolic pathway, galactose metabolism, argentines and proline metabolism, retinol metabolism, glycine and serine metabolism, galactose metabolism, and amino sugar and nucleotide sugar metabolism. In conclusion, substantial differences in metabolic characteristics were detected between BMSCs and ADSCs, which may be associated with the different efficacies of atherosclerosis therapies employing these cell types.

目的 对比紫绀型先心病(cyanotic congenital heart disease,C-CHD)和非紫绀型先心病(acyanotic congenital heart disease,A-CHD)患儿来源的骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)在乏氧环境下的抗凋亡能力,并探讨其发生机制.方法 hMSCs取自C-CHD(C组)和A-CHD(A组)患儿(男女不限,0～5岁),于体外乏氧环境下(1％O2,5％ CO2,94％ N2)培养.以Annexin V/PI双染结合流式细胞技术对比两组细胞抵御缺血缺氧诱导细胞凋亡的能力;以Western blot法检测两组细胞中B细胞淋巴瘤-2基因(B-cell lymphoma-2,Bcl-2),Bcl-2相关X蛋白(Bax)以及半胱氨酸天冬氨酸蛋白酶-3(caspase-3)的含量.结果 Annexin V/PI双染结合流式细胞检测结果显示:在缺血缺氧环境下,C组细胞的早期凋亡率(P＜0.01)和总体凋亡率(P＜0.05)低于A组细胞.Western-blot检测结果显示:C组细胞的凋亡抑制基因Bcl-2的含量高于A组细胞(P＜0.05),抗凋亡抑制基因Bax (P＜0.05)以及caspase-3 (P＜0.05)的含量低于A组细胞.结论 C-CHD患者来源的hMSCs具有更好的抗缺血缺氧诱导的总凋亡及早期凋亡能力,这可能与其天然乏氧诱导的Bcl-2表达上调,Bax,caspase-3表达下调有关.