目的:旨在比较自体来源脂肪干细胞(ADSCs)与骨髓间充质干细胞(BMSCs)构建工程化心肌组织(EHTs)修补大鼠左心室室壁瘤,为EHTs提供合适的种子细胞.方法:实验共分四组(n=6),分别为年轻脂肪干细胞组(YA),年轻骨髓干细胞组(YB),老年脂肪干细胞组(OA),老年骨髓干细胞组(OB).于左胸第三肋间结扎大鼠前降支,2周后行心脏功能检测,选取心脏梗死面积为25％～ 30％的大鼠进行实验.将ADSCs与BMSCs(1.0×106/片)种植于经EDC(24 mg/mL)及NHS(60 mg/mL)活化、同时共价结合VEGF(1μg/mL)及bFGF(1 μg/mL)的胶原蛋白补片,3d后行左心室室壁瘤修补.术后7、14及28 d测定心脏超声检测左心室室壁瘤重建术后心脏功能;固定距离(35 cm)下测定补片面积大小及补片厚度;同时测定了新生血管生成情况及内皮细胞覆盖.结果:EHTs植人体内28 d后,OA组大鼠心脏功能在EF值、FS值高于OB组,OA组LVIDd及LVIDs值低于OB组,而YA组与YB组,差异无统计学意义(P＞0.05).此外,OA组大鼠心脏补片面积低于OB组(P＜0.05),OA组补片厚度高于OB组(P＜0.05),而YA组与YB组差异无统计学意义(P＞0.05).同时,在细胞存活及新生血管生成方面,OA组明显高于OB组(P＜0.05),而YA组与YB组,差异无统计学意义(P＞0.05).结论:老年患者来源的ADSCs更加适合作为种子细胞构建EHTs进行细胞移植.

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.

Objective To evaluate the effects of TIMP-1 and Ang-1 gene-modified BMSCs transplantation on the left ventricular function of rats with myocardial infarction.Methods The rat BMSCs were.transfected with eukaryotic expression plasmid encoding TIMP-1 or/and Ang-1 gene by liposome.Acute myocardial infarction was made in male rats by ligation of the left anterior descending (LAD) coronary artery.BMSCs carrying TIMP-1 or/and Ang-1 gene were injected into the ischemic myocardium after LAD ligatior.Four weeks after the administration,cardiac function was assessed by echocardiography and the hearts were harvested and sectioned for immunohistochemistry to examine the apoptosis,the collagen content and angiogenesis density.Results TIMP-1 and Ang-1 genemodified BMSCs transplantation significantly improved the cardiac function,myocardial apoptosis was alleviated,collagen content decreased and the angiogenesis density in border-zone was increased significantly (P＜0.05).Conclusions The results suggest that the combination of TIMP-1 and Ang-1-gene modified BMSCs transplantation can improve the cardiac function of rats with myocardial infarction.The increase of the blood supply,the alleviation of myocardial apoptosis and ventricle remolding after myocardial infarction possibly play important roles in the mechanism.

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.