ABSTRACT
PURPOSE: Adipogenic differentiation of adipose tissue-derived mesenchymal stem cells (AMSCs) is critical to many disease-related disorders, such as obesity and diabetes. Studies have demonstrated that miRNA-138 (miR-138) is closely involved in adipogenesis. However, the mechanisms affected by miR-138 remain unclear. This work aimed to investigate interactions between miR-138 and lipoprotein lipase (LPL), a key lipogenic enzyme, in AMSCs. MATERIALS AND METHODS: Human AMSCs (hAMSCs) isolated from human abdomen tissue were subjected to adipogenic differentiation medium. Quantitative real-time polymerase chain reaction and Western blot assay were applied to measure the expressions of miR-138, LPL, and the two adipogenic transcription factors cytidine-cytidine-adenosine-adenosine-thymidine enhancer binding protein alpha (C/EBPα) and peroxisome proliferator-activated receptor gamma (PPARγ). The relationship between miR-138 and LPL was predicted utilizing the miRTarBase database and validated by dual luciferase reporter assay. RESULTS: Showing increases in C/EBPα and PPARγ expression levels, hAMSCs were induced into adipogenic differentiation. During adipogenesis of hAMSCs, miR-138 expression was significantly downregulated. Overexpression of miR-138 by transfection inhibited hAMSCs adipogenic differentiation in vitro. Mechanically, LPL was a target of miR-138. LPL expression was upregulated during adipogenesis of hAMSCs, and this upregulation was reversed by miR-138 overexpression. Functionally, silencing of LPL by transfection exerted similar inhibition of the expressions of C/EBPα and PPARγ. Meanwhile, LPL ectopic expression was able to partly abolish the suppressive effect of miR-138 overexpression on adipogenic differentiation of hAMSCs. CONCLUSION: Upregulation of miR-138 inhibits adipogenic differentiation of hAMSCs by directly downregulating LPL.
Subject(s)
Humans , Abdomen , Adipogenesis , Blotting, Western , Carrier Proteins , Ectopic Gene Expression , In Vitro Techniques , Lipoprotein Lipase , Lipoproteins , Luciferases , Mesenchymal Stem Cells , Obesity , PPAR gamma , Real-Time Polymerase Chain Reaction , Transcription Factors , Transfection , Up-RegulationABSTRACT
BACKGROUND: Whether the differences exist between adipose-derived stem cells isolated from different parts of rats when cultured in vitro has been poorly understood. OBJECTIVE: To compare the growth characteristics and adipogenic ability of adipose-derived stem cells isolated from different parts of rats. METHODS: Freshly isolated adipose-derived stem cells were obtained from 5 mL inguinal groove and greater omentum adipose tissue of F344 rats using type Ⅰ col agenase digestion method. Then, adipose-derived stem cells were counted and cultured in vitro. Morphological and growth characteristics of adipose-derived stem cells derived from the two sites were observed. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was utilized to examine the doubling time of adipose-derived stem cells from different parts. The passage 2 adipose-derived stem cells were induced adipogenical y. Fourteen days after being induced, the differentiated cells were stained with oil red O and the positive cells were counted. The adipogenic differentiation ability of adipose-derived stem cells from the different parts was assessed. RESULTS AND CONCLUSION: (1) The number of adipose-derived stem cells from the greater omentum fat tissue in the same group was (281±10)×107/L, which was significantly higher than that from the inguinal groove fat tissue [(85±10)×107/L] (P < 0.01). Adipose-derived stem cells from the greater omentum and inguinal groove fat tissue achieved the exponential growth period on days 5 and 6, respectively, and achieved the platform period on days 9 and 10, respectively. The corresponding doubling time was 50 hours and 60 hours, respectively. After being passaged, adipose-derived stem cells grew in fibroblast-like shape actively. The adipogenic differentiation rate of adipose-derived stem cells from the greater omentum fat tissue was higher than that from the inguinal groove fat tissue [(38.90±2.86)% vs. (35.30±3.29)%, P < 0.01]. This shows that the number and the adipogenic differentiation ability of adipose-derived stem cells from different parts of the same F344 rat are different.
ABSTRACT
Objective To introduce a new method for correcting prominent malar complex deformity. Methods Through an intraoral incision, the highest area of zygomatic body marked preoperatively was grinded. Then an L shape incomplete osteotomy of the zygomatic body was performed with a reciprocating saw, and a complete osteotomy just 1 cm anterior to the articular tubercle of the zygomatic arch was made. Light pressure on the posterior part of the arch produced a greenstick fracture of the anterior osteotomy site, resulting in posterior-inward repositioning of the malar complex. Internal fixation was unnecessary. Results Operative procedures for reductive malar complex plasty were performed in 650 cases, which included 60 males and 590 females whose age ranged from 19 to 39 years.Incisions of all cases healed well. One case had maxillary sinusitis 2 weeks postoperatively, and recovered after 1 week by using antibiotics and drainage. There was 1 case with skin necrosis about 1 cm in diameter in the area of zygomatic body because of local liposuction, and the wound was healed by changing dressing. The forehead wrinkle of one side had disappeared in 1 case 1 week postoperatively, but had recovered 2 weeks later. Postoperative follow-up for 2-24 months showed satisfactory results.Conclusions This modified method has many advantages, such as simplicity, without internal fixation, short operation and recovery time, and little complications. The authors conclude that this technique is an effective and safe method of reduction malarplasty.
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BACKGROUND:Cadherin-11 which was used to modify bone marrow mesenchymal stem cells aimed to promote cell adhesion ability;while,core binding factor α1 (cbfα1) which was used to modify bone marrow mesenchymal stem cells aimed to realize ossification of multipotential stem cells and play a signalization role in cascade effect.OBJECTIVE:To construct cadherin-11 and cbfα1 gene adenovirus expression vectors so as to modify bone marrow mesenchymal stem cells,and observe the expressions of the two genes.DESIGN,TIME AND SETTING:Cell-genetics experiment was performed at Central Laboratory of Zhujiang Hospital in August 2008.MATERIALS:Bone marrow mesenchymal stem cells were extracted from a male patient with thoracic vertebral fracture re from Zhujiang Hospital.The patient provided the informed consent.Full length cadherin-11 cDNA plasmid was provided by Professor Takeichi,Riken Molecular Organism Center,Japan;full length cbfα1 gene plasmid was provided by Professor Ducy,Baylor Medical College,USA;pAdeasy adenovirus system was provided by Professor Bert,McKuslck-Nathans Institute of Genetic Medicine,USA.METHODS:Human bone marrow mesenchymal stem cells were separated using Percoll density gradient centrifugation combined with adherence method.Cadherin-11 and cbfα1 genes were obtained by PCR,and then they were inserted into shuttle vector of adenovirus;thereafter,the recombinant adenovirus plasmids were gained by homologous recombination.Finally,the two plasmids were re-packed to obtain recombinant adenovirus venom,and cadherin-11 and cbfα1 genes were transfected in bone marrow mesenchymal stem cells by adenovirus.MAIN OUTCOME MEASURES:The expressions of cadherin-11 and cbfα1 genes were detected by Western biotting.RESULTS:The adenovirus venom carrying the cadherin-11 and cbfα1 gene was successfully obtained.Western blotting showed that the expressions of the cadherin-11 and cbfα1 genes in bone marrow mesenchymal stem cells were remarkably increased by infection viral.CONCLUSION:The bone marrow mesenchymal stem cells may express high level of cedherin-11 and cbfα1 by gene modification of adenovirus.
ABSTRACT
Regeneration of injured peripheral nerve is one of the recent research focuses. Many inspiring achievements have been made in such fields as choice of seeding cell source and proliferation in vitro. It is hopeful that nerve regeneration may be enhanced by combining the new techniques with microsurgery. In this paper, design and fabrication principles, key technologies, various applications, and new directions in the field of tissue engineered nerve are introduced. Much of this paper is devoted to the discussion of tissue engineered nerve in repair of peripheral nerve injury.
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Objective Tostudythecellapoptosisandp5 3geneexpressionoffibroblastsisolatedfromdifferenttissues .Methods Thefibroblastsfrom 8casesofnormalskin ,8hypertrophicscarsand 9keloids wereculturedrespectivelyinvitro.Flowcytometryanddotblotanalysisofp5 3withcDNAprobewereappliedandtheirbiologicalpropertieswerecomparedinthisstudy .Results Apoptoticcellsofthreegroupswereasfollows:(10 .8? 1.2 ) % ,(10 .6? 1.8) %and (5 .5? 0 .8) % ,respectively .Theresultsrevealedthatthere werefewerapoptoticcellsinkeloidthanthoseinhypertrophicscarandnormalskin (P