title: [Effect of different temperatures on system of in vitro physiological environment fostering limbs].
abstract: OBJECTIVE
  To investigate the effects of different temperatures on the system of in vitro physiological environment fostering limbs.
  
  
  METHODS
  Twenty-four limbs were harvested from 6 adult Bama mini pigs and were randomly divided into 4 groups (n=6) according to different temperatures: limbs were placed in in vitro physiological environment fostering limbs at 26 degrees C (group A), 4 degrees C (group B), 10 degrees C (group C), and 18 degrees C (group D). After 12 hours of perfusion, the morphology observation was done for the structure and ultrastructure changes of the skeletal muscle by light microscope and transmission electron microscope. The mRNA levels of tumor necrosis factor n (TNF-alpha) and interleukin 10 (IL-1beta) were detected by real-time fluorescent quantitative PCR (RT-qPCR).
  
  
  RESULTS
  Histological results showed that the skeletal muscle exhibited mild edema, integrity of the sarcolemma, and occasional perivascular inflammatory cell infiltration in groups B, C, and D, meanwhile, the cells of group C had normal morphology; however, muscle fibers degenerated, muscle cells were seriously damaged, a great number of inflammatory cells infiltrated in the fractured muscle fibers in group A. Transmission electron microscope results showed as follows: the muscle fibers arranged in disorder, and many focal solubility necrosis occurred in group A; the muscle fibers arranged in order relatively and sarcolemma was still intact, with mild swelling and flocculent degenerative mitochondria in group B; a large number of muscle fibers arranged in order and regularity with clear sarcomere in group C; and the muscle fibers arranged in disorder and irregularity and partly dissolved in group D. RT-qPCR results showed that the expressions of inflammatory factor TNF-alpha and IL-1beta mRNA in group A were significantly higher than those in groups B, C, and D (P < 0.05); the expressions were significantly lower in groups B and C than in group D, and in group C than in group B (P < 0.05).
  
  
  CONCLUSIONS
  In the system of in vitro physiological environment fostering limbs, temperature plays an important role in the preservation of amputated limbs. It is suggested that 10 degrees C can significantly attenuate the reperfusion-induced skeletal muscle cell injuries in this system.
authors: Graduate School, Dalian University, Dalian Liaoning, 116622, PR China.
authors: Yegeng
authors: Y
authors: Yin
authors: Jiangning
authors: J
authors: Wang
authors: Lei
authors: L
authors: Gao
authors: Ning
authors: N
authors: Liu
authors: Xiaoyong
authors: X
authors: Xia
authors: Xinyuan
authors: X
authors: Qin
authors: Youwei
authors: Y
authors: Zuo
mesh_terms: D000673:Amputation, Traumatic; D000818:Animals; D015925:Cryopreservation; D005121:Extremities; D066298:In Vitro Techniques; D008297:Male; D018485:Muscle Fibers, Skeletal / Q000378:metabolism / Q000473:pathology*; D009926:Organ Preservation / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D013552:Swine; D013556:Swine, Miniature; D013696:Temperature; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha