Prostacyclin Analogue-Loaded Nanoparticles Attenuate Myocardial Ischemia/Reperfusion Injury in Rats.

Intravenously injected ONO-1301-containing nanoparticles (ONO-1301NPs), unlike an ONO-1301 solution, selectively accumulated in the ischemia/reperfusion (I/R)-injured myocardium of rats and contributed to the prolonged retention of ONO-1301 in the targeted myocardial tissue. In the ischemic area, proangiogenic cytokines were up-regulated and inflammatory cytokines were down-regulated upon ONO-1301NP administration. Consequently, ONO-1301NP-injected rats exhibited a smaller infarct size, better-preserved capillary networks, and a better-preserved myocardial blood flow at 24 h after I/R injury, compared with those in vehicle-injected or ONO-1301 solution-injected rats. ONO-1301NPs attenuate the myocardial I/R injury via proangiogenic and anti-inflammatory effects of the drug.

Development of a vitrification method for preserving human myoblast cell sheets for myocardial regeneration therapy.

BACKGROUND: Tissue-engineered cardiac constructs have potential in the functional recovery of heart failure; however, the preservation of these constructs is crucial for the development and widespread application of this treatment. We hypothesized that tissue-engineered skeletal myoblast (SMB) constructs may be preserved by vitrification to conserve biological function and structure. RESULTS: Scaffold-free cardiac cell-sheet constructs were prepared from SMBs and immersed in a vitrification solution containing ethylene glycol, sucrose, and carboxyl poly-L-lysine. The cell sheet was wrapped in a thin film and frozen rapidly above liquid nitrogen to achieve vitrification (vitrification group, n = 8); fresh, untreated SMB sheets (fresh group, n = 8) were used as the control. The cryopreserved SMB sheets were thawed at 2 days, 1 week, 1 month, and 3 months after cryopreservation for assessment. Thawed, cryopreserved SMB sheets were transplanted into rat hearts in a myocardial infarction nude rat model, and their effects on cardiac function were evaluated. Cell viability in the cardiac constructs of the vitrification group was comparable to that of the fresh group, independent of the period of cryopreservation (p > 0.05). The structures of the cell-sheet constructs, including cell-cell junctions such as desmosomes, extracellular matrix, and cell membranes, were maintained in the vitrification group for 3 months. The expression of cytokine genes and extracellular matrix proteins (fibronectin, collagen I, N-cadherin, and integrin α5) showed similar levels in the vitrification and fresh groups. Moreover, in an in vivo experiment, the ejection fraction was significantly improved in animals treated with the fresh or cryopreserved constructs as compared to that in the sham-treated group (p < 0.05). CONCLUSIONS: Overall, these results show that the vitrification method proposed here preserves the functionality and structure of scaffold-free cardiac cell-sheet constructs using human SMBs after thawing, suggesting the potential clinical application of this method in cell-sheet therapy.

The simultaneous induction of tumorigenesis and Cre-loxP recombination in mice.

To investigate the role of Rac1 for tumorigenesis, we generated inducible transgenic (Tg) mice that simultaneously express polyomavirus middle T antigen (mT) and Cre recombinase under the control of mouse mammary tumor virus long terminal repeat (MMTV-LTR) promoter (MMTV-LTR-tTA/mT-TRE-cre Tg). MMTV-LTR-tTA/mT- TRE-cre Tg mice formed tumors in the subcutaneous tissue and developed lung metastasis. We examined tumor latency and types in rac1 deficient (rac1(flox/-)) and control (rac1(+/+), rac1(+/-) or rac1(flox/+)) MMTV-LTR-tTA/mT-TRE-cre Tg mice and found that formation of cutaneous appendage tumor was suppressed although tumor latency in these mice was not affected by loss of Rac1. These results suggested that Rac1 may play a pivotal role in induction and growth of the mT-mediated epithelial tumors. MMTV-LTR-tTA/mT-TRE-cre Tg mice would provide a versatile animal model to investigate genetic interaction in the tumorigenesis.