We have reported earlier that peripheral blood

We have reported earlier that peripheral blood cells expressing the tyrosine kinase receptors VEGFR-2 or Tie-2 define functionally competent cell populations capable of re-endothelialization and differentiation into smooth muscle cells, the numbers of which are ≈0.8% and 2% respectively in PWB (Nowak et al., 2004). In the present study enumeration of VEGFR-2+ cells after perfusion with 25ml blood from various donors showed an average cell count of 7.65±1.47×104cells/cm2 in RV which was similar to the numbers reported by Ranjan AK et al. who used 6.6×104 EPC for endothelialization of small diameter vascular prosthesis (Ranjan et al., 2009). The volume of 25ml blood for recellularization was chosen for the following two reasons, a) this was the minimum amount required for perfusion through the vein and the bioreactor system and b) consideration for pediatric patients with fragile blood vessels. Larger volumes of blood did not result in reduced time needed for repopulation (unpublished observations), although this finding needs to be verified in a larger study. In the present and other currently ongoing studies we have successfully recellularized veins using blood from individuals and patients in the age range of 4–55years. However, it is reported that the numbers of circulating stem/precursor cells is decreased in patients with diabetes (van Ark et al., 2012) and end-stage renal diseases (Westerweel et al., 2007). So it remains to be tested whether this method would work in such patients. We did not detect any HLA topoisomerase inhibitors after transplantation indicating satisfactory decellularization of the blood vessels. The patients have been followed for one year and Nine months and one year and Seven months respectively. Although patient 2 had to be re-operated and problems with the anastomoses could be noted, this was evident already at the primary operation. Small children or children with inferior intrahepatic portal vein system may require sequential surgery. The second intervention however, opened up of the obstructed portal bed and diminished the initial gradient over the anastomosis. Taking into account that the alternative treatment for these patients is liver transplantation including lifelong immunosuppression, a revision must be seen as a minor drawback. The following are the supplementary data related to this article.
Conflicts of Interest
Author Contributions SSH conceived, designed and oversaw all of the in vitro studies, collection of results, interpretation of the data, and writing of the manuscript. SSH is also responsible for the new recellularization protocol. MO is the senior surgeon who did the surgery and perioperative care. MO was also responsible for collection, interpretation and writing of the clinical data. VKK did the all preclinical work, and supervised the whole procedure and wrote the report. GT assisted in the development of the decellularization process for matrix creation, the seeding procedure, and did immunofluorescence histology. PBP designed the bioreactor. HBä designed and performed all the biomechanical studies and co-wrote the paper. RS and HBo performed patient screening and follow-up. AJ supplied blood vessels for the study and co-wrote the paper. All authors have seen and approved the final version of the report to be published. Funding: Swedish Government. The sponsor of the study has no role in the study design, data collection, data analysis, data interpretation, or writing of the report.
Acknowledgments This study was financed by the Swedish Government LUA ALF grants to SSH and MO, The Swedish Heart and Lung Foundation20130505, The Swedish Research CouncilK2013-65X-22347-01-3 and The Inga Britt and Arne Lundberg\'s Foundation2009-362 to SSH.
Introduction Although DCCT, UKPDS and our Kumamoto Study (Shichiri et al., 2000) have shown that strict glycemic control could prevent microvascular complications, the increase of diabetes is still an important issue worldwide. The increase of type 2 diabetes mellitus (T2DM) is associated with excess visceral adiposity, which is tightly linked to metabolic syndrome (MS). MS is recognized as a cluster of cardiovascular risk factors such as hyperglycemia, dyslipidemia, elevated blood pressure and chronic inflammation (Alberti et al., 2005). Visceral fat has been demonstrated to express more pro-inflammatory cytokines than subcutaneous fat in obese states (Ohman et al., 2009). Inflammatory markers such as C-reactive protein (CRP) (Tamakoshi et al., 2003) and tumor necrosis factor (TNF)-α have been linked to MS.