title: Reconstruction of lymph vessel by lymphatic endothelial cells combined with polyglycolic acid scaffolds: a pilot study.
abstract: Restoration of lymphatic drainage using lymph vessels or tissue grafting is becoming an efficient method for alleviating obstructive lymphedema. However, the lack of ideal lymphatic grafts is the key problem that limits the application of lymphatic transplantation, but now that may be resolved with tissue-engineered lymph vessels. In this study, the feasibility of reconstructing lymph vessels was explored using lymphatic endothelial cells (LECs) combined with polyglycolic acid (PGA) scaffolds. The highly purified human dermal LECs can be isolated from human dermis by immunomagnetic bead sorting and multiplied in culture. The viability and growth potential of subcultured LECs make it possible to obtain large amount of cells in vitro. Light and scanning electron microscopy (SEM) showed that the prefabricated PGA scaffolds, with three-dimensional structure, can support cell adhesion, growth and spreading. The constructs formed with LECs combined with PGA scaffolds were cultured in vitro for 10 days and then implanted subcutaneously into nude mice. Six weeks after implantation, the portions of implanted tubules were harvested. Gross and histological observation demonstrated that the tubular structure still remained in the experimental groups but not in the control groups. Immunohistochemical staining and RT-PCR assay of the implanted vessels revealed positive staining in experimental groups for the lymphatic specific markers Podoplanin, VEGFR-3 and LYVE-1. The results indicate that LECs can serve as seed cells and be successfully combined with PGA scaffolds, and the tissue-engineered tubular structure using implanted LECs-PGA compounds showed preliminary characteristics of lymph vessels. A gap between the nearly normal or functional lymph vessel still exists as we have only the endothelial cell-lined duct, but this study demonstrates that it is feasible to construct tissue-engineered lymph vessels using LECs combined with a biodegradable material.
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Zhizaoju Road 639, Shanghai 200011, China.
authors.forename: Ting ting
authors.initials: Tt
authors.lastname: Dai
authors.forename: Zhao hua
authors.initials: Zh
authors.lastname: Jiang
authors.forename: Sheng li
authors.initials: Sl
authors.lastname: Li
authors.forename: Guang dong
authors.initials: Gd
authors.lastname: Zhou
authors.forename: James D
authors.initials: JD
authors.lastname: Kretlow
authors.forename: Wei gang
authors.initials: Wg
authors.lastname: Cao
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Yi lin
authors.initials: Yl
authors.lastname: Cao
mesh_terms: D000818:Animals; D001807:Blood Vessel Prosthesis; D002470:Cell Survival; D042783:Endothelial Cells / Q000166:cytology*; D052816:Foreskin / Q000166:cytology; D006651:Histocytochemistry; D006801:Humans; D042601:Lymphatic Vessels / Q000166:cytology*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008855:Microscopy, Electron, Scanning; D010865:Pilot Projects; D011100:Polyglycolic Acid / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D011100:Polyglycolic Acid