title: [Complex gene-cell therapy in treatment of critical lower limbs ischemia].
abstract: OBJECTIVE
  The paper presents the results of our hybrid gene-cell therapy in case of critical lower limb ischemia. Eighteen patients with critical limb ischemia were enrolled to the study. The bone marrow from each patient was harvested. It had been incubated with VEGF165 gene plasmid for two hours before it was administrated intramuscularly into patient's lower limb.
  
  
  METHODS
  In the study we evaluated: safety, clinical outcomes of the therapy, venous blood VEGF protein concentration before and after (at: 7th, 14th, 28th and 90th day) administration of the stem cells. We obtained samples of muscles from the patients who underwent the limb amputation, which were examined histological and by PCR (Polymerase Chain Reaction) for detection of plasmid genes.
  
  
  RESULTS
  We observed very good clinical outcomes of the therapy. In ten of eighteen patients (-55%) critical lower limb ischemia symptoms subsided--they saved their limbs. The serum VEGF concentration was higher than in healthy controls (p < 0.05). We observed the increase of the concentration of the cytokine at the 14th day an decrease at the 90th day after administration of cells with plasmid. We found expression of plasmid VEGF in transfected stem cells and in tissues taken from amputated limbs. However histological examination did not reveal any sings of new blood vessels formation in the samples taken from ischemic limbs.
  
  
  CONCLUSIONS
  The therapy is safe and effective. We observed significant improvement in patient's clinical state. The therapy needs further investigations.
authors.affiliation: Akademia Medyczna we Wrocławiu, Katedra i Klinika Chirurgii Naczyniowej, Ogólnej i Transplantacyjnej. janskora@wp.pl
authors.forename: Jan
authors.initials: J
authors.lastname: Skóra
authors.forename: Artur
authors.initials: A
authors.lastname: Pupka
authors.forename: Piotr
authors.initials: P
authors.lastname: Barć
authors.forename: Przemysław
authors.initials: P
authors.lastname: Szyber
authors.forename: Wojciech
authors.initials: W
authors.lastname: Polak
authors.forename: Piotr
authors.initials: P
authors.lastname: Szyber
mesh_terms: D000368:Aged; D000671:Amputation, Surgical; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D003422:Critical Care; D005260:Female; D005500:Follow-Up Studies; D015870:Gene Expression / Q000187:drug effects; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007273:Injections, Intramuscular / Q000379:methods; D007511:Ischemia / Q000000981:diagnostic imaging / Q000209:etiology / Q000628:therapy*; D007866:Leg / Q000098:blood supply* / Q000000981:diagnostic imaging / Q000601:surgery; D008297:Male; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000378:metabolism; D016491:Peripheral Vascular Diseases / Q000150:complications / Q000000981:diagnostic imaging / Q000628:therapy*; D010957:Plasmids; D011859:Radiography; D033581:Stem Cell Transplantation; D014182:Transplantation, Autologous / Q000379:methods; D017211:Treatment Failure; D016896:Treatment Outcome; D042461:Vascular Endothelial Growth Factor A / Q000097:blood / Q000235:genetics* / Q000494:pharmacology
chemical_list: D042461:Vascular Endothelial Growth Factor A