title: Therapy with autologous adipose-derived regenerative cells for the care of chronic ulcer of lower limbs in patients with peripheral arterial disease.
abstract: BACKGROUND
  An ulcer is a trophic lesion with loss of tissue that often has a multifactorial genesis. It typically diverges from the physiologic processes of regeneration because it rarely tends to heal spontaneously. In this study, we used purified adipose-derived stem and regenerative cells (ADRCs) extracted from autologous fat, for the care of chronic ulcers of the lower limbs of arteriopathic patients. The primary objective of this study was complete re-epithelization of chronic ulcers; the secondary objective was a decrease in diameter and depth.
  
  
  METHODS
  From January 2010 to January 2012, 20 patients with peripheral arterial disease, with an ankle-brachial index between 0.30-0.40, in the age range 60-70 y (14 men and six women), with chronic ulcers of the lower limb, were involved in the study. Only 10 arteriopathic patients (seven men and three women) with chronic ulcers of the lower limb were surgically treated. Using the Celution system, we isolated a solution of ADRCs in about 150 min. The isolated cells were injected through a 10-mL syringe into the edges of the ulcer, taking care to spread it in all directions. Using a small amount of Celution extract, we performed cell characterization by flow cytometry analysis and cell viability assay.
  
  
  RESULTS
  We monitored patients treated with ADRC or untreated at 4, 10, 20, 60, and 90 d. In all cases treated with ADRC, we found a reduction in both diameter and depth of the ulcer, which led to a decrease in pain associated with the ulcer process. In six of 10 cases there was complete healing of the ulcer. Characterization of the cells by FACS clearly showed that the ADRC cells contained adipose-derived stem cells. Viability assays demonstrated that partial or total closure of the ulcer was attributable exclusively to ADRC cells present in the Celution extract, and not to growth factors extracted during the process of purification of the Celution and injected together with the cells.
  
  
  CONCLUSIONS
  For the first time, the Celution method has been applied for the care of chronic ulcers in the lower extremity of patients with peripheral arterial disease. Our results demonstrate that the technique is feasible for autologous cell application and is not associated with adverse events. Moreover, the transplantation of autologous stem cells extracted with Celution may represent a valuable method for the treatment of chronic ulcers in lower limbs of arteriopathic patients.
authors.affiliation: Laboratory of Applied Biotechnology, Department of Anesthesiology, Surgery, and Emergency Sciences, Second University of Naples, Naples, Italy. Electronic address: gerardo.marino@unina2.it.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Marino
authors.forename: Marco
authors.initials: M
authors.lastname: Moraci
authors.forename: Emilia
authors.initials: E
authors.lastname: Armenia
authors.forename: Consiglia
authors.initials: C
authors.lastname: Orabona
authors.forename: Renato
authors.initials: R
authors.lastname: Sergio
authors.forename: Gabriele
authors.initials: G
authors.lastname: De Sena
authors.forename: Vincenza
authors.initials: V
authors.lastname: Capuozzo
authors.forename: Manlio
authors.initials: M
authors.lastname: Barbarisi
authors.forename: Francesco
authors.initials: F
authors.lastname: Rosso
authors.forename: Giovanni
authors.initials: G
authors.lastname: Giordano
authors.forename: Francesco
authors.initials: F
authors.lastname: Iovino
authors.forename: Alfonso
authors.initials: A
authors.lastname: Barbarisi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000368:Aged; D055109:Ankle Brachial Index; D002908:Chronic Disease; D003131:Combined Modality Therapy; D005260:Female; D005434:Flow Cytometry; D006085:Graft Survival; D006801:Humans; D006931:Hyperbaric Oxygenation; D007871:Leg Ulcer / Q000209:etiology* / Q000601:surgery / Q000628:therapy*; D008297:Male; D008875:Middle Aged; D058729:Peripheral Arterial Disease / Q000150:complications*; D012038:Regeneration; D033581:Stem Cell Transplantation / Q000379:methods*; D014182:Transplantation, Autologous; D016896:Treatment Outcome
keywords: Adipose stem cells; Arterial disease; Chronic ulcer