Therapy with autologous adipose-derived regenerative cells for the care of chronic ulcer of lower limbs in patients with peripheral arterial disease



      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.


      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.


      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.


      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.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Surgical Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Gentzkow G.D.
        • Iwasaki S.D.
        • Hershon K.S.
        • et al.
        Use of dermagraft, a cultured human dermis, to treat diabetic foot ulcers.
        Diabetes Care. 1996; 19: 350
        • Hanft J.R.
        • Surprenant M.S.
        Healing of chronic foot ulcers in diabetic patients treated with a human fibroblast derived dermis.
        J Foot Ankle Surg. 2002; 41: 291
        • Akita S.
        • Yoshimoto H.
        • Akino K.
        • et al.
        Early experiences with stem cells in treating chronic wounds.
        Clin Plast Surg. 2012; 39: 281
        • Mizuno H.
        • Nambu M.
        Adipose-derived stem cells for skin regeneration.
        Methods Mol Biol. 2011; 702: 453
        • Lin K.
        • Matsubara Y.
        • Masuda Y.
        • et al.
        Characterization of adipose tissue-derived cells isolated with the Celution system.
        Cytotherapy. 2008; 10: 417
        • Xu D.
        • Zou L.
        • Xing Y.
        • et al.
        Diagnostic value of ankle-brachial index in peripheral arterial disease: a meta-analysis.
        Can J Cardiol. 2013; 29: 492
        • Alfano C.
        • Angelisanti M.
        • Calzoni C.
        • Somma F.
        • Chiummariello S.
        Treatment of ulcer and difficult wounds of the lower limbs: our experience.
        Ann Ital Chir. 2012; 83: 135
        • Wagner W.
        • Wein F.
        • Seckinger A.
        • et al.
        Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood.
        Exp Hematol. 2005; 33: 1402
        • Mitchell J.
        • Micintosh K.
        • Zavonic S.
        • et al.
        Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell–associated markers.
        Stem Cells. 2006; 24: 376
        • Akita S.
        • Akino K.
        • Hirano A.
        • Ohtsuru A.
        • Yamashita S.
        Noncultured autologous adipose-derived stem cells therapy for chronic radiation injury.
        Stem Cells Int. 2010; 2010: 532704
        • Planat-Benard V.
        • Silvestre J.S.
        • Cousin B.
        • et al.
        Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives.
        Circulation. 2004; 109: 656
        • Marino G.
        • Rosso F.
        • Ferdinando P.
        • et al.
        Growth and endothelial differentiation of adipose stem cells on polycaprolactone.
        J Biomed Mater Res A. 2012; 100: 543
        • Rigotti G.
        • Marchi A.
        • Galiè M.
        • et al.
        Clinical treatment of radiotherapy tissue damage by lipoaspirate transplant: a healing process mediated by adipose-derived adult stem cells.
        Plast Reconstr Surg. 2007; 119: 1409
        • Boquest A.C.
        • Shahdadfar A.
        • Fronsdal K.
        • et al.
        Isolation and transcription profiling of purified uncultured human stromal stem cells: alteration of gene expression after in vitro cell culture.
        Mol Biol Cell. 2005; 16: 1131