title: Metformin Induces Cell Cycle Arrest, Reduced Proliferation, Wound Healing Impairment In Vivo and Is Associated to Clinical Outcomes in Diabetic Foot Ulcer Patients.
abstract: BACKGROUND
  Several epidemiological studies in diabetic patients have demonstrated a protective effect of metformin to the development of several types of cancer. The underlying mechanisms of such phenomenon is related to the effect of metformin on cell proliferation among which, mTOR, AMPK and other targets have been identified. However, little is known about the role that metformin treatment have on other cell types such as keratinocytes and whether exposure to metformin of these cells might have serious repercussions in wound healing delay and in the development of complications in diabetic patients with foot ulcers or in their exacerbation.
  
  
  METHODS
  HaCaT Cells were exposed to various concentrations of metformin and cell viability was evaluated by a Resazurin assay; Proliferation was also evaluated with a colony formation assay and with CFSE dilution assay by flow cytometry. Cell cycle was also evaluated by flow cytometry by PI staining. An animal model of wound healing was used to evaluate the effect of metformin in wound closure. Also, an analysis of patients receiving metformin treatment was performed to determine the effect of metformin treatment on the outcome and wound area. Statistical analysis was performed on SPSS v. 18 and GraphPad software v.5.
  
  
  RESULTS
  Metformin treatment significantly reduces cell proliferation; colony formation and alterations of the cell cycle are observed also in the metformin treated cells, particularly in the S phase. There is a significant increase in the area of the wound of the metformin treated animals at different time points (P<0.05). There is also a significant increase in the size and wound area of the patients with diabetic foot ulcers at the time of hospitalization. A protective effect of metformin was observed for amputation, probably associated with the anti inflammatory effects reported of metformin.
  
  
  CONCLUSIONS
  Metformin treatment reduces cell proliferation and reduces wound healing in an animal model and affects clinical outcomes in diabetic foot ulcer patients. Chronic use of this drug should be further investigated to provide evidence of their security in association with DFU.
authors.affiliation: Unidad de Investigación Biomédica de Zacatecas, Instituto Mexicano del Seguro Social (IMSS), Zacatecas, Zac., México.|Área de ciencias de la salud, Universidad Autónoma de Zacatecas. Zacatecas, Zac., México.
authors.forename: Fatima
authors.initials: F
authors.lastname: Ochoa-Gonzalez
authors.affiliation: Unidad Académica de Ciencias Químicas, Área de Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, México.
authors.forename: Alberto R
authors.initials: AR
authors.lastname: Cervantes-Villagrana
authors.affiliation: Unidad de Investigación Biomédica de Zacatecas, Instituto Mexicano del Seguro Social (IMSS), Zacatecas, Zac., México.
authors.forename: Julio C
authors.initials: JC
authors.lastname: Fernandez-Ruiz
authors.affiliation: Unidad de Investigación Biomédica de Zacatecas, Instituto Mexicano del Seguro Social (IMSS), Zacatecas, Zac., México.
authors.forename: Hilda S
authors.initials: HS
authors.lastname: Nava-Ramirez
authors.affiliation: Unidad Académica de Ciencias Químicas, Área de Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, México.
authors.forename: Adriana C
authors.initials: AC
authors.lastname: Hernandez-Correa
authors.affiliation: Unidad de Investigación Biomédica de Zacatecas, Instituto Mexicano del Seguro Social (IMSS), Zacatecas, Zac., México.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Enciso-Moreno
authors.affiliation: Unidad de Investigación Biomédica de Zacatecas, Instituto Mexicano del Seguro Social (IMSS), Zacatecas, Zac., México.
authors.forename: Julio E
authors.initials: JE
authors.lastname: Castañeda-Delgado
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D002460:Cell Line; D003921:Diabetes Mellitus, Experimental / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D003924:Diabetes Mellitus, Type 2 / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D017719:Diabetic Foot / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D005260:Female; D006801:Humans; D008297:Male; D008687:Metformin / Q000008:administration & dosage*; D008875:Middle Aged; D051381:Rats; D017208:Rats, Wistar; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D008687:Metformin