title: Critical-size bone defect repair using amniotic fluid stem cell/collagen constructs: effect of oral ferutinin treatment in rats.
abstract: OBJECTIVE
  This study aims to evaluate the bone regeneration in a rat calvarias critical size bone defect treated with a construct consisting of collagen type I and human amniotic fluid stem cells (AFSCs) after oral administration of phytoestrogen ferutinin.
  
  
  METHODS
  In 12 week old male rats (n=10), we performed two symmetric full-thickness cranial defects on each parietal region, and a scaffold was implanted into each cranial defect. The rats were divided into four groups: 1) collagen scaffold, 2) collagen scaffold+ferutinin at a dose of 2mg/kg/5 mL, 3) collagen scaffold + AFSCs, and 4) collagen scaffold + AFSCs + ferutinin. The rats were sacrificed after 4 weeks, and the calvariae were removed, fixed, embedded in paraffin and cut into 7 μm thick sections. Histomorphometric measures, immunohistochemical and immunofluorescence analyses were performed on the paraffin sections.
  
  
  RESULTS
  The histomorphometric analysis on H&E stained sections showed a significant increase in the regenerated area of the 4th group compared with the other groups. Immunohistochemistry performed with a human anti-mitochondrial antibody showed the presence of AFSCs 4 weeks after the transplant. Immunofluorescence analysis revealed the presence of osteocalcin and estrogen receptors (ERα and GPR30) in all groups, with a greater expression of all markers in samples where the scaffold was treated with AFSCs and the rats were orally administered ferutinin.
  
  
  CONCLUSIONS
  Our results demonstrated that the oral administration of ferutinin is able to improve the bone regeneration of critical-size bone defects in vivo that is obtained with collagen-AFSCs constructs.
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplants, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy. Electronic address: manuela.zavatti@unimore.it.
authors.forename: Manuela
authors.initials: M
authors.lastname: Zavatti
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplants, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Bertoni
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplants, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Tullia
authors.initials: T
authors.lastname: Maraldi
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplants, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Resca
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplants, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Beretti
authors.affiliation: EURAC Research, Center for Biomedicine, Bolzano, Italy.
authors.forename: Marianna
authors.initials: M
authors.lastname: Guida
authors.affiliation: Unit of Obstetrics & Gynecology, IRCCS-Arcispedale Santa Maria Nuova, Reggio Emilia, Italy.
authors.forename: Giovanni B
authors.initials: GB
authors.lastname: La Sala
authors.affiliation: Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplants, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Anto
authors.initials: A
authors.lastname: De Pol
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D001565:Benzoates / Q000494:pharmacology*; D001846:Bone Development / Q000187:drug effects*; D001643:Bridged Bicyclo Compounds / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology*; D003508:Cycloheptanes / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000378:metabolism; D006801:Humans; D008297:Male; D015675:Osteocalcin / Q000378:metabolism; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D012717:Sesquiterpenes / Q000494:pharmacology*; D033581:Stem Cell Transplantation
chemical_list: D001565:Benzoates; D001643:Bridged Bicyclo Compounds; D003508:Cycloheptanes; D047628:Estrogen Receptor alpha; C520305:Gper1 protein, rat; D043562:Receptors, G-Protein-Coupled; D012717:Sesquiterpenes; D015675:Osteocalcin; C111068:4-oxy-6-(4-oxybezoyloxy)dauc-8,9-en; D003094:Collagen
keywords: AFSCs; Bone regeneration; Collagen scaffold; Ferutinin; Rat