title: Hepatocyte growth factor incorporated chitosan nanoparticles augment the differentiation of stem cell into hepatocytes for the recovery of liver cirrhosis in mice.
abstract: BACKGROUND
  Short half-life and low levels of growth factors in the niche of injured microenvironment necessitates the exogenous and sustainable delivery of growth factors along with stem cells to augment the regeneration of injured tissues.
  
  
  METHODS
  Here, recombinant human hepatocyte growth factor (HGF) was incorporated into chitosan nanoparticles (CNP) by ionic gelation method and studied for its morphological and physiological characteristics. Cirrhotic mice received either hematopoietic stem cells (HSC) or mesenchymal stemcells (MSC) with or without HGF incorporated chitosan nanoparticles (HGF-CNP) and saline as control. Biochemical, histological, immunostaining and gene expression assays were carried out using serum and liver tissue samples. One way analysis of variance was used for statics application
  
  
  RESULTS
  Serum levels of selected liver protein and enzymes were significantly increased in the combination of MSC and HGF-CNP (MSC+HGF-CNP) treated group. Immunopositive staining for albumin (Alb) and cytokeratin 18 (CK18), and reverse transcription-polymerase chain reaction (RT-PCR) for Alb, alpha fetoprotein (AFP), CK18, cytokeratin 19 (CK19) ascertained that MSC-HGF-CNP treatment could be an effective combination to repopulate liver parenchymal cells in the liver cirrhosis. Zymogram and western blotting for matrix metalloproteinases 2 and 9 (MMP2 and MMP9) revealed that MMP2 actively involved in the fibrolysis of cirrhotic tissue. Immunostaining for alpha smooth muscle actin (αSMA) and type I collagen showed decreased expression in the MSC+HGF-CNP treatment. These results indicated that HGF-CNP enhanced the differentiation of stem cells into hepatocytes and supported the reversal of fibrolysis of extracellular matrix (ECM).
  
  
  CONCLUSIONS
  Bone marrow stem cells were isolated, characterized and transplanted in mice model. Biodegradable biopolymeric nanoparticles were prepared with the pleotrophic protein molecule and it worked well for the differentiation of stem cells, especially mesenchymal phenotypic cells. Transplantation of bone marrow MSC in combination with HGF-CNP could be an ideal approach for the treatment of liver cirrhosis.
authors.affiliation: Department of Biotechnology, Central Leather Research Institute, Adyar, Chennai-600020, India.
authors.forename: Sivasami
authors.initials: S
authors.lastname: Pulavendran
authors.forename: Chellan
authors.initials: C
authors.lastname: Rose
authors.forename: Asit Baran
authors.initials: AB
authors.lastname: Mandal
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000418:Albumins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D048271:Chitosan / Q000008:administration & dosage*; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D017228:Hepatocyte Growth Factor / Q000627:therapeutic use*; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D053538:Keratin-18 / Q000235:genetics / Q000378:metabolism; D053539:Keratin-19 / Q000235:genetics / Q000378:metabolism; D008103:Liver Cirrhosis / Q000139:chemically induced / Q000628:therapy*; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000187:drug effects; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage*; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D000418:Albumins; D024042:Collagen Type I; C066855:HGF protein, human; D053538:Keratin-18; D053539:Keratin-19; D000509:alpha-Fetoproteins; D017228:Hepatocyte Growth Factor; D048271:Chitosan; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9