Clara Sanjurjo-Rodríguez1, Adela Helvia Martínez-Sánchez2, Tamara Hermida-Gómez3, Isaac Fuentes-Boquete4, Silvia Díaz-Prado5, Francisco J Blanco6. 1. Cell Therapy and Regenerative Medicine Unit, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), Department of Medicine, Faculty of Health Sciences, University of A Coruña, A Coruña, Spain. clara.sanjurjo@udc.es. 2. Tissue Bioengineering and Cell Therapy Unit (GBTTC-CHUAC), Rheumatology Group, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), A Coruña, Spain. bioams01@udc.es. 3. Tissue Bioengineering and Cell Therapy Unit (GBTTC-CHUAC), Rheumatology Group, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), A Coruña, Spain. tamara.hermida.gomez@sergas.es. 4. Cell Therapy and Regenerative Medicine Unit, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), Department of Medicine, Faculty of Health Sciences, University of A Coruña, A Coruña, Spain. isaac.fuentes.boquete@sergas.es. 5. Cell Therapy and Regenerative Medicine Unit, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), Department of Medicine, Faculty of Health Sciences, University of A Coruña, A Coruña, Spain. s.diaz1@udc.es. 6. Tissue Bioengineering and Cell Therapy Unit (GBTTC-CHUAC), Rheumatology Group, Institute of Biomedical Research of A Coruña (INIBIC), University Hospital Complex A Coruña (CHUAC), Galician Health Service (SERGAS), University of Santiago de Compostela, A Coruña, Spain. fblagar@sergas.es.
Abstract
AIM: The aim of this study was to evaluate proliferation and chondrogenic differentiation of human bone-marrow mesenchymal stromal cells (hBMSCs) cultured on collagen biomaterials. MATERIALS AND METHODS: hBMSCs were seeded on five different collagen (Col) sponges: C1C2 (types I and II Col), C1C2HS (types I and II Col plus heparan sulphate (HS)), C1C2CHS (types I and II Col plus chondroitin sulphate (CHS)), C1-OLH3 (type I Col plus low molecular weight heparin) and C1CHS (type I Col plus CHS). The resulting constructs were analyzed by histological and immunohistochemical staining, molecular biology and electron microscopy. Col released into culture media was measured by a dye-binding method Results: hBMSCs on biomaterials C1C2, C1C2HS and C1C2CHS had more capacity to attach, proliferate and synthesize Col II and proteoglycans in the extracellular matrix (ECM) than on C1-OLH3 and C1CHS. The presence of aggrecan was detected only at the gene level. Total Col liberated by the cells in the supernatants in all scaffold cultures was detected. The level of Col I in the ECM was lower in C1-OLH3 and that of Col II was highest in C1C2 and C1C2HS. Electron microscopy showed differently shaped cells, from rounded to flattened, in all constructs. Col fibers in bundles were observed in C1C2CHS by transmission electron microscopy. CONCLUSIONS: The results show that Col I and Col II (C1C2, C1C2HS and C1C2CHS) biomaterials allowed cell proliferation and chondrogenic-like differentiation of hBMSCs at an early stage. Constructs cultured on C1C2HS and C1C2CHS showed better cartilage-like phenotype than the other ones.
AIM: The aim of this study was to evaluate proliferation and chondrogenic differentiation of human bone-marrow mesenchymal stromal cells (hBMSCs) cultured on collagen biomaterials. MATERIALS AND METHODS: hBMSCs were seeded on five different collagen (Col) sponges: C1C2 (types I and II Col), C1C2HS (types I and II Col plus heparan sulphate (HS)), C1C2CHS (types I and II Col plus chondroitin sulphate (CHS)), C1-OLH3 (type I Col plus low molecular weight heparin) and C1CHS (type I Col plus CHS). The resulting constructs were analyzed by histological and immunohistochemical staining, molecular biology and electron microscopy. Col released into culture media was measured by a dye-binding method Results: hBMSCs on biomaterials C1C2, C1C2HS and C1C2CHS had more capacity to attach, proliferate and synthesize Col II and proteoglycans in the extracellular matrix (ECM) than on C1-OLH3 and C1CHS. The presence of aggrecan was detected only at the gene level. Total Col liberated by the cells in the supernatants in all scaffold cultures was detected. The level of Col I in the ECM was lower in C1-OLH3 and that of Col II was highest in C1C2 and C1C2HS. Electron microscopy showed differently shaped cells, from rounded to flattened, in all constructs. Col fibers in bundles were observed in C1C2CHS by transmission electron microscopy. CONCLUSIONS: The results show that Col I and Col II (C1C2, C1C2HS and C1C2CHS) biomaterials allowed cell proliferation and chondrogenic-like differentiation of hBMSCs at an early stage. Constructs cultured on C1C2HS and C1C2CHS showed better cartilage-like phenotype than the other ones.
Authors: Clara Sanjurjo-Rodríguez; Rocío Castro-Viñuelas; Tamara Hermida-Gómez; Isaac Manuel Fuentes-Boquete; Francisco Javier de Toro; Francisco Javier Blanco; Silvia María Díaz-Prado Journal: Int J Med Sci Date: 2017-09-28 Impact factor: 3.738