In the last decade, the production and use of decellularized tissues in tissue engineering applications is receiving considerable attention, as the natural extracellular matrix (ECM) can provide necessary physical cues supporting the restoration and development of functional tissues. In relation to bone and osteochondral tissue engineering, we are interested in producing decellularized Wharton’s jelly matrix (DWJM) from human umbilical cord to be used as a scaffold to restore the bone/ osteochondral microenvironment. WJ is a mucous connective tissue that surrounds the umbilical cord vessels and is covered by a layer of simple amniotic epithelium. This is a very peculiar tissue with several unique biomechanical characteristics related to its biochemical structure. We have explored the composition of human WJ extracellular matrix and developed a strategy for decellularization based on a previous protocol set up in our lab with reagents preserving ECM structure and composition. Decellularization is a process that allows the DNA removal and should preserve morphology and chemical factors of the processed tissue. In this work, Wharton’s jelly decellularization was performed through different methods, including the chemical Latrunculin and the detergent enzymatic strategy (sodium dehoxycholate-SDC- /DNAse) demonstrating that the second method was more efficient in balancing both preservation of tissue structure, composition and cell removal. Masson’s trichrome and Alcian blue staining were performed to measure both the efficacy of cells removal and the percentage of glycosaminoglycans (GAGs) content; DNA quantification was carried out to assess the elimination of genetic material. We are now planning cell culture experiments to investigate the potentiality of DWJM in regenerative medicine applications, and improve the properties of the cells for bone and cartilage tissue repair. For this purpose, the attachment, survival and differentiation ability of human mesenchymal stem cells, osteoblasts and chondrocytes will be tested (a) in X3 Hypoxia Hood and Culture Combo – Xvivo System with 1-3% oxygen tension in order to mimic the physiological conditions, and (b) in the absence of differentiating agents, relying on the potential of the components of the DWJM in supporting the behavior of the cells.
Decellularized Wharton’s Jelly from human umbilical cord: a novel scaffold for tissue engineering
Lambertini E.;Scussel Bergamin L.;Piva R.;Penolazzi L.
2018
Abstract
In the last decade, the production and use of decellularized tissues in tissue engineering applications is receiving considerable attention, as the natural extracellular matrix (ECM) can provide necessary physical cues supporting the restoration and development of functional tissues. In relation to bone and osteochondral tissue engineering, we are interested in producing decellularized Wharton’s jelly matrix (DWJM) from human umbilical cord to be used as a scaffold to restore the bone/ osteochondral microenvironment. WJ is a mucous connective tissue that surrounds the umbilical cord vessels and is covered by a layer of simple amniotic epithelium. This is a very peculiar tissue with several unique biomechanical characteristics related to its biochemical structure. We have explored the composition of human WJ extracellular matrix and developed a strategy for decellularization based on a previous protocol set up in our lab with reagents preserving ECM structure and composition. Decellularization is a process that allows the DNA removal and should preserve morphology and chemical factors of the processed tissue. In this work, Wharton’s jelly decellularization was performed through different methods, including the chemical Latrunculin and the detergent enzymatic strategy (sodium dehoxycholate-SDC- /DNAse) demonstrating that the second method was more efficient in balancing both preservation of tissue structure, composition and cell removal. Masson’s trichrome and Alcian blue staining were performed to measure both the efficacy of cells removal and the percentage of glycosaminoglycans (GAGs) content; DNA quantification was carried out to assess the elimination of genetic material. We are now planning cell culture experiments to investigate the potentiality of DWJM in regenerative medicine applications, and improve the properties of the cells for bone and cartilage tissue repair. For this purpose, the attachment, survival and differentiation ability of human mesenchymal stem cells, osteoblasts and chondrocytes will be tested (a) in X3 Hypoxia Hood and Culture Combo – Xvivo System with 1-3% oxygen tension in order to mimic the physiological conditions, and (b) in the absence of differentiating agents, relying on the potential of the components of the DWJM in supporting the behavior of the cells.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
