title: Decellularized bone matrix grafts for calvaria regeneration.
abstract: Decellularization is a promising new method to prepare natural matrices for tissue regeneration. Successful decellularization has been reported using various tissues including skin, tendon, and cartilage, though studies using hard tissue such as bone are lacking. In this study, we aimed to define the optimal experimental parameters to decellularize natural bone matrix using 0.5% sodium dodecyl sulfate and 0.1% NH4OH. Then, the effects of decellularized bone matrix on rat mesenchymal stem cell proliferation, osteogenic gene expression, and osteogenic differentiations in a two-dimensional culture system were investigated. Decellularized bone was also evaluated with regard to cytotoxicity, biochemical, and mechanical characteristics in vitro. Evidence of complete decellularization was shown through hematoxylin and eosin staining and DNA measurements. Decellularized bone matrix displayed a cytocompatible property, conserved structure, mechanical strength, and mineral content comparable to natural bone. To study new bone formation, implantation of decellularized bone matrix particles seeded with rat mesenchymal stem cells was conducted using an orthotopic in vivo model. After 3 months post-implantation into a critical-sized defect in rat calvaria, new bone was formed around decellularized bone matrix particles and also merged with new bone between decellularized bone matrix particles. New bone formation was analyzed with micro computed tomography, mineral apposition rate, and histomorphometry. Decellularized bone matrix stimulated mesenchymal stem cell proliferation and osteogenic differentiation in vitro and in vivo, achieving effective bone regeneration and thereby serving as a promising biological bone graft.
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Dong Joon
authors.initials: DJ
authors.lastname: Lee
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Shannon
authors.initials: S
authors.lastname: Diachina
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Yan Ting
authors.initials: YT
authors.lastname: Lee
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Lixing
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Rui
authors.initials: R
authors.lastname: Zou
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Na
authors.initials: N
authors.lastname: Tang
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Han
authors.initials: H
authors.lastname: Han
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.affiliation: Oral and Craniofacial Health Sciences Research, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA; Department of Orthodontics, UNC School of Dentistry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Ching-Chang
authors.initials: CC
authors.lastname: Ko
keywords: Decellularized bone matrix; histomorphometry; mesenchymal stem cell; mineral apposition rate; orthotopic