title: Human fibroblast-derived extracellular matrix constructs for bone tissue engineering applications.
abstract: We exploited the biomimetic approach to generate constructs composed of synthetic biphasic calcium phosphate ceramic and extracellular matrix (SBC-ECM) derived from adult human dermal fibroblasts in complete xeno-free culture conditions. The construct morphology and composition were assessed by scanning electron microscopy, histology, immunohistochemistry, Western blot, glycosaminoglycan, and hydroxyproline assays. Residual DNA quantification, endotoxin testing, and local inflammatory response after implantation in a rat critical-sized calvarial defect were used to access the construct biocompatibility. Moreover, in vitro interaction of human mesenchymal stem cells (hMSCs) with the constructs was studied. The bone marrow- and adipose tissue-derived mesenchymal stem cells were characterized by flow cytometry and tested for osteogenic differentiation capacity prior seeding onto SBC-ECM, followed by alkaline phosphatase, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, and real-time quantitative polymerase chain reaction to assess the osteogenic differentiation of hMSCs after seeding onto the constructs at different time intervals. The SBC-ECM constructs enhanced osteogenic differentiation of hMSCs in vitro and exhibited excellent handling properties and high biocompatibility in vivo. Our results highlight the ability to generate in vitro fibroblast-derived ECM constructs in complete xeno-free conditions as a step toward clinical translation, and the potential use of SBC-ECM in craniofacial bone tissue engineering applications.
authors.affiliation: Department of Dental Medicine, Karolinska Institutet, 14104 Huddinge, Sweden.
authors.forename: Gregory
authors.initials: G
authors.lastname: Tour
authors.forename: Mikael
authors.initials: M
authors.lastname: Wendel
authors.forename: Ion
authors.initials: I
authors.lastname: Tcacencu
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000328:Adult; D000818:Animals; D040761:Biomimetic Materials / Q000494:pharmacology; D001842:Bone and Bones / Q000187:drug effects / Q000502:physiology*; D002113:Calcification, Physiologic / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002516:Ceramics / Q000494:pharmacology; D020405:Dermis / Q000166:cytology; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D006882:Hydroxyapatites / Q000494:pharmacology; D008297:Male; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000235:genetics; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019919:Prosthesis Implantation; D051381:Rats; D012886:Skull / Q000098:blood supply / Q000473:pathology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D006882:Hydroxyapatites; C074950:hydroxyapatite-beta tricalcium phosphate
keywords: biomimetic materials; bone regeneration; stem cells; tissue engineering; xeno-free culture