Decellularized Cartilage Extracellular Matrix Incorporated Silk Fibroin Hybrid Scaffolds for Endochondral Ossification Mediated Bone Regeneration

Vivek Jeyakumar, Nedaa Amraish, Eugenia Niculescu-Morsza, Christoph Bauer, Dieter Pahr, Stefan Nehrer

Research output: Journal article (peer-reviewed)Journal article

10 Citations (Scopus)


Tissue engineering strategies promote bone regeneration for large bone defects by stimulating the osteogenesis route via intramembranous ossification in engineered grafts, which upon implantation are frequently constrained by insufficient integration and functional anastomosis of vasculature from the host tissue. In this study, we developed a hybrid biomaterial incorporating decellularized cartilage extracellular matrix (CD-ECM) as a template and silk fibroin (SF) as a carrier to assess the bone regeneration capacity of bone marrow-derived mesenchymal stem cells (hBMSC's) via the endochondral ossification (ECO) route. hBMSC's were primed two weeks for chondrogenesis, followed by six weeks for hypertrophy onto hybrid CD-ECM/SF or SF alone scaffolds and evaluated for the mineralized matrix formation in vitro. Calcium deposition biochemically determined increased significantly from 4-8 weeks in both SF and CD-ECM/SF constructs, and retention of sGAG's were observed only in CD-ECM/SF constructs. SEM/EDX revealed calcium and phosphate crystal localization by hBMSC's under all conditions. Compressive modulus reached a maximum of 40 KPa after eight weeks of hypertrophic induction. μCT scanning at eight weeks indicated a cloud of denser minerals in groups after hypertrophic induction in CD-ECM/SF constructs than SF constructs. Gene expression by RT-qPCR revealed that hBMSC's expressed hypertrophic markers VEGF, COL10, RUNX2, but the absence of early hypertrophic marker ChM1 and later hypertrophic marker TSBS1 and the presence of osteogenic markers ALPL, IBSP, OSX under all conditions. Our data indicate a new method to prime hBMSC'S into the late hypertrophic stage in vitro in mechanically stable constructs for ECO-mediated bone tissue regeneration.

Original languageEnglish
Article number4055
JournalInternational Journal of Molecular Sciences
Issue number8
Publication statusPublished - 02 Apr 2021


  • Adipose Tissue/growth & development
  • Animals
  • Bone Regeneration/genetics
  • Bone and Bones/metabolism
  • Cartilage/growth & development
  • Cell Differentiation/genetics
  • Chondrocytes/metabolism
  • Chondrogenesis/genetics
  • Extracellular Matrix/genetics
  • Fibroins/chemistry
  • Humans
  • Mesenchymal Stem Cells/cytology
  • Osteogenesis/genetics
  • Tissue Engineering
  • Tissue Scaffolds


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