Role of Endothelial Differentiated Adipose-derived Stem Cells in Repairing Calvarial Critical Size Defects in the Laboratory Rat (Rattus norvegicus)
Abstract
Objectives: Calvarial bone defects are amongst the most common combat injuries. The treatment of large defects is difficult due to donor site limitations. Our aim in this study was to evaluate in vivo bone engineering in poly lactic-co-glycolic acid (PLGA) scaffolds seeded with endothelial and osteogenic differentiated adipose-derived stem cells (ASCs). Methods: Rat ASCs were induced into endothelial (ASC-endo) and osteogenic (ASC osteo) lineages. The optimal duration of endothelial cell differentiation was evaluated with flow cytometry analysis. Osteogenic differentiation was confirmed with alizarin red staining. Critical size (8 mm) defects were created in the calvaria of Lewis rats. The defects were treated with blank PLGA scaffolds (group I), PLGA scaffolds with undifferentiated ASCs (group II), PLGA scaffolds with ASC-osteo (group III), or PLGA scaffolds with ASC-endo (group IV). Bone healing in the defects with evaluated at 8 weeks postsurgery with micro-CT scans and histological staining with hematoxylin-eosin and Masson's trichrome stains). Results: Micro-CT analyses of calvarial defects showed the highest bone mineral density in the ASC-osteo group, but there was no statistically significant difference between treatments and control (p = 0.56). Photometric analysis of histology slides suggested a trend towards more bone formation in the ASC-osteo group, but there was no significant difference between treatments and control (p = 0.13). Conclusions: We were able to successfully differentiate ASCs into endothelial and osteogenic lineages and confirmed this using gene expression, protein expression, and histology. PLGA scaffold was a suitable medium for cell seeding. However, we were unsuccessful in producing significant new bone formation when osteocyte or endothelial cell differentiated ASCs were seeded separately on scaffolds in rat critical sized calvarial defects.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 16, 2014
- Accession Number
- ADA608955
Entities
People
- Chris Gold
- David Sahar
Organizations
- David Grant USAF Medical Center