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Tissue-engineered Neogenesis of Human-shaped Mandibular Condyle from Rat Mesenchymal Stem Cells

Tissue Engineering Laboratory, Rm. 237, Departments of Orthodontics (MC 841), Bioengineering, and Anatomy and Cell Biology, University of Illinois at Chicago, 801 S. Paulina Street, Chicago, IL 60612-7211, USA;

View larger version (69K): [in a new window]   Figure 1. Tissue-engineered neogenesis of human-shaped mandibular condyle from rat mesenchymal stem cells. (A) Recovery process of a tissue-engineered mandibular condyle after eight-week in vivo implantation in immunodeficient mouse. (B,C) Harvested osteochondral construct retained the shape and size of the cadaver human mandibular condyle mold. (D) Acrylic model of a cadaver human mandibular condyle. (E) Polyurethane mold used to load the cell/polymer suspensions.

View larger version (98K): [in a new window]   Figure 2. Photomicrographs of histologic phenotypes of a representative tissue-engineered mandibular condyle following 8 wks of in vivo implantation. (A) Von Kossa silver-stained section showing the interface between chondral and osseous layers. Multiple mineralization nodules were present in the osseous layer (lower half of the photomicrograph), but absent in the chondral layer (upper half of the photomicrograph). (B) Positive safranin O staining of the chondrogenic layer was represented by intense red, indicating the synthesis of negatively charged cartilage-specific glycosaminoglycans in the extracellular matrix. (C) H&E-stained section of the osteogenic layer showing a representative island structure consisting of MSC-differentiated osteoblast-like cells on the surface and in the center. (D) Positive toluidine blue staining of an island structure in the osseous layer.

View larger version (142K): [in a new window]   Figure 3. Chondrogenesis driven by MSC-derived chondrogenic cells in ex vivo samples. (A) Positive reaction of MSC-derived chondrogenic cells to safranin O in monolayer culture following four-week treatment with chondrogenic medium containing TGF-ß1. (B) Monolayer culture of MSCs from the same population as in (A), cultured for 4 wks with DMEM/FBS but without TGF-ß1, showed no positive reaction to safranin O. (C) Positive reaction of PEG hydrogel encapsulating MSC-derived chondrogenic cells to safranin O, demonstrating the presence of cartilage-specific glycosaminoglycans after four-week incubation in chondrogenic medium containing TGF-ß1. (D) PEG hydrogel encapsulating the same population of MSCs as in (C), but without exposure to TGF-ß1, showed negative reaction to safranin O.

View larger version (150K): [in a new window]   Figure 4. Osteogenesis driven by MSC-derived chondrogenic cells in ex vivo samples. (A) Positive reactions of MSC-derived osteogenic cells to alkaline phosphatase (white arrow) and von Kossa silver stain (green arrow) following four-week incubation in oseteogenic medium. (B) Monolayer culture of MSCs from the same population as in (A), cultured for 4 wks with DMEM/FBS but without osteoinduction factors, showed no positive reaction to either alkaline phosphatase or von Kossa silver stains. (C) Von Kossa silver-stained section of PEG-hydrogel encapsulating MSC-derived osteogenic cells showing mineral nodules. (D) The same population of MSCs encapsulated in the PEG-hydrogel construct without exposure to osteogenic medium showed no evidence of mineralization by von Kossa silver staining.