J Vet Sci.  2015 Dec;16(4):397-404. 10.4142/jvs.2015.16.4.397.

Effect of serum-derived albumin scaffold and canine adipose tissue-derived mesenchymal stem cells on osteogenesis in canine segmental bone defect model

• Yoon D
• Kang BJ
• Kim Y
• Lee SH
• Rhew D
• Kim WH
• Kweon OK

• KMID: 2133621
• DOI: http://doi.org/10.4142/jvs.2015.16.4.397

Abstract

Composite biological and synthetic grafts with progenitor cells offer an alternative approach to auto- or allografts for fracture repair. This study was conducted to evaluate osteogenesis of autologous serum-derived albumin (ASA) scaffolds seeded with canine adipose tissue-derived mesenchymal stem cells (Ad-MSCs) in a canine segmental bone defect model. ASA scaffold was prepared with canine serum using cross-linking and freeze-drying procedures. Beta-tricalcium phosphate (beta-TCP) was mixed at the cross-linking stage. Ad-MSCs were seeded into the scaffold and incubated for one day before implantation. After 16 weeks, the grafts were harvested for histological analysis. The dogs were divided into five groups: control, ASA scaffolds with and without Ad-MSCs, and ASA scaffolds including beta-TCP with and without Ad-MSCs. ASA scaffolds with Ad-MSCs had a significantly larger area of increased opacity at the proximal and distal host cortex-implant interfaces in radiographs 16 weeks after implantation compared to the groups with beta-TCP (p < 0.05). Histomorphometric analysis showed that ASA scaffolds with Ad-MSCs had significantly greater new bone formation than other groups (p < 0.05). These results suggest that Ad-MSCs seeded into ASA scaffolds enhanced osteogenesis in the bone defect model, but that beta-TCP in the ASA scaffold might prevent penetration of the cells required for bone healing.

Keyword

adipose tissue-derived mesenchymal stem cells; bone defect; serum-derived albumin scaffold

MeSH Terms

Allografts
Animals
Dogs
Mesenchymal Stromal Cells*
Osteogenesis*
Stem Cells
Transplants

Figure

• 

  Fig. 1 The orthotopic implantation procedure. (A) Scaffold preparation: Left, autologous serum-derived albumin (ASA) scaffold; Right, ASA scaffold mixed with β-tricalcium phosphate (β-TCP). (B) Segmental defect in the ulnar diaphysis. (C) Filling the bone defect with the scaffold; Left, ASA scaffold mixed with β-TCP; Right, ASA scaffold. (D) Implant harvested after 16 weeks.

• 

  Fig. 2 Mediolateral radiographs of the treated defects obtained immediately after the operation and 4, 8, and 16 weeks after surgery. The radiopacity area near the proximal and distal host-implant interfaces in each group changes over time. Canine groups: (A) control; (B) Alb; (C) Alb + Ad-MSCs; (D) Alb + β-TCP; and (E) Alb + β-TCP + Ad-MSCs. The β-TCP remained in the middle section of the defect (D and E). Alb, albumin; Ad-MSC, adipose tissue-derived mesenchymal stem cells.

• 

  Fig. 3 Histological findings in longitudinal sections of the segmental bone defects 16 weeks after implantation. The merged picture of the entire defect area, including the adjacent normal bone areas in each group, is shown: (A) Control, (B) Alb, (C) Alb + Ad-MSCs, (D) Alb + β-TCP, and (E) Alb + β-TCP + Ad-MSCs. The proximal portion of the longitudinal section is shown in the left panels (A-E). All experimental groups showed increased bone formation compared to the control group. H&E stain. ncb, native cortical bone area; nba, newly formed bone area; f, fibrous tissue; ts, ASA scaffold mixed with β-TCP.

• 

  Fig. 4 Histological findings in the longitudinal sections of the interfacial area between bone defect and host bone 16 weeks after implantation. Canine groups (A) Alb + Ad-MSCs, (B) Alb + Ad-MSCs + β-TCP groups. (A2 and B2) Merged picture of the defect area including interfacial areas and implanted scaffold. (A3) The remaining ASA scaffold, solid frame of A2. (A4) Chondrocytic change and new woven bone formation, dot frame of A2. (B3) Fibrous tissue, solid frame of B2. (B4) The remaining ASA scaffold mixed with β-TCP, dot frame of B2. H&E staining (A1 and B1), Masson's trichrome staining (A2-4 and B2-4). Magnification: 100× (A3 and 4), 40× (B3 and 4). Scale bars = 200 µm (A3 and 4), 400 µm (B3 and 4).

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