J Korean Med Sci.  2011 Apr;26(4):482-491. 10.3346/jkms.2011.26.4.482.

Establishment of Efficacy and Safety Assessment of Human Adipose Tissue-Derived Mesenchymal Stem Cells (hATMSCs) in a Nude Rat Femoral Segmental Defect Model

• Choi HJ
• Kim JM
• Kwon E
• Che JH
• Lee JI
• Cho SR
• Kang SK
• Ra JC
• Kang BC

• KMID: 1777877
• DOI: http://doi.org/10.3346/jkms.2011.26.4.482

Abstract

Human adipose tissue-derived mesenchymal stem cell (hATMSC) have emerged as a potentially powerful tool for bone repair, but an appropriate evaluation system has not been established. The purpose of this study was to establish a preclinical assessment system to evaluate the efficacy and safety of cell therapies in a nude rat bone defect model. Segmental defects (5 mm) were created in the femoral diaphyses and transplanted with cell media (control), hydroxyapatite/tricalcium phosphate scaffolds (HA/TCP, Group I), hATMSCs (Group II), or three cell-loading density of hATMSC-loaded HA/TCP (Group III-V). Healing response was evaluated by serial radiography, micro-computed tomography and histology at 16 weeks. To address safety-concerns, we conducted a GLP-compliant toxicity study. Scanning electron microscopy studies showed that hATMSCs filled the pores/surfaces of scaffolds in a cell-loading density-dependent manner. We detected significant increases in bone formation in the hATMSC-loaded HA/TCP groups compared with other groups. The amount of new bone formation increased with increases in loaded cell number. In a toxicity study, no significant hATMSC-related changes were found in body weights, clinical signs, hematological/biochemical values, organ weights, or histopathological findings. In conclusion, hATMSCs loaded on HA/TCP enhance the repair of bone defects and was found to be safe under our preclinical efficacy/safety hybrid assessment system.

Keyword

Safety; Efficacy; Human Adipose Tissue-Derived Mesenchymal Stem Cells; HA/TCP Scaffold; Bone Defect Model; Nude Rats

MeSH Terms

Adipose Tissue/*cytology
Animals
Biocompatible Materials/therapeutic use
Bone Diseases/pathology/radiography/*therapy
Bone Regeneration/physiology
Calcium Phosphates/therapeutic use
Diaphyses/radiography/surgery/ultrastructure
Disease Models, Animal
Durapatite/therapeutic use
Femur/*pathology/radiography/surgery
Humans
Male
*Mesenchymal Stem Cell Transplantation
Mesenchymal Stem Cells/*cytology
Rats
Rats, Nude
Tissue Engineering
Tomography, X-Ray Computed
Transplantation, Heterologous

Figure

• 

  Fig. 1 Procedures for surgical implantation in the femoral segmental defect model from implant preparation to closure. (A, B) Preparation of the hATMSC-loaded implants, (C) femoral segmental defects, (D, E) transplantation of test items into the defects, (F) closure.

• 

  Fig. 2 SEM observations of hATMSC-loaded HA/TCP scaffolds with different cell densities. (A-C) HA/TCP scaffold, (D-L) varying density of hATMSC-loaded HA/TCP (D-F, 7.5 × 105 cells/mL; G-I, 7.5 × 106 cells/mL; J-L, 7.5 × 107 cells/mL). Arrow, attached cells.

• 

  Fig. 3 Radiographs of bone defects treated with hATMSC-loaded HA/TCP scaffolds. (A) Serial radiographs at postoperative 0, 4, 8, 12, and 16 weeks. (B) The percentage areas occupied by newly formed bone were scored from 1 to 4 (*P < 0.05 compared with control).

• 

  Fig. 4 Micro-computed tomography of bone defects at 16 weeks after surgery. (A) Control, (B) Group I, (C) Group II, (D) Group III, (E) Group IV, and (F) Group V. The dotted box approximately indicates the original defect zone.

• 

  Fig. 5 Histolgical and histomorphometric analysis of bone defects treated with hATMSC-loaded HA/TCP scaffolds at 16 weeks after surgery. (A) H&E stain, (B) The graphs demonstrate the percentages of connective tissues, residual HA/TCP scaffolds and new bone (*P < 0.05 compared with control, †P < 0.05 compared with Group I).

• 

  Fig. 6 Body weight changes in nude rats treated with hATMSC-loaded HA/TCP scaffolds. Data are shown as mean ± SD.

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