Yonsei Med J.  2016 Mar;57(2):441-448. 10.3349/ymj.2016.57.2.441.

Transplantation of a Scaffold-Free Cartilage Tissue Analogue for the Treatment of Physeal Cartilage Injury of the Proximal Tibia in Rabbits

• Lee SU
• Lee JY
• Joo SY
• Lee YS
• Jeong C

• KMID: 2374051
• DOI: http://doi.org/10.3349/ymj.2016.57.2.441

Abstract

PURPOSE
The purpose of this study was to investigate the effects of transplantation of an in vitro-generated, scaffold-free, tissue-engineered cartilage tissue analogue (CTA) using a suspension chondrocyte culture in a rabbit growth-arrest model.
MATERIALS AND METHODS
We harvested cartilage cells from the articular cartilage of the joints of white rabbits and made a CTA using a suspension culture of 2x107 cells/mL. An animal growth plate defect model was made on the medial side of the proximal tibial growth plate of both tibias of 6-week-old New Zealand white rabbits (n=10). The allogenic CTA was then transplanted onto the right proximal tibial defect. As a control, no implantation was performed on the left-side defect. Plain radiographs and the medial proximal tibial angle were obtained at 1-week intervals for evaluation of bone bridge formation and the degree of angular deformity until postoperative week 6. We performed a histological evaluation using hematoxylin-eosin and Alcian blue staining at postoperative weeks 4 and 6.
RESULTS
Radiologic study revealed a median medial proximal tibial angle of 59.0degrees in the control group and 80.0degrees in the CTA group at 6 weeks. In the control group, statistically significant angular deformities were seen 3 weeks after transplantation (p<0.05). On histological examination, the transplanted CTA was maintained in the CTA group at 4 and 6 weeks postoperative. Bone bridge formation was observed in the control group.
CONCLUSION
In this study, CTA transplantation minimized deformity in the rabbit growth plate injury model, probably via the attenuation of bone bridge formation.

Keyword

Cartilage tissue analogue; chondrocyte; growth plate injury; angular deformity; bone bridge; surgical treatment

MeSH Terms

Animals
*Bone Transplantation
Cartilage/anatomy & histology
Cell Culture Techniques
Cells, Cultured
Chondrocytes/*cytology/transplantation
Growth Plate/anatomy & histology/*surgery
*Mesenchymal Stem Cell Transplantation
Rabbits
Tibia/*surgery
Tissue Engineering
Transplantation, Autologous/methods
Transplantation, Homologous

Figure

• 

  Fig. 1 (A) Intraoperative photograph showing an experimental model of growth arrest created by excising the growth plate at the proximal medial side of the tibia (4-mm diameter and 3-mm depth). (B) After sterile saline water irrigation, cartilage tissue analogue (black *) was implanted on the proximal tibial defect.

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  Fig. 2 The medial proximal tibial angle was defined as the angle between the long axis of the tibia and tibial plateau in the anteroposterior lower limb. MPTA, medial proximal tibial angle.

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  Fig. 3 Histological view of the representative cartilage tissue analogue grown for 8 weeks, stained with Alcian blue (A, ×200) and immunofluorescence (B, ×400).

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  Fig. 4 (A) Gross and (B and C) cross-sectional views of the rabbit tibia at 6 weeks after surgery showing angular deformity of the proximal tibia on the left side (Lt, control group). However, the right side had no deformity (Rt, cartilage tissue analogue group).

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  Fig. 5 Serial radiographs for both groups revealing the attenuation and progression of angular deformity for 6 weeks (A). The cartilage tissue analogue group (Rt) showed more attenuation of the angular deformity than the control group did (Lt). (B) Another serial radiograph showing the same result. Rt, right tibia; Lt, left tibia; W, week.

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  Fig. 6 Radiographic evaluation of growth plate regeneration. Compared with the control group, the CTA group showed a higher average medial proximal tibia angle, with the difference gradually increasing with statistical significance at 3 postoperative weeks (*p<0.05). CTA, cartilage tissue analogue; MPTA, medial proximal tibial angle.

• 

  Fig. 7 Histological evaluation. (A and C) Histological evaluation of the 4-week-old specimens revealed that new bone was formed (black *) in the excised epiphyseal plate area and it partially closed the growth plate in the control group. (B and D) The CTA group showed regeneration of the growth plate and slight bone bridge formation, that is, longitudinally columnar chondrocytes (arrowheads; black arrows: normal epiphyseal plate). (E) Histological evaluation of the 6-week-old specimens revealed bone bridge formation that replaced the area of the excised epiphyseal plate in the control group. (F) The CTA group revealed regeneration of the epiphyseal plate and some bone bridge formation (A and B: hematoxylin-eosin staining, ×40; C and D: Alcian blue staining, ×40; E and F: Alcian blue staining, ×40). CTA, cartilage tissue analogue.

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