Adipose-derived Stem Cell Sheet Promote Meniscus Regeneration in Rabbit Model
Author(s):
Takata Y. (Japan)
,
Takata Y. (Japan)
Affiliations:
Nakase J.
,
Nakase J.
Affiliations:
Shimozaki K.
,
Shimozaki K.
Affiliations:
Asai K.
,
Asai K.
Affiliations:
Tsuchiya H.
Tsuchiya H.
Affiliations:
ESSKA Academy. Takata Y. 05/09/18; 209308; P02-721 Topic: Basic Science
Yasushi Takata
Yasushi Takata
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Abstract
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Objectives: Meniscal regeneration is one of the most challenging aspects of orthopedic surgery. Various procedures and factors, including stem cells, have been studied in meniscal treatment, but to date, no significant healing effect could be achieved.
We have focused on adipose-derived stem cells (ADSC), which can be harvested easily in large amounts, and can be processed into sheets by culturing them. The ADSC sheet is characteristic, in that, it can be transplanted while maintaining a sheet-like form by collagen secreted from the cells, and does not require a scaffold at the time of transplantation.
The purpose of this study was to investigate the meniscal regeneration-promoting effect of ADSC sheets in a rabbit model with meniscal defect.

Methods: Twenty mature female Japanese white rabbits (aged 15-17 weeks) weighing 3.0 to 3.5 kg (mean, 3.2 kg) were selected for the study. ADSCs were extracted from the adipose tissue of the interscapular fat pad. Isolation of ADSCs was performed according to the method described by Zuk et al., with some modifications. It was imperative to confirm that the cells reached confluence at the 3rd passage and the culture medium changed to the one, to which, ascorbic acid was added. In about one week, the cells in the dish became peelable in sheet form, which was then used as an ADSC sheet.
Medial parapatellar surgical approach was used to expose medial meniscus, which was sharply cut off at the posterior edge of the Medial collateral ligament (MCL), and the anterior half of the meniscus was removed. An autologous ADSC sheet was transplanted to one knee and the wound was closed to sew with the joint capsule (transplant side). The other side was closed as it was (control side).
They were euthanized 4 and 12 weeks after transplantation to harvest the entire medial menisci. The area of the meniscal tissue and the transverse diameter on the inside of the MCL were measured using Image J software. In addition, the histological score was evaluated according to the report of Hatsushika et al. The average values obtained for the transplant side and the control side were compared using the t-test.

Results: The area and transverse diameter of regenerated tissue were larger in the transplanted side than in the control side for both 4 weeks (area: transplanted side 42.5±7.66mm², control side 31.7±10.1mm², P=0.02; transverse diameter: transplanted side 1.87±0.38mm, control side 1.21±0.47mm, P=0.01) and 12 weeks (area: transplanted side 47.2±8.08mm², control side 33.7±8.49mm², P=0.001; transverse diameter: transplanted side 1.78±0.32mm, control side 1.27±0.47mm, P<0.001). Also, the histological score was significantly higher in the transplanted side than in the control side for both (transplanted side 7.3±2.9, control side 4.5±3.0, P=0.047) and 12 weeks (transplanted side 9.0±3.8, control side 4.5±3.5, P=0.034).

Conclusions: In conclusion, it can be suggested that ADSC can promote meniscal regeneration after meniscus excision in rabbit model.

Keywords:
Adipose-derived Stem Cell, Meniscus, Rabbit
Objectives: Meniscal regeneration is one of the most challenging aspects of orthopedic surgery. Various procedures and factors, including stem cells, have been studied in meniscal treatment, but to date, no significant healing effect could be achieved.
We have focused on adipose-derived stem cells (ADSC), which can be harvested easily in large amounts, and can be processed into sheets by culturing them. The ADSC sheet is characteristic, in that, it can be transplanted while maintaining a sheet-like form by collagen secreted from the cells, and does not require a scaffold at the time of transplantation.
The purpose of this study was to investigate the meniscal regeneration-promoting effect of ADSC sheets in a rabbit model with meniscal defect.

Methods: Twenty mature female Japanese white rabbits (aged 15-17 weeks) weighing 3.0 to 3.5 kg (mean, 3.2 kg) were selected for the study. ADSCs were extracted from the adipose tissue of the interscapular fat pad. Isolation of ADSCs was performed according to the method described by Zuk et al., with some modifications. It was imperative to confirm that the cells reached confluence at the 3rd passage and the culture medium changed to the one, to which, ascorbic acid was added. In about one week, the cells in the dish became peelable in sheet form, which was then used as an ADSC sheet.
Medial parapatellar surgical approach was used to expose medial meniscus, which was sharply cut off at the posterior edge of the Medial collateral ligament (MCL), and the anterior half of the meniscus was removed. An autologous ADSC sheet was transplanted to one knee and the wound was closed to sew with the joint capsule (transplant side). The other side was closed as it was (control side).
They were euthanized 4 and 12 weeks after transplantation to harvest the entire medial menisci. The area of the meniscal tissue and the transverse diameter on the inside of the MCL were measured using Image J software. In addition, the histological score was evaluated according to the report of Hatsushika et al. The average values obtained for the transplant side and the control side were compared using the t-test.

Results: The area and transverse diameter of regenerated tissue were larger in the transplanted side than in the control side for both 4 weeks (area: transplanted side 42.5±7.66mm², control side 31.7±10.1mm², P=0.02; transverse diameter: transplanted side 1.87±0.38mm, control side 1.21±0.47mm, P=0.01) and 12 weeks (area: transplanted side 47.2±8.08mm², control side 33.7±8.49mm², P=0.001; transverse diameter: transplanted side 1.78±0.32mm, control side 1.27±0.47mm, P<0.001). Also, the histological score was significantly higher in the transplanted side than in the control side for both (transplanted side 7.3±2.9, control side 4.5±3.0, P=0.047) and 12 weeks (transplanted side 9.0±3.8, control side 4.5±3.5, P=0.034).

Conclusions: In conclusion, it can be suggested that ADSC can promote meniscal regeneration after meniscus excision in rabbit model.

Keywords:
Adipose-derived Stem Cell, Meniscus, Rabbit
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