삼인산 칼슘 혼합 하이드로젤 지지체와 중간엽줄기세포로구성된 복합화합물의 신생골조직 형성능 (The Role of Beta-Tricalcium Phosphate-Hydrogel Scaffold and Mesenchymal Stem Cells on Neogenic Bone Formation)

서지정보

· 발행기관 : 대한정형외과학회
· 수록지 정보 : 대한정형외과학회지 / 53권 / 2호 / 143 ~ 151페이지
· 저자명 : 이다름, 강용구

초록

목적: 중간엽줄기세포와 베타 3인산 칼슘염(beta-tricalcium phosphate, β-TCP), hydrogel로 구성된 복합화합물의 세포 지지 및 신생조직 형성능에 대하여 알아보고자 하였다.
대상 및 방법: 중간엽줄기세포와 β-TCP를 hydrogel에 혼합하여 복합화합물을 제조하고, 제조한 복합화합물을 3일간 배양하여 신생골 형성을 위한 유전자 발현도를 분석하였다. 또한 복합화합물을 흰쥐의 피하에 주사하고 4주 후 형성된 신생조직을 채취해 조직학적 염색을 시행하였다.
결과: 일반 2차원 배양과 복합화합물에서 배양한 중간엽줄기세포의 유전자 발현 양상을 비교할 때 복합화합물의 세포에서 RUNX2 1.26배, osteopontin 1.23배, transforming growth factor-β 2.12배, osterix 1.07배, type I collagen 1.3배, fibronectin 1.3배로 신생조직의 골 형성과 관련된 유전자 발현도가 높아진 것을 확인하였다(p＜0.05). 동물실험에서도 신생조직이 관찰되었으며 조직염색을 통하여 복합화합물이 중간엽줄기세포의 이탈을 막고 신생조직을 형성한 것을 확인하였으며 osterix 면역염색을 통해 조직 내 골분화세포를 관찰하였다. 대조군의 경우 이식한 중간엽줄기세포가 모두 소실되고 신생조직 형성이 없었다.
결론: β-TCP와 hydrogel로 구성된 지지체와 중간엽줄기세포를 혼합한 복합화합물은 중간엽줄기세포를 지지하고 신생조직을 형성하는 데 유익한 환경을 제공하는 것으로 판단한다.

영어초록

Purpose: The purpose of this paper was to determine the ability of a mixture consisting of mesenchymal stem cells, beta-tricalcium phosphate b-TCP), and hydrogel, to support cells and form new tissue.
Materials and Methods: A composite was produced by adding b-TCP to hydrogel, and mesenchymal stem cells were cultivated in the composite. Then, reverse transcription polymerase chain reaction (RT-PCR) was conducted to measure the level of gene expression for the new bone formation in the cells. Moreover, a composite in which the mesenchymal stem cells were added was injected into the subcutaneous fat of sprague-dawley rats. After four weeks, H&E, Masson trichrome, silver nitrate staining, and osterix immunohistochemical staining were conducted by taking the tissue to evaluate whether the composite supported mesenchymal stem cells and formed new tissue.
Results: By using RT-PCR, we found that the level of gene expression became significantly higher in 3-dimensional gel culture with RUNX2 by 1.26 times, with osteopontin by 1.23 times, transforming growth factor-b by 2.12 times, osterix by 1.07 times, type I collagen by 1.3 times, and fibronectin by 1.3 times. In the animal experiment in which a composite was transplanted into the subcutaneous fat, newly formed tissue was observed. Also, it was found that the composite prevented mesenchymal stem cells from leaving and formed new tissue.
Osteogenic differentiation cells in the tissue was observed through osterix immunostaining.
Conclusion: It was identified that the composite prevented mesenchymal stem cells dispersal and contributed to the formation of neogenic tissue. Therefore we conclude that the composite plays a role of a scaffold to support the implanted cells and form neogenic tissue more effectively.

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