Fu W-M, Yang L, Wang B-J, Xu J-K, Wang J-L, Qin L, Zhao D-W
Southern Medical University, Guangzhou, Guangdong, China.
Eur Rev Med Pharmacol Sci. 2016 Aug;20(16):3490-9.
Bone marrow mesenchymal stem cells (BMMSCs) have been widely applied in osteonecrosis. However, lack of biomechanical support limited application of BMMSCs. And porous tantalum (PTA) has been identified as a cell-friendly scaffold for bone regeneration. Herein, we aimed to investigate the efficacy of PTA seeded with BMMSCs in the treatment of osteonecrosis.
After the production of PTA seeded with BMMSCs, MTT and GFP were performed to identify the proliferation and adhesion of BMMSCs respectively, which was further examined by scanning electron microscopy (SEM). And real-time PCR was also used to determine mRNA level of osteogenic markers, including Alp, OCN, OPN, Col I and Runx-2 in BMMSCs. Nineteen adult rabbits were applied for building steroid-associated osteonecrosis (SAON) models. Bone formation rate (BFR) and mineral apposition rate (MAR) were determined. And Goldner Trichrome Staining was used in these SAON models, which further confirmed the efficacy of PTA seeded with BMMSCs in SAON.
PTA seeded with BMMSCs showed excellent biocompatibility. Additionally, SEM assay showed that BMMSCs adhered tightly and spread fully in the pores of PTA. Next, the expression of ALP and OPN mRNA in BMMSCs were significantly (p < 0.05) higher in the PTA-treated group compared to those in the PTA-untreated group. Furthermore, compared to those treated by only PTA, the dynamic bone formation in rabbits treated by PTA seeded with BMMSCs was significantly increased (p < 0.001) at both week 3rd and week 6th.
The product, PTA seeded with BMMSCs, was successfully produced, and was determined as high efficacy for treatment of steroid-associated osteonecrosis. PTA seeded with BMMSCs may afford a promising option for treating osteonecrosis.
骨髓间充质干细胞（BMMSCs）已广泛应用于骨坏死的治疗。然而，缺乏生物力学支持限制了BMMSCs的应用。而多孔钽（PTA）已被确定为一种有利于细胞生长的骨再生支架。在此，我们旨在研究接种BMMSCs的PTA治疗骨坏死的疗效。
制备接种BMMSCs的PTA后，分别进行MTT和绿色荧光蛋白（GFP）检测以鉴定BMMSCs的增殖和黏附情况，并用扫描电子显微镜（SEM）进一步观察。同时采用实时定量聚合酶链反应（real-time PCR）检测BMMSCs中成骨标志物碱性磷酸酶（Alp）、骨钙素（OCN）、骨桥蛋白（OPN）、I型胶原（Col I）和Runx-2的mRNA水平。选用19只成年兔建立类固醇相关骨坏死（SAON）模型，测定骨形成率（BFR）和矿物质沉积率（MAR）。对这些SAON模型进行戈德纳三色染色，进一步证实接种BMMSCs的PTA对SAON的治疗效果。
接种BMMSCs的PTA表现出良好的生物相容性。此外，SEM检测显示BMMSCs紧密黏附并充分铺展于PTA的孔隙中。接下来，与未处理PTA组相比，PTA处理组BMMSCs中ALP和OPN mRNA的表达显著升高（p < 0.05）。此外，与仅用PTA处理的兔子相比，接种BMMSCs 的PTA处理的兔子在第3周和第6周时的动态骨形成均显著增加（p < 0.001）。
成功制备了接种BMMSCs的PTA产品，并确定其对类固醇相关骨坏死具有高效治疗作用。接种BMMSCs的PTA可能为治疗骨坏死提供一个有前景的选择。
