Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.
Zainab Panjwani Memorial Hospital, Mohammadali Habib Road, Numaish Karachi, 74800, Pakistan.
Mol Cell Biochem. 2021 Aug;476(8):3191-3205. doi: 10.1007/s11010-021-04155-9. Epub 2021 Apr 17.
Intervertebral disc (IVD) degeneration is an asymptomatic pathophysiological condition and a strong causative factor of low back pain. There is no cure available except spinal fusion and pain management. Stem cell-based regenerative medicine is being considered as an alternative approach to treat disc diseases. The current study aimed to differentiate human umbilical cord-mesenchymal stem cells (hUC-MSCs) into chondrocyte-like cells and to elucidate their feasibility and efficacy in the degenerated IVD rat model. Chondrogenic induction medium was used to differentiate hUC-MSCs into chondroprogenitors. Rat tail IVD model was established with three consecutive coccygeal discs. qPCR was performed to quantify the molecular markers of pain and inflammation. Histological staining was performed to evaluate the degree of regeneration. Induced chondroprogenitors showed the expression of chondrogenic genes, SOX9, TGF-β1, ACAN, BMP2, and GDF5. Immunocytochemical staining showed positive expression of chondrogenic proteins SOX9, TGF-β1, TGF-β2, and Collagen 2. In in vivo study, transplanted chondroprogenitors showed better survival, homing, and distribution in IVD as compared to normal MSCs. Expression of pain and inflammatory genes at day 5 of cell transplantation modulated immune response significantly. The transplanted labeled MSCs and induced chondroprogenitors differentiated into functional nucleus pulposus (NP) cells as evident from co-localization of red (DiI) and green fluorescence for SOX9, TGF-β1, and TGF-β2. Alcian blue and H & E staining showed standard histological features, indicating better preservation of the NP structure and cellularity than degenerated discs. hUC-MSCs-derived chondroprogenitors showed better regeneration potential as compared to normal MSCs. The pain and inflammation genes were downregulated in the treated group as compared to the degenerated IVD.
椎间盘（IVD）退变是一种无症状的病理生理状态，也是腰痛的一个强烈致病因素。除了脊柱融合和疼痛管理外，目前尚无其他治疗方法。基于干细胞的再生医学被认为是治疗椎间盘疾病的一种替代方法。本研究旨在将人脐带间充质干细胞（hUC-MSCs）分化为软骨细胞样细胞，并阐明其在退变的大鼠 IVD 模型中的可行性和疗效。采用软骨诱导培养基将 hUC-MSCs 分化为软骨祖细胞。通过三个连续的尾骨椎间盘建立大鼠尾椎 IVD 模型。通过 qPCR 定量测定疼痛和炎症的分子标志物。通过组织学染色评估再生程度。诱导的软骨祖细胞表达软骨基因 SOX9、TGF-β1、ACAN、BMP2 和 GDF5。免疫细胞化学染色显示软骨蛋白 SOX9、TGF-β1、TGF-β2 和 Collagen 2 的阳性表达。在体内研究中，与正常 MSCs 相比，移植的软骨祖细胞在 IVD 中表现出更好的存活、归巢和分布。细胞移植后第 5 天，疼痛和炎症基因的表达显著调节免疫反应。标记的 MSC 和诱导的软骨细胞向功能性髓核（NP）细胞分化，如 SOX9、TGF-β1 和 TGF-β2 的红色（DiI）和绿色荧光共定位所示。阿尔辛蓝和 H&E 染色显示出标准的组织学特征，表明 NP 结构和细胞密度的保存优于退变的椎间盘。与正常 MSCs 相比，hUC-MSCs 衍生的软骨祖细胞具有更好的再生潜力。与退变的 IVD 相比，治疗组的疼痛和炎症基因表达下调。
