间充质干细胞和胶原贴剂在前交叉韧带修复中的应用。
Mesenchymal stem cells and collagen patches for anterior cruciate ligament repair.
机构信息
Benjamin Gantenbein, Neha Gadhari, Samantha CW Chan, Sufian S Ahmad, Tissue and Organ Mechanobiology, Institute for Surgical Technology and Biomechanics, University of Bern, CH-3014 Bern, Switzerland.
出版信息
World J Stem Cells. 2015 Mar 26;7(2):521-34. doi: 10.4252/wjsc.v7.i2.521.
AIM
To investigate collagen patches seeded with mesenchymal stem cells (MSCs) and/or tenocytes (TCs) with regards to their suitability for anterior cruciate ligament (ACL) repair.
METHODS
Dynamic intraligamentary stabilization utilizes a dynamic screw system to keep ACL remnants in place and promote biological healing, supplemented by collagen patches. How these scaffolds interact with cells and what type of benefit they provide has not yet been investigated in detail. Primary ACL-derived TCs and human bone marrow derived MSCs were seeded onto two different types of 3D collagen scaffolds, Chondro-Gide(®) (CG) and Novocart(®) (NC). Cells were seeded onto the scaffolds and cultured for 7 d either as a pure populations or as "premix" containing a 1:1 ratio of TCs to MSCs. Additionally, as controls, cells were seeded in monolayers and in co-cultures on both sides of porous high-density membrane inserts (0.4 μm). We analyzed the patches by real time polymerase chain reaction, glycosaminoglycan (GAG), DNA and hydroxy-proline (HYP) content. To determine cell spreading and adherence in the scaffolds microscopic imaging techniques, i.e., confocal laser scanning microscopy (cLSM) and scanning electron microscopy (SEM), were applied.
RESULTS
CLSM and SEM imaging analysis confirmed cell adherence onto scaffolds. The metabolic cell activity revealed that patches promote adherence and proliferation of cells. The most dramatic increase in absolute metabolic cell activity was measured for CG samples seeded with tenocytes or a 1:1 cell premix. Analysis of DNA content and cLSM imaging also indicated MSCs were not proliferating as nicely as tenocytes on CG. The HYP to GAG ratio significantly changed for the premix group, resulting from a slightly lower GAG content, demonstrating that the cells are modifying the underlying matrix. Real-time quantitative polymerase chain reaction data indicated that MSCs showed a trend of differentiation towards a more tenogenic-like phenotype after 7 d.
CONCLUSION
CG and NC are both cyto-compatible with primary MSCs and TCs; TCs seemed to perform better on these collagen patches than MSCs.
目的
研究包被间充质干细胞（MSCs）和/或腱细胞（TCs）的胶原贴片在治疗前交叉韧带（ACL）损伤方面的适用性。
方法
动态韧带内稳定利用动态螺钉系统将 ACL 残端固定在适当位置，并促进生物愈合，同时辅以胶原贴片。这些支架与细胞的相互作用以及它们提供的益处类型尚未详细研究。原代 ACL 衍生的 TC 和人骨髓来源的 MSC 接种到两种不同类型的 3D 胶原支架上，即 Chondro-Gide（®）（CG）和 Novocart（®）（NC）。细胞接种到支架上，在纯培养或 1:1 TC 与 MSC 混合培养条件下培养 7 天。此外，作为对照，细胞单层接种并在多孔高密度膜插入物（0.4μm）两侧共培养。通过实时聚合酶链反应、糖胺聚糖（GAG）、DNA 和羟脯氨酸（HYP）含量分析贴片。为了确定细胞在支架中的扩展和粘附，应用了微观成像技术，即共聚焦激光扫描显微镜（cLSM）和扫描电子显微镜（SEM）。
结果
CLSM 和 SEM 成像分析证实了细胞在支架上的粘附。代谢细胞活性表明，贴片促进了细胞的粘附和增殖。CG 接种 TC 或 1:1 细胞预混物的样本中，绝对代谢细胞活性的增加最为显著。DNA 含量分析和 cLSM 成像也表明，MSC 在 CG 上的增殖不如 TC 好。由于 GAG 含量略低，HYP 与 GAG 的比值显著改变，表明细胞正在修饰基底基质。实时定量聚合酶链反应数据表明，MSC 在 7d 后表现出向更腱样表型分化的趋势。
结论
CG 和 NC 均与原代 MSC 和 TC 细胞具有细胞相容性；TC 在这些胶原贴片上的表现优于 MSC。
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