Trauma Department, Hanover Medical School (MHH), OE 6230, Carl-Neuberg-Straße 1, 30625 Hannover, Germany.
Knee Surg Sports Traumatol Arthrosc. 2012 Feb;20(2):223-31. doi: 10.1007/s00167-011-1600-3. Epub 2011 Jul 13.
The purpose of this study was to investigate the influence of continuous perfusion and mechanical stimulation on bone marrow stromal cells seeded on a collagen meniscus implant.
Bone marrow aspirates from 6 donors were amplified in vitro. 10(6) human BMSC were distributed on a collagen meniscus implant. Scaffolds were cultured under static conditions (control) or placed into a bioreactor system where continuous perfusion (10 ml/min) or perfusion and mechanical stimulation (8 h of 10% cyclic compression at 0.5 Hz) were administered daily. After 24 h, 7 and 14 days, cell proliferation, synthesis of procollagen I and III peptide (PIP, PIIIP), histology, and the equilibrium modulus of the constructs were analyzed.
Proliferation demonstrated a significant increase over time in all groups (p < 0.001). PIP synthesis was found to increase from 0.1 ± 0.0 U/ml/g protein after 24 h to 2.0 ± 0.5 (perfusion), 3.8 ± 0.3 (mechanical stimulation), and 1.8 ± 0.2 U/ml/g protein (static control, lower than perfusion and mechanical stimulation, p < 0.05). These differences were also evident after 2 weeks (2.7 ± 0.3, 4.0 ± 0.6, and 1.8 ± 0.2 U/ml/g protein, p < 0.01); PIIIP synthesis was found to increase from 0.1 ± 0.0 U/ml/g protein after 24 h to 2.9 ± 0.7 (perfusion), 3.1 ± 0.9 (mechanical stimulation), and 1.6 ± 0.3 U/ml/g protein (controls) after 1 week and remained significantly elevated under the influence of perfusion and mechanical stimulation (p < 0.01) after 2 weeks. Mechanical stimulation increased the equilibrium modulus more than static culture and perfusion after 2 weeks (24.7 ± 7.6; 12.3 ± 3.7; 15.4 ± 2.6 kPa; p < 0.02).
Biomechanical stimulation and perfusion have impact on collagen scaffolds seeded with BMSCs. Cell proliferation can be enhanced using continuous perfusion and differentiation is fostered by mechanical stimulation.
本研究旨在探讨连续灌注和机械刺激对骨髓基质细胞接种胶原半月板植入物的影响。
从 6 名供体中抽取骨髓抽吸物，在体外扩增。将 10(6)个人骨髓基质细胞分布在胶原半月板植入物上。支架在静态条件下培养（对照）或置于生物反应器系统中，每天进行连续灌注（10ml/min）或灌注和机械刺激（0.5Hz 时 10%循环压缩 8 小时）。在 24 小时、7 天和 14 天后，分析细胞增殖、原胶原 I 和 III 肽（PIP、PIIIP）的合成、组织学和构建物的平衡模量。
所有组的增殖均随时间显著增加（p<0.001）。PIP 合成从 24 小时后的 0.1±0.0U/ml/g 蛋白增加至 2.0±0.5（灌注）、3.8±0.3（机械刺激）和 1.8±0.2U/ml/g 蛋白（静态对照，低于灌注和机械刺激，p<0.05）。2 周后也有明显差异（2.7±0.3、4.0±0.6 和 1.8±0.2U/ml/g 蛋白，p<0.01）；PIIIP 合成从 24 小时后的 0.1±0.0U/ml/g 蛋白增加至 2.9±0.7（灌注）、3.1±0.9（机械刺激）和 1.6±0.3U/ml/g 蛋白（对照），在灌注和机械刺激的影响下，1 周后仍显著升高（p<0.01），2 周后仍显著升高（p<0.01）。2 周后，机械刺激比静态培养和灌注增加了平衡模量（24.7±7.6；12.3±3.7；15.4±2.6kPa；p<0.02）。
生物力学刺激和灌注对骨髓基质细胞接种胶原支架有影响。连续灌注可增强细胞增殖，机械刺激促进分化。
