Yao Jeffrey, Korotkova Tatiana, Riboh Jonathan, Chong Alphonsus, Chang James, Smith R Lane
Department of Orthopaedic Surgery, Stanford University Medical Center, Palo Alto, CA 94304, USA.
J Hand Surg Am. 2008 Nov;33(9):1558-64. doi: 10.1016/j.jhsa.2008.06.010.
Pluripotential embryonic cells may be seeded onto sutures intended for tendon repair. These cells may be influenced to adhere to suture material using adhesion substrates, and furthermore, these cells may remain in culture attached to those sutures. These cell-impregnated sutures may be useful for promoting healing of tendon repairs.
Ten-centimeter segments of 4-0 sutures (FiberWire) were coated overnight with 10 microg/mL fibronectin, 10 microg/mL poly-l-lysine, or phosphate-buffered saline. The sutures were placed in dishes and covered with a suspension of C3H10T1/2 cells at concentrations of 1 x 10(6), 2 x 10(6), or 4 x 10(6) cells for 24 hours. The sutures were then placed into low adhesion polypropylene tubes with Dulbecco's modified Eagle's medium and 10% fetal bovine serum for 7 days. The presence of viable cells on these sutures was assessed by the colorimetric Alamar blue cell proliferation assay. Spectrophotometry was used to quantify the relative amount of cell proliferation across the experimental groups. The sutures were also visually inspected using phase-contrast light microscopy.
Our results show that at all seeding densities (1 x 10(6), 2 x 10(6), and 4 x 10(6) cells), the suture segments coated with poly-l-lysine and fibronectin showed a significant increase in C3H10T1/2 cell adhesion. Coating the suture with poly-l-lysine increased the adherent cell number to 17% of the initial seeding concentration compared with 2% for the control. Fibronectin coating increased the number of adherent viable cells present to 6.6%.
Pluripotential embryonic cells may be seeded onto sutures, adhere, and survive in culture. Coating sutures with poly-l-lysine and fibronectin offers significant improvement in retention of viable cells. This technique may be a useful adjunct for future tendon healing studies.
可将多能胚胎细胞接种到用于肌腱修复的缝合线上。使用黏附底物可促使这些细胞黏附于缝合材料，此外，这些细胞可在培养中保持附着于那些缝合线上。这些细胞浸渍的缝合线可能有助于促进肌腱修复的愈合。
将4-0缝合线（FiberWire）的10厘米片段用10微克/毫升纤连蛋白、10微克/毫升聚-L-赖氨酸或磷酸盐缓冲盐水包被过夜。将缝合线置于培养皿中，并用浓度为1×10⁶、2×10⁶或4×10⁶个细胞的C3H10T1/2细胞悬液覆盖24小时。然后将缝合线放入含有杜氏改良 Eagle 培养基和10%胎牛血清的低黏附聚丙烯管中培养7天。通过比色法Alamar蓝细胞增殖测定评估这些缝合线上活细胞的存在情况。使用分光光度法量化各实验组细胞增殖的相对量。还使用相差光学显微镜对缝合线进行目视检查。
我们的结果表明，在所有接种密度（1×10⁶、2×10⁶和4×10⁶个细胞）下，用聚-L-赖氨酸和纤连蛋白包被的缝合线片段显示C3H10T1/2细胞黏附显著增加。用聚-L-赖氨酸包被缝合线使黏附细胞数增加至初始接种浓度的17%，而对照组为2%。纤连蛋白包被使存在的黏附活细胞数增加至6.6%。
多能胚胎细胞可接种到缝合线上，黏附并在培养中存活。用聚-L-赖氨酸和纤连蛋白包被缝合线可显著提高活细胞的保留率。该技术可能是未来肌腱愈合研究有用的辅助手段。
