Tissue engineering of flexor tendons: Optimization of tenocyte proliferation using growth factor supplementation
Tissue engineering of flexor tendons: Optimization of tenocyte proliferation using growth factor supplementation
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DOI：
10.1089/ten.2006.12.1937
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发表时间：
2006-07-01
影响因子：
--
通讯作者：
Chang, James
中科院分区：
文献类型：
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作者：
Costa, Melinda Aliza;Wu, Cindy;Chang, James
屈肌腱修复的一个重要问题是缺乏合适的移植材料进行重建。屈肌腱移植物构建体的离体生产需要原代细胞的扩增。已知生长因子，如血小板衍生生长因子-BB（PDGF-BB）、胰岛素样生长因子-1（IGF-1）和碱性成纤维细胞生长因子（bFGF）促进肌腱愈合和肌腱细胞增殖。这些实验的目的是使用生长因子补充剂优化3种肌腱细胞群中的肌腱细胞增殖。从兔趾深屈肌腱中分离滑膜鞘、腱外膜和腱内膜细胞并保持培养。细胞培养物补充有单独和组合的IGF-1、PDGF-BB和bFGF。用于个体生长因子补充的条件为IGF-1（10、50和100 ng/mL）、PDGF-BB（1、10和50 ng/mL）和bFGF（0.5、1和5 ng/mL）。用于生长因子组合的条件为IGF-1 + PDGF-BB（分别为50 + 10和100 + 50 ng/mL）和IGF-1 + PDGF-BB + bFGF（分别为50 + 10 + 1; 50 + 10 + 5; 100 + 50 + 1;和100 + 50 + 5 ng/mL）。对于所有3个肌腱细胞群，在72小时的增殖是更大的个体生长因子的存在下，与对照组相比。补充PDGF-BB（50 ng/mL）后，S细胞的平均吸光度值增加了97%（0.57至1.13），E细胞增加了37%（0.51至0.70），T细胞增加了33%（0.33至0.44）（p < 0.001）。此外，观察到协同效应。与最大剂量的单个生长因子相比，生长因子的组合导致更大的增殖。在补充有IGF-1（100 ng/mL）+ PDGF-BB（50 ng/mL）的培养物中，S细胞的平均吸光度增加114%（0.57至1.22），E细胞增加63%（0.51至0.831），T细胞增加47%（0.33至0.48）（p < 0.001）。IGF-1（100 ng/mL）+ PDGF-BB（50 ng/mL）+ bFGF（5 ng/mL）导致所有3种肌腱细胞群的最大量的细胞增殖。在S细胞中，平均增殖率增加251%，在E细胞中增加98%，在T细胞中增加106%（p < 0.001）。总之，IGF-1、PDGF-BB和bFGF可以组合使用以使腱细胞增殖最大化。生长因子之间的协同作用可能为促进肌腱工程提供一种手段。
A significant problem in flexor tendon repair is the lack of suitable graft material for reconstruction. The ex vivo production of flexor tendon graft constructs requires the expansion of primary cells. Growth factors, such as platelet-derived growth factor-BB (PDGF-BB), insulin-like growth factor-1 (IGF-1), and basic fibroblast growth factor (bFGF), are known to promote tendon healing and tendon cell proliferation. The purpose of these experiments was to optimize tenocyte proliferation in 3 tendon cell populations using growth factor supplementation. Cells of the synovial sheath, epitenon, and endotenon were isolated from rabbit flexor digitorum profundus tendons and maintained in culture. Cell cultures were supplemented with IGF-1, PDGF-BB, and bFGF alone and in combination. The conditions used for individual growth factor supplementation were IGF-1 (10, 50, and 100 ng/mL), PDGF-BB (1, 10, and 50 ng/mL), and bFGF (0.5, 1, and 5 ng/mL). The conditions used for combinations of growth factors were IGF-1 + PDGF-BB (50 + 10 and 100 + 50 ng/mL, respectively) and IGF-1 + PDGF-BB + bFGF (50 + 10 + 1; 50 + 10 + 5; 100 + 50 + 1; and 100 + 50 + 5 ng/mL, respectively). For all 3 tendon cell populations, proliferation at 72 h was greater in the presence of individual growth factors as compared to controls. With PDGF-BB (50 ng/mL) supplementation, mean absorbance values increased 97% (0.57 to 1.13) in S cells, 37% (0.51 to 0.70) in E cells, and 33% (0.33 to 0.44) in T cells (p < 0.001). In addition, a synergistic effect was observed. The combination of growth factors resulted in greater proliferation as compared to maximal doses of individual growth factors. In cultures supplemented with IGF-1 (100 ng/mL) + PDGF-BB (50 ng/mL), mean absorbance increased 114% (0.57 to 1.22) in S cells, 63% (0.51 to 0.831) in E cells, and 47% (0.33 to 0.48) in T cells (p < 0.001). IGF-1 (100 ng/mL) + PDGF-BB (50 ng/mL) + bFGF (5 ng/mL) resulted in the greatest amount of cell proliferation for all 3 tendon cell populations. The mean absorbances increased 251% in S cells, 98% in E cells, and 106% in T cells (p < 0.001). In summary, IGF-1, PDGF-BB, and bFGF can be used in combination to maximize tenocyte proliferation. Synergism among growth factors may provide a means to facilitate tendon engineering.