The combination of insulin-like growth factor 1 and osteogenic protein 1 promotes increased survival of and matrix synthesis by normal and osteoarthritic human articular chondrocytes
The combination of insulin-like growth factor 1 and osteogenic protein 1 promotes increased survival of and matrix synthesis by normal and osteoarthritic human articular chondrocytes
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DOI：
10.1002/art.11209
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发表时间：
2003-08-01
影响因子：
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通讯作者：
Chubinskaya, S
中科院分区：
文献类型：
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作者：
Loeser, RF;Pacione, CA;Chubinskaya, S
Objective.虽然生长因子疗法可能是刺激受损软骨基质修复的有吸引力的方法，但有证据表明，随着年龄的增长和/或骨关节炎（OA）的发展，关节软骨细胞可能对生长因子刺激无反应。本研究的目的是比较胰岛素样生长因子1（IGF-1）和成骨蛋白I（OP-1）单独和联合刺激培养的人正常和OA软骨细胞的能力。将通过酶消化软骨而分离的软骨细胞在无血清培养基中的藻酸盐珠中培养，所述软骨得自经受OA膝关节置换的受试者（n = 6）或得自组织供体的正常踝关节（n = 7），并用100 ng/ml IGF-1、100 ng/ml OP-1或两者处理21天。对照组仅用溶媒处理。通过DNA分析评价培养物的细胞存活率、细胞数量、颗粒排阻试验评价基质产生、二甲基亚甲基蓝试验评价蛋白多糖积累水平。在无血清藻酸盐培养21天后，来自OA软骨的细胞的存活率为65 +/- 2%（平均值+/- SEM），而来自正常软骨的细胞的存活率显著更高（82 +/- 3%）。单独使用IGF-1或OP-1治疗可最小程度地改善存活率，而IGF + OP组合可显著改善存活率，OA细胞为87 +/- 2%，正常细胞为95 +/- 1%。仅在IGF + OP组中观察到细胞增殖;这对于正常和OA细胞都是显著的（类似于DNA水平增加2倍）。通过颗粒排除和蛋白多糖积累评估的基质产生在正常和OA培养物中用IGF + OP处理的细胞中最大。当蛋白聚糖水平根据细胞数量校正时（μ g蛋白聚糖/ng DNA），在正常和OA培养物中，单独OP-1和IGF + OP组均显著高于对照组，但单独IGF-1组则无显著差异，联合组水平最高（高于对照组3倍）。OP-1比IGF-1更有效地刺激正常和OA细胞中蛋白多糖的产生。然而，最好的结果是用组合获得的，这表明IGF-1和OP-1的组合疗法可能是治疗OA软骨损伤的有效策略。
Objective. Although growth factor therapy could be an attractive method for stimulating the repair of damaged cartilage matrix, there is evidence that With aging and/or with the development of osteoarthritis (OA), articular chondrocytes may become unresponsive to growth factor stimulation. The aim of the current study was to compare the ability of insulin-like growth factor 1 (IGF-1) and osteogenic protein I (OP-1), alone and in combination, to stimulate human normal and OA chondrocytes in culture.Methods. Chondrocytes isolated by enzymatic digestion of cartilage obtained from subjects undergoing knee replacement for OA (n = 6) or from normal ankle joints of tissue donors (n = 7) were cultured in alginate beads in serum-free medium and treated for 21 days with 100 ng/ml IGF-1, 100 ng/ml OP-1, or both. Controls were treated with vehicle alone. The cultures were evaluated for cell survival, cell number by DNA analysis, matrix production by particle exclusion assay, and level of accumulated proteoglycan by dimethylmethylene blue assay.Results. After 21 days in serum-free alginate culture, survival of cells from OA cartilage was 65 +/- 2% (mean +/- SEM), while survival of cells from normal cartilage was significantly greater (82 +/- 3%). Treatment with either IGF-1 or OP-1 alone minimally improved survival, while the combination IGF + OP significantly improved survival, to 87 +/- 2% for OA cells and 95 +/- 1% for normal cells. Cell proliferation was noted only in the IGF + OP group; this was significant for both normal and OA cells (similar to2-fold increase in DNA levels). Matrix production, assessed by particle exclusion and by proteoglycan accumulation, was greatest in the cells treated with IGF + OP in both normal and OA cultures. When proteoglycan levels were corrected for cell numbers (mug proteoglycan/ng DNA), a significant increase over control was noted with OP-1 alone and IGF + OP, but not IGF-1 alone, in both normal and OA cultures, with the greatest levels in the combination group (3-fold increase over control).Conclusion. OP-1 was more potent than IGF-1 in stimulating proteoglycan production in both normal and OA cells. However, the best results were obtained with the combination, suggesting that combined therapy with IGF-1 and OP-1 may be an effective strategy for treating OA cartilage damage.