Chondroitin-6-sulphate incorporated into collagen gels for the growth of human keratinocytes: The effect of cross-linking agents and diamines
Chondroitin-6-sulphate incorporated into collagen gels for the growth of human keratinocytes: The effect of cross-linking agents and diamines
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DOI：
10.1016/0142-9612(96)81414-1
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发表时间：
1996-04-01
期刊：
影响因子：
14
通讯作者：
Grant, MH
中科院分区：
文献类型：
--
作者：
Hanthamrongwit, M;Reid, WH;Grant, MH
本研究证明了糖胺聚糖、透明质酸和 6-硫酸软骨素 (Ch6SO(4))、二胺和碳二亚胺交联剂对胶原凝胶上人类表皮细胞生长的影响。 Ch6SO(4) 掺入胶原蛋白凝胶中可刺激细胞生长速度，但发现效果并不一致。我们发现，在培养的前 3 天后，凝胶中掺入的 Ch6SO(4) 大约有 50% 浸出到生长培养基中，这被认为导致细胞生长反应不一致。为了最大限度地减少Ch6SO(4)从凝胶中的洗脱，从而最大化其对角质形成细胞生长的影响，在培养基中添加1-100μg ml(-1) Ch6SO(4)。结果表明，培养基中这些浓度的 Ch6SO(4) 不会刺激普通胶原凝胶或含有 20% Ch6SO(4) 的凝胶上的细胞生长。作为替代策略，使用 1-乙基-3-(3-二甲基氨基丙基) 碳二亚胺和二胺（腐胺或二氨基己烷）将 Ch6SO(4) 固定到胶原凝胶上并使凝胶交联。交联过程部分阻止了 Ch6SO(4) 从凝胶中洗脱。有趣的是，只有腐胺，而不是二氨基己烷，促进了交联普通胶原凝胶上角质形成细胞的生长。我们提议开发一种含有腐胺作为人类表皮细胞生长因子的人造皮肤替代品。
This study demonstrates the effect of the glycosaminoglycans, hyaluronic acid and chondroitin-6-sulphate (Ch6SO(4)), diamines and a carbodiimide cross-linking agent on the growth of human epidermal cells on collagen gels. Ch6SO(4) incorporated into collagen gels stimulated cell growth rate, but the effect was found to be inconsistent. We found that approximately 50% of the incorporated Ch6SO(4) in the gels leached out into the growth medium after the first 3 d in culture, and this is thought to lead to the inconsistent cell growth response. In order to minimize the elution of Ch6SO(4) from the gels and thereby maximize its effect on the growth of the keratinocytes, 1-100 mu g ml(-1) Ch6SO(4) was added in the medium. The results showed that Ch6SO(4) at these concentrations in the medium did not stimulate the cell growth on either plain collagen gels or gels containing 20% Ch6SO(4). As an alternative strategy, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and diamines (putrescine or diaminohexane) were used to immobilize Ch6SO(4) onto the collagen gels and to cross-link the gels. The cross-linking process partially prevented the elution of Ch6SO(4) from the gels. Interestingly, only putrescine, not diaminohexane, promoted the growth of keratinocytes on the cross-linked plain collagen gels. We proposed to develop an artificial skin substitute containing putrescine as a growth factor for the human epidermal cells.