Kao W J, Hubbell J A
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Biotechnol Bioeng. 1998 Jul 5;59(1):2-9.
Polyethyleneglycol-based networks were employed as substrates to graft bioactive peptides to study macrophage interactions with materials. Our overall objective was to utilize biologically active factors to stimulate certain macrophage function on materials suitable for implantation in connective tissues. In this study, we sought to explore the bioactivity of several peptides derived from extracellular matrix adhesion proteins and macrophage-active proteins that are normally soluble. The candidate peptides examined corresponded to residues 63 to 77 of complement component C3a (C3a(63-77)), residues 178 to 207 of interleukin-1 beta (IL1beta(178-207)), residues 1615 to 1624 of fibronectin (FN(1615-1624)), endothelial-macrophage activating polypeptide II, complement component C5a inhibitory sequence, macrophage inhibitory peptide, and YRGDG; materials lacking peptides were used as negative controls. An established murine cell-line IC-21 was employed as a macrophage model, and human dermal fibroblasts were used for comparison. Our results showed that the substrates without grafted peptides were free from artifactual cell adhesion associated with the adsorption of serum or cellularly secreted proteins for long duration of culture. Of all grafted samples, IL1beta(178-207)- and C3a(63-77)-grafted surfaces supported higher adherent macrophage densities. C3a(63-77)- and FN(1615-1624)-grafted surfaces supported higher adherent fibroblast densities. From competitive inhibition studies, cell adhesion was determined to occur in a receptor-peptide specific manner. The presence of grafted YRGDG in addition to IL1beta(178-207), C3a(63-77), or FN(1615-1624) synergistically increased macrophage and fibroblast adhesion. Materials grafted with IL1beta(178-207) or C3a(63-77) co-grafted with or without YRGDG did not support the formation of multinucleated giant cells from the fusion of adherent macrophages in vitro.
以聚乙二醇为基础的网络被用作底物来接枝生物活性肽，以研究巨噬细胞与材料的相互作用。我们的总体目标是利用生物活性因子在适合植入结缔组织的材料上刺激某些巨噬细胞功能。在本研究中，我们试图探索几种源自细胞外基质粘附蛋白和通常可溶的巨噬细胞活性蛋白的肽的生物活性。所检测的候选肽对应于补体成分C3a的63至77位残基（C3a(63 - 77)）、白细胞介素-1β的178至207位残基（IL1β(178 - 207)）、纤连蛋白的1615至1624位残基（FN(1615 - 1624)）、内皮巨噬细胞激活多肽II、补体成分C5a抑制序列、巨噬细胞抑制肽以及YRGDG；缺乏肽的材料用作阴性对照。已建立的小鼠细胞系IC - 21被用作巨噬细胞模型，人皮肤成纤维细胞用于比较。我们的结果表明，在长时间培养中，未接枝肽的底物不存在与血清或细胞分泌蛋白吸附相关的人为细胞粘附。在所有接枝样品中，接枝了IL1β(178 - 207)和C3a(63 - 77)的表面支持更高的贴壁巨噬细胞密度。接枝了C3a(63 - 77)和FN(1615 - 1624)的表面支持更高的贴壁成纤维细胞密度。通过竞争性抑制研究，确定细胞粘附以受体 - 肽特异性方式发生。除了IL1β(178 - 207)、C3a(63 - 77)或FN(1615 - 1624)之外，接枝YRGDG可协同增加巨噬细胞和成纤维细胞的粘附。接枝有IL1β(178 - 207)或C3a(63 - 77)且共接枝或未共接枝YRGDG的材料不支持体外贴壁巨噬细胞融合形成多核巨细胞。
