Design of biomimetic habitats for tissue engineering with P-15, a synthetic peptide analogue of collagen
Design of biomimetic habitats for tissue engineering with P-15, a synthetic peptide analogue of collagen
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DOI：
10.1089/ten.1999.5.53
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发表时间：
1999-03-01
影响因子：
--
通讯作者：
Smith, N
中科院分区：
文献类型：
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作者：
Bhatnagar, RS;Qian, JJ;Smith, N
在组织中，胶原蛋白形成细胞附着和迁移的支架，并通过介导化学和机械刺激的通量来调节细胞分化和形态发生。我们通过将胶原蛋白衍生的高亲和力细胞结合肽P-15固定在三维（3-D）模板中来构建仿生环境。预期细胞结合肽可以抵抗机械力。在其生理环境中，牙周膜成纤维细胞（PDLF）受到显著的机械力。我们研究了人PDLF在P-15包被的颗粒状牛无机骨矿物质（ABM-P-15）上培养的行为。与单独的ABM相比，与ABM-P-15相关的细胞数量更多。与ABM培养物相比，ABM-P-15上放射性标记前体在DNA和蛋白质中的掺入水平更高，这与存在更大量的细胞一致。扫描电子显微镜检查表明，ABM-P-15上的培养物产生高度定向的细长细胞的3-D集落，并形成大量的纤维以及膜基质，使人联想到韧带结构。在ABM上培养的PDLF形成稀疏的单层，几乎没有秩序和贫乏的基质。茜素红染色的颗粒相关的细胞和颗粒间的细胞桥P-15相关的文化，表明矿化的基质。三维集落形成和有序的细胞沿着增加矿化表明，耦合的细胞ABM矩阵通过P-15可以提供一个仿生环境，允许细胞分化和形态发生。我们的研究表明，ABM-P-15模板作为骨内移植物可能是有效的，并且，当与PDLF一起接种时，这些基质可以作为自体骨移植物的组织工程替代物。
In tissues, collagen forms the scaffold for cell attachment and migration, and it modulates cell differentiation and morphogenesis by mediating the flux of chemical and mechanical stimuli. We are constructing biomimetic environments by immobilizing a collagen-derived high-affinity cell-binding peptide P-15 in three-dimensional (3-D) templates. The cell-binding peptide can be expected to transduce mechanical forces. In their physiological environment, periodontal ligament fibroblasts (PDLF) are subject to significant mechanical forces. We have examined the behavior of human PDLF in culture on particulate bovine anorganic bone mineral (ABM) coated with P-15 (ABM-P-15). Greater numbers of cells associated with ABM-P-15 compared to ABM alone. Higher levels of incorporation of radiolabeled precursors in DNA and protein were consistent with the presence of larger numbers of cells on ABM-P-15 compared to ABM cultures. Scanning electron microscopic examination showed that cultures on ABM-P-15 generated highly oriented 3-D colonies of elongated cells and formed copious amounts of fibrous as well as membranous matrix reminiscent of ligamentous structures. PDLF cultured on ABM formed sparse monolayers with little order and a meager matrix. Alizarin Red stained the matrix of particle associated cells and inter-particle cellular bridges in P-15-associated cultures, indicating mineralization. 3-D colony formation and ordering of cells along with increased mineralization suggests that the coupling of cells to the ABM matrix through P-15 may provide a biomimetic environment permissive for cell differentiation and morphogenesis. Our studies suggest that ABM-P-15 templates may be effective as endosseous grafts, and, when seeded with PDLF, these matrices may serve as tissue engineered substitutes for autologous bone grafts.