Dual-functioning peptides discovered by phage display increase the magnitude and specificity of BMSC attachment to mineralized biomaterials.
Dual-functioning peptides discovered by phage display increase the magnitude and specificity of BMSC attachment to mineralized biomaterials.
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DOI：
10.1016/j.biomaterials.2017.04.034
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发表时间：
2017-07
期刊：
影响因子：
14
通讯作者：
Kohn DH
中科院分区：
文献类型：
--
作者：
Ramaraju H;Miller SJ;Kohn DH
用于基于细胞的治疗的生物材料的设计需要向细胞呈现特定的物理和化学线索，类似于由天然细胞外基质（ECM）提供的线索。我们先前使用噬菌体展示鉴定了对磷灰石具有高亲和力的肽序列（VTKHLNQISQSY，VTK）。本研究的目的是通过噬菌体展示技术筛选人骨髓基质细胞特异性短肽序列，并将其与磷灰石特异性短肽序列进行联合收割机结合，验证所构建的双功能短肽对磷灰石和人骨髓基质细胞的特异性。在这项研究中，组合噬菌体展示鉴定了细胞结合序列（DPIYALSWSGMA，DPI），其与矿物质结合序列组合以产生双肽DPI-VTK。与VTK、磷酸化VTK（VTKphos）、DPI-VTKphos、RGD-VTK和无肽磷灰石表面相比，DPI-VTK表现出对磷灰石表面的显著更大的结合亲和力（1/KD）（p < 0.01），同时显著增加hBMSC粘附强度（τ50，p < 0.01）。与其他细胞类型相比，MSC表现出对DPI-VTK的显著更大的粘附强度，而MC 3 T3前成骨细胞和鼠成纤维细胞的附着是有限的（p < 0.01）。与前成骨细胞和成纤维细胞相比，DPI-VTK包被表面上的MSC也表现出增加的扩散。与对照相比，在DPI-VTK涂覆的磷灰石膜上培养的MSC表现出显著更大的增殖（p < 0.001）。此外，与对照组相比，DPI-VTK涂层磷灰石膜上的早期和晚期成骨分化标志物升高。总之，噬菌体展示可以识别非明显的细胞和材料特异性肽，以增加人MSC对特定生物材料表面的粘附强度，并随后增加细胞增殖和分化。这些新的肽扩展了用于骨缺损的基于细胞的再生的生物材料设计方法。这种结合细胞和物质的噬菌体展示衍生肽的策略广泛适用于需要特定细胞群的靶向粘附的各种系统，并且可以推广到任何粘附表面的工程化。
Design of biomaterials for cell-based therapies requires presentation of specific physical and chemical cues to cells, analogous to cues provided by native extracellular matrices (ECM). We previously identified a peptide sequence with high affinity towards apatite (VTKHLNQISQSY, VTK) using phage display. The aims of this study were to identify a human MSC-specific peptide sequence through phage display, combine it with the apatite-specific sequence, and verify the specificity of the combined dual-functioning peptide to both apatite and human bone marrow stromal cells. In this study, a combinatorial phage display identified the cell binding sequence (DPIYALSWSGMA, DPI) which was combined with the mineral binding sequence to generate the dual peptide DPI-VTK. DPI-VTK demonstrated significantly greater binding affinity (1/KD) to apatite surfaces compared to VTK, phosphorylated VTK (VTKphos), DPI-VTKphos, RGD-VTK, and peptide-free apatite surfaces (p < 0.01), while significantly increasing hBMSC adhesion strength (τ50, p < 0.01). MSCs demonstrated significantly greater adhesion strength to DPI-VTK compared to other cell types, while attachment of MC3T3 pre-osteoblasts and murine fibroblasts was limited (p < 0.01). MSCs on DPI-VTK coated surfaces also demonstrated increased spreading compared to pre-osteoblasts and fibroblasts. MSCs cultured on DPI-VTK coated apatite films exhibited significantly greater proliferation compared to controls (p < 0.001). Moreover, early and late stage osteogenic differentiation markers were elevated on DPI-VTK coated apatite films compared to controls. Taken together, phage display can identify non-obvious cell and material specific peptides to increase human MSC adhesion strength to specific biomaterial surfaces and subsequently increase cell proliferation and differentiation. These new peptides expand biomaterial design methodology for cell-based regeneration of bone defects. This strategy of combining cell and material binding phage display derived peptides is broadly applicable to a variety of systems requiring targeted adhesion of specific cell populations, and may be generalized to the engineering of any adhesion surface.