National & Regional United Engineering Lab of Tissue Engineering, Department of Orthopedics, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China; Center of Regenerative and Reconstructive Engineering Technology in Chongqing City, Chongqing, China.
Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA; Center of Regenerative and Reconstructive Engineering Technology in Chongqing City, Chongqing, China; Department of Orthopedic Surgery, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Acta Biomater. 2019 Feb;85:106-116. doi: 10.1016/j.actbio.2018.12.018. Epub 2018 Dec 14.
Among various bone tissue engineering strategies, selective cell retention (SCR) technology has been used as a practical clinical method for bone graft manufacturing in real time. The more mesenchymal stem cells (MSCs) are retained, the better the osteoinductive microenvironment provided by the scaffold, which in turn promotes the osteogenesis of the SCR-fabricated bone grafts. Integrin receptors are crucial to cell-matrix adhesion and signal transduction. We designed a collagen-binding domain (CBD)-containing IKVAV-cRGD peptide (CBD-IKVAV-cRGD peptide) to complement the collagen-based demineralized bone matrix (DBM) with a functionalized surface containing multiple integrin ligands, which correspond to the highly expressed integrin subtypes on MSCs. This DBM/CBD-IKVAV-cRGD composite exhibited superior in vitro adhesion capacity to cultured MSCs, as determined by oscillatory cell adhesion assay, centrifugal cell adhesion assay and mimetic SCR. Moreover, it promoted the retention of MSC-like CD271 cells and MSC-like CD90/CD105 cells in the clinical SCR method. Furthermore, the DBM/CBD-IKVAV-cRGD composite induced robust MSC osteogenesis, coupled with the activation of the downstream FAK-ERK1/2 signaling pathway of integrins. The SCR-prepared DBM/CBD-IKVAV-cRGD composite displayed superior in vivo osteogenesis, indicating that it may be potentially utilized as a biomaterial in SCR-mediated bone transplantation. STATEMENT OF SIGNIFICANCE: Selective cell retention technology (SCR) has been utilized in clinical settings to manufacture bioactive bone grafts. Specifically, demineralized bone matrix (DBM) is a widely-used SCR clinical biomaterial but it displays poor adhesion performance and osteoinduction. Improvements of the DBM that promote cell adhesion and osteoinduction will benefit SCR-prepared implants. In this work, we developed a novel peptide that complements the DBM with a functionalized surface of multiple integrin ligands, which are corresponding to integrin subtypes available on human bone marrow-derived mesenchymal stem cells (MSCs). Our results indicate this novel functionalized bioscaffold greatly increases SCR-mediated MSC adhesion and in vivo osteogenesis. Overall, this novel material has promising SCR applications and may likely provide highly bioactive bone implants in clinical settings.
在各种骨组织工程策略中，选择细胞保留（SCR）技术已被用作实时制造骨移植物的实用临床方法。保留的间充质干细胞（MSCs）越多，支架提供的成骨诱导微环境越好，从而促进 SCR 制造的骨移植物的成骨作用。整联蛋白受体对于细胞-基质粘附和信号转导至关重要。我们设计了一种含有胶原结合域（CBD）的 IKVAV-cRGD 肽（CBD-IKVAV-cRGD 肽），以补充具有包含多种整联蛋白配体的功能化表面的基于胶原的脱矿骨基质（DBM），这些配体对应于 MSCs 上高度表达的整联蛋白亚型。通过振荡细胞粘附测定、离心细胞粘附测定和模拟 SCR，该 DBM/CBD-IKVAV-cRGD 复合材料表现出对培养的 MSCs 的优异体外粘附能力。此外，它促进了 MSC 样 CD271 细胞和 MSC 样 CD90/CD105 细胞在临床 SCR 方法中的保留。此外，DBM/CBD-IKVAV-cRGD 复合材料诱导了强大的 MSC 成骨作用，并伴有整联蛋白下游 FAK-ERK1/2 信号通路的激活。SCR 制备的 DBM/CBD-IKVAV-cRGD 复合材料在体内表现出优异的成骨作用，表明它可能作为 SCR 介导的骨移植中的生物材料具有潜在用途。
选择性细胞保留技术（SCR）已在临床环境中用于制造生物活性骨移植物。具体来说，脱矿骨基质（DBM）是一种广泛用于 SCR 临床的生物材料，但它表现出较差的粘附性能和成骨诱导能力。改善 DBM 以促进细胞粘附和成骨诱导将有益于 SCR 制备的植入物。在这项工作中，我们开发了一种新型肽，该肽用多种整联蛋白配体的功能化表面补充 DBM，这些配体对应于人骨髓来源的间充质干细胞（MSCs）上可用的整联蛋白亚型。我们的结果表明，这种新型功能化生物支架大大增加了 SCR 介导的 MSC 粘附和体内成骨作用。总的来说，这种新型材料具有很有前途的 SCR 应用前景，并且可能在临床环境中提供高生物活性的骨植入物。
