Steplewski Andrzej, Fertala Jolanta, Beredjiklian Pedro, Wang Mark L, Fertala Andrzej
1 Division of Orthopaedic Research, Department of Orthopaedic Surgery, Sidney Kimmel Medical College, Thomas Jefferson University , Philadelphia, Pennsylvania.
Tissue Eng Part A. 2015 Apr;21(7-8):1207-16. doi: 10.1089/ten.tea.2014.0401. Epub 2015 Jan 13.
Biomedical strategies for tissue engineering and repair utilize specific cells, scaffolds, and growth factors to reconstruct elements of damaged tissue. The cellular element of these strategies is limited, however, by poor efficiency of delivery and retention of therapeutic cells in target sites. We propose that the presence of a cellular anchor that is able to specifically bind a defined element of target tissue will facilitate efficient binding and retention of therapeutic cells, thereby promoting repair of the target site. To do so, we engineered an artificial collagen-specific anchor (ACSA) that is able to specifically bind collagen I. The ACSA was engineered by creating a construct comprising rationally designed consecutive domains. The binding specificity of the ACSA was achieved by employing variable regions of a monoclonal antibody that recognizes a unique epitope present in human collagen I. Meanwhile, cell membrane localization of the ACSA was provided by the presence of a transmembrane domain. We determined that the ACSA was localized within cell membranes and interacted with its intended target, that is, collagen I. We have demonstrated that, in comparison to the control, the cells expressing the ACSA attached better to collagen I and exhibited improved retention in sites of seeding. We have also demonstrated that the presence of the ACSA did not interfere with cell proliferation, the biosynthesis of endogenous collagen I, or the biological functions of native collagen receptors. Since the presented cell delivery system utilizes a common characteristic of major connective tissues, namely the presence of collagen I, the findings described here could have a broad positive impact for improving the repair processes of tendon, ligament, bone, intervertebral disc, skin, and other collagen I-rich connective tissues. If successful, the ACSA approach to deliver cells will serve as an outline for developing cell delivery methods that target other elements of extracellular matrices, including other collagen types, laminins, and fibronectins.
用于组织工程和修复的生物医学策略利用特定的细胞、支架和生长因子来重建受损组织的成分。然而,这些策略中的细胞成分受到治疗性细胞在靶位点递送和保留效率低下的限制。我们提出,存在一种能够特异性结合靶组织特定成分的细胞锚定物将有助于治疗性细胞的有效结合和保留,从而促进靶位点的修复。为此,我们设计了一种人工胶原蛋白特异性锚定物(ACSA),它能够特异性结合I型胶原蛋白。ACSA是通过构建一个包含合理设计的连续结构域的构建体来设计的。ACSA的结合特异性是通过使用单克隆抗体的可变区来实现的,该可变区识别存在于人I型胶原蛋白中的独特表位。同时,ACSA的细胞膜定位是由跨膜结构域的存在提供的。我们确定ACSA定位于细胞膜内并与其预期靶点即I型胶原蛋白相互作用。我们已经证明,与对照相比,表达ACSA的细胞与I型胶原蛋白的附着更好,并且在接种部位的保留率提高。我们还证明,ACSA的存在不会干扰细胞增殖、内源性I型胶原蛋白的生物合成或天然胶原蛋白受体的生物学功能。由于所提出的细胞递送系统利用了主要结缔组织的共同特征,即I型胶原蛋白的存在,这里描述的发现可能对改善肌腱、韧带、骨骼、椎间盘、皮肤和其他富含I型胶原蛋白的结缔组织的修复过程产生广泛的积极影响。如果成功,ACSA递送细胞的方法将为开发针对细胞外基质其他成分(包括其他胶原蛋白类型、层粘连蛋白和纤连蛋白)的细胞递送方法提供一个框架。