Tumor Immunology and Immunotherapy Group, Advanced Centre for Treatment, Research and Education in Cancer, Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
The Ritchie Centre, Hudson Institute of Medical Research, Clayton VIC Australia 3168.
Biomacromolecules. 2020 Dec 14;21(12):4888-4903. doi: 10.1021/acs.biomac.0c01134. Epub 2020 Nov 2.
There is an increasing need for bone substitutes for reconstructive orthopedic surgery following removal of bone tumors. Despite the advances in bone regeneration, the use of autologous mesenchymal stem cells (MSC) presents a significant challenge, particularly for the treatment of large bone defects in cancer patients. This study aims at developing new chemokine-based technology to generate biodegradable scaffolds that bind pharmacologically active proteins for regeneration/repair of target injured tissues in patients. Primary MSC were cultured from the uninvolved bone marrow (BM) of cancer patients and further characterized for "stemness". Their ability to differentiate into an osteogenic lineage was studied in 2D cultures as well as on 3D macroporous PLGA scaffolds incorporated with biomacromolecules bFGF and homing factor chemokine stromal-cell derived factor-1 (SDF1). MSC from the uninvolved BM of cancer patients exhibited properties similar to that reported for MSC from BM of healthy individuals. Macroporous PLGA discs were prepared and characterized for pore size, architecture, functional groups, thermostability, and cytocompatibility by ESEM, FTIR, DSC, and CCK-8 dye proliferation assay, respectively. It was observed that the MSC+PLGA+bFGF+SDF1 construct cultured for 14 days supported significant cell growth, osteo-lineage differentiation with increased osteocalcin expression, alkaline phosphatase secretion, calcium mineralization, bone volume, and soluble IL6 compared to unseeded PLGA and PLGA+MSC, as analyzed by confocal microscopy, biochemistry, ESEM, microCT imaging, flow cytometry, and EDS. Thus, chemotactic biomacromolecule SDF1-guided tissue repair/regeneration ability of MSC from cancer patients opens up the avenues for development of "off-the-shelf" pharmacologically active construct for optimal repair of the target injured tissue in postsurgery cancer patients, bone defects, damaged bladder tissue, and radiation-induced skin/mucosal lesions.
在骨肿瘤切除后，重建骨科手术对骨替代物的需求不断增加。尽管骨再生技术取得了进展，但使用自体间充质干细胞（MSC）仍然是一个重大挑战，特别是对于癌症患者的大骨缺损治疗。本研究旨在开发新的趋化因子基技术，以生成可生物降解的支架，将药理活性蛋白结合起来，用于再生/修复患者目标受损组织。原代 MSC 从癌症患者未受累骨髓（BM）中培养，并进一步对其“干性”进行特征分析。研究了它们在 2D 培养物以及掺入生物大分子 bFGF 和归巢因子趋化因子基质细胞衍生因子-1（SDF1）的 3D 大孔 PLGA 支架上向成骨谱系分化的能力。来自癌症患者未受累 BM 的 MSC 表现出与健康个体 BM 来源 MSC 报道的相似特性。通过 ESEM、FTIR、DSC 和 CCK-8 染料增殖测定法分别对大孔 PLGA 圆盘进行了孔径、结构、官能团、热稳定性和细胞相容性的表征。观察到，与未接种 PLGA 和 PLGA+MSC 的细胞相比，培养 14 天的 MSC+PLGA+bFGF+SDF1 构建体支持显著的细胞生长、成骨谱系分化，骨钙素表达增加、碱性磷酸酶分泌、钙矿化、骨体积和可溶性 IL6，通过共聚焦显微镜、生物化学、ESEM、microCT 成像、流式细胞术和 EDS 进行分析。因此，癌症患者 MSC 趋化生物大分子 SDF1 引导组织修复/再生能力为开发“现成”的药理活性构建体开辟了道路，用于优化手术后癌症患者、骨缺损、受损膀胱组织和辐射诱导的皮肤/黏膜损伤等目标受损组织的修复。
