Objective. Bone defects or atrophy may arise as a consequence of injury, inflammation of various etiologies, and neoplastic or traumatic processes or as a result of surgical procedures. Sometimes the regeneration process of bone loss is impaired, significantly slowed down, or does not occur, e.g., in congenital defects. For the bone defect reconstruction, a piece of the removed bone from ala of ilium or bone transplantation from a decedent is used. Replacement of the autologous or allogenic source of the bone-by-bone substitute could reduce the number of surgeries and time in the pharmacological coma during the reconstruction of the bone defect. Application of mesenchymal stem cells in the reconstruction surgery may have positive influence on tissue regeneration by secretion of angiogenic factors, recruitment of other MSCs, or differentiation into osteoblasts. Materials and Methods. Mesenchymal stem cells derived from the umbilical cord (Wharton’s jelly (WJ-MSC)) were cultured in GMP-grade DMEM low glucose supplemented with heparin, 10% platelet lysate, glucose, and antibiotics. In vitro WJ-MSCs were seeded on the bone substitute Bio-Oss Collagen® and cultured in the StemPro® Osteogenesis Differentiation Kit. During the culture on the 1st, 7th, 14th, and 21st day (day in vitro (DIV)), we analyzed viability (confocal microscopy) and adhesion capability (electron microscopy) of WJ-MSC on Bio-Oss scaffolds, gene expression (qPCR), and secretion of proteins (Luminex). In vivo Bio-Oss® scaffolds with WJ-MSC were transplanted to trepanation holes in the cranium to obtain their overgrowth. The computed tomography was performed 7, 14, and 21 days after surgery to assess the regeneration. Results. The Bio-Oss® scaffold provides a favourable environment for WJ-MSC survival. WJ-MSCs in osteodifferentiation medium are able to attach and proliferate on Bio-Oss® scaffolds. Results obtained from qPCR and Luminex® indicate that WJ-MSCs possess the ability to differentiate into osteoblast-like cells and may induce osteoclastogenesis, angiogenesis, and mobilization of host MSCs. In animal studies, WJ-MSCs seeded on Bio-Oss® increased the scaffold integration with host bone and changed their morphology to osteoblast-like cells. Conclusions. The presented construct consisted of Bio-Oss®, the scaffold with high flexibility and plasticity, approved for clinical use with seeded immunologically privileged WJ-MSC which may be considered reconstructive therapy in bone defects.

中文翻译：

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使用源自脐带的间充质干细胞支持的骨替代物修复骨缺损。

客观的. 骨缺损或萎缩可能是由于损伤、各种病因的炎症、肿瘤或创伤过程或外科手术的结果而引起的。有时，骨质流失的再生过程会受到损害、显着减慢或不发生，例如在先天性缺陷中。对于骨缺损重建，使用取自髂骨翼或死者骨移植的一块骨。在骨缺损重建过程中，替代自体或同种异体来源的骨替代物可以减少手术次数和药理学昏迷时间。间充质干细胞在重建手术中的应用可能通过分泌血管生成因子、募集其他 MSCs、材料和方法。来自脐带的间充质干细胞（Wharton's jelly (WJ-MSC)）在补充有肝素、10% 血小板裂解物、葡萄糖和抗生素的 GMP 级 DMEM 低葡萄糖中培养。将体外WJ-MSCs 接种在骨替代物 Bio-Oss Collagen® 上，并在 StemPro® 成骨分化试剂盒中培养。在第 1、7、14 和 21 天（体外日（DIV））培养期间，我们分析了 WJ-MSC 在 Bio-Oss 支架上的活力（共聚焦显微镜）和粘附能力（电子显微镜）、基因表达（ qPCR）和蛋白质分泌（Luminex）。体内将带有 WJ-MSC 的 Bio-Oss® 支架移植到颅骨的钻孔中以使其过度生长。手术后 7、14 和 21 天进行计算机断层扫描以评估再生情况。结果。Bio-Oss® 支架为 WJ-MSC 的生存提供了有利的环境。骨分化培养基中的 WJ-MSC 能够在 Bio-Oss® 支架上附着和增殖。从 qPCR 和 Luminex® 获得的结果表明，WJ-MSCs 具有分化成成骨细胞样细胞的能力，并可能诱导破骨细胞生成、血管生成和宿主 MSCs 的动员。在动物研究中，接种在 Bio-Oss® 上的 WJ-MSCs 增加了支架与宿主骨的整合，并将其形态改变为成骨细胞样细胞。结论. 所提出的构建体由 Bio-Oss® 组成，该支架具有高柔韧性和可塑性，已批准用于临床使用具有免疫特权的 WJ-MSC 种子，可被视为骨缺损的重建疗法。