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Cultured bone on biomaterial substrates : a tissue engineering approach to treat bone defects

机译：生物材料基质上的培养骨：用于治疗骨缺陷的组织工程方法

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In the present thesis, a tissue engineering approach to treat bone defects was investigated. Such strategy was based on the use of patient own cultured bone marrow stromal cells (BMSCs) in association with biomaterials to produce autologous living bone equivalents. When engineering such implants, three main factors had to be taken into account: (i) the cells, (ii) the culture technology and (iii) the biomaterial scaffolds. The capacity of BMSCs to proliferate, differentiate along the osteogenic lineage and form a bone like tissue was demonstrated in various in vitro assays making use of biochemical,udimmunological, microscopic and gene expression techniques. The ability of the cells to produce bone in vivo was established using an ectopic (extra osseous) implantation model.udResults indicated that BMSC cultures were composed of a heterogeneous populationudcontaining a subpopulation of cells with high proliferative capacity and with potential to differentiate into bone forming cells. Both the growth and the differentiation pattern of these cells could be manipulated, to a certain degree, through the use of bioactive factors during culture. After implantation, the bone forming capacity of the cultures proved to be related to the amount of early osteoprogenitors and precursors cells that could be induced into starting the osteogenic differentiation process. In bone marrow aspirates, this subpopulation appeared to decrease with donor age and to be strongly dependent on the donor, indicating that the aspiration procedure plays an important role in the obtained bone marrow cell population. In order to evaluate the in vivo bone formation capacity of BMSC cultures prior to implantation, an experimental method was developed in which the amount of earlyudosteoprogenitors and precursors cells could be quantified. With regard to the technology design, data indicated that the culture of cells on the biomaterial scaffolds prior to implantation resulted in implants with faster in vivo boneudforming ability as compared to scaffolds implanted after cell seeding. In addition, two biodegradable polymeric systems were proposed as scaffolds to be used in the described bone engineering approach after evaluating their ability to support bone marrow cell growth, differentiation and in vivo bone formation.udIn summary, although the complete knowledge of the factors controlling BMSC rowth and osteogenic differentiation still needs to be further expanded, the obtained results suggestudthat the bone tissue engineering approach described in this thesis presents a great potentialudfor the repair of bone defects and will become an advantageous alternative to the traditionaludautologous bone grafting.
机译：本文研究了一种组织工程治疗骨缺损的方法。这种策略是基于将患者自身培养的骨髓基质细胞（BMSC）与生物材料结合使用，以产生自体活骨等效物。在设计此类植入物时，必须考虑三个主要因素：（i）细胞，（ii）培养技术和（iii）生物材料支架。利用生化，免疫，显微镜和基因表达技术的各种体外试验证明了BMSCs沿成骨细胞系增殖，分化和形成骨样组织的能力。 ud结果表明，BMSC培养物由异质性种群组成，其中含有高增殖能力的亚细胞群，并具有分化为成骨细胞的潜能。骨形成细胞。通过在培养过程中使用生物活性因子，可以在一定程度上控制这些细胞的生长和分化模式。植入后，培养物的骨形成能力证明与可被诱导开始成骨分化过程的早期骨祖细胞和前体细胞的数量有关。在骨髓抽吸物中，该亚群似乎随着供体的年龄而减少，并且强烈依赖于供体，这表明抽吸过程在获得的骨髓细胞群中起着重要的作用。为了评估BMSC培养物在植入之前的体内骨形成能力，开发了一种实验方法，其中可以量化早期 uudosteo祖细胞和前体细胞的数量。关于技术设计，数据表明，与植入细胞后植入的支架相比，植入前在生物材料支架上的细胞培养导致植入物具有更快的体内骨形成能力。此外，在评估了它们支持骨髓细胞生长，分化和体内骨骼形成的能力后，提出了两种可生物降解的聚合物系统作为支架，用于所述的骨工程学方法。 BMSC的行和成骨分化仍需进一步扩大，结果表明，本文所描述的骨组织工程方法具有修复骨缺损的巨大潜力，将成为传统自体骨的有利替代物。嫁接。

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