Oliveira Joaquim M, Sousa Rui A, Kotobuki Noriko, Tadokoro Mika, Hirose Motohiro, Mano João F, Reis Rui L, Ohgushi Hajime
3B's Research Group-Biomaterials, Biodegradables and Biomimetics, Univ. Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, S. Cláudio de Barco, 4806-909 Taipas, Guimarães, Portugal.
Biomaterials. 2009 Feb;30(5):804-13. doi: 10.1016/j.biomaterials.2008.10.024. Epub 2008 Nov 25.
There is an increasing interest in developing novel macromolecular vehicles for the intracellular and controlled delivery of bioactive molecules, since they can allow modulation of the cellular functions in a more effective manner ex vivo, and maintain the cellular phenotype in vivo upon re-implantation. The present study was designed to investigate the effect of combining novel dexamethasone-loaded carboxymethylchitosan/poly(amidoamine) dendrimer (Dex-loaded CMCht/PAMAM) nanoparticles and, both HA and SPCL scaffolds (3D system) on the proliferation and osteogenic differentiation of rat bone marrow stromal cells (RBMSCs) in vitro. A luminescent cell viability assay using RBMSCs was performed for screening cytotoxicity of the developed HA and SPCL scaffolds. Results corroborated previous ones which have demonstrated in vitro, the superior performance of the HA and SPCL scaffolds on supporting cells adhesion and proliferation. Furthermore, this work showed that RBMSCs seeded onto the surface of both HA and SPCL scaffolds differentiate into osteoblasts when cultured in the presence of 0.01 mg ml(-1) Dex-loaded CMCht/PAMAM dendrimer nanoparticles. In addition, results demonstrated that Dex-loaded CMCht/PAMAM dendrimer nanoparticles combined with the HA enhance osteogenesis by increasing ALP activity and mineralization of the extra-cellular matrix. The pre-incubation of stem cells with these kinds of nanoparticles allows the delivery of Dex inside the cells and directly influences their cellular fate, being a promising new tool to be used in cells and tissue engineering strategies.
开发新型大分子载体用于生物活性分子的细胞内和可控递送的兴趣与日俱增,因为它们能够以更有效的方式在体外调节细胞功能,并在重新植入后在体内维持细胞表型。本研究旨在探讨新型载地塞米松的羧甲基壳聚糖/聚(酰胺胺)树枝状大分子(载地塞米松的CMCht/PAMAM)纳米颗粒与HA和SPCL支架(三维系统)相结合对大鼠骨髓基质细胞(RBMSCs)体外增殖和成骨分化的影响。使用RBMSCs进行发光细胞活力测定以筛选所开发的HA和SPCL支架的细胞毒性。结果证实了先前的研究,这些研究在体外证明了HA和SPCL支架在支持细胞粘附和增殖方面的优越性能。此外,这项工作表明,当在含有0.01 mg ml(-1)载地塞米松的CMCht/PAMAM树枝状大分子纳米颗粒的情况下培养时,接种到HA和SPCL支架表面的RBMSCs会分化为成骨细胞。此外,结果表明,载地塞米松的CMCht/PAMAM树枝状大分子纳米颗粒与HA结合可通过增加碱性磷酸酶活性和细胞外基质矿化来增强成骨作用。用这类纳米颗粒对干细胞进行预孵育可使地塞米松在细胞内递送并直接影响其细胞命运,这是一种有望用于细胞和组织工程策略的新工具。