将一种顺序性骨形态发生蛋白-2/骨形态发生蛋白-7递送系统整合到用于骨组织工程的壳聚糖基支架中。

Incorporation of a sequential BMP-2/BMP-7 delivery system into chitosan-based scaffolds for bone tissue engineering.

作者信息

Yilgor Pinar, Tuzlakoglu Kadriye, Reis Rui L, Hasirci Nesrin, Hasirci Vasif

机构信息

METU, BIOMAT, Department of Biotechnology, 06531 Ankara, Turkey.

出版信息

Biomaterials. 2009 Jul;30(21):3551-9. doi: 10.1016/j.biomaterials.2009.03.024. Epub 2009 Apr 9.

Abstract

The aim of this study was to develop a 3-D construct carrying an inherent sequential growth factor delivery system. Poly(lactic acid-co-glycolic acid) (PLGA) nanocapsules loaded with bone morphogenetic protein BMP-2 and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanocapsules loaded with BMP-7 made the early release of BMP-2 and longer term release of BMP-7 possible. 3-D fiber mesh scaffolds were prepared from chitosan and from chitosan-PEO by wet spinning. Chitosan of 4% concentration in 2% acetic acid (CHI4-HAc2) and chitosan (4%) and PEO (2%) in 5% acetic acid (CHI4-PEO2-HAc5) yielded scaffolds with smooth and rough fiber surfaces, respectively. These scaffolds were seeded with rat bone marrow mesenchymal stem cells (MSCs). When there were no nanoparticles the initial differentiation rate was higher on (CHI4-HAc2) scaffolds but by three weeks both the scaffolds had similar alkaline phosphatase (ALP) levels. The cell numbers were also comparable by the end of the third week. Incorporation of nanoparticles into the scaffolds was achieved by two different methods: incorporation within the scaffold fibers (NP-IN) and on the fibers (NP-ON). It was shown that incorporation on the CHI4-HAc2 fibers (NP-ON) prevented the burst release observed with the free nanoparticles, but this did not influence the total amount released in 25 days. However NP-IN for the same fibers revealed a much slower rate of release; ca. 70% released at the end of incubation period. The effect of single, simultaneous and sequential delivery of BMP-2 and BMP-7 from the CHI4-HAc2 scaffolds was studied in vitro using samples prepared with both incorporation methods. The effect of delivered agents was higher with the NP-ON samples. Delivery of BMP-2 alone suppressed cell proliferation while providing higher ALP activity compared to BMP-7. Simultaneous delivery was not particularly effective on cell numbers and ALP activity. The sequential delivery of BMP-2 and BMP-7, on the other hand, led to the highest ALP activity per cell (while suppressing proliferation) indicating the synergistic effect of using both growth factors holds promise for the production of tissue engineered bone.

摘要

本研究的目的是开发一种带有内在顺序生长因子递送系统的三维构建体。负载骨形态发生蛋白BMP - 2的聚乳酸 - 乙醇酸共聚物(PLGA)纳米胶囊和负载BMP - 7的聚(3 - 羟基丁酸酯 - 共 - 3 - 羟基戊酸酯)(PHBV)纳米胶囊使得BMP - 2的早期释放和BMP - 7的长期释放成为可能。通过湿法纺丝由壳聚糖和壳聚糖 - 聚环氧乙烷制备三维纤维网状支架。在2%乙酸(CHI4 - HAc2)中浓度为4%的壳聚糖以及在5%乙酸(CHI4 - PEO2 - HAc5)中的壳聚糖(4%)和聚环氧乙烷(2%)分别产生具有光滑和粗糙纤维表面的支架。用大鼠骨髓间充质干细胞(MSCs)接种这些支架。当没有纳米颗粒时,在(CHI4 - HAc2)支架上初始分化率较高,但到三周时,两个支架的碱性磷酸酶(ALP)水平相似。到第三周结束时细胞数量也相当。通过两种不同方法将纳米颗粒掺入支架中:掺入支架纤维内(NP - IN)和在纤维上(NP - ON)。结果表明,掺入CHI4 - HAc2纤维上(NP - ON)可防止游离纳米颗粒观察到的突发释放,但这并不影响25天内的总释放量。然而,对于相同的纤维,NP - IN显示出慢得多的释放速率;在孵育期结束时约释放70%。使用两种掺入方法制备的样品在体外研究了从CHI4 - HAc2支架中单次、同时和顺序递送BMP - 2和BMP - 7的效果。NP - ON样品中递送剂的效果更高。单独递送BMP - 2抑制细胞增殖,同时与BMP - 7相比提供更高的ALP活性。同时递送对细胞数量和ALP活性不是特别有效。另一方面,BMP - 2和BMP - 7的顺序递送导致每个细胞的ALP活性最高(同时抑制增殖),表明使用两种生长因子的协同效应有望用于组织工程骨的生产。

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