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孔径大小调节细胞和组织与用于骨工程的 PLGA-CaP 支架的相互作用。

Pore size regulates cell and tissue interactions with PLGA-CaP scaffolds used for bone engineering.

机构信息

Cell Culture Laboratory, School of Dentistry of Ribeirao Preto, University of Sao Paulo, Brazil.

出版信息

J Tissue Eng Regen Med. 2012 Feb;6(2):155-62. doi: 10.1002/term.422. Epub 2011 Mar 28.

Abstract

A common subject in bone tissue engineering is the need for porous scaffolds to support cell and tissue interactions aiming at repairing bone tissue. As poly(lactide-co-glycolide)-calcium phosphate (PLGA-CaP) scaffolds can be manufactured with different pore sizes, the aim of this study was to evaluate the effect of pore diameter on osteoblastic cell responses and bone tissue formation. Scaffolds were prepared with 85% porosity, with pore diameters in the ranges 470-590, 590-850 and 850-1200 µm. Rat bone marrow stem cells differentiated into osteoblasts were cultured on the scaffolds for up to 10 days to evaluate cell growth, alkaline phosphatase (ALP) activity and the gene expression of the osteoblast markers RUNX2, OSX, COL, MSX2, ALP, OC and BSP by real-time PCR. Scaffolds were implanted in critical size rat calvarial defects for 2, 4, and 8 weeks for histomorphometric analysis. Cell growth and ALP activity were not affected by the pore size; however, there was an increase in the gene expression of osteoblastic markers with the increase in the pore sizes. At 2 weeks all scaffolds displayed a similar amount of bone and blood vessels formation. At 4 and 8 weeks much more bone formation and an increased number of blood vessels were observed in scaffolds with pores of 470-590 µm. These results show that PLGA-CaP is a promising biomaterial for bone engineering. However, ideally, combinations of larger (-1000 µm) and smaller (-500 µm) pores in a single scaffold would optimize cellular and tissue responses during bone healing.

摘要

在骨组织工程中,一个常见的课题是需要多孔支架来支持细胞和组织相互作用,以修复骨组织。由于聚(乳酸-共-乙醇酸)-磷酸钙(PLGA-CaP)支架可以制造出不同孔径的多孔支架,因此本研究的目的是评估孔径对成骨细胞反应和骨组织形成的影响。支架的孔隙率为 85%,孔径分别为 470-590 µm、590-850 µm 和 850-1200 µm。将大鼠骨髓干细胞分化为成骨细胞,在支架上培养长达 10 天,以评估细胞生长、碱性磷酸酶(ALP)活性以及成骨细胞标志物 runt 相关转录因子 2(RUNX2)、成骨特异性转录因子(OSX)、Ⅰ型胶原(COL)、肌肉特异性同源盒 2(MSX2)、ALP、骨钙素(OC)和骨桥蛋白(BSP)的基因表达情况,通过实时 PCR 进行检测。将支架植入大鼠颅骨临界大小缺损中 2、4 和 8 周,进行组织形态计量学分析。细胞生长和 ALP 活性不受孔径的影响;然而,成骨细胞标志物的基因表达随着孔径的增加而增加。在 2 周时,所有支架都显示出相似数量的骨和血管形成。在 4 周和 8 周时,在孔径为 470-590 µm 的支架中观察到更多的骨形成和更多的血管形成。这些结果表明 PLGA-CaP 是一种有前途的骨工程生物材料。然而,理想情况下,在单个支架中组合较大(-1000 µm)和较小(-500 µm)的孔将优化骨愈合过程中的细胞和组织反应。

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