Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type
Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type
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DOI：
10.1038/sj.gt.3302006
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发表时间：
2003-08-01
期刊：
影响因子：
5.1
通讯作者：
Davis, AR
中科院分区：
文献类型：
--
作者：
Gugala, Z;Olmsted-Davis, EA;Davis, AR
本研究的目的是分析和比较具有内在不同成骨潜能的各种人类细胞类型在用编码骨形态发生蛋白2型（BMP 2）的两种不同腺病毒载体离体转导后诱导异位骨形成的能力。所述细胞包括原代人骨髓间充质干细胞（BM-MSC）、原代人皮肤成纤维细胞（SF）和人二倍体胎肺细胞系（MRC-5）。所述载体包括5型腺病毒或具有35型腺病毒的纤维基因的嵌合5型腺病毒（Ad 5 F35-BMP 2），两者在体外均显示出显著不同的BMP 2表达。实验组由用两种腺病毒载体中的每一种转导的三种人细胞类型组成。采用非肥胖型糖尿病重症联合免疫缺陷（NOD/SCID）小鼠，在体外腺病毒转导后肌内注射转导细胞，通过X线和组织学观察异位骨形成的性质和程度。在注射后14天，在注射Ad 5 F35-BMP 2转导的细胞的小鼠中形成了丰富的、高度矿化的骨，与细胞类型无关。如从双平面平片X线片上的骨表面积评估的，在用Ad 5 F35-BMP 2转导的BM-MSC、SF和MRC-5细胞之间形成的骨的量没有统计学显著差异。在注射用Ad 5-BMP 2转导的细胞的小鼠中可以检测到显著更少量的骨或没有骨。免疫组织化学分析证实，早在分娩后2天，肌肉中就存在人细胞;然而，在注射后6-7天，在周围肌肉或异位骨中无法检测到转导的细胞，表明新形成的骨的宿主来源。该研究的结果表明，用编码BMP 2的相同类型的腺病毒离体转导的BM-MSC、SF和MRC-5细胞之间的骨诱导性质没有显著差异。BMP 2的表达水平似乎是决定异位骨形成程度的关键因素，并且受到所使用的腺病毒类型的显着影响。在所研究的细胞类型中，Ad 5 F35-BMP 2在提供足够水平的BMP 2用于有效的骨诱导方面比Ad 5 BMP 2更有效。
The objective of the study was to analyze and compare the abilities of various human cell types with inherently dissimilar osteogenic potentials to induce heterotopic bone formation following ex vivo transduction with two distinct adenoviral vectors encoding bone morphogenetic protein type 2 (BMP2). The cells comprised primary human bone marrow mesenchymal stem cells (BM-MSCs), primary human skin fibroblasts (SFs), and a human diploid fetal lung cell line (MRC-5). The vectors included adenovirus type 5 or a chimeric adenovirus type 5 with the fiber gene of adenovirus type 35 (Ad5F35-BMP2), both demonstrating significantly different expression of BMP2 in vitro. The experimental groups consisted of the three human cell types transduced with each of the two adenoviral vectors. Using nonobese diabetic severe combined immunodeficiency (NOD/SCID) mice, the transduced cells were injected intramuscularly following ex vivo adenoviral transduction, The nature and extent of heterotopic bone formation were analyzed radiographically and histologically. At 14 days postinjection, abundant, highly mineralized bone was formed in mice injected with Ad5F35-BMP2-transduced cells irrespective of the cell type, There was no statistically significant difference in the amount of bone formed between BM-MSCs, SFs, and MRC-5 cells transduced with Ad5F35-BMP2, as assessed from bone surface area on biplanar plain radiography. Substantially lesser amounts or no bone could be detected in mice injected with cells transduced with Ad5-BMP2. Immunohistochemical analysis confirmed the presence of human cells in muscle as early as 2 days postdelivery; however, at 6-7 days after injection, the transduced cells could not be detected in surrounding muscle, or in the heterotopic bone, indicating the host origin of the newly formed bone. The results of the study demonstrate no significant difference in osteoinductive properties between BM-MSCs, SFs, and MRC-5 cells transduced ex vivo with the same type of adenovirus encoding BMP2. The level of BMP2 expression appears to be a crucial factor determining the extent of heterotopic bone formation and was significantly affected by the type of adenovirus used. In the cell types studied, Ad5F35-BMP2 was more efficacious than Ad5BMP2 in providing adequate levels of BMP2 for efficient osteoinduction.