Stem cells from human fat as cellular delivery vehicles in an athymic rat posterolateral spine fusion model
Stem cells from human fat as cellular delivery vehicles in an athymic rat posterolateral spine fusion model
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DOI：
10.2106/jbjs.g.00292
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发表时间：
2008-05-01
影响因子：
5.3
通讯作者：
Lieberman, Jay R.
中科院分区：
文献类型：
--
作者：
Hsu, Wellington K.;Wang, Jeffrey C.;Lieberman, Jay R.
背景：源自人类吸脂抽吸物的间充质干细胞，称为加工的吸脂细胞，已被用作组织工程和基因治疗策略中诱导骨形成的细胞递送载体。在这项研究中，我们试图评估产生骨形态发生蛋白（BMP）-2的脂肪源性干细胞在无胸腺大鼠模型中诱导后外侧脊柱融合的功效。方法：在五组无胸腺大鼠（每组八只）中尝试使用分型胶原基质进行单水平（L4-L5）横突间脊柱关节融合术。 I组用5 X 106个脂肪干细胞进行处理，这些干细胞用含有BMP-2基因的腺病毒载体转导； II组，用成骨培养基和1μg/mL重组BMP-2（rhBMP-2）处理的5×106脂肪干细胞； III组，10μg rhBMP-2； IV组，1μg rhBMP-2； V 组，仅使用 5 x 106 个脂肪干细胞。显示愈合迹象的动物在细胞植入后四周被处死，并通过平片、手动触诊、微型计算机断层扫描和组织学分析进行检查。结果：术后四周，第一组中的所有八只动物均表现出成功的脊柱融合，并具有较大的融合肿块。此外，I 组标本一致显示头侧水平（L3 和 L4）存在脊柱融合，其中未通过手术准备融合床。相比之下，尽管在体外测量到大量的 BMP-2 产生，但第二组动物即使在植入后八周也表现出最小的骨形成。在单独应用rhBMP-2治疗的组中，接受相对高剂量的组（第III组）的融合率（在所有八个标本中都可见）比接受低剂量的组（第IV组，其中在八个标本中的四个中看到融合）更高。手术后八周，V 组动物（仅用脂肪干细胞治疗）均未表现出成功的脊柱融合。结论：脂肪干细胞有望作为诱导骨形成的基因转导靶标，从而在生物学严格的环境中增强脊柱融合。临床相关性：脂肪干细胞显示出作为区域基因治疗策略中传递重组蛋白的细胞载体的潜力，以增强脊柱融合。然而，在基于细胞的策略中专门使用脂肪干细胞成为常规之前，有必要对生物潜力、不同载体之间的相互作用以及宿主生物学的影响进行更广泛的研究。
Background: Mesenchymal stem cells derived from human liposuction aspirates, termed processed lipoaspirate cells, have been utilized as cellular delivery vehicles for the induction of bone formation in tissue engineering and gene therapy strategies. In this study, we sought to evaluate the efficacy of bone morphogenetic protein (BMP)-2-producing adipose-derived stem cells in inducing a posterolateral spine fusion in an athymic rat model.Methods: Single-level (L4-L5) intertransverse spinal arthrodesis was attempted with use of a typed collagen matrix in five groups of athymic rats, with eight animals in each group. Group I was treated with 5 X 106 adipose-derived stem cells transduced with an adenoviral vector containing the BMP-2 gene; group II, with 5 x 106 adipose-derived stem cells treated with osteogenic media and 1 mu g/mL of recombinant BMP-2 (rhBMP-2); group III, with 10 mu g of rhBMP-2; group IV, with 1 mu g of rhBMP-2; and group V, with 5 x 106 adipose-derived stem cells alone. The animals that showed radiographic evidence of healing were killed four weeks after cell implantation and were examined with plain radiographs, manual palpation, microcomputed tomography scanning, and histological analysis.Results: All eight animals in group I demonstrated successful spinal fusion, with a large fusion mass, four weeks postoperatively. Furthermore, group-I specimens consistently revealed spinal fusion at the cephalad level (L3 and L4), where no fusion bed had been prepared surgically. In contrast, despite substantial BMP-2 production measured in vitro, group-II animals demonstrated minimal bone formation even eight weeks after implantation. Of the groups treated with the application of rhBMP-2 alone, the one that received a relatively high dose (group III) had a higher rate of fusion (seen in all eight specimens) than the one that received the low dose (group IV, in which fusion was seen in four of the eight specimens). None of the group-V animals (treated with adipose-derived stem cells alone) demonstrated successful spine fusion eight weeks after the surgery.Conclusions: Adipose-derived stem cells show promise as gene transduction targets for inducing bone formation to enhance spinal fusion in biologically stringent environments.Clinical Relevance: Adipose-derived stem cells demonstrate potential as cellular vehicles for the delivery of recombinant protein in a regional gene therapy strategy to enhance spinal fusion. However, before the exclusive use of adipose-derived stem cells in a cell-based strategy can become routine, more extensive study of the biological potential, the interactions between different carriers, and the influence of host biology is necessary.