Department of Orthopaedic Surgery, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Department of Joint and Trauma Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Biomed Pharmacother. 2020 May;125:110034. doi: 10.1016/j.biopha.2020.110034. Epub 2020 Feb 25.
Osteosarcoma is the most common bone sarcoma in adolescents. Decorin (DCN) has been proposed to be a new anti-osteosarcoma therapeutic strategy. Our previous study has loaded decorin on titanium (Ti) surface by polydopamine (DOPA) as an anchor to enhance osseointegration. In this study, we investigated the effect of decorin-coated Ti substrates (TI-DOPA-DCN) on the oncogenic potential of osteosarcoma cells SAOS-2. The substrates were placed in 24-well plates for cell culture. Cell viability was determined by Cell Counting Kit-8 (CCK8) assay. Apoptosis was evaluated by DAPI staining and Annexin V-FITC/PI double staining analysis. Cell cycle was analyzed by flow cytometry. Cell migration and invasion were evaluated by Transwell assay. For co-culture, the pre-osteogenic cells MEC3T3-E1 and osteosarcoma cells SAOS-2 were stained with cell membrane fluorescent dyes, and then mixed (1:1) for co-culture. The cells were observed under a fluorescence microscope at four time points of 24, 48, 72, and 96 h. The results showed that TI-DOPA-DCN substrate can selectively inhibit cell proliferation of osteosarcoma cells but not pre-osteoblasts. However, the cell cycle of SAOS-2 was not affected by TI-DOPA-DCN substrates. Both DAPI staining and Annexin V-FITC/PI double staining analysis revealed that TI-DOPA-DCN substrates induced apoptosis of osteosarcoma cells. Transwell assay showed that TI-DOPA-DCN substrates inhibited invasion and migration of osteosarcoma cells. Moreover, TI-DOPA-DCN substrates inhibited the growth of osteosarcoma cells but promoted that of pre-osteoblasts in the coculture system. Taken together, these findings suggested that decorin coating on Ti surface simultaneously inhibited the oncogenic potential of osteosarcoma cells but enhanced cell growth of pre-osteoblasts, which could be applied to surface modification of Ti orthopedic implant.
成骨肉瘤是青少年中最常见的骨肉瘤。现已提出核心蛋白聚糖（DCN）是一种新的抗成骨肉瘤治疗策略。我们之前的研究已经通过聚多巴胺（DOPA）将核心蛋白聚糖负载到钛（Ti）表面作为锚定物，以增强骨整合。在这项研究中，我们研究了核心蛋白聚糖涂层钛基底（TI-DOPA-DCN）对成骨肉瘤细胞 SAOS-2 的致癌潜能的影响。将基底放置在 24 孔板中进行细胞培养。通过 Cell Counting Kit-8（CCK8）测定细胞活力。通过 DAPI 染色和 Annexin V-FITC/PI 双重染色分析评估细胞凋亡。通过流式细胞术分析细胞周期。通过 Transwell 测定评估细胞迁移和侵袭。对于共培养，将预成骨细胞 MEC3T3-E1 和成骨肉瘤细胞 SAOS-2 用细胞膜荧光染料染色，然后混合（1:1）进行共培养。在 24、48、72 和 96 h 四个时间点，在荧光显微镜下观察细胞。结果表明，TI-DOPA-DCN 基底可选择性抑制成骨肉瘤细胞但不抑制前成骨细胞的增殖。然而，SAOS-2 的细胞周期不受 TI-DOPA-DCN 基底的影响。DAPI 染色和 Annexin V-FITC/PI 双重染色分析均表明 TI-DOPA-DCN 基底诱导成骨肉瘤细胞凋亡。Transwell 测定表明 TI-DOPA-DCN 基底抑制成骨肉瘤细胞的侵袭和迁移。此外，TI-DOPA-DCN 基底抑制共培养体系中成骨肉瘤细胞的生长，但促进前成骨细胞的生长。综上所述，这些发现表明，Ti 表面的核心蛋白聚糖涂层同时抑制成骨肉瘤细胞的致癌潜能，同时增强前成骨细胞的细胞生长，可应用于 Ti 骨科植入物的表面改性。
