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聚乙二醇脂质体阿霉素作为犬转移性骨肉瘤的潜在治疗药物及研究。

PEG-liposomal doxorubicin as a potential agent for canine metastatic osteosarcoma - and studies.

作者信息

Walewska Magdalena, Małek Anna, Taciak Bartosz, Wojtalewicz Anna, Wilk Sylwia, Wojtkowska Agata, Zabielska-Koczywąs Katarzyna, Lechowski Roman

机构信息

Department of Small Animal Diseases and Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, 02-787 Warsaw, Poland.

Department of Cancer Biology, Institute of Biology, Warsaw University of Life Sciences, 02-787 Warsaw, Poland.

出版信息

J Vet Res. 2023 Jun 16;67(2):297-305. doi: 10.2478/jvetres-2023-0026. eCollection 2023 Jun.

Abstract

INTRODUCTION

Appendicular osteosarcoma (OSA) is a highly aggressive and metastatic primary bone tumour in dogs. Standard therapy is amputation and adjuvant chemotherapy (. with doxorubicin). Liposomal drug delivery may augment therapeutic efficacy and reduce negative side effects. Polyethylene glycol (PEG)-liposomal doxorubicin treats human metastatic cancers effectively. The study aimed was to evaluate PEG-liposomal doxorubicin's inhibitory effect on canine metastatic proliferation and migration . It also aimed to appraise the drug's extravasation inhibition using the human medicine-proven chick embryo chorioallantoic membrane model.

MATERIAL AND METHODS

The canine D-17 OSA cell line was cultured and inoculated with decreasing concentrations of PEG-liposomal doxorubicin and conventional doxorubicin in a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) test of cell viability, proliferation and cytotoxicity. Flow cytometry with Annexin V and Draq 7 staining confirmed the MTT test results, indicating dead, early and late apoptotic, and live cells. The inhibitory effect of the two preparations on cancer cell migration was investigated with a wound-healing assay. Culture plates seeded with cells were prepared. The cell monolayer was scratched and images of cells migrating to the scratch were captured at 0 h, 12 h and 24 h. Also, embryos were removed from three-day-incubated fertilised chicken eggs. On the 12 day, labelled D-17 cells were injected into each embryo. Embryos in one group received 100 μL of phosphate-buffered saline as controls, those in another group 30 μg/mL of PEG-liposomal doxorubicin, and those in the last group 6 μg/mL of conventional doxorubicin. The effectiveness of the intravascular administration of the D-17 cells was confirmed under a microscope.

RESULTS

PEG-liposomal doxorubicin inhibited the migration of canine OSA cells more effectively than conventional doxorubicin (P ≤ 0.05). The model showed that both drugs had similar impacts on canine metastatic OSA.

CONCLUSION

The liposomal form of the drug may be considered a potentially effective compound in canine metastatic OSA; nevertheless, further studies are essential to confirm this hypothesis.

摘要

引言

附肢骨肉瘤(OSA)是犬类一种具有高度侵袭性和转移性的原发性骨肿瘤。标准治疗方法是截肢和辅助化疗(使用阿霉素)。脂质体药物递送可能会增强治疗效果并减少负面副作用。聚乙二醇(PEG)-脂质体阿霉素能有效治疗人类转移性癌症。本研究旨在评估PEG-脂质体阿霉素对犬类转移性增殖和迁移的抑制作用。同时,还旨在使用已在人类医学中得到验证的鸡胚绒毛尿囊膜模型来评估该药物对血管外渗的抑制作用。

材料与方法

培养犬类D-17骨肉瘤细胞系,并在3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)细胞活力、增殖和细胞毒性试验中接种浓度逐渐降低的PEG-脂质体阿霉素和传统阿霉素。采用膜联蛋白V和Draq 7染色的流式细胞术证实了MTT试验结果,表明存在死亡、早期和晚期凋亡以及活细胞。通过伤口愈合试验研究了这两种制剂对癌细胞迁移的抑制作用。制备接种有细胞的培养板。刮伤细胞单层,并在0小时、12小时和24小时拍摄细胞向刮痕迁移的图像。此外,从孵化三天的受精鸡蛋中取出胚胎。在第12天,将标记的D-17细胞注射到每个胚胎中。一组胚胎接受100μL磷酸盐缓冲盐水作为对照,另一组接受30μg/mL的PEG-脂质体阿霉素,最后一组接受6μg/mL的传统阿霉素。在显微镜下确认D-17细胞血管内给药的有效性。

结果

PEG-脂质体阿霉素比传统阿霉素更有效地抑制犬类骨肉瘤细胞的迁移(P≤0.05)。该模型表明,两种药物对犬类转移性骨肉瘤的影响相似。

结论

该药物的脂质体形式可能被认为是犬类转移性骨肉瘤中一种潜在有效的化合物;然而,需要进一步的研究来证实这一假设。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b5df/10541662/5821127dcd9c/j_jvetres-2023-0026_fig_001.jpg

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