A bifronted cure system for osteosarcoma, a common aggressive bone tumor, is highly in demand to prevail the postsurgical adversities in connection with systemic chemotherapy and repair of critical-size bone defects. The hierarchically porous therapeutic scaffolds presented here are synthesized by free radical-initiated copolymerization of hydroxyethyl methacrylate and methyl methacrylate [HEMA/MMA 80:20 and 90:10 mM, H₂O/NaCl porogen], which are further surface-phosphorylated [P-PHM] and transformed to bifunctional by impregnating doxorubicin (DOX) [DOXP-PHM]. The P-PHM scaffolds exhibited porous microarchitecture analogous to native cancellous bone (scanning electron microscopy analysis), while X-ray photoelectron spectroscopy analysis authenticated surface phosphorylation. Based on pore characteristics, swelling attributes and slow-pace degradation, P-PHM9163 and P-PHM8263 (HEMA/MMA 90:10 and 80:20 with H₂O/NaCl: 60/3.0 weight %, respectively) were chosen from the series and evaluated for osteoinductive efficacy in vitro. Both P-PHM9163 and P-PHM8263 invoked calcium phosphate mineralization in simulated physiological conditions (day 14) with Ca/P ratios of 1.58 and 1.66 respectively, comparable to human bone (1.67). Early biomineralization (Alizarin Red S and von Kossa staining) was evidenced at day 7, while osteoblast differentiation was verified by time-dependent expression of the typical late marker, osteocalcin, at day 14 and 21 in rat bone marrow mesenchymal cells. DOX-loaded P-PHM9163 (DOXP-PHM9163) exhibited pH-responsive (tumor analogous pH; 6.5) sustained release of DOX for prolonged time (up to 45 days) and invoked cellular alterations by cortical stress fiber formation and DNA fragmentation in human osteosarcoma cells leading to early apoptosis (24 h), validated by annexin V/PI staining (FACS) and immunostaining (F-actin/DAPI). Subsequent to DOX release tenure, the scaffold induced the formation of well-organized, porous post-release Ca-P apatite coating (Ca/P is 1.3) in simulated body fluid (day 14) which further endorses the dual functionality of the system. Altogether, the results accentuate that DOXP-PHM9163 is a potential bifunctional therapeutic scaffold capable of extended localized chemotherapeutic delivery in-line with inherent osteogenesis for efficient bone cancer treatment.

中文翻译：

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多孔多孔成骨诱导聚（甲基丙烯酸羟乙酯-甲基丙烯酸甲酯）共固位骨肉瘤治疗和骨缺损修复的支架。

对于骨肉瘤（一种常见的侵袭性骨肿瘤）的双向治疗系统，迫切需要在全身化疗和关键尺寸骨缺损修复中战胜术后逆境。本文介绍的分级多孔治疗支架是通过自由基引发的甲基丙烯酸羟乙酯和甲基丙烯酸甲酯的共聚反应合成的[HEMA / MMA 80:20和90:10 mM，H ₂O / NaCl致孔剂]，将其进一步表面磷酸化[P-PHM]，并通过注入阿霉素（DOX）[DOXP-PHM]使其转变为双功能。P-PHM支架表现出类似于天然松质骨的多孔微结构（扫描电子显微镜分析），而X射线光电子能谱分析证实了表面磷酸化。根据孔隙特征，溶胀属性和慢速降解，从P-PHM9163和P-PHM8263（HEMA / MMA 90:10和80：20，H ₂ O / NaCl分别为60 / 3.0重量％）中选择。系列并评估体外骨诱导功效。P-PHM9163和P-PHM8263在模拟的生理条件下（第14天）都激活了磷酸钙矿化作用，钙/磷比分别为1.58和1.66，可与人体骨骼（1.67）相比。在第7天证实了早期生物矿化（茜素红S和von Kossa染色），而在大鼠骨髓间充质细胞中第14天和第21天典型的晚期标记物骨钙素的时间依赖性表达证实了成骨细胞的分化。载有DOX的P-PHM9163（DOXP-PHM9163）表现出pH响应（肿瘤类似的pH; 6.5）长时间（最多45天）持续释放DOX，并通过人骨肉瘤中的皮质应激纤维形成和DNA断裂引起细胞改变通过膜联蛋白V / PI染色（FACS）和免疫染色（F-actin / DAPI）验证了导致早期凋亡（24 h）的细胞。在DOX释放后，支架在模拟体液（第14天）中诱导了组织良好的多孔Ca-P磷灰石后释放磷灰石涂层的形成（Ca / P为1.3），这进一步认可了系统的双重功能。总而言之，结果突显了DOXP-PHM9163是一种潜在的双功能治疗支架，能够与固有的成骨作用一致地扩展局部化学治疗的传递，从而有效地治疗骨癌。