Long-term sustained-released in situ gels of a water-insoluble drug amphotericin B for mycotic arthritis intra-articular administration: preparation,in vitro and in vivo evaluation

Amphotericin B (AMB) was often used in intra-articular injection administration for fungal arthritis, because it could often bring a satisfactory therapeutic efficacy and a minimum systemic toxic side effect. However, because of the multiple operations and the frequent injections, the compliance of the patients was bad. Therefore, to develop a long-term sustained-released preparation of AMB for mycotic arthritis intra-articular administration is of great significance. The purpose of present study was to develop a long-term sustained-released in situ gel of a water-insoluble drug AMB for mycotic arthritis intra-articular administration. Based on the evaluations of the in vitro properties of the formulations, the formulation containing 10% (w/w) ethanol, 15% (w/w) PG, 0.75% (w/w) HA, 5% (w/w) purified soybean oil, 0.03% (w/w) α-tocopherol, 15% (w/w) water and 55% (w/w) glyceryl monooleate was selected as a suitable intra-articular injectable in situ gel drug delivery system for water-insoluble drug AMB. Furthermore, the results of the in vivo study on rabbits showed that the selected formulation was a safe and effective long-term sustained-released intra-articular injectable AMB preparation. Therefore, the presented in situ AMB gel could reduce the frequency of the administration in the AMB treatment of fungal arthritis, and then would get a good patient compliance.     

Versatile Hypoxic Extracellular Vesicles Laden in an Injectable and Bioactive Hydrogel for Accelerated Bone Regeneration

Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have emerged as an appealing alternative to cell therapy in regenerative medicine. Unlike bone marrow MSCs (BMSCs) cultured in vitro with normoxia, bone marrow in vivo is exposed to a hypoxic environment. To date, it remains unclear whether hypoxia preconditioning can improve the function of BMSC-derived EVs and be more conducive to bone repair. Herein, it is found that hypoxia preconditioned BMSCs secrete more biglycan (Bgn)-rich EVs via proteomics analysis, and these hypoxic EVs (Hypo-EVs) significantly promote osteoblast proliferation, migration, differentiation, and mineralization by activating the phosphatidylinositide 3-kinase/protein kinase B pathway. Subsequently, an injectable bioactive hydrogel composed of poly(ethylene glycol)/polypeptide copolymers is developed to improve the stability and retention of Hypo-EVs in vivo. The Hypo-EVs-laden hydrogel shows continuous liberation of Hypo-EVs for 3 weeks and substantially accelerates bone regeneration in 5-mm rat cranial defects. Finally, it is confirmed that Bgn in EVs is a pivotal protein regulating osteoblast differentiation and mineralization and exerts its effects through paracrine mechanisms. Therefore, this study shows that hypoxia stimulation is an effective approach to optimize the therapeutic effects of BMSC-derived EVs and that injectable hydrogel-based EVs delivery is a promising strategy for tissue regeneration.     

注射用药用辅料甘露醇细菌内毒素检查方法研究

为了建立注射用药用辅料甘露醇的细菌内毒素检查方法,根据《中华人民共和国药典二部（2010年版）》中的“细菌内毒素检查法及指导原则”进行试验。将甘露醇溶解稀释至100mg·mL-1,用标示灵敏度A为0．125EU·mL-1的鲎试剂检测其细菌内毒素。结果表明,100mg·mL-1的甘露醇及稀释液对细菌内毒素检查无干扰。说明鲎试剂方法用于甘露醇的细菌内毒素检查是可行、有效的。     

An Injectable Hydrogel for Simultaneous Photothermal Therapy and Photodynamic Therapy with Ultrahigh Efficiency Based on Carbon Dots and Modified Cellulose Nanocrystals

The convenience of injectable hydrogels that can provide high loading of diverse phototherapy agents and further long-time retention at the tumor site has attracted tremendous interest in simultaneous photothermal and photodynamic cancer therapies. However, to incorporate the phototherapy agents into hydrogels, complex modifications are generally unavoidable. Moreover, these phototherapy agents usually suffer from low efficiency and work at different irradiation wavelengths outside the near infrared windows. Hence, a method for the fabrication of an injectable hydrogel for simultaneous photothermal therapy and photodynamic therapy, through the Schiff-base reaction between amido modified carbon dots (NCDs) and aldehyde modified cellulose nanocrystals is proposed. The NCDs act as both phototherapy agents and crosslinkers to form hydrogels. Significantly, the NCDs demonstrate an extremely high photothermal conversion efficiency of 77.6% which is among the highest levels for photothermal agents and a high singlet quantum yield of 0.37 under a single 660 nm light-emitting diode irradiation. The hydrogels are examined through in vitro and in vivo animal experiments which show nontoxic and effectively tumor inhibition. Thus, the strategy of direct reaction of phototherapy agents and the matrix not only provides new strategies for injectable hydrogel fabrication but paves a new road for advanced tumor treatment.     

基于壳聚糖/银离子的可逆温敏水凝胶的制备与性能

以壳聚糖(CS)和Ag NO₃为主要原料,通过CS与Ag⁺的超分子络合作用制备了一系列可逆温敏CS-Ag水凝胶,采用FTIR、XRD、XPS、SEM和EDS对CS-Ag水凝胶的结构和化学状态进行了表征。结果表明,通过同时调节Ag NO₃用量和体系p H可制备具有相同凝胶温度的CS-Ag水凝胶。当CS用量为0.60 g、Ag NO₃用量为0.19 g、体系总体积为21 m L、保持水凝胶p H=4.24时,制备的CS-Ag-19水凝胶凝胶温度为50℃,具有最佳的热可逆性能,至少可循环相变5次,具有室温避光保存5个月的稳定性,并表现出对大肠杆菌良好的抗菌性能。CS-Ag-19的溶胶-凝胶转换是由CS的氨基在不同温度下的超分子相互作用控制,H⁺和Ag⁺在温度、浓度变化的驱动下,对游离氨基的竞争反应(质子化和络合作用)之间的平衡关系使水凝胶具有温敏可逆性。CS-Ag-19水凝胶的可注射和温敏凝胶性能使其具有作为水凝胶抗菌敷料的应用潜力。     

Capillary alginate gel (Capgel™) for the treatment of full-thickness dermal wounds in a hypoxic mouse model

Abstract

In this study, capillary alginate gel (Capgel^?), a collagen and alginate based self-assembling biomaterial, was used as a cellular scaffold for the treatment of ischemic, full-thickness dermal wounds in mice. Capgel^? was synthesized using copper sulfate to form the initial sol before rinsing and stabilizing the patent capillary structures with carbodiimide chemistry. This crosslinked hydrogel was then injected into ischemic, full-thickness dermal wounds and analyzed via histology after 7 and 10 days to assess wound contracture, granulation bed tissue and vascular structures. Capgel^? showed good resorbability and was well-invested with infiltrating host cells and vascular structures during and after resorption.