硫酸钙人工骨负载唑来膦酸的体外药物释放研究

发布时间：2018-03-03 21:09

本文选题：硫酸钙人工骨　切入点：唑来膦酸　出处：《郑州大学》2014年硕士论文　论文类型：学位论文

【摘要】：背景富含巨细胞病变中含有丰富的破骨型多核巨细胞，该种细胞反应活跃、具有低度侵袭性。在良性或恶性骨肿瘤病变中均出现这类细胞，如继发性棕色瘤（Recklinghausen病）、巨细胞修复性肉芽肿(GCRG)、动脉瘤样骨囊肿（ABC）、软骨母细胞瘤、巨细胞骨肉瘤和良、恶性骨巨细胞瘤等。骨巨细胞瘤(GCT)是临床中常见的原发性骨肿瘤之一，瘤体主要由破骨型多核巨细胞和单核基质细胞组成。WHO骨肿瘤分类将GCT描述为“一种侵袭性的潜在恶性病变”。中国的发病率较西方国家高，占骨肿瘤的14％～16％，发病高峰在20～50岁，治疗不当易造成劳动资源的浪费。病变多侵犯长骨，常见的部位为股骨下端和胫骨上端，其他部位见于桡骨远端、腓骨小头、股骨近端、肱骨近端、脊柱、骨盆、胸骨、肋骨、颅骨及跟骨等。骨巨细胞瘤的治疗原则为治疗或控制局部病变，尽量保留患肢功能。几十年前，广泛切除是常用术式，复发率较低。然而，广泛切除后可对临近关节面造成损害，随之而来的重建也较复杂，出现较多的并发症。目前骨巨细胞瘤常用囊内刮除植骨术，但单纯的刮除植骨术难以彻底清除肿瘤组织，微量隐藏在病灶陷窝中的肿瘤细胞即可导致术后复发。多种辅助措施用于处理瘤腔，如液氮、酒精、石碳酸、双氧水等冲洗，磨钻打磨等被广泛研究和应用。辅助治疗使病灶边缘产生近似广泛切除的区域，同时未破坏周围骨性结构，既降低了肿瘤的复发率，又极大地保存患肢功能。但目前各种辅助治疗措施的效果尚未达到完全肯定和统一。二磷酸盐类药物具有一定的抗破骨细胞活性作用，是一种人工合成的焦磷酸盐类似物。在人体中不能被代谢，对骨骼有较强的吸附作用，能够抑制破骨细胞的形成，促进凋亡，，抑制骨吸收，同时可以诱导瘤体中的间质细胞凋亡。二磷酸盐不仅对破骨细胞起到调控作用，同时对于成骨细胞有一定的调节作用。二磷酸盐类药物可诱导GCT中的多核巨细胞和单核基质细胞凋亡，抑制破骨活性。近年来，该类药物被应用于破骨细胞介导的骨肿瘤，如骨巨细胞瘤、溶骨性骨转移瘤、多发性骨髓瘤、骨肉瘤等引发的骨相关事件，同时临床用于预防骨巨细胞瘤复发。目前已经发展了三代药物，其中第三代代表药物为唑来膦酸、阿伦膦酸钠等。静脉滴注或口服二磷酸盐类药物后，可能会出现一过性流感样综合征、下颌骨坏死等诸多副作用，可累及消化、神经、骨骼肌肉等系统。二磷酸盐类药物在人体内不经过代谢，以原形经肾排出，对药物造成了一定的浪费。假如唑来膦酸能够局部缓释，不但能够减少副作用，同时可以提高生物利用率。目的观察不同浓度的唑来膦酸/硫酸钙人工骨复合物体外释放规律。方法制备0.0%、1.3%、2.6%、5.2%、10.4%五个浓度梯度组的唑来膦酸/硫酸钙人工骨复合物标本，记录体外凝结时间。从每梯度组中随机选取重量为0.900±0.012g3个标本，以空白对照组作为A实验组，其余依据浓度梯度依次编为B、C、D、E实验组，使用高效液相色谱法(HPLC)检测药物溶出浓度，研究药物体外的释放规律。结果未使用烘干、加强空气流通等促凝手段，室温下A~E组的凝固时间依次为10.6分钟、20.9分钟、32.8分钟，45.5分钟，58.2分钟。统计学分析凝固时间的差异具有统计学意义（P＜0．05）。体外释放实验B～E组0至24h有明显突释相，累计释放度均值分别达19.11%、23.45%、35.93%、26.88%，各组释放度在同一时间点有统计学差异（P＜0．05），48h以后出现缓释效应，C、D、E组3～21d缓释相溶出率曲线出现波动，D、E组波动明显。结论随着唑来膦酸载药量的增加，硫酸钙结构逐渐不稳定。载药量不超过硫酸钙人工骨最大载药量时，唑来膦酸在硫酸钙人工骨释放体系中能够持续、稳定的释放。
[Abstract]:background
With the broken bone type is rich in giant cell lesions in multinucleated giant cells, the activity is low. This type of invasive cells in benign or malignant bone tumor lesions, such as secondary brown tumor (Recklinghausen disease), giant cell reparative granuloma (GCRG), aneurysmal bone cyst (ABC), chondroblastoma and giant cell osteosarcoma and benign, malignant giant cell tumor of bone. Bone giant cell tumor (GCT) is a common clinical primary bone tumors, tumor type is mainly composed of osteoclast like multinucleated giant cells and mononuclear stromal cells.WHO bone tumor classification described GCT as "malignant potential" an aggressive disease. The incidence rate of China compared with western countries, accounting for 14% ~ 16% of bone tumors, the peak incidence at the age of 20~50, easily lead to improper treatment of labor resources waste. The lesions invaded long bones, common site of distal femur and tibia In the end, other parts of the distal radius, fibula, proximal femur, proximal humerus, spine, pelvis, sternum, ribs, skull and calcaneus. The principle of treatment of giant cell tumor of bone for the treatment or control of local disease, try to keep the limb function. A few decades ago, extensive resection is a common operation, recurrence rate low. However, after extensive resection can cause damage to the adjacent articular surface, the reconstruction is more complex, more complications. The giant cell tumor of bone used intracapsular curettage and bone grafting, but the simple curettage and bone grafting is difficult to completely remove the tumor tissue, tumor cells can trace hidden in the lesions of lacuna the cause of postoperative recurrence. Various auxiliary measures for the treatment of tumor cavity, such as liquid nitrogen, alcohol, phenol, hydrogen peroxide and other washing, drill grinding is widely researched and applied. The adjuvant treatment of wide excision of the lesion edge approximate area, and not Destruction of the surrounding bony structure not only reduces the recurrence rate of the tumor, but also greatly preserves the function of the affected limb.
Two phosphate drugs has certain anti osteoclast activity, is a synthetic analogue of pyrophosphate. Cannot be metabolized in the body, has the stronger adsorption on bone, can inhibit osteoclast formation, promote apoptosis, inhibit bone resorption and induce tumor stroma cell apoptosis. Two phosphate not only affects osteoclast, also have a regulatory role for osteoblasts. Two phosphate drugs can induce multinucleated giant cells and mononuclear stromal cells apoptosis in GCT, inhibition of osteoclast activity. In recent years, the drugs of bone tumor were used in osteoclasts mediated, such as giant cell tumor of bone, osteolytic bone metastasis, multiple myeloma, bone related events triggered by osteosarcoma, and clinical for the prevention of recurrence of giant cell tumor of bone. There are three generations of drugs, the representative of the third generation The drug for zoledronic acid, Allen alendronate. Intravenous or oral two phosphate drugs, there may be a flu like syndrome, and other side effects of osteonecrosis of the jaw, involving the digestive, nervous, skeletal muscle system. Two phosphate drugs in the human body without metabolism. In order to prototype excreted by kidney caused a waste of drug. If zoledronic acid can local release, not only can reduce the side effect, and can improve the bioavailability.
objective
The external release of zoledronic acid / calcium sulphate artificial bone in different concentrations was observed.
Method
The preparation of 0%, 1.3%, 2.6%, 5.2%, 10.4% and five concentration gradient group zoledronic acid / calcium sulfate artificial bone composite specimens were recorded in vitro. The setting time from each group were randomly selected for gradient weight 0.900 + 0.012g3 specimens, the control group was taken as the experimental group A, the other based on concentration gradient in order for the B, C, D, E in the experimental group, using high performance liquid chromatography (HPLC) detection of the concentration of dissolved drug release of drug in vitro.
Result
Without the use of drying, enhance air circulation to promote coagulation, coagulation time of group A~E at room temperature in 10.6 minutes, 20.9 minutes, 32.8 minutes, 45.5 minutes, 58.2 minutes. The difference between the solidification time was statistically significant (P < 0.05). The in vitro release experiment of B ~ E were 0 to 24h significant burst release phase, cumulative release rate value of 19.11%, respectively, 23.45%, 35.93%, 26.88%, each release at the same time there were significant differences (P < 0.05), 48h after C, D, sustained release effect, E group of 3 ~ 21d release phase dissolution rate curve fluctuations, D, E group fluctuated significantly.
conclusion
With the increase of loading dose of zoledronic acid, the structure of calcium sulfate is gradually unstable. When loading dose is not greater than that of calcium sulfate artificial bone, zoledronic acid can release continuously and steadily in calcium sulfate artificial bone release system.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943

【共引文献】