Fever after zoledronic acid administration is due to increase in TNF-alpha and IL-6.

The most common adverse event typically associated with bisphosphonate therapy is transient fever. The aim of this study was to define the role of the main cytokines of the acute-phase reaction interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) involved in the pathogenesis of zoledronic acid-induced fever. Eighteen consecutive cancer patients with bone metastases were treated, for the first time, with a single dose of 4 mg zoledronic acid infusion. They were prospectively evaluated for circulating TNF-alpha, interferon-gamma (IFN-gamma), and IL-6 levels at different times, just before and 1, 2, 7, and 21 days after diphosphonate infusion. Clinical and standard laboratory parameters were recorded at the same time points. TNF-alpha circulating levels increased significantly 1 and 2 days after zoledronic acid infusion (respectively, p = 0.002 and p < 0.001) and then decreased to levels similar to the basal levels. IL-6 levels increased significantly 1 day after the infusion (p = 0.007), returning to values similar to the median basal values 2 days after zoledronic acid administration. Moreover, in patients who experienced fever, the TNF-alpha and IL-6 increases were higher than in patients without fever. No statistically significant differences in IFN-gamma were identified at different time points in patients with and without fever. Our results show that zoledronic acid induces transient TNF-alpha and IL-6 increases and that these increases are higher in patients who have developed fever, suggesting that these cytokines could be responsible for fever pathogenesis. The sharp reduction in serum calcium levels observed in patients with fever may be related to zoledronic acid pharmacokinetic modifications.     

A prospective, multicenter, open-label trial of zoledronic acid in patients with hormone refractory prostate cancer

PURPOSE: The short-term safety and efficacy of zoledronic acid for the treatment of skeletal metastasis was evaluated in patients with hormone-refractory prostate cancer. PATIENTS AND METHODS: A total of 19 hormone-refractory prostate cancer patients with bone metastases were enrolled. All patients received up to six infusions of zoledronic acid (4 mg, given intravenously over 15 minutes, every 3-4 weeks). Safety was assessed by monitoring adverse events and serum creatinine levels. Efficacy was assessed by monitoring skeletal-related events, brief pain inventory score, quality of life score, type of pain medication, and analgesic score. Mean age of patients was 67.3 years (46-86 years), mean time from diagnosis of bone metastases was 27.6 months (0-117 months), and mean time from diagnosis of hormone-refractory disease was 7.5 months (0-26 months). RESULTS: There was no clinically significant change in serum creatinine levels. Eleven adverse events (musculoskeletal disorders and systemic disorders) in 8 patients were classed as having a possible relationship to study drug. Fifteen patients completed six courses of zoledronic acid infusion. There were no significant changes in the brief pain inventory composite scores, quality of life questionnaire scores or analgesic score. No new skeletal-related events developed during the treatment period. CONCLUSION: Zoledronic acid administered in this study as a 15-minute infusion demonstrated an acceptable and well-known safety profile in patients with refractory prostate cancer with bone metastases. However, prospective placebo- controlled clinical trials are required to elucidate the efficacy of zoledronic acid.     

Adipose-Derived Mesenchymal Stem Cell Protects Kidneys against Ischemia-Reperfusion Injury through Suppressing Oxidative Stress and Inflammatory Reaction

Tumour cells communicate with the cells of their microenvironment via a series of molecular and cellular interactions to aid their progression to a malignant state and ultimately their metastatic spread. Of the cells in the microenvironment with a key role in cancer development, tumour associated macrophages (TAMs) are among the most notable. Tumour cells release a range of chemokines, cytokines and growth factors to attract macrophages, and these in turn release numerous factors (e.g. VEGF, MMP-9 and EGF) that are implicated in invasion-promoting processes such as tumour cell growth, flicking of the angiogenic switch and immunosuppression. TAM density has been shown to correlate with poor prognosis in breast cancer, suggesting that these cells may represent a potential therapeutic target. However, there are currently no agents that specifically target TAM's available for clinical use. Bisphosphonates (BPs), such as zoledronic acid, are anti-resorptive agents approved for treatment of skeletal complication associated with metastatic breast cancer and prostate cancer. These agents act on osteoclasts, key cells in the bone microenvironment, to inhibit bone resorption. Over the past 30 years this has led to a great reduction in skeletal-related events (SRE's) in patients with advanced cancer and improved the morbidity associated with cancer-induced bone disease. However, there is now a growing body of evidence, both from in vitro and in vivo models, showing that zoledronic acid can also target tumour cells to increase apoptotic cell death and decrease proliferation, migration and invasion, and that this effect is significantly enhanced in combination with chemotherapy agents. Whether macrophages in the peripheral tumour microenvironment are exposed to sufficient levels of bisphosphonate to be affected is currently unknown. Macrophages belong to the same cell lineage as osteoclasts, the major target of BPs, and are highly phagocytic cells shown to be sensitive to bisphosphonates in model studies; In vitro, zoledronic acid causes increased apoptotic cell death; in vivo the drug has been shown to inhibit the production of pro-angiogenic factor MMP-9, as well as most recent evidence showing it can trigger the reversal of the TAMs phenotype from pro-tumoral M2 to tumoricidal M1. There is thus accumulating evidence supporting the hypothesis that effects on TAMs may contribute to the anti-tumour effect of bisphosphonates. This review will focus in detail on the role of tumour associated macrophages in breast cancer progression, the actions of bisphosphonates on macrophages in vitro and in tumour models in vivo and summarise the evidence supporting the potential for the targeting of tumour macrophages with bisphosphonates.     

辛伐他汀对合并高胆固醇血症非手术治疗老年髋臼骨折愈合的影响

目的 调查口服辛伐他汀是否可以改善骨密度、促进髋臼骨折愈合.方法 收集2017年1月～2019年12月冀中能源峰峰集团有限公司总医院诊断为髋臼骨折共98例患者的信息:年龄、性别、肝功能、肾功能、血糖、血脂、凝血功能、心电图、骨密度.随机将患者分别对照组和实验组:对照组给予唑来膦酸钠注射液静脉滴注5mg;实验组给予唑来膦...     

Preventive effects of zoledronic acid on bone metastasis in mice injected with human breast cancer cells

Bisphosphonates are used routinely to reduce bone-related events in breast cancer patients with bone metastasis. We evaluated the effects of zoledronic acid, a third generation, nitrogen-containing bisphosphonate, to prevent bone metastasis in breast cancer. Zoledronic acid or vehicle alone was administered to nude mice either simultaneously or after intracardiac injection of human breast cancer MDA-MB-231 cells. Nude mice treated with zoledronic acid at early time points showed a lower incidence of bone metastases than did vehicle-treated nude mice, but these differences were not statistically significant. Only 37.5% of mice treated with zoledronic acid at the time of tumor cell inoculation developed bone metastases compared to over 51.8% of mice receiving vehicle alone (P = 0.304). Cell count of apoptosis confirmed by immunohistochemical staining in metastatic bone tissue significantly increased in the zoledronic acid-treated groups compared to non-treated group (1,018.3 vs 282.0; P = 0.046). However, metastatic tumor cells, which invade soft tissue around the bone, did not show extensive apoptosis; there were no differences between the zoledronic acid-treated and control groups. These results suggest that zoledronic acid increases apoptosis of metastatic breast tumor cells in the bone and could therefore reduce metastatic tumor burden. These results support the use of zoledronic acid to reduce the incidence of bone metastasis in breast cancer.     

A randomized controlled trial to compare the efficacy of bisphosphonates in the management of painful bone metastasis

Introduction:

The prospective interventional single-institution randomized control study was carried out to compare the pain relieving efficacy among different bisphosphonates at the cost of incidence of skeletal-related events (SRE).

Materials and Methods:

During June 2008 and May 2011, 256 patients with painful bone metastasis in solid tumors with a pain score of at least 5 were randomized into three arms: zoledronic acid (4 mg, i.v.), ibandronate (6 mg, i.v.) and pamidronate (90 mg, i.v.). Radiation was given to all patients, either 800 cGy single fraction or 20 Gy in five fractions. The ANOVA test was used for analysis. The Pearson test was used to correlate pain scores with proportions of responders as statistical estimation of pain relief.

Results:

With a mean baseline pain score of 6.5 ± 1.2, there was no difference in pain scores among the three treatment arms, assessed at 3 months and at the end of the study. However, the pain scores at 6 months were statistically reduced in zoledronic acid-receiving patients (1.5 ± 0.4) unlike the scores in patients receiving ibandronate (3.1 ± 0.5) and pamidronate (2.3 ± 0.4), with a P-value of 0.024. The response was statistically significant at 6 months (0.039) and at the end of the study (0.023), in favor of zoledronic acid. Pearson''s correlation demonstrated a statistically significant positive correlation between pain scores and response rates. There were no statistical differences in the narcotic scores among the treatment arms during the study period. The overall duration of pain relief was not different in any of treatment arms. The time of detection of hypercalcemia was no different; however, the incidence of hypercalcemia was significantly less in the zoledronic acid arm (28.3%) against 44.6% and 50% in ibandronate and pamidronate arms, respectively, with a P-value of 0.041.

Conclusion:

The use of bisphosphonates for 6 months or more results in a statistical significant improvement in bone pain, more so with zoledronic acid. Hypercalcemia, an SRE, was significantly less in the zoledronic acid arm.     

唑来膦酸对髋关节置换后假体柄周围骨丢失的预防效果

背景：无论是股骨侧还是髋臼侧在髋关节置换后1年内均会出现不同程度的骨丢失,对假体的长期稳定性和骨强度造成严重影响。因而减少全髋关节置换后假体柄周围骨丢失量对延长假体使用时间、预防假体周围骨折具有重要意义。 目的：观察分析唑来膦酸对髋关节置换后假体柄周围骨丢失的预防效果。 方法：对照组患者不给予,两组其余用药相同。对两组置换前和置换后1年的髋部骨密度变化进行测量;观察两组置换前、用药后3 d和用药后1年的血清钙、磷水平及碱性磷酸酶活性变化;对观察组患者用药期间出现的不良反应进行记录。  结果与结论：置换后1年两组患者股骨假体周围的平均骨密度均显著下降,与置换前比较差异有显著性意义(P < 0.05)。置换后1年观察组患者的平均骨密度显著高于对照组(P < 0.05)。用药后3 d两组患者的钙、磷水平均较置换前显著下降,差异均有显著性意义(P < 0.05)。用药后1年钙、磷水平维持在置换前水平。置换后短期内两组患者的碱性磷酸酶活性均稍微下降,但与置换前比较差异无显著性意义(P > 0.05)。用药后1年,观察组患者的碱性磷酸酶活性较低,与置换前、对照组比较差异均有显著性意义(P < 0.05)。用药后1年,对照组患者的碱性磷酸酶活性与置换前差异无显著性意义(P > 0.05)。用药两三天内观察组有9例患者出现不同程度的肌肉酸痛和发热,给予乙酰氨基酚后症状得到缓解。提示全髋关节置换后给予唑来膦酸注射液能够有效预防患者置换后早期假体柄周围骨丢失,但是置换后1周内可能会出现发热的症状,因此建议在置换完成1周后给予患者唑来膦酸注射液,若出现发热等症状,可给予乙酰氨基酚进行缓解。 中国组织工程研究杂志出版内容重点：人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程     

Infusion of zymosan-activated plasma affects neutrophils in peripheral blood and bone marrow in sheep.

Intravenous infusion of zymosan-activated plasma (ZAP) in sheep results in acute lung injury mediated in part by free radical release from stimulated neutrophils that are retained in increased numbers in the pulmonary microcirculation. ZAP infusion is also associated with long-term reduction in elicited superoxide anion generation from segmented neutrophils in the circulating and bone marrow pools for at least 24 h after the infusion. The purpose of our study was to investigate the effect of ZAP infusion on leukocyte counts and neutrophil granule-associated enzyme activity in the circulating and bone marrow pools. Cytochemical methods were used to characterize enzyme activity in primary granules (acid phosphatase and myeloperoxidase) and secondary granules (alkaline phosphatase). During the infusion period, neutrophil differential counts decreased less in venous blood than in matched arterial blood. Release from bone marrow stores was evident as increased numbers of circulating band neutrophils during and after the infusion. Neutrophils in venous and arterial blood smears were degranulated acutely during and 1-3 h after the infusion was stopped. Band and segmented neutrophils in bone marrow also appeared slightly degranulated 1-2 h after the infusion. In vitro experiments showed that band and segmented neutrophils in bone marrow degranulated in response to ZAP incubation. Immature polymorphonuclear leukocytes did not degranulate. Five to 24 h after ZAP infusion, enzyme activity in circulating and bone marrow neutrophils was at baseline levels, suggesting that new cells were being released into the circulating pool because degranulated neutrophils do not synthesize new granules. Another indication of a long-term effect in bone marrow were slight decreases in the percentage of immature polymorphonuclear leukocytes and band neutrophils and a significant decrease in the percentage of eosinophils, all of which coincided with a significant increase in the percentage of segmented neutrophils. Our results demonstrate that circulating complement anaphylatoxins affect neutrophils in bone marrow as well as the vascular space.     

Calcium and vitamin D supplementation during bisphosphonate administration may increase osteoclastic activity in patients with bone metastasis

Bone metastasis are a frequent complication of cancer, occurring in up to 70% of patients with advanced breast or prostate cancer. The consequences of bone metastasis are often devastating. Osteolytic metastasis can cause different kinds of skeletal related events including severe pain, pathologic fractures, life-threatening hypercalcemia, spinal cord compression, and other nerve-compression syndromes. These skeletal-related events are the result of the resorption of mineralized bone by osteoclasts. Bisphosphonates are synthetic analogues of naturally occurring pyrophosphate compounds that inhibit bone resorption. Potent bisphosphonates, pamidronate and, more importantly zoledronic acid may cause hypocalcemia, but mostly asymptomatic, mild, transient in most cases. Sufficient calcium and vitamin D intake needs to be ensured in patients with malignancy who have borderline or low levels of calcium when commencing treatment with bisphosphonates. Vitamin D itself induce the formation of osteoclasts by increasing the expression of RANKL on marrow stromal cells. Local calcium also promotes tumor growth and the production of parathyroid hormone-related peptide which in turn stimulates bone resorption. Vitamin D and calcium supplementation during bisphosphonate administration for the purpose of elimination of the side effects related to hypocalcemia in patients with bone metastasis may increase the bone resorption and decrease the efficacy of bisphosphonates. Therefore, vitamin D and calcium supplementation must not be routinely recommended during bisphosphonate administration.     

Hyaluronan synthesis inhibitor supplements the inhibitory effects of zoledronic acid on bone metastasis of lung cancer

Hyaluronan is known to have pivotal roles in the growth, migration and invasion of malignant tumors. Bone metastases are critical lesions greatly impairing the quality of patients with malignancies. We investigated whether hyaluronan synthesis inhibitor supplements the inhibitory effects of zoledronic acid, which is a conventional therapeutic agent for bone metastasis. We examined the effects of methylumbelliferone, an inhibitor of hyaluronan synthesis and/or ZA on the tumorigenicity of one murine lung carcinoma and two human (A549, SK-MES-1) lung cancer cell lines in vitro. The interaction between methylumbelliferone and zoledronic acid was analyzed using Calcucyn software. With a murine bone metastasis model of lung cancer in vivo, we investigated the inhibitory effects and interaction of the two drugs on the progression of metastatic bone lesions. Methylumbelliferone or zoledronic acid treatment individually suppressed proliferation, migration and invasion of 3 cell lines, and combination treatment showed synergistic effects. Although methylumbelliferone as a single agent did not enhance apoptotic activity, it showed additive effects on apoptotic activity to those of zoledronic acid. Co-localization of CD44 and ezrin, which might be a pathway of hyaluronan signaling, was abrogated by methylumbelliferone treatment. Combination therapy showed additive inhibitory effects on metastatic bone lesions in vivo, which paralleled the inhibition of hyaluronan accumulation by methylumbelliferone, and inhibition of osteoclastogenesis. Although the detailed mechanisms underlying the synergistic or additive inhibitory effects of these two drugs should be further analyzed, inhibition of hyaluronan synthesis by methylumbelliferone is a promising novel therapeutic candidate for bone metastasis of lung cancer in addition to zoledronic acid.     

Cytokines and Insulin Resistance after Zoledronic Acid-Induced Acute Phase Response

Zoledronic acid is known to induce a transient acute phase response (APR). The aim of the study was to investigate whether an APR caused by zoledronic acid administration can induce insulin resistance in post-menopausal osteoporotic women and the potential involvement of different inflammatory markers, cytokines and adipokines to this response. Insulin resistance was calculated by the homeostasis model assessment (HOMA-IR). APR symptoms appeared in 30 post-menopausal osteoporotic women within 24?h and attenuated on day 3 after zoledronic acid infusion. Twenty-eight age- and body mass index-matched, patients without an APR following zoledronic acid administration, served as a control group. In patients with APR, concurrently with a significant increase in serum high sensitive C-reactive protein (hsCRP), interleukin-6 (hsIL-6), tumour necrosis factor-alpha (hsTNF-α) and cortisol levels on days one and two, serum insulin was also significantly elevated, resulting in an increased HOMA-IR. Leptin and resistin significantly increased on day two in contrast to adiponectin which declined, though not statistically significant. The alterations in HOMA-IR were mainly associated to the increase of hsCRP and leptin. In conclusion, zoledronic acid induces an acute, short term insulin resistance, due to an APR, by altering the levels of various adipokines and cytokines.     

胰岛素联合唑来膦酸治疗糖尿病骨质疏松症对患者血清指标的影响

目的 分析胰岛素联合唑来膦酸治疗对糖尿病骨质疏松（DOP）患者血糖血脂以及骨代谢指标的影响。方法 选择2016年12月~2019年4月于我院诊治的DOP患者共136例进行研究,采用随机数字表法分为对照组和实验组,每组68例,对照组采用二甲双胍联合唑来膦酸治疗,实验组采用胰岛素联合唑来膦酸治疗,比较两组干预后血脂水平（TG、TC、LCL-C、HDL-C）、血糖指标（空腹、餐后2h、糖化血红蛋白）以及骨代谢相关指标（BAP、TRAP-5b、BGP以及s-CTX）。结果 治疗后实验组空腹、餐后2h血糖及HbA1c均低于对照组（P＜0.05）;实验组治疗后TG、TC、LCL-C、HDL-C水平均低于对照组（P＜0.05）;实验组BAP、BGP水平均高于对照组（P＜0.05）;实验组TRAP-5b水平均低于对照组（P＜0.05）;两组s-CTX指标比较,差异无统计学意义（P＞0.05）。结论 对DOP患者使用胰岛素与唑来膦酸联合治疗方案,有助于改善患者的血脂以及血糖水平,同时利于患者骨骼骨质的沉积,值得临床应用。     

RANK expression on breast cancer cells promotes skeletal metastasis

RANK ligand (RANKL), acting through its cognate receptor RANK, is a key factor for bone remodeling and metastasis by regulating the differentiation, survival and activation of osteoclasts. RANKL is also crucial for the development of mouse mammary glands during pregnancy and has been recently linked to the etiology of breast cancer via its direct activity on RANK-expressing normal or transformed breast epithelial cells, leading to increased mitogenesis, enhanced regenerative potential of mammary stem cells, and increased invasion and migration. We demonstrate that higher RANK expression in MDA-MB-231 breast cancer cells (MDA-231-RANK cells) is sufficient to confer a significantly greater metastatic growth rate in the bone compared with MDA-MB-231 cells which do not express high levels of RANK. Blockade of osteoclastic bone resorption, achieved with treatment by either RANKL inhibition or zoledronic acid, did reduce skeletal tumor progression of MDA-231-RANK cells suggesting that the vicious cycle contributes to metastatic growth. However, RANKL inhibition reduced skeletal growth of MDA-231-RANK tumors to a significantly greater extent than zoledronic acid, indicating that skeletal growth of RANK-positive tumors is also driven by direct RANKL effects. RANKL stimulated the expression of multiple genes associated with cell invasive behavior, including several matrix metalloproteinases and other genes previously defined as part of a bone metastasis gene signature. These data indicate that RANKL provokes breast cancer bone metastases via two distinct, but potentially overlapping mechanisms: stimulation of tumor-associated osteoclastogenesis and stimulation of RANK-expressing tumor cells.     

Expression and prognostic relevance of vascular endothelial growth factor (VEGF) and its receptor (FLT-1) in nephroblastoma

AIMS: To investigate the prognostic relevance of vascular endothelial growth factor (VEGF) and its receptor Flt-1 in nephroblastoma and whether tumour microvessel density (MVD) immunoreactivity, determined by the CD31 antigen, is related to the expression of VEGF and Flt-1. METHODS: The expression of VEGF and Flt-1 and MVD were investigated by means of immunohistochemical analysis in 62 Wilms's tumours. Patients were treated preoperatively with chemotherapy and had a mean follow up of 5.7 years. RESULTS: In general, VEGF and Flt-1 were expressed in normal kidney parenchyma and to a variable extent in the three main components of Wilms's tumour, namely: the blastemal, epithelial, and stromal cells. In tumour tissue, 52% and 47% of blastemal cells were positive for VEGF and Flt-1, respectively. A non-significant correlation was found between the expression of VEGF and Flt-1 in blastemal and epithelial cells and the clinicopathological stage. MVD was significantly higher in VEGF and Flt-1 positive tumours than in VEGF and Flt-1 negative tumours. Univariate analysis showed that the expression of VEGF and Flt-1 in blastemal cells was indicative of clinical progression and tumour specific survival. In addition, MVD expression was indicative of clinical progression. Epithelial staining was of no prognostic value. In a multivariate analysis, VEGF protein expression by blastemal cells was an independent prognostic marker for clinical progression. CONCLUSIONS: These results indicate that VEGF and Flt-1 protein expression are closely related to MVD and seem to be an important predictor for poor prognosis in treated patients with Wilms's tumour. Therefore, the expression of these molecules in primary Wilms's tumour may be useful in identifying those patients at high risk of tumour recurrence and in guiding antiangiogenic treatment.     

An interferon-gamma-related cytokine storm in SARS patients

Fourteen cytokines or chemokines were analyzed on 88 RT-PCR-confirmed severe acute respiratory syndrome (SARS) patients. IFN-gamma, IL-18, TGF-beta, IL-6, IP-10, MCP-1, MIG, and IL-8, but not of TNF-alpha, IL-2, IL-4, IL-10, IL-13, or TNFRI, were highly elevated in the acute phase sera of Taiwan SARS patients. IFN-gamma was significantly higher in the Ab(+) group than in the Ab(-) group. IFN-gamma, IL-18, MCP-1, MIG, and IP-10 were already elevated at early days post fever onset. Furthermore, levels of IL-18, IP-10, MIG, and MCP-1 were significantly higher in the death group than in the survival group. For the survival group, IFN-gamma and MCP-1 were inversely associated with circulating lymphocytes count and monocytes count, but positively associated with circulating neutrophils count. It is concluded that an interferon-gamma-related cytokine storm was induced post SARS coronavirus infection, and this cytokine storm might be involved in the immunopathological damage in SARS patients.     

低剂量唑来膦酸对去势拔牙大鼠破骨及成骨细胞的影响

文题释义：去势大鼠：即通过去势法建立骨质疏松动物模型,该模型建造方法主要有3种：去势法、维甲酸灌胃法和糖皮质激素肌注法,而去势法是目前最常用、最成熟的造模方法,主要是通过去除大鼠双侧卵巢至少3个月,构建骨量少、骨显微结构退化而引起骨脆性增加及骨折发生率升高的一种全身性骨病,即骨质疏松疾病模型。 TUNEL细胞凋亡检测：用于检测细胞在凋亡过程中细胞核DNA的断裂情况,其原理是生物素标记的dUTP在脱氧核糖核苷酸末端转移酶的作用下,可以连接到凋亡细胞中断裂DNA的3’-OH末端,并可与连接辣根过氧化酶的链霉亲和素特异结合,在显色剂DAB的存在下,产生很强的颜色反应(呈深棕色),特异准确定位正在凋亡的细胞。 背景：目前唑来膦酸虽能有效地预防绝经后妇女的骨丢失,但其对下颌骨的影响及其机制尚不清楚。 目的：观察去势大鼠经低剂量唑来膦酸作用后下颌骨组织病理学改变,探讨RANKL/RANK/OPG信号系统在唑来膦酸抑制骨吸收过程中的调控效应与机制。 方法：取36只雌性SD大鼠随机分为对照组、模型组和治疗组,后2组大鼠行两侧卵巢切除术,建立骨质疏松模型;对照组行假手术去除同等质量卵巢周围的脂肪。去卵巢3个月后,治疗组皮下一次性注射唑来膦酸20 μg/kg,对照组和模型组皮下注射相应剂量的盐水;用药1周后拔除大鼠左侧下颌磨牙,拔牙后4周麻醉动物取出拔牙侧下颌骨组织。通过影像学X 射线大致观察大鼠拔牙窝剩余牙槽骨状况,苏木精-伊红染色检测下颌骨皮质和骨松质的病理结构改变,TUNEL凋亡实验检测凋亡的成骨细胞数量,利用免疫组织化学技术检测下颌牙槽骨内细胞核因子κB受体活化因子配基(RANKL)、骨保护素、细胞核转录因子(NF-κB)的表达情况,最后Western blot检测核因子κB受体活化因子配基、细胞核转录因子蛋白的表达。 结果与结论：①拔牙4周后,治疗组相较模型组牙槽骨骨吸收减少,拔牙窝底有新骨形成;②苏木精-伊红染色可见,模型组的骨皮质变薄,骨小梁结构变细,甚至出现断裂,大量骨吸收陷窝,成骨细胞较少;治疗组骨皮质增厚,骨小梁结构正常,只有少量骨吸收陷窝,成骨细胞增多;③治疗组的成骨细胞凋亡数目显著低于模型组(P < 0.001);④免疫组织化学染色可见,治疗组RANKL蛋白、NF-KB p65蛋白表达显著低于模型组(P < 0.001,P < 0.002),骨保护素蛋白表达显著高于模型组(P < 0.001);⑤Western Blot结果显示,与对照组相比较,模型组的RANKL、NF-κB蛋白高表达,治疗组其表达量显著降低(均P < 0.001);⑥结果说明,唑来膦酸可通过下调细胞核转录因子信号通路抑制破骨细胞的分化,同时调控成骨细胞的凋亡。 ORCID: 0000-0002-0300-0460(程余婷) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Effect of zoledronic acid and an antibody against bone sialoprotein II on MDA-MB-231GFP breast cancer cells in vitro and on osteolytic lesions induced in vivo by this cell line in nude rats

The aim of this study was to investigate the combined effect of zoledronic acid and an antibody against bone sialoprotein II (BSPII) on proliferation and osteolytic activity of MDA-MB-231^GFP breast cancer cells. For this purpose, the cells were exposed to zoledronic acid (10–20 μg/ml [25–50 μM]) and an anti-BSPII IgY (10–100 μg/ml) for up to 5 days alone or in combination. The combined treatment showed synergistic antiproliferative effects at the higher dose of zoledronic acid. Following inoculation of 1 × 10⁵ MDA-MB-231 ^GFP breast cancer cells into a branch of the femoral artery of nude rats, lytic lesions developed in the tibia, femur or fibula of the injected hind leg after approximately 30 days. The appearance and development of these lesions were monitored radiographically. Rats with lytic lesions were treated with zoledronic acid (60 μg/kg/week sc × 8; n = 10), zoledronic acid and an anti-BSPII IgY antibody (60 μg/kg/week sc × 8 + 10 mg/kg/week sc × 8; n = 10), or left untreated (n = 20). In addition, rats were treated for 4 weeks (n = 10) with both regimens starting right after tumor cell inoculation. Finally, ten rats were treated with zoledronic acid for 2 weeks before tumor cell inoculation (60 μg/kg/week sc × 2). The antiosteolytic effect of zoledronic acid was high as shown by inhibition of osteolytic growth. Addition of the anti-BSPII IgY further decreased the incidence of femoral osteolytic lesions (40% reduction), indicating remineralization, and reduced periosteal defects of cortical bone (20% reduction). These observations favor using the IgY-antibody in addition to zoledronic acid in order to stimulate osteoblast-induced remineralization.     

Two-year postoperative use of zoledronic acid prevents secondary fractures following percutaneous kyphoplasty北大核心

BACKGROUND: Percutaneous kyphoplasty is the main treatment for senile osteoporotic thoracolumbar fractures, but increasing number of patients who have not been treated with anti-osteoporosis therapy after operation develop secondary fractures due to decreased bone mineral density and changes of stress. OBJECTIVE: To investigate the feasibility of consecutive 2-year zoledronic acid treatment following percutaneous kyphoplasty for preventing secondary vertebral fracture. METHODS: 186 elderly patients with thoracolumbar compressive fractures were divided into experimental (n=84) and control (n=102) groups based on their willingness to receive zoledronic acid treatment or not after percutaneous kyphoplasty. The experimental group was treated with calcium and alfacalcidol followed by 2 years of zoledronic acid treatment, while only calcium and alfacalcidol treatment was done in the control group. The bone mineral density, pain and function were respectively assessed by dual energy X-ray absorptiometry, visual analogue scale and Oswestry disability index, and the number of refractures was calculated at baseline and at the 2nd year after the second injection of zoledronic acid. RESULTS AND CONCLUSION: The bone mineral density, visual analogue scale and Oswestry disability index scores at 2 years after treatment in both two groups were significantly superior to those at baseline (P < 0.05). The number of refractures in the experimental group (n=1) was significantly less than that in the control group (n=9) (P < 0.05). These results suggest that zoledronic acid can prevent secondary fractures after percutaneous kyphoplasty in the elderly with osteoporotic thoracolumbar fractures, improve long-term function and clinical effectiveness. © 2017, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.     

Immune cells regulate VEGF signalling via release of VEGF and antagonistic soluble VEGF receptor‐1

Vascular endothelial growth factor (VEGF) is an important regulator of physiological and pathological angiogenesis. Besides malignant and stromal cells, local immune cells shape VEGF signalling in the tumour microenvironment. Aminobisphosphonates such as zoledronic acid (Zol) are drugs known to inhibit osteoclast activity and bone resorption, but also have immunomodulatory and anti‐tumour effects. These properties have been linked previously to the down‐regulation of VEGF and interference with tumour neo‐angiogenesis. It was therefore surprising to find that treatment with Zol in combination with low‐dose interleukin (IL)‐2 increased serum VEGF levels in cancer patients. In this study we aimed to characterize the effect of Zol and IL‐2 on VEGF signalling of blood‐derived immune cells in vitro. Upon stimulation with IL‐2, T cells and natural killer (NK) cells increase production of VEGF consecutively to the release of proinflammatory interferon (IFN)‐γ, and Zol accelerates this response specifically in γδ T cells. VEGF can, in turn, be antagonized by soluble VEGF receptor (sVEGFR)‐1, which is released depending on stimulatory conditions and the presence of monocytes. Additionally, malignant cells represented by leukaemia and lymphoma cell lines produce VEGF and some release sVEGFR‐1 simultaneously. Our findings indicate a mechanism by which the VEGF and the sVEGFR‐1 production by immune cells regulates local VEGF signalling. Therefore, immunotherapeutic interventions may enable both pro‐ as well as anti‐tumour effects via immune cell‐mediated alterations of VEGF homeostasis.     

局部应用唑来膦酸与纳米银对即刻种植骨结合影响的比较

背景:有研究表明局部应用唑来膦酸和局部应用纳米银都可促进骨形成,但两者效果的差异尚未见研究报道。目的:观察局部应用唑来膦酸和局部应用纳米银对兔拔牙窝即刻种植钛螺纹钉骨结合的影响差异。方法:将24只新西兰大白兔随机分为唑来膦酸和纳米银组,每组12只,在拔除兔上下颌4个切牙后,分别在2组的拔牙窝内填唑来膦酸-纳米羟基磷灰石复合物和纳米银-纳米羟基磷灰石混合物,并均同期植入钛螺纹钉。术后4,8,12周取兔上、下颌骨离体标本,通过一般形态观察、扭矩测试、骨密度检测及组织形态学观察。实验于2015年12月经河北医科大学第三医院医学伦理委员会批准(批准号:Z2015-021-1)。结果与结论:①扭矩力学实验测试结果显示,两组钛钉扭矩峰值的平均值随着时间延长而增加(扭矩峰值平均值数值),均在12周时达到最大(扭矩峰值平均值数值),术后4,8,12周,唑来膦酸组钛螺纹钉扭矩峰值平均值均略高于同期纳米银组,唑来膦酸组钛螺纹钉稳定性略优于纳米银组,但差异无显著性意义(P>0.05);②一般形态和组织学观察发现,术后4,8,12周唑来膦酸组种植体周围骨硬度和结构均明显优于同期纳米银组;③骨密度分析结果显示,术后4,8,12周,唑来膦酸组钛螺纹钉周围骨组织灰度值略高于同期纳米银组,但差异无显著性意义(P>0.05);④结果证实,从植入材料对影响种植体周围骨替代材料成骨的效果来看,用唑来膦酸纳米羟基磷灰石复合物进行即刻种植骨结合的效果略优于纳米银与纳米羟基磷灰石混合植骨。