破骨细胞分化因子诱导小鼠骨髓细胞形成破骨细胞的实验研究

目的:探讨破骨细胞分化因子(ODF)和巨噬细胞集落刺激因子(M-CSF)联合应用进行体外诱导小鼠骨髓细胞形成破骨细胞(OC)的能力。方法:收集5～6周小鼠骨髓细胞,在含M-CSF(10 ng/ml)的α-MEM全培养液中培养24h,然后将悬浮生长的细胞接种到24孔培养板,并加入不同浓度的ODF和M-CSF。,通过观察抗酒石酸盐酸性磷酸酶(TRAP)染色的阳性细胞和能否在骨磨片上形成吸收陷窝来鉴定OC形成情况。结果:在只加入ODF或M-CSF一种细胞因子时,未见有TRAP阳性或CTR阳性细胞形成,同时加入ODF和M-CSF,可形成典型的OC。TRAP阳性多核细胞的数目和培养液中钙离子浓度的增加与ODF的浓度呈正相关。结论:小鼠骨髓来源的单核细胞在ODF和M-CSF共同作用下可形成具有骨吸收功能的OC,为体外研究OC的分化发育和功能调节提供了一种新的方法。     

颏部骨移植修复牙槽裂的临床研究

１６例单侧唇腭裂伴牙槽裂和１例双侧唇腭裂伴牙槽裂患者，用下颌正中联合区骨移植修得牙槽裂，术后随访时间平均１８个月，术后６个月，咬合Ｘ线片证明移植骨完全愈合，再造牙槽突的形态满意且无牙周并发症，手术后６个月Ｘ线片见供区愈合，下颌骨正中联合骨移植有并发症少，住院时间短，手术时间短而且可避免供区瘢痕等优点，本研究结果表明下颌正中联合区骨可用作为整复牙槽裂的供区。     

Localization of interleukin-1 (IL-1) mRNA-expressing macrophages in human inflamed gingiva and lL-1 activity in gingival crevicular fluid

The exact cell type and site(s) involved in interleukin-1 (lL-1) production during gingival inflammation was determined by combining immunohistochemistry and in situ hybridization. IL-1 messenger RNA (mRNA)-expressing cells in human inflamed gingiva were identified as macrophages. The rate of IL-α mRNA expression in these macrophages was the same as IL-1 β mRNA expression. The rate of IL-1 mRNA expression was higher in connective tissue furthest from the pocket epithelium, although more macrophages were present at the connective tissue subjacent to the pocket epithelium. The IL-1 activity in gingival crevicular fluid (GCF) obtained from inflamed gingiva was higher than that from healthy gingiva and decreased after periodontal therapy. The IL-1 activity in GCF was almost completely abolished by the addition of anti-IL-1α antibody but not by anti-IL-1 β antibody, indicating that IL-1α is the predominant form in GCF. However, the IL-1 activity in GCF was unrelated to the number of IL-1 mRNA-exprerssing macrophages in the same gingival site where the GCF was obtained at the same time. The results suggest that macrophages in the connective tissue subjacent to the oral epithelium contribute to the production of IL-1 but those in connective tissue subjacent to the pocket epithelium play a different role in the generation of gingival inflammation.     

拔牙位点保留后骨胶原联合自体软组织瓣植入对前牙拔牙后美学满意度的影响

目的:探讨拔牙位点保留后骨胶原联合自体软组织瓣植入对前牙拔牙后美学满意度的影响。方法:选择2018年1月-2019年7月笔者医院就诊的84例有前牙拔除需求的患者为研究对象,采用随机数字表法分为42例对照组(Bio-Oss Collagen植入),42例观察组(Bio-Oss Collagen+自体软组织瓣植入)。比较两组患者术后拔牙位点愈合情况及两组患者术前、术后6个月牙槽骨高度、牙槽骨宽度,记录两组患者术前、术后6个月牙槽骨密度、垂直向骨吸收量、唇腭向骨吸收量,术后美学效果、满意度,随访1年,观察植后种植体周围骨高度、宽度吸收情况。结果:术后2周拆线,观察组可见拔牙位点愈合良好,无感染、坏死情况,硬腭带蒂结缔组织移植的成活率为100%,对照组拔牙位点愈合良好,但牙槽嵴呈现不同程度的萎缩。两组患者术前牙槽骨宽度、高度、密度比较,差异无统计学意义(P>0.05);术后6个月,两组牙槽骨宽度、高度、密度均低于术前,观察组牙槽骨高度、密度、宽度均高于对照组,差异均有统计学意义(P<0.05)。术后6个月,观察组垂直向骨吸收量、唇腭向骨吸收量均低于对照组,差异有统计学意义(P<0.05)。术后6个月,观察组美学效果、满意度评分较对照组高,随访1年,观察组种植体周围骨吸收低于对照组,差异有统计学意义(P<0.05)。结论:骨胶原联合自体软组织瓣植入可减缓拔牙后牙槽骨高度、宽度及骨密度的下降,在保留骨量方面具有一定的临床价值,能够有效改善患者牙槽美学效果。     

Pyridoxamine ameliorates methylglyoxal-induced macrophage dysfunction to facilitate tissue repair in diabetic wounds

Methylglyoxal (MGO) is a highly reactive dicarbonyl compound formed during hyperglycaemia. MGO combines with proteins to form advanced glycation end products (AGEs), leading to cellular dysfunction and organ damage. In type 2 diabetes mellitus (T2DM), the higher the plasma MGO concentration, the higher the lower extremity amputation rate. Here, we aimed to identify the mechanisms of MGO-induced dysfunction. We observed that the accumulation of MGO-derived AGEs in human diabetic wounds increased, whereas the expression of glyoxalase 1 (GLO1), a key metabolic enzyme of MGO, decreased. We show for the first time that topical application of pyridoxamine (PM), a natural vitamin B6 analogue, reduced the accumulation of MGO-derived AGEs in the wound tissue of type-2 diabetic mice, promoted the influx of macrophages in the early stage of tissue repair, improved the dysfunctional inflammatory response, and accelerated wound healing. In vitro, MGO damaged the phagocytic functions of M1-like macrophages induced by lipopolysaccharide (LPS), but not those of M0-like macrophages induced by PMA or of M2-like macrophages induced by interleukins 4 (IL-4) and 13 (IL-13); the impaired phagocytosis of M1-like macrophages was rescued by PM administration. These findings suggest that the increase in MGO metabolism in vivo might contribute to macrophage dysfunction, thereby affecting wound healing. Our results indicate that PM may be a novel therapeutic approach for treating diabetic wounds. MGO forms protein adducts that cause macrophage dysfunction. These adducts cause cell and organ dysfunction that is common in diabetes. Pyridoxamine scavenges MGO to ameliorate this dysfunction, promoting wound healing. Pyridoxamine could be used therapeutically to treat non-healing diabetic wounds.     

不同加力时间下静止期牙周病大鼠牙槽骨骨微结构的变化

目的建立静止期牙周病动物模型,观察不同加力时间下其牙槽骨骨微结构的变化。方法选取156只28周龄SD雄性大鼠,随机分为对照组(50只)和实验组(106只),实验组建立静止期牙周病动物模型,再随机分为牙周治疗组和牙周+正畸治疗组(各50只),对牙周+正畸治疗组动物模型上颌右侧第一磨牙加力,力值为50 g。各组于加力后0、7、14、21、28 d各处死10只动物,取材,行Micro-CT检察。结果①加力0、7、14、21、28 d,实验组动物模型右侧上颌第一磨牙牙槽骨骨密度(bone mineral density,BMD)、骨体积分数(BV/TV)、骨小梁厚度(Tb·Th)均较对照组降低,骨小梁数量(Tb·N)、骨小梁分离距离(Tb·Sp)均较对照组升高;②相较牙周治疗组,加力14 d时牙周+正畸治疗组动物模型右侧上颌第一磨牙牙槽骨BMD、BV/TV、Tb·Th降低,Tb·N、Tb·Sp升高;加力28 d时,BV/TV升高,Tb·N、Tb·Sp降低;③加力14 d,牙周+正畸治疗组动物模型右侧上颌第一磨牙牙槽骨BMD、BV/TV、Tb·Th降至最低,Tb·N、Tb·Sp升至最高,但至28 d时各项指标已恢复至未加力时状态。结论①静止期牙周病动物模型牙槽骨骨质疏松且对力学刺激的反应变缓慢;②突然受力后静止期牙周病动物模型牙槽骨以吸收为主,但随着加力装置力量的减弱和力值的稳定,牙槽骨可逐渐恢复到加力前的状况;③较小的力值刺激有利于静止期牙周病动物模型牙槽骨的修复。     

The first case of combined heart-liver transplantation in a patient with alveolar echinococcosis

We report a rare case of liver alveolar echinococcosis with an invasion of the hepaticocaval confluence, inferior vena cava, pericardium, right atrium, atrial septum, and superior vena cava, and its successful treatment by combined heart-liver transplantation.     

Trained immunity in organ transplantation

Consistent induction of donor‐specific unresponsiveness in the absence of continuous immunosuppressive therapy and toxic effects remains a difficult task in clinical organ transplantation. Transplant immunologists have developed numerous experimental treatments that target antigen‐presentation (signal 1), costimulation (signal 2), and cytokine production (signal 3) to establish transplantation tolerance. While promising results have been obtained using therapeutic approaches that predominantly target the adaptive immune response, the long‐term graft survival rates remain suboptimal. This suggests the existence of unrecognized allograft rejection mechanisms that contribute to organ failure. We postulate that trained immunity stimulatory pathways are critical to the immune response that mediates graft loss. Trained immunity is a recently discovered functional program of the innate immune system, which is characterized by nonpermanent epigenetic and metabolic reprogramming of macrophages. Since trained macrophages upregulate costimulatory molecules (signal 2) and produce pro‐inflammatory cytokines (signal 3), they contribute to potent graft reactive immune responses and organ transplant rejection. In this review, we summarize the detrimental effects of trained immunity in the context of organ transplantation and describe pathways that induce macrophage training associated with graft rejection.     

Effect of micro-roughening of poly(ether ether ketone) on bone marrow derived stem cell and macrophage responses,and osseointegration

Poly(ether ether ketone) (PEEK) has emerged as a candidate to replace metal implants because of its satisfactory mechanical properties, radiolucency, and lack of metal allergy. However, PEEK lacks osseointegration ability limiting its clinical applications. To overcome this problem, we prepared PEEK with a micro-rough surface using the sandblast method to modulate its osseointegration property; the sandblast method is simple, cost-effective, and is already applied to clinical metal implants. The surface roughness of the sandblasted PEEK was about 2.3 μm, whereas that of mirror-polished PEEK was 0.06 μm. Rat bone marrow-derived mesenchymal stem cells (RMSCs) showed higher proliferation, osteocalcin (OC) expression and bone-like nodule formation on micro-roughened PEEK compared with those cultured on mirror-polished PEEK, suggesting that micro-roughening facilitated RMSCs proliferation and differentiation. The micro-roughened surface slightly mitigated secretion of inflammatory C-C motif chemokine 2 (CCL-2) from lipopolysaccharide (LPS)-stimulated macrophages, but not of tumor necrosis factor α (TNFα) and interleukin-6 (IL-6). Finally, to compare osseointegration, specimens were implanted in rat femur bone marrow cavities, and then the pull-out force was measured. The pull-out force of micro-roughened PEEK was about four times higher than that of the mirror-polished PEEK. These results showed that micro-roughening of PEEK using the sandblast method was able to improve osseointegration, partly through elevating proliferation and differentiation of RMSCs.     

Immune-enhancing effects of a polysaccharide PRG1-1 from Russula griseocarnosa on RAW264.7 macrophage cells via the MAPK and NF-κB signalling pathways

Polysaccharides are one of many bioactive compounds found in edible mushrooms. Edible mushrooms have become attractive as “health foods” and as source materials for immunomodulators. The aim of this project was to study the immunoregulatory effects of a purified polysaccharide derived from wild Russula griseocarnosa (PRG1-1) on macrophages. Our data showed that in RAW264.7 macrophage cells, PRG1-1 increased expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Furthermore, PRG1-1 increased the production of nitric oxide (NO) and cytokines, including interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-α). Western blotting demonstrated that the regulation of NO and cytokines was mediated through the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signalling pathways. Therefore, PRG1-1 has the capacity to activate macrophages via the NF-κB and MAPK pathways. These findings helped to elucidate the immune-modulatory properties of the polysaccharide from R. griseocarnosa.