Preparation,characterization and antioxidant activity analysis of three Maillard glycosylated bone collagen hydrolysates from chicken,porcine and bovine

Bone collagen hydrolysates(peptides)derived from byproduct of animal product processing have been used to produce commercially valuable products due to their potential antioxidant activity.Maillard glycosylated reaction is considered as a promising method to enhance the antioxidant activity of peptides.Hence,this research aims at investigating the Maillard glycosylation activity and antioxidant activity of bone collagen hydrolysates from different sources.In this study,3 glycosylated bone collagen hydrolysates were prepared and characterized,and cytotoxicity and antioxidant activity were analyzed and evaluated.The free amino groups loss,browning intensity,and fluorescence intensity of G-Cbcp(glycosylated chicken bone collagen hydrolysates(peptides))were the heaviest,followed by G-Pbcp(glycosylated porcine bone collagen hydrolysates(peptides))and G-Bbcp(glycosylated bovine bone collagen hydrolysates(peptides)).The results of amino acid analysis showed that amino acid composition of different bone collagen hydrolysates was significantly different and the amino acid decreased to different degrees after Maillard glycosylated reaction,which may lead to differences in Maillard glycosylated reaction activity.Furthermore,the 3 glycosylated hydrolysates showed no significant cytotoxicity.The results showed that glycosylation process significantly increased the antioxidant activity of bone collagen hydrolysates,and G-Cbcp showed the strongest antioxidant activity,followed by G-Pbcp and G-Bbcp.Therefore,compared with the bone collagen hydrolysates,3 glycosylated hydrolysates showed significant characteristic and structural changes,and higher antioxidant activity.

Supplementation with yak (Bos grunniens) bone collagen hydrolysate altered the structure of gut microbiota and elevated short-chain fatty acid production in mice

In this study, yak bone collagen hydrolysate(YBCH)was produced by mixed proteases and provided to standard-diet mice at a different dose(low dose(LD), medium dose(MD), and high dose(HD))to investigate its effects on the composition of gut microbiota and short-chain fatty acids(SCFA)production. It was found that YBCH was mainly composed of small molecular peptides whose molecular weight below 2 000 Da. Notably, supplementation with different doses of YBCH could significantly downregulate the ratio of Firmicutes to Bacteroidetes in the fecal microbiota. At the family level, the Lachnospiraceae abundance was significantly reduced in the YBCH gavage groups(mean reduction ratio 41.7 %, 35.2%, and 36.4% for LD, MD, and HD group, respectively). The predicted functions of gut microbes in the MD group were significantly increased at “lipid metabolism” and “glycan biosynthesis and metabolism”. Moreover, the SCFA production in the YBCH groups was elevated. Especially, the concentration of acetic acid, propionic acid, and butyric acid in the MD group was separately increased 79.7%, 89.2%, and 78.8% than that in the NC group. These results indicated that YBCH might be applied in the development of functional food for intestinal microecological regulation.

The characterization of acid and pepsin soluble collagen from ovine bones (Ujumuqin sheep)

Ovine bones are the major by-products after slaughtered. The present study was conducted to extract and characterize acid soluble collagens （ASC） and pepsin soluble collagens （PSC） from ovine bones （Ujumuqin sheep）. Ovine bones collagen were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and liquid chromatography-tan- dem mass spectrometry （LC-MS/MS） as type I collagen. The results of Fourier transform infrared （FTIR） spectra analysis testified the existence of triple superhelical structure in both ASC and PSC, showing pepsin did not disrupt the triple helical structure of ovine bones collagen. Glycine, accounting for one-third of total amino acids, was the major amino acid for ovine bones collagen. Higher imino acid content was responsible for higher thermal denaturation temperature of ovine bones collagen compared to fish collagens. The isoelectric point of ASC was lower than PSC due to the higher content of acidic amino acids. Therefore, this study provides the potential reference for collagen extraction and application of ovine bones by-procduct.

Real-time-guided bone regeneration around standardized critical size calvarial defects using bone marrow-derived mesenchymal stem cells and collagen membrane with and without using tricalcium phosphate: an in vivo microcomputed tomographic and histologic e

The aim of the present real time in vivo micro-computed tomography （pCT） and histologic experiment was to assess the efficacy of guided bone regeneration （GBR） around standardized calvarial critical size defects （CSD） using bone marrow-derived mesenchymal stem cells （BMSCs）, and collagen membrane （CM） with and without tricalcium phosphate （TCP） graft material. In the calvaria of nine female Sprague-Dawley rats, full-thickness CSD （diameter 4.6 mm） were created under general anesthesia. Treatment-wise, rats were divided into three groups. In group 1, CSD was covered with a resorbable CM; in group 2, BMSCs were filled in CSD and covered with CM; and in group 3, TCP soaked in BMSCs was placed in CSD and covered with CM. All defects were closed using resorbable sutures. Bone volume and bone mineral density of newly formed bone （NFB） and remaining TCP particles and rate of new bone formation was determined at baseline, 2, 4, 6, and 10 weeks using in vivo pCT. At the lOth week, the rats were killed and calvarial segments were assessed histologically. The results showed that the hardness of NFB was similar to that of the native bone in groups I and 2 as compared to the NFB in group 3. Likewise, values for the modulus of elasticity were also significantly higher in group 3 compared to groups 1 and 2. This suggests that TCP when used in combination with BMSCs and without CM was unable to form bone of significant strength that could possibly provide mechanical ＂lock＂ between the natural bone and NFB. The use of BMSCs as adjuncts to conventional GBR initiated new bone formation as early as 2 weeks of treatment compared to when GBR is attempted without adiunct BMSC therapy.

Comparative study of chitosan/fibroin–hydroxyapatite and collagen membranes for guided bone regeneration in rat calvarial defects: micro-computed tomography analysis

This study aimed to utilize micro-computed tomography （micro-CT） analysis to compare new bone formation in rat calvarial defects using chitosan/fibroin-hydroxyapatite （CFB-HAP） or collagen （Bio-Gide） membranes. Fifty-four （54） rats were studied. A circular bony defect （8 mm diameter） was formed in the centre of the calvaria using a trephine bur. The CFB-HAP membrane was prepared by thermally induced phase separation. In the experimental group （n= 18）, the CFB-HAP membrane was used to cover the bony defect, and in the control group （n= 18）, a resorbable collagen membrane （Bio-Gide） was used. In the negative control group （n= 18）, no membrane was used. In each group, six animals were euthanized at 2, 4 and 8 weeks after surgery. The specimens were then analysed using micro-CT. There were significant differences in bone volume （BV） and bone mineral density （BMD） （P〈O.05） between the negative control group and the membrane groups. However, there were no significant differences between the CFB-HAP group and the collagen group. We concluded that the CFB-HAP membrane has significant potential as a guided bone regeneration （GBR） membrane.

Chitosan-collagen porous scaffold and bone marrow mesenchymal stem cell transplantation for ischemic stroke

In this study, we successfully constructed a composite of bone marrow mesenchymal stem cells and a chitosan-collagen scaffold in vitro, transplanted either the composite or bone marrow mesenchymal stem cells alone into the ischemic area in animal models, and compared their effects. At 14 days after co-transplantation of bone marrow mesenchymal stem cells and the hi- tosan-collagen scaffold, neurological function recovered noticeably. Vascular endothelial growth factor expression and nestin-labeled neural precursor cells were detected in the iscbemic area, surrounding tissue, hippocampal dentate gyrus and subventricular zone. Simultaneously, a high level of expression of glial fibrillary acidic protein and a low level of expression of neuron-spe- cific enolase were visible in BrdU-labeled bone marrow mesenchymal stem cells. These findings suggest that transplantation of a composite of bone marrow mesenchymal stem cells and a chi- tosan-collagen scaffold has a neuroprotective effect following ischemic stroke.

Thermal,infrared spectroscopy and molecular modeling characterization of bone:An insight in the apatite-collagen type I interaction

An insight into the interaction of collagen type I with apatite in bone tissue was performed by using differential scanning calorimetry, Fourier transform infrared spectroscopy, and molecular modeling. Scanning electron microscopy shows that bone organic content incinerate gradually through the different temperatures studied. We suggest that the amide regions of the type I collagen molecule (mainly C=O groups of the peptide bonds) will be important in the control of the interactions with the apatite from bone. The amide I infrared bands of the collagen type I change when interacting to apatite, what might confirm our assumption. Bone tissue results in a loss of thermal stability compared to the collagen studied apart, as a consequence of the degradation and further combustion of the collagen in contact with the apatite microcrystals in bone. The thermal behavior of bone is very distinctive. Its main typical combustion temperature is at 360°C with a shoulder at 550°C compared to the thermal behavior of collagen, with the mean combustion peak at ca. 500°C. Our studies with molecular mechanics (MM+ force field) showed different interaction energies of the collagen-like molecule and different models of the apatite crystal planes. We used models of the apatite (100) and (001) planes;additional two planes (001) were explored with phosphate-rich and calcium-rich faces;an energetic preference was found in the latter case. We preliminary conclude that the peptide bond of collagen type I is modified when the molecule interacts with the apatite, producing a decrease in the main peak from ca. 500°C in collagen, up to 350°C in bone. The combustion might be related to collagen type I, as the ΔH energies present only small variations between mineralized and non-mineralized samples. The data obtained here give a molecular perspective into the structural properties of bone and the change in collagen properties caused by the interaction with the apatite. Our study can be useful to understand the biological synthesis of minerals as well as the organic-inorganic interaction and the synthesis of apatite implant materials.

Collagen-chitosan scaffold impregnated with bone marrow mesenchymal stem cells for treatment of traumatic brain injury

Combinations of biomaterials and cells can effectively target delivery of cells or other therapeutic factors to the brain to rebuild damaged nerve pathways after brain injury.Porous collagen-chitosan scaffolds were prepared by a freeze-drying method based on brain tissue engineering.The scaffolds were impregnated with rat bone marrow mesenchymal stem cells.A traumatic brain injury rat model was established using the 300 g weight free fall impact method.Bone marrow mesenchymal stem cells/collagen-chitosan scaffolds were implanted into the injured brain.Modified neurological severity scores were used to assess the recovery of neurological function.The Morris water maze was employed to determine spatial learning and memory abilities.Hematoxylin-eosin staining was performed to measure pathological changes in brain tissue.Immunohistochemistry was performed for vascular endothelial growth factor and for 5-bromo-2-deoxyuridine(BrdU)/neuron specific enolase and BrdU/glial fibrillary acidic protein.Our results demonstrated that the transplantation of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds to traumatic brain injury rats remarkably reduced modified neurological severity scores,shortened the average latency of the Morris water maze,increased the number of platform crossings,diminished the degeneration of damaged brain tissue,and increased the positive reaction of vascular endothelial growth factor in the transplantation and surrounding areas.At 14 days after transplantation,increased BrdU/glial fibrillary acidic protein expression and decreased BrdU/neuron specific enolase expression were observed in bone marrow mesenchymal stem cells in the injured area.The therapeutic effect of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds was superior to stereotactic injection of bone marrow mesenchymal stem cells alone.To test the biocompatibility and immunogenicity of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds,immunosuppressive cyclosporine was intravenously injected 12 hours before transplantation and 1-5 days after transplantation.The above indicators were similar to those of rats treated with bone marrow mesenchymal stem cells and collagen-chitosan scaffolds only.These findings indicate that transplantation of bone marrow mesenchymal stem cells in a collagen-chitosan scaffold can promote the recovery of neuropathological injury in rats with traumatic brain injury.This approach has the potential to be developed as a treatment for traumatic brain injury in humans.All experimental procedures were approved by the Institutional Animal Investigation Committee of Capital Medical University,China(approval No.AEEI-2015-035)in December 2015.

Synthesis and Characterization of a Polymeric Material Blended to Bone Forming Elements

A synthetizing material blended with two distinct proteins (collagen and casein) and mineral mixture, was developed in order to evaluate their properties suitable for possible applications in the biomedical such as inducing the regeneration of damaged bone, either due to an accident or illness. Samples were evaluated by 1) Mechanical properties tests under the bending, 2) Scanning electronic microscopy and 3) Infrared spectroscopy were carried out. The results showed that the developed material has breaking strength and structure characteristics associated with the protein used in their composition. This fact suggests that the used protein determines the resistance of the material, in such a way according to the required use, being able to choose appropriate strength and duration either short or long time. The material composition for specific use, in order to find the most suitable mixture for bone replacement, or induce bone recovery, according to the required properties similar to those of damaged living tissue.

大西洋鳕鱼骨胶原蛋白肽的抗氧化活性及稳定性研究

该研究以大西洋鳕鱼骨胶原蛋白肽为研究对象,通过测定其自由基清除能力和还原力系统评价大西洋鳕鱼骨胶原蛋白肽的抗氧化活性,并考察温度、酸碱度、模拟胃肠道消化等因素对其稳定性的影响。结果表明,大西洋鳕鱼骨胶原蛋白肽具有良好的自由基清除能力和显著的还原能力,其清除DPPH自由基、羟自由基、超氧阴离子自由基和ABTS阳离子自由基的IC_(50)值分别为33.48、9.68、6.86、17.78 mg/mL,其还原力呈浓度依赖效应。大西洋鳕鱼骨胶原蛋白肽的抗氧化活性在温度25~100℃条件下较为稳定;在pH 6.0~8.0条件下可保持较高抗氧化活性,但在强酸或强碱条件下抗氧化活性显著下降(P<0.05);经体外模拟胃肠道消化后仍能保持原有抗氧化活性的80.26%以上。综合分析,大西洋鳕鱼骨胶原蛋白肽具有较好的抗氧化活性,其相关产品的加工贮藏应避免强酸、强碱条件的影响。研究结果可为鳕鱼骨资源高值利用以及多肽类抗氧化产品开发提供科学指导。

大鲵骨酸溶性和胃蛋白酶溶性胶原蛋白的提取与表征

大鲵为一种珍贵的两栖动物,营养丰富。为了充分且有效利用其加工副产物,分离并表征了来自大鲵骨的酸溶性胶原蛋白(ASC)和胃蛋白酶溶解性胶原蛋白(PSC)。大鲵骨ASC和PSC的产率分别为10.71%和13.11%。通过傅里叶红外光谱揭示胶原蛋白稳定的三螺旋结构。SDS-PAGE电泳图显示两个样品均为I型胶原蛋白,同时具有高水平的亚氨基酸(179~187/1000个残基)。样品在pH 6时溶解度最高。热变性温度(45.78~46.51℃)显著高于海洋和淡水鱼物种。在SEM下,胶原蛋白显示出致密的片状和不规则的孔洞状。这些结果表明大鲵骨胶原蛋白特性与哺乳动物胶原蛋白相似,具有成为其替代品的潜力,为其后续的研究和开发利用提供了基础数据。

鮰鱼骨胶原蛋白肽-钙螯合物促进钙转运和成骨细胞分化作用

以实验室前期制备的鮰鱼骨胶原蛋白肽-钙螯合物(F3-Ca螯合物)为原料,采用Caco-2细胞模型评价其对钙转运和吸收的影响,再分析其对hFoB1.19成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)活性以及分化关键基因和蛋白表达的影响。结果发现F3-Ca螯合物能使Caco-2细胞的钙转运量提高到0.31 mg/mL,并促进成骨细胞的矿化。当F3-Ca螯合物的质量浓度为100μg/mL时,成骨细胞ALP的活性提高了53.68%。反转录聚合酶链式反应和蛋白质免疫印迹分析表明F3-Ca螯合物能提高成骨细胞中骨保护素(osteoprotegerin,OPG)、骨钙素和Runt相关转录因子2的基因和蛋白表达水平,降低核因子受体激活因子-κB配体(receptor activator for nuclear factor-κB ligand,RANKL)的基因和蛋白表达水平,增大OPG/RANKL的比值,从而激活OPG/RANKL/核因子受体激活因子-κB信号通路,促进成骨细胞的增殖、分化和矿化,达到加强钙吸收和抗骨质疏松的作用。本研究可为鮰鱼骨的高值化利用和新型钙补充剂的开发提供科学依据。

羧甲基壳聚糖对牛骨胶原蛋白微观结构、热稳定性及自组装性质的影响

构建具备良好热稳定性、自组装性质及生物相容性的可食性细胞外基质(extracellular matrix,ECM)类似物支架对于制造结构化细胞培养肉制品至关重要。将羧甲基壳聚糖(carboxymethyl chitosan,CMCS)引入牛骨胶原蛋白(bovine bone collagen,BBC)体系中,通过光谱分析(紫外、红外、荧光光谱)发现BBC与CMCS的相互作用随着引入CMCS添加量的增加而增强,但并未影响BBC的三螺旋结构。差示扫描量热法/热重分析结果表明,CMCS的引入增强了BBC体系的热稳定性。浊度试验及扫描电子显微镜/透射电子显微镜观察结果证实了CMCS引入后胶原蛋白纤维形成度呈上升趋势,聚集行为更明显且自组装速率产生变化,呈现出更疏松扭曲的三维结构以及更大的纤维直径及更广泛的直径分布。但CMCS的引入并未明显影响BBC的D-周期性结构(胶原纤维自组装过程中形成的特征性明暗交替的周期性横纹结构)形成及其长度,且CMCS引入前后体系的细胞相容性也未呈现显著性差异。随着引入CMCS添加量增加,CMCS和BBC之间的静电作用力可能较共价相互作用和氢键更占优势。这些结果表明,CMCS的引入不影响BBC三螺旋结构完整性和生物相容性,并改善了BBC的热稳定性及体外自组装性质。这为开发新型优良可食性胶原蛋白基ECM仿生支架在细胞培养肉领域的应用以及畜禽骨副产物高值化精深加工利用提供了参考信息。

胶原水解物抗骨质疏松疗效功能的研究

解决骨质疏松导致的骨微观结构被破坏、骨密度下降的问题是目前全球老龄化背景下的一个研究热点。为了缓解骨质疏松,本研究使用来自细胞外基质的胶原水解物促进成骨和成血管,进而缓解骨质疏松。通过细胞活性分析,发现胶原水解物可以同时促进成骨细胞MC3T3-E1和内皮细胞EA.hy926的增殖,分别可达到141%、116%(P<0.05),通过碱性磷酸酶和茜素红S染色,发现胶原水解物能促进成骨细胞分化和矿化。建立了去势小鼠骨质疏松模型,micro CT表征发现胶原水解物能较为显著的抑制骨质疏松引起的骨小梁结构破坏,从而缓解骨质疏松症状。本研究表明胶原水解物在促进成骨、抑制骨质疏松等领域具有较大的应用潜力。

干细胞工程化双面异性静电纺丝膜促进硬脊膜修复的体外实验

背景:目前临床上采用自体组织或明胶海绵等材料对硬脊膜进行修补,但都存在不同程度的缺陷,因此亟需一种可以促进硬脊膜修复的生物材料。目的:利用定向静电纺丝及胶原自组装技术构建双面异性静电纺丝膜,并将其负载骨髓间充质干细胞构建人工硬脊膜,探讨该人工硬脊膜的各项理化性能以及生物特性。方法:采用静电纺丝技术及胶原自组装技术制备有序聚乳酸静电纺丝纤维的双面(一面为胶原蛋白,另一面为聚乳酸)异性静电纺丝膜(胶原组)、无序的聚乳酸静电纺丝膜(无序纤维组)、有序定向的聚乳酸静电纺丝膜(有序纤维组),通过扫描电镜、力学拉伸、水接触角测试、降解实验等来表征静电纺丝膜的理化性能。将胶原组(在胶原蛋白面接种骨髓间充质干细胞,即得到干细胞工程化静电纺丝膜)、无序纤维组、有序纤维组静电纺丝膜分别与骨髓间充质干细胞共培养,采用CCK-8法和活/死染色评估静电纺丝膜的生物相容性,整合素β1免疫荧光染色评估静电纺丝膜的黏附特性。将干细胞工程化静电纺丝膜及胶原组静电纺丝膜分别与骨髓巨噬细胞共培养,通过诱导型一氧化氮合酶(M1型)、CD206(M2型)免疫荧光染色及qRT-PCR检测炎症相关基因的表达来评估免疫调控性能。结果与结论:①定向静电纺丝纤维膜可模拟天然硬脊膜的纵向排列结构,胶原蛋白加入后,纤维膜的亲水性提高约2倍,力学性能提升了1.2倍;②与骨髓间充质干细胞共培养时,CCK-8和活/死染色显示,胶原组静电纺丝膜的细胞生物活性明显高于无序纤维组、有序纤维组;免疫荧光染色显示,胶原组整合素β1表达约为无序纤维组、有序纤维组的2.6倍,且细胞铺展形态良好;③与骨髓巨噬细胞共培养时,免疫荧光染色显示,干细胞工程化静电纺丝膜组M1型巨噬细胞荧光强度低于胶原组(P<0.01),M2型巨噬细胞荧光强度高于胶原组(P<0.01);qRT-PCR检测显示,干细胞工程化静电纺丝膜组促炎性基因肿瘤坏死因子α和白细胞介素1βmRNA表达低于胶原组(P<0.001),抑炎性基因白细胞介素10和转化生长因子βmRNA表达高于胶原组(P<0.001);④结果表明,干细胞工程化的两面异性人工硬脊膜可模拟正常硬脊膜的定向结构,内表面利于细胞生长黏附,外表面避免组织粘连,同时通过间充质干细胞旁分泌组分促进巨噬细胞向M2亚型极化,调控局部的炎症微环境。

血清BALP、BGP、β-CTx联合FRAX预测绝经后骨质疏松症患者髋部骨折风险的价值

目的探讨血清骨碱性磷酸酶(BALP)、骨钙素(BGP)、β-Ⅰ型胶原羧基端肽(β-CTx)联合骨折风险评估工具(FRAX)预测绝经后骨质疏松症髋部骨折风险的价值。方法选取2020年1月至2022年12月该院收治的绝经后骨质疏松症患者132例纳入骨质疏松组,另选择同期在该院诊断为骨量下降的123例和骨量正常的120例绝经后女性分别纳入骨量下降组和对照组。比较各组血清BALP、BGP、β-CTx水平及FRAX检测的髋部骨折风险。对绝经后骨质疏松症患者进行4~36个月的随访,根据骨质疏松症患者髋部是否骨折将其分为骨折组和未骨折组。采用多因素Logistic回归分析绝经后骨质疏松症患者髋部骨折的影响因素,绘制受试者工作特征(ROC)曲线分析血清BALP、BGP、β-CTx与FRAX单独及联合对绝经后骨质疏松症患者髋部骨折的预测价值。结果骨质疏松组血清BALP、BGP、β-CTx水平和FRAX检测的髋部骨折风险均高于骨量下降组和对照组,差异有统计学意义(P<0.05),骨量下降组血清BALP、BGP、β-CTx水平和FRAX检测的髋部骨折风险均高于对照组,差异有统计学意义(P<0.05)。132例绝经后骨质疏松症患者随访过程中,共有44例发生髋部骨折,发生率为33.33%。骨折组BALP、BGP、β-CTx水平和FRAX检测的髋部骨折风险均高于未骨折组,差异有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,父母有髋部骨折史,有脆性骨折史、长期糖皮质激素治疗史、类风湿关节炎史,无规律补钙、缺乏阳光照射及血清BALP、BGP、β-CTx水平升高均是绝经后骨质疏松症患者髋部骨折的危险因素(P<0.05),骨密度(BMD)升高、体育锻炼则是绝经后骨质疏松症患者髋部骨折的保护因素(P<0.05)。ROC曲线分析结果显示,血清BALP、BGP、β-CTx联合FRAX预测绝经后骨质疏松症患者髋部骨折的灵敏度、曲线下面积(AUC)分别为95.45%、0.954,均高于单独预测(P<0.05)。结论血清BALP、BGP、β-CTx及FRAX均对绝经后骨质疏松症患者髋部骨折发生风险具有一定的预测价值,且4项指标或方法联合检测的预测价值更高。

骨质疏松症患者PINP、25-(OH)VitD_(3)、BMP-2与中医辨证分型的相关性研究

目的:分析骨质疏松症患者血清总I型胶原氨基末端前肽(PINP)、25-羟维生素D3[25-(OH)VitD_(3)]及骨形态发生蛋白2(BMP-2)与其中医辨证分型的相关性。方法:收集我院2020年8月~2023年8月医院收治的157例骨质疏松症患者的一般资料及中医四诊资料进行回顾性分析,并统计骨质疏松症患者中医辨证分型结果,对不同症型患者基本资料、PINP、25-(OH)VitD_(3)、BMP-2水平进行比较。结果:157例骨质疏松症患者中医辨证分型属肾阳虚证34例,脾肾阳虚证43例,肝肾阴虚证49例,血瘀气滞证31例。不同中医辨证分型的骨质疏松患者血清PINP、25-(OH)VitD_(3)及BMP-2水平整体相比,差异有统计学意义(P<0.05);其中,血瘀气滞组PINP水平显著高于肾阳虚组、脾肾阳虚组及肝肾阴虚组患者(P<0.05),而其余3组两两间相比,差异无统计学意义(P>0.05);脾肾阳虚证患者25-(OH)VitD_(3)水平明显低于肾阳虚组、肝肾阴虚组及血瘀气滞组(P<0.05),且肾阳虚组低于肝肾阴虚组及血瘀气滞组(P<0.05),而其余两组相比差异无统计学意义(P>0.05);血瘀气滞组BMP-2水平显著低于肾阳虚组、脾肾阳虚组及肝肾阴虚组患者(P<0.05),而其余3组两两间相比,差异无统计学意义(P>0.05);二分类logistics回归分析结果显示,血瘀气滞与PINP呈正相关(P<0.05),与BMP-2呈负相关(P<0.05);脾肾阳虚与25-(OH)VitD_(3)呈负相关(P<0.05)。结论:骨质疏松症患者的血清PINP、25-(OH)VitD_(3)、BMP-2水平与其中医辨证分型具有一定相关性,可作为评估患者中医辨证分型的参考指标。

TGF-β_1、BMP-2和TypeⅡ Collagen在黄韧带中的表达及意义

目的研究TGF-1β、BM P-2和typeⅡco llagen在退行性腰椎滑脱(degenerative lum bar spondy lo listhes is,DLS)和腰椎间盘突出症(lum bar d isc hern iation,LDH)黄韧带中的表达及其意义。方法37例手术切除的腰椎椎板间部黄韧带标本分为3组,第1组为退行性腰椎滑脱组(DLS)10例;第2组为腰椎间盘突出症组(LDH)17例,第3组为正常对照组10例,其中7例取自腰椎骨折手术病人,3例取自意外死亡者。应用EnV is ion二步免疫组化的方法检测其TGF-1β、BM P-2和typeⅡco llagen的表达情况,普通光镜观察,计算出各标本的表达阳性率和表达强度,数据以x-±s标准差及表达强度表示,结果分别用Spss统计软件和R id it进行分析。结果TGF-1β、BM P-2和typeⅡco llagen的阳性表达产物见于成纤维细胞、成软骨细胞和软骨细胞中,而Ⅱ型胶原染色还可同时见于基质。TGF-1β、BM P-2和typeⅡco llagen在DLS组中的表达明显高于LDH组和正常组(P<0.01或P<0.05),Ⅱ型胶原基质染色明显深于LDH组和对照组。LDH组的TGF-1β和typeⅡco llagen的表达阳性率和表达强度与正常组之间差异无显著性(P>0.05),而BM P-2的表达阳性率和表达强度在LDH组与正常组之间具有统计学意义(P<0.01)。结论黄韧带所受到的异常机械牵张力可以增加TGF-1β在黄韧带细胞中的合成,而TGF-1β则促进退行性腰椎滑脱黄韧带中的Ⅱ型胶原合成,导致黄韧带的退变和肥厚。BM P-2在退变黄韧带中的表达异常增高,可能与黄韧带的软骨化倾向有关。

成纤维细胞生长因子受体1 抑制剂对胶原诱导关节炎模型大鼠骨破坏的影响

背景:课题组前期的研究表明靶向成纤维细胞生长因子受体1(fibroblast growth factor receptor 1,FGFR1)可能是治疗类风湿性关节炎的有效靶点。目的:探讨FGFR1抑制剂(PD173074)对胶原诱导关节炎模型大鼠骨破坏的影响。方法:将25只雌性SD大鼠随机分为5组,正常对照组、模型组、甲氨蝶呤组、PD173074低剂量组、PD173074高剂量组。除正常对照组外,其余各组大鼠建立Ⅱ型胶原诱导关节炎模型。造模成功后正常组及模型组大鼠腹腔注射无菌PBS,甲氨蝶呤组药物注射剂量为1.04 mg/kg,PD173074低剂量组和高剂量组药物注射剂量分别为5,20 mg/kg,1次/周。给药4周后取材,观察大鼠临床症状以及关节肿胀情况,踝关节Micro-CT三维重建及分析,观察踝关节病理变化,检测关节周围血管生成情况及核因子κB受体活化因子配体的表达,检测关节滑膜中p-FGFR1、血管内皮生长因子A、抗酒石酸酸性磷酸酶的表达,观察肝、脾、肾病理变化并计算肝、脾、肾指数。结果与结论:①PD173074能够减轻模型大鼠踝关节临床症状及关节肿胀,延缓骨质丢失,改善骨结构,减轻关节滑膜侵袭以及软骨骨侵蚀,降低关节周围破骨细胞数量,抑制关节滑膜组织中的血管生成,降低核因子κB受体活化因子配体的表达,抑制FGFR1磷酸化蛋白、抗酒石酸酸性磷酸酶和血管内皮生长因子A的蛋白表达。②大鼠肝、脾、肾病理观察表明经过PD173074治疗后无明显的毒副作用。③研究证明了FGFR1抑制剂能够延缓Ⅱ型胶原诱导关节炎模型大鼠关节炎症及骨破坏的进展,并抑制血管的生成。初步验证了PD173074在Ⅱ型胶原诱导关节炎模型中的治疗作用,其可能是通过抑制FGFR1磷酸化发挥作用,为寻找类风湿性关节炎新的治疗靶点提供了方向。

超声骨刀微创拔牙法联合医用胶原蛋白海绵在下颌阻生牙拔除中的应用效果

目的 探究超声骨刀微创拔牙法联合医用胶原蛋白海绵应用于下颌阻生牙拔除的效果。方法 选取2020年12月至2022年12月四川省南充市中心医院收治的150例下颌阻生牙患者为研究对象,采用随机数字表法将其分为两组,各75例。患者均采用超声骨刀微创拔牙法拔除患牙,对照组术中予以常规治疗,试验组术中联合医用胶原蛋白海绵治疗。术后30 min比较两组口腔出血情况;术后24、72 h,评估患者的口腔疼痛程度[视觉模拟评分法(VAS)];术后72 h评估两组开口受限程度;记录术后并发症。结果 试验组术后出血情况优于对照组(P<0.05)。两组不同时间点VAS评分比较,差异有统计学意义(P<0.05);术后24、72 h,试验组VAS评分低于对照组(P<0.05)。试验组术后开口受限严重程度优于对照组(P<0.05)。试验组术后并发症发生率均低于对照组(P<0.05)。结论 超声骨刀微创拔牙法联合医用胶原蛋白海绵应用于下颌阻生牙拔除的治疗中,更有助于降低患者术后的疼痛程度、出血率及并发症发生风险。