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Brain Derived Neurotrophic Factor Levels in Aged Rats Post-Systemic Human Mesenchymal Stem Cell Administration 人间充质干细胞注射后老龄大鼠脑源性神经营养因子水平的变化
Pub Date : 2019-02-23 DOI: 10.20473/JSCRTE.V2I2.11895
Adisti Dwijayanti
Brain-Derived Neurotrophic Factor (BDNF) levels were affected by aging. Brain BDNF levels were known to decrease along with advanced age thus correlated with any diseases such as cognitive impairment and Alzheimer. Mesenchymal Stem Cell (MSC) is one of the potential modalities actively investigated against age-related diseases. This study evaluated the effect of human MSC administration to brain BDNF levels in aged rats. Intravenous injection of 10 million per body weight human MSC were given four times in 3 months interval to 22-24 months old female and male Spraque–Dawley rats. As control group, aged rats were injected by normal saline at the same volume and frequencies. Moreover, young 3-6 months rats also examined as negative control.  By the end of the experiment, we analyzed three rats from each group. Brain BDNF levels were measured by enzyme-linked immunosorbent assay and normalize to the protein levels. One-way ANOVA and LSD post hoc analysis was performed to compare the differences between groups. BDNF levels in male appeared similar between young, aged, and MSC treated groups. Meanwhile, control aged female groups had significantly lower BDNF levels compared to young (p = 0.019) and MSC-treated aged rats (p = 0.001). There was no difference of BDNF levels between young and MSC-treated aged in female rats (p = 0,068). Both sex had similar BDNF levels (p = 0.249) in control-aged groups. In contrast, female young and MSC-treated aged rats achieved significantly higher BDNF levels (p = 0.009 and p <0.001) compared to the male groups, respectively. These results suggest that human mesenchymal stem cell intravenous injection can increase brain BDNF levels in female aged rats.
脑源性神经营养因子(BDNF)水平受年龄影响。大脑BDNF水平随着年龄的增长而下降,因此与认知障碍和阿尔茨海默病等疾病有关。间充质干细胞(MSC)是被积极研究的对抗年龄相关疾病的潜在方式之一。本研究评估了人骨髓间充质干细胞给药对老龄大鼠脑BDNF水平的影响。22-24月龄雌、雄Spraque-Dawley大鼠静脉注射人骨髓间充质干细胞每体重1000万个,间隔3个月4次。老龄大鼠按相同体积和频率注射生理盐水作为对照组。3 ~ 6月龄大鼠作为阴性对照。实验结束时,我们每组分析3只大鼠。采用酶联免疫吸附法测定脑BDNF水平,并将其归一化至蛋白水平。采用单因素方差分析和LSD事后分析比较组间差异。男性BDNF水平在年轻、年老和MSC治疗组之间相似。同时,对照组老年雌性大鼠的BDNF水平明显低于年轻大鼠(p = 0.019)和msc处理的老年大鼠(p = 0.001)。雌性大鼠的BDNF水平在年轻和msc处理的老年大鼠之间无差异(p = 0.068)。在对照组中,男女的BDNF水平相似(p = 0.249)。相比之下,雌性年轻大鼠和msc处理的老年大鼠的BDNF水平分别显著高于雄性组(p = 0.009和p <0.001)。这些结果提示静脉注射人间充质干细胞可增加雌性老龄大鼠脑BDNF水平。
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引用次数: 1
COLLAGEN ANALYSIS OF GRAFT IN BONE TUNNEL MODEL ANTERIOR CRUCIATE LIGAMENT (ACL) RECONSTRUCTION WITH INTRATUNNEL ALLOGENIC BONE MARROW MESENCHYMAL STEM CELLS (MSCs) AND VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) 同种异体骨髓间充质干细胞(MSCs)和血管内皮生长因子(VEGF)重建骨隧道模型前交叉韧带(ACL)的胶原蛋白分析
Pub Date : 2018-09-18 DOI: 10.20473/JSCRTE.V2I1.9259
petrasama petrasama
ACL reconstruction measures aim to obtain stable knees, and reduce the risk of further injury to the meniscus and joint surfaces. Acceleration of the integration process between the tendon graft and the bone tunnel will improve the final result of ACL reconstruction. The purpose of this study was to investigate the collagen composition of the bone tunnel graft model of anterior cruciate ligament reconstruction with intravenous allogenic bone marrow mesenchymal stem cells and vascular endothelial growth factor in experimental animals. The design of this study was Post-test only Control Group Design using 20 rabbits divided into treatment group and control group. Collagen immunohistochemical evaluation was performed at weeks 3 and 6. Evaluation at week 3 obtained the area of collagen type-1 in the higher treatment group at treatment (p <0.001). In the 6th week evaluation, it was found that the area of collagen type-1 in the treatment group was higher (p <0.05). Type-1 collagen at week 6 did not differ significantly with week 3 (p> 0.05). Provision of allogenic bone marrow mesenchymal stem cells and intratonal vascular endothelial growth factor in ACL reconstruction enhanced the formation of collagen type-1 which is the acceleration of incorporation of the graft tendon process with bone tunnel.Keywords : Anterior Cruciate Ligament, allogenic bone marrow mesenchymal stemcells, vascular endothelial growth factor, graft and collagen.
ACL重建措施的目的是获得稳定的膝关节,并减少进一步损伤半月板和关节表面的风险。加速肌腱移植物与骨隧道之间的融合过程将改善ACL重建的最终结果。本研究旨在探讨静脉注射同种异体骨髓间充质干细胞和血管内皮生长因子重建前交叉韧带骨隧道移植模型的胶原组成。本研究采用后验组设计,将20只家兔分为治疗组和对照组。在第3周和第6周进行胶原免疫组化评价。第3周评价高剂量组治疗时1型胶原面积(p < 0.05)。同种异体骨髓间充质干细胞和血管内皮生长因子在ACL重建中促进了1型胶原的形成,加速了移植物肌腱突与骨隧道的融合。关键词:前十字韧带,同种异体骨髓间充质干细胞,血管内皮生长因子,移植物,胶原
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引用次数: 0
INTRATUNNEL THE EFFECT OF ADMINISTRATION OF BONE MARROW MESENCHYMAL STEM CELLS (BM-MSCs) AND VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) TENDON-BONE TO INTERFACE HISTOLOGICAL GRAFT ANTERIOR CRUCIATE LIGAMENT APPEARANCE AFTER RECONSTRUCTION IN RABBITS 骨髓间充质干细胞(bmmscs)和血管内皮生长因子(VEGF)对兔重建后组织移植前交叉韧带外观的影响
Pub Date : 2018-09-18 DOI: 10.20473/jscrte.v2i1.9258
A. Abirama
The success of the Anterior Cruciate Ligament (ACL) reconstruction using a tendon graft is determined by integration in the bone tendon-graft interface on the bone tunnel. The use of stem cells and growth factors proved to accelerate the healing of the bone tendon-graft interface. The aim of this study was to inveestigate the difference of histology picture in the tendon-bone tunnel model after ACL reconstruction with intratunnel intravenous allogenic bone marrow mesenchymalstemcells (BM-MSCs) and intratunnel vascular endothelial growth factor (VEGF). This research used Post-Test Only Control Group design with 20 rabbits divided into treatment group and control group. Each group performed histologic image evaluation (thickness of collagen fiber or sharpey fiber) at week 3 and 6. Evaluation of histology overview at week 3 and week 6 showed a significantly thicker thickness of collagen fiber or sharpey fiber in treatment group compared with control group (p <0.05). Intravenous administration of BM-SCs and VEGF after ACL reconstruction can speed healing of the bone tunnel significantly from week 3 and 6. The study by Faridyan et al has concluded that intravenous BM-SCs + VEGF increased ultimate tension strength in the bone-tendon interface significantly. In this study, intravenous administration of BM-SCs and VEGF gave histologic images showing acceleration of bone tunnel healing.Keywords:Anterior cruciate ligament reconstruction, allogenic bone marrow mesenchymal stem cells, vascular endothelial growth factor, graft tunnel healing, and Sharpey fiber.
使用肌腱移植重建前交叉韧带(ACL)的成功与否取决于骨隧道上的骨肌腱-移植物界面的整合。干细胞和生长因子的使用被证明可以加速骨肌腱-移植物界面的愈合。本研究旨在探讨应用同种异体骨髓间充质干细胞(bmscs)和血管内皮生长因子(VEGF)重建前交叉韧带后肌腱-骨隧道模型组织学图像的差异。本研究采用后验对照组设计,将20只家兔分为治疗组和对照组。各组于第3周和第6周进行组织学图像评估(胶原纤维或锐纤维厚度)。第3周和第6周组织学评价显示,治疗组胶原纤维或锐纤维厚度显著高于对照组(p <0.05)。ACL重建后静脉给药BM-SCs和VEGF可在第3周和第6周显著加速骨隧道愈合。Faridyan等人的研究表明,静脉注射BM-SCs + VEGF可显著提高骨-肌腱界面的极限拉伸强度。在这项研究中,静脉给药BM-SCs和VEGF的组织学图像显示骨隧道愈合加速。关键词:前交叉韧带重建,同种异体骨髓间充质干细胞,血管内皮生长因子,移植物隧道愈合,Sharpey纤维。
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引用次数: 0
THE EFFECT OF INJECTION OF INTRA ARTICULAR ALLOGENIC BONE MARROW MESENCHYMAL STEM CELL-PLATELET CELL RICH PLASMA (BMSCs-PRP) ON FULL-THICKNESS ARTICULAR CARTILAGE DEFFECT REGENERATION IN RABBIT 关节内注射同种异体骨髓间充质干细胞-富血小板细胞血浆(BMSCs-PRP)对兔全层关节软骨缺损再生的影响
Pub Date : 2018-09-18 DOI: 10.20473/JSCRTE.V2I1.9260
S. Taufik, D. Utomo
The joint cartilage defectfullthickness is still a problem today because its currenttreatment still has not delivered maximum results. Current treatment uses cartilage enginering using mesenchymal stem cells alone and or combining growth factor. The aim of this study was to investigate the effect of intra-articular injection of Allogenic bone marrow mesenchymal stem cell - Platelet rich plasma (BMSCs-PRP) on regeneration of cartilage defect fullthickness in rabbits. The design of this study was a post-test only control group design using 36 New Zealand white rabbits divided into three groups. Each group were treated with PRP, BMSCs and BMSCs-PRP. Results were evaluated after 10 weeks. In the evaluation, macroscopic images showed the best healing in the BMSCs-PRP group. Histopathologic examination showed that in the MSCs-PRP group, there was a significant increase in the number of chondrocytes (p = 0,000), cartilage area (p = 0,000), as well as the number of Agecoprogenitorexpec- tion cells (p = 0,000) and collagen type 2 (p = 0,000). BMSCs were able to differentiate into condroblasts which then synthesize aggressive and collagen type 2. Platelet rich plasma (PRP) contains growth factor BMP, TGF, FGF and IGF which can accelerate the occurrence of MSCs differentiation. Intra-articular injections Allogenic bone marrow mesenchymal stem cell (BMSCs-PRP) is able to regenerate and cure full-thickness joint cartilage defects through differentiation of MSCs into condroblasts.Keyword: Allogenic, Bone marrow Mesenchymal stem cell, Cartilage, Platelet rich plasma, Full-thickness.
关节软骨缺损厚度至今仍是一个问题,因为目前的治疗仍未达到最大效果。目前的治疗方法是软骨工程,使用单独的间充质干细胞和或联合生长因子。本研究旨在探讨同种异体骨髓间充质干细胞-富血小板血浆(BMSCs-PRP)对兔软骨缺损全层再生的影响。本研究采用后验纯对照组设计,36只新西兰大白兔分为3组。各组分别采用PRP、BMSCs和BMSCs-PRP治疗。10周后评估结果。在评价中,宏观图像显示BMSCs-PRP组的愈合效果最好。组织病理学检查显示,MSCs-PRP组软骨细胞数量(p = 0000)、软骨面积(p = 0000)、agecoprogenrepec细胞数量(p = 0000)和2型胶原蛋白数量(p = 0000)均显著增加。骨髓间充质干细胞能够分化成成软骨细胞,然后合成侵袭性和2型胶原。富血小板血浆(PRP)中含有BMP、TGF、FGF、IGF等生长因子,可加速MSCs分化的发生。同种异体骨髓间充质干细胞(BMSCs-PRP)通过向成软骨细胞的分化,能够再生和治愈全层关节软骨缺损。关键词:同种异体,骨髓间充质干细胞,软骨,富血小板血浆,全层。
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引用次数: 0
INFLUENCE OF LOW OXYGEN CONDITION OF BONE MARROW MESENCHYMAL STEM CELL 低氧条件对骨髓间充质干细胞的影响
Pub Date : 2018-09-18 DOI: 10.20473/JSCRTE.V2I1.9267
Nugroho Setyowardoyo
The state of low oxygen levels known as hypoxia in humans is considered a dangerous condition is apparently a normal physiological condition and required by the stem cells as they are in the body. Mesencyimal Stem Cells (MSCs) require physiologically optimal conditions of low O2 tension of 1-3% in the bone marrow. The purpose of this study was to reveal the difference between in vitro culture of MSC in normoxia condition (20% O 2 concentration) with hypoxia condition (1% O2 content) especially in terms of viability, pluripotent properties, and MSC proliferation ability of the culture it produces. This research is an explorative laboratory research invitro on Bone Marrow Mesenchymal Stem Cells (BMSCs) culture using hypoxia condition. The study design used Randomized Control Group Post-Test Only Design. This research was conducted for 2 months. There was a significant difference in mean slow proliferation based on the number of Least-like CFU-Cs between the control group, treatment group 1 and treatment group 2, the mean percentage of the number of cells expressing the OCT4 coding gene on immunofluorosense examination between the control group, 1 and the treatment group 2 and the mean percentage of cell numbers expressing the OCT4 coding gene on the immunofluorosense examination between the control group, the treatment group 1 and the treatment group 2 showing p <0.01. There was a significant difference of percentage of non-absorbing color cell number of trypsin blue (viable cells) between control group, treatment group 1, and treatment group 2 showing p value <0.05. This suggests that the precondition of culture with normoxia provides an opportunity for cells to adapt and proliferate before being conditioned in hypoxic cultures. Cultures with hypoxic conditions and preconditions of normoxia are the best culture conditions because they produce cells that are capable of maintaining pluripotency properties while still having better proliferation and viability capability compared with direct hypoxia conditions.Keywords: Hypoxia, normoxia, bone marrow, mesenchymal stem cells.
人体的低氧状态,即缺氧,被认为是一种危险的状态,显然是一种正常的生理状态,是干细胞在体内所需要的。间充质干细胞(MSCs)需要骨髓中1-3%的低氧张力的生理最佳条件。本研究的目的是揭示常氧条件下(20% O2浓度)与缺氧条件下(1% O2含量)间充质干细胞体外培养的差异,特别是在培养物的活力、多能性和MSC增殖能力方面。本研究为缺氧条件下骨髓间充质干细胞(BMSCs)体外培养的探索性实验室研究。研究设计采用随机对照组测试后设计。本研究历时2个月。以对照组、治疗组1和治疗组2之间的least -样CFU-Cs的数量计算,对照组、治疗组1和治疗组2之间在免疫荧光检查中表达OCT4编码基因的细胞数量的平均百分比,对照组和治疗组之间在免疫荧光检查中表达OCT4编码基因的细胞数量的平均百分比,对照组之间在免疫荧光检查中表达OCT4编码基因的细胞数量的平均百分比,治疗1组与治疗2组比较p <0.01。对照组、处理1组和处理2组胰蛋白酶蓝非吸收色细胞数(活细胞)百分比差异有统计学意义(p值<0.05)。这表明,常氧培养的前提条件为细胞在缺氧培养之前适应和增殖提供了机会。低氧条件和常氧条件下的培养是最好的培养条件,因为它们产生的细胞能够保持多能性,同时与直接缺氧条件相比仍具有更好的增殖和生存能力。关键词:缺氧,常氧,骨髓,间充质干细胞。
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引用次数: 1
BIOMECHANIC STUDY OF GRAFT BONE TUNNEL MODEL IN ANTERIOR CRUCIATE LIGAMENT RECONSTRUCTION USING INTRATUNNEL ALLOGENIC BONE MARROW MESENCHYMAL STEM CELLS (BM-MSCs) AND VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) 同种异体骨髓间充质干细胞(bmscs)和血管内皮生长因子(VEGF)重建前交叉韧带移植骨隧道模型的生物力学研究
Pub Date : 2018-09-18 DOI: 10.20473/jscrte.v2i1.9262
brian vicky faridyan
Successful anterior cruciate ligament (ACL) reconstruction using tendon graft requires good and rapid integration between the tendon graft and the bone tunnel. The strength of the tendon-bone tunnel graft in the initial phase is very important to facilitate aggressive rehabilitation and as early as possible to support rapid recovery to normal activities. The objective of this study was to determine ultimate tension strength (UTS) on the femoral tendon-bone tunnel graft model after reconstruction of anterior cruciate ligament (ACL) by administering allogenic bone marrow mesenchymal stemcells (BM-MSCs) and vascular endothelial growth factor (VEGF) intratunnel in experimental animals. The design of this research was Post-Test Only Control Group Design using 24 rabbits divided into treatment and control group. Biomechanical evaluation was done at week 3 and 6. Evaluation at week 3 found ultimate tension strength of treatment group significantly higher than control (p <0,05). In the 6th week evaluation, Ultimate tension strength was found that the treatment group significantly higher than the control group (p <0.05). Ultimate tension strength at week 3 did not differ significantly with week 6 (p> 0.05). Intravenous administration of BM-MSCs and VEGF on ACL reconstruction increased ultimate tension strength in graft-bone tunnel significantly since week 3. The study of Ferdiansis et al using BM-MSCs and VEGF intraarticular, only showed a significant increase in ultimate tension strength in graft-bone tunnel since week 6. Comparison of this method indicates acceleration in incorporation of tendon graft with bone tunnel on intratunnel method better thaninvitro intraarticular method.Keywords : Anterior cruciate ligament, allogenic bone marrow mesenchymal stem cells, vascular endothelial growth factor and biomechanic study.
成功的前交叉韧带(ACL)重建需要肌腱移植物和骨隧道之间良好和快速的融合。肌腱-骨隧道移植物在初始阶段的强度对于促进积极康复和尽早支持快速恢复正常活动非常重要。本研究的目的是通过给药同种异体骨髓间充质干细胞(bmmscs)和血管内皮生长因子(VEGF)在实验动物的前交叉韧带(ACL)重建后股骨肌腱-骨隧道移植模型的极限张力(UTS)。本研究采用验后对照组设计,将24只家兔分为实验组和对照组。在第3周和第6周进行生物力学评价。第3周时,治疗组抗拉强度显著高于对照组(p < 0.05)。自第3周起,静脉给药BM-MSCs和VEGF重建ACL可显著增加移骨隧道的极限拉伸强度。Ferdiansis等使用BM-MSCs和VEGF在关节内进行的研究仅显示从第6周开始移植物骨隧道的极限拉伸强度显著增加。结果表明,隧道内法比体外关节内法更有利于肌腱移植物与骨隧道的结合。关键词:前交叉韧带,同种异体骨髓间充质干细胞,血管内皮生长因子,生物力学研究
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引用次数: 0
Injectable Bone Substitute Paste Based on Hydroxyapatite, Gelatin and Streptomycin for Spinal Tuberculosis 以羟基磷灰石、明胶和链霉素为基础的可注射骨替代物治疗脊柱结核
Pub Date : 2015-11-02 DOI: 10.4172/2165-7939.1000266
H. Maulida, D. Hikmawati, A. S. Budiatin
World Health Organization (WHO) in 2005 reported that cases of tuberculosis (TB) in the world occur more than 8 million annually and 5-10% were attacked in spine. The most effective treatment of spinal TB is evacuation of infected bone segments and fill with bone graft. It has been synthesized and characterized of injectable bone substitute (IBS) paste based on hydroxyapatite, gelatin and streptomycin. IBS paste synthesized by mixing hydroxyapatite and gelatin 20% w/v with 75:25, 70:30, 65:35 and 60:40 ratio and streptomycin 10 wt%. The mixture was then added with hydroxypropyl methylcellulose (HPMC) 4% w/v as suspending agent. In vitro characterization performed include acidity (pH), injectability test, setting time, cytotoxicity (MTT assay) and microbacterium test. Acidity test results indicate a fourth variation of the samples had pH values approaching normal body pH (7,3 to 7,6) and is able to maintain stability when measured in 7 days. Injectability test results indicate IBS paste is injectable with the highest percentage of the injectability value at 97,74% ± 0,19%. IBS paste has been setting within 30 minutes to 1 hour when injected on hydroxyapatite scaffold that resembles the bone cavity and is able to cover the pore scaffold seen from the Scanning Electron Microscope (SEM). Scaffold pore size is smaller from range of 780,8 to 835,4 μm into 225,2 μm. MTT assay results showed that IBS paste is not toxic and experiencing proliferation (viability >100%) that are expected to trigger osteoblast cell growth when applied. Microbacterium test results showed that IBS paste is an antibacterial seen from inhibition zone diameter of Staphylococcus aureus and has a high strength-sensitive antibacterial. Thus, hydroxyapatite, gelatin and streptomycin composites had qualified as injectable bone substitute which applied in cases of spinal tuberculosis.
世界卫生组织(WHO) 2005年报告说,全世界每年发生的结核病病例超过800万例,其中5-10%的病例发生在脊柱。脊髓结核最有效的治疗方法是清除感染的骨节段并用骨移植物填充。合成了以羟基磷灰石、明胶和链霉素为基材的可注射性骨替代物(IBS)浆料,并对其进行了表征。将羟基磷灰石和明胶按75:25、70:30、65:35和60:40的比例和链霉素10 wt%混合,w/v为20%,合成IBS膏体。然后加入羟丙基甲基纤维素(HPMC) 4% w/v作为悬浮剂。体外鉴定包括酸度(pH)、注射性试验、凝固时间、细胞毒性(MTT试验)和微细菌试验。酸度测试结果表明,第四种样品的pH值接近正常身体pH值(7,3至7,6),并且能够在7天内保持稳定。注射性试验结果表明,IBS膏体是可注射的,注射性值的最高百分比为97.74%±0.19%。从扫描电镜(SEM)可以看出,IBS浆料注射在类似于骨腔的羟基磷灰石支架上,并能够覆盖孔支架,在30分钟至1小时内凝固。支架孔径在780、8 ~ 835、4 μm ~ 225、2 μm范围内变小。MTT试验结果显示,IBS膏体无毒且具有增殖能力(生存能力>100%),应用后有望引发成骨细胞生长。微菌试验结果表明,从对金黄色葡萄球菌的抑制带直径来看,IBS膏是一种抗菌药物,具有高强度敏感性的抗菌药物。因此,羟基磷灰石、明胶和链霉素复合材料可作为注射用骨替代物应用于脊柱结核病例。
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引用次数: 18
期刊
Journal of Stem Cell Research and Tissue Engineering
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