骨髓间充质干细胞来源外泌体对炎症微环境中巨噬细胞表型及软骨细胞的调控作用

目的 研究骨髓间充质干细胞来源外泌体（BMSCs-Exo）在炎症微环境中对巨噬细胞表型极化及软骨修复的调控作用。方法 差速离心法提取BMSCs-Exo,透射电镜观察外泌体形态,Western blot检测Alix、TSG101表达;使用不同浓度的BMSCs-Exo（10 μg/mL 和50 μg/mL）处理M1巨噬细胞及IL-1β刺激的软骨细胞,CCK-8测定BMSCs-Exo促巨噬细胞增殖能力,Western blot和qRT-PCR验证BMSCs-Exo对M1型巨噬细胞表型极化的影响,炎症相关基因IL-1β、IL-6的表达水平,以及成软骨相关因子COL II、MMP-13、TGF-β1的表达水平。结果 BMSCs-Exo为圆形双层囊泡,表达Alix、TSG101,可促进巨噬细胞增殖。与对照组比较,M1型巨噬细胞经BMSCs-Exo处理后,M1型极化标记物（iNOS）的表达水平降低,M2型极化标记物（Arg-1）的表达水平升高（P<0.05）;炎症相关基因IL-1β、IL-6的表达水平显著降低（P<0.05）,炎症软骨细胞中的COL II、TGF-β1表达升高,MMP-13表达水平降低（P<0.05）。结论 BMSCs-Exo可调控M1型巨噬细胞向M2型巨噬细胞极化,降低炎症因子表达水平,促进炎症微环境中软骨修复。     

Bone marrow mesenchymal stem cells-derived exosomes for penetrating and targeted chemotherapy of pancreatic cancer

Pancreatic ductal adenocarcinoma (PDAC) is one of the most intractable malignancy, with an only 6% 5-year relative survival rate. The dismal therapeutic effect is attributed to the chemotherapy resistance and unique pathophysiology with abundant inflammatory cytokines and abnormal hyperplasia of extracellular matrix (ECM). Based on the theory that bone marrow mesenchymal stem cells (BM-MSCs) can influence the tumorous microenvironment and malignant growth of PDAC, we employed exosomes (Exos) derived from BM-MSCs as PDAC-homing vehicles to surpass the restrictions of pathological ECM and increase the accumulation of therapeutics in tumor site. To overcome chemoresistance of PDAC, paclitaxel (PTX) and gemcitabine monophosphate (GEMP)—an intermediate product of gemcitabine metabolism—were loaded in/on the purified Exos. In this work, the Exo delivery platform showed superiorities in homing and penetrating abilities, which were performed on tumor spheroids and PDAC orthotopic models. Meanwhile, the favorable anti-tumor efficacy in vivo and in vitro, plus relatively mild systemic toxicity, was found. Loading GEMP and PTX, benefitting from the naturally PDAC selectivity, the Exo platform we constructed performs combined functions on excellent penetrating, anti-matrix and overcoming chemoresistance (Scheme 1). Worth expectantly, the Exo platform may provide a prospective approach for targeted therapies of PDAC.     

人脐带间充质干细胞来源的外泌体调控血管内皮生长因子对小鼠胫骨骨折的修复作用研究

目的探讨人脐带间充质干细胞( huc-MSC)来源外泌体( Exo)对小鼠胫骨骨折的修复作用及可能机制。方法于 2018年 12月至 2022年 1月,体外分离、培养 hucMSCs,超速离心法收集 Exo并鉴定。 24只 8周龄雄性 C57BL/6J小鼠行胫骨骨折,并采用随机数字表法分为模型对照组、磷酸缓冲盐溶液( PBS)对照组及 huc-MSC-Exo组,每组各 8只,其中 huc-MSC-Exo组小鼠骨髓腔内注射 huc-MSC-Exo(100 μg溶解于 100 μL PBS),PBS对照组小鼠注射等量 PBS,模型对照组小鼠不进行处理。 21 d后, X线观察骨折愈合情况;双能 X线检测骨矿物质密度( BMD);苏木精 -伊红( HE)染色观察胫骨组织学变化;免疫组织化学染色检测骨折区骨组织中血管内皮生长因子 A(VEGFA)阳性表达情况;蛋白质印迹法检测骨折区骨组织中 VEGFA、Runt相关转录因子 2(RUNX2)、骨形成蛋白 2(BMP-2)和骨桥蛋白( OPN)蛋白表达水平。结果经鉴定,成功提取 huc-MSC-Exo;模型对照组和 PBS对照组小鼠胫骨骨折断端明显、骨膜增厚,骨折带的两侧可见大量未分化的间充质细胞及部分软骨细胞,骨痂的软骨内骨区域软骨细胞形态肥大,而 huc-MSC-Exo组小鼠胫骨骨折区愈合明显,外侧骨皮质连续性良好,部分软骨细胞已被吸收,膜内成骨区域出现骨痂;另与模型对照组(VEGFA 0.23±0.02)比较, PBS对照组(VEGFA 0.24±0.02)无明显变化(P>0.05)huc-MSC-Exo组小鼠胫骨骨折断端恢复良好,骨折线基本消失,同时骨折愈合评分、 BMD、VEGFA IRS评分、 VEGFA(0.48±0.04,)、 RUNX2、 BMP-2和 OPN蛋白相对表达水平均升高( P<0.05)。结论 huc-MSC-Exo可加快小鼠胫骨骨折修复,其作用机制可能与提高 VEGFA表达水平,促进血管生成有关。     

人脐带间充质干细胞旁分泌物质的作用及对肝功能衰竭的影响

脐带间充质干细胞(umbilical cord mesenchymalstem cell, UC-MSC)是从人脐带华尔通胶(Wharton jelly)中分离出的细胞,它可以表达间充质干细胞的表面标志,其生物学特性与骨髓间充质干细胞相似,可以诱导分化为多种细胞.最近国内外学者对UC-MSC的移植及诱导分化治疗进行了大量研究,如在心肌梗死、肾脏及血液系统疾病中的研究.但对UC-MSC旁分泌物质的研究却不多见.此文就UC-MSC的分泌、分化及免疫调节作用、旁分泌物质的作用及对肝功能衰竭的影响和应用前景作一综述.     

Multifunctional icariin and tanshinone IIA co-delivery liposomes with potential application for Alzheimer’s disease

The blood–brain barrier (BBB) is a protective barrier for brain safety, but it is also a major obstacle to the delivery of drugs to the cerebral parenchyma such as the hippocampus, hindering the treatment of central nervous system diseases such as Alzheimer’s disease (AD). In this work, an anti-AD brain-targeted nanodrug delivery system by co-loading icariin (ICA) and tanshinone IIA (TSIIA) into Aniopep-2-modified long-circulating (Ang2-ICA/TSIIA) liposomes was developed. Low-density lipoprotein receptor-related protein-1 (LRP1) was a receptor overexpressed on the BBB. Angiopep-2, a specific ligand of LRP1, exhibited a high binding efficiency with LRP1. Additionally, ICA and TSIIA, drugs with neuroprotective effects are loaded into the liposomes, so that the liposomes not only have an effective BBB penetration effect, but also have a potential anti-AD effect. The prepared Ang2-ICA/TSIIA liposomes appeared narrow dispersity and good stability with a diameter of 110 nm, and a round morphology. Cell uptake observations, BBB models in vitro, and imaging analysis in vivo showed that Ang2-ICA/TSIIA liposomes not only penetrate the BBB through endocytosis, but also accumulate in N2a cells or brain tissue. The pharmacodynamic analysis in vivo demonstrated that Ang2-ICA/TSIIA liposomes could improve AD-like pathological features in APP/PS1 mice, including inhibiting neuroinflammation and oxidative stress, reducing apoptosis, protecting neurons, and improving cognitive function. Therefore, Ang2-ICA/TSIIA liposomes are considered a potentially effective therapeutic strategy for AD.     

Bone marrow mesenchymal stem cells-derived exosomes improve injury of hippocampal neurons in rats with depression by upregulating microRNA-26a expression

ObjectiveBone marrow mesenchymal stem cells (BMSCs)-derived exosomes have been widely applied in disease therapies. However, the role of BMSCs-derived exosomes in depression remains obscure. This study aims to explore the mechanism of BMSCs-derived exosomal microRNA-26a (miR-26a) on hippocampal neuronal injury of depressed rats.MethodsBMSCs and their exosomes were obtained and identified. Rat models of depression were established by corticosterone injection, then injected with BMSCs-derived exosomes. The contents of superoxide dismutase (SOD), imalondialdehyde (MDA), lactate dehydrogenase (LDH), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β) in rats’ serum, hippocampal tissues and neurons were determined. The expression of miR-26a in hippocampal tissues and neurons was detected by RT-qPCR. The injury models of rat hippocampal neurons were established to figure out the role of BMSCs-derived exosomes and miR-26a in neuron apoptosis and proliferation.ResultsIn hippocampal tissues of depressed rats, miR-26a was lowly expressed, and BMSCs-derived exosomes upregulated miR-26a expression. BMSCs-derived exosomes restrained apoptosis in hippocampal tissues of depressed rats. BMSCs-derived exosomes and upregulated miR-26a elevated SOD level, lessened MDA, LDH, TNF-α and IL-1β levels, boosted hippocampal neuron proliferation and suppressed apoptosis in depressed rats.ConclusionCollectively, our study reveals that miR-26a is lowly expressed in depressed rats, and BMSCs-derived exosomes improve hippocampal neuron injury of rat with depression by upregulating miR-26a.     

Regenerative medicine through mesenchymal stem cells for bone and cartilage repair

Bone and cartilage defects are common features of bone fracture and joint diseases, such as rheumatoid arthritis or osteoarthritis, that have great social and economic impact on the aging occidental population. Despite progress in orthopedic surgery, bone and cartilage repair is a major challenge as large defects will not spontaneously heal. Recent investigations on the stromal mesenchymal stem cell (MSC) offer a new perspectivefor bone and cartilage tissue engineering. However, the standard offull healing is extremely demanding and may be achieved through the engineering of MSCs combined with scaffolds and growth factors as recombinant proteins, or using a gene therapy approach.     

VEGF基因转染间充质干细胞移植与单纯间充质干细胞移植对大鼠心肌梗死的疗效对比

目的探讨VEGF基因转染骨髓间充质干细胞与单纯骨髓间充质干细胞移植对大鼠心肌梗死的疗效。方法将SD大鼠建立心肌缺血模型,1周后随机取30只采用随机分组法分为3组,A组为联合移植组,B组为单纯干细胞移植组,C组为对照组,通过一系列检测,对比3组的疗效。结果 A组的疗效优于B组和C组,B组的疗效优于C组（P〈0.05）。结论 VEGF基因转染骨髓间充质干细胞移植比单纯骨髓间充质干细胞移植的综合疗效好。     

山药及干细胞对肾缺血再灌注损伤的保护作用

目的 研究山药灌胃预处理及移植骨髓间充质干细胞(MSCs)对大鼠肾缺血再灌注损伤的保护作用.方法 4～6周龄雌性SD大鼠60只.随机分正常对照(A)组(n=6),再灌注损伤(B)组(n=18),再灌注损伤移植MSCs(C)组(n=18)和再灌注损伤山药灌胃移植MSCs(Ⅰ))组(n=18).D组损伤前山药灌胃5 d,C、D组再灌注1 h输汴BrdU标记的MSCs,B、C、D组再灌注6、48 h及2周各采集6只鼠的血并取肾.4组标本进行血清尿素氮(BUN)及肌酐(SCr)、肾匀浆内皮素(ET)测定,常规病理切片,免疫组化检测增殖细胞核抗原(PCNA)、5-溴脱氧尿嘧啶核苷(BrdU)标记MSCs、Ⅷ因子相关抗原(FⅧ-RAg)阳件细胞情况.结果 再灌注6 h,C、D组BUN、SCr低于B组,D 组低于C组;再灌注48 h,仅B组高于A组;2周4组间无统计学意义.再灌注6 h,B、C组ET较A组高.D组与A组无统计学意义,C、D组较B组低;48h 4组间无统计学意义.再灌注6、48hC、D组于肾小球、肾间质微血管见散在BrdU标记的MSCs,2周则见较多.再灌注6、48 h及2周PCNA及FⅧ-RAg阳性细胞,C、D组较B组多,D组较C组多.结论 肾缺血冉灌注损伤时移植MSCs有保护作用,山药灌胃预处理联合MSCs移植有协同治疗作用.     

脐带间充质干细胞移植联合负压封闭引流技术治疗糖尿病足溃疡的疗效观察

目的观察脐带间充质干细胞移植( UCMSCs)联合负压封闭引流技术( VSD)治疗糖尿病足溃疡( DFU)的疗效。方法纳入 2020年 5月至 2022年 5月邯郸市中心医院收治的 86例 DFU病人,根据随机数字表法分为引流组( 43例)和联合组( 43例)。其中引流组给予 VSD,联合组给予 UCMSCs联合 VSD。比较两组创面愈合时间及创面愈合率,足部相关指标［足背动脉血管内径、血流速度、踝肱指数( ABI)］,周围血管恢复情况［密歇根神经体征评分( MNSI)、 CT血管造影( CTA)检查］,炎症指标［C反应蛋白(CRP)、白细胞( WBC)中性粒细胞百分比(NEU%)］,肾功能(尿酸、肌酐)及临床疗效。结果联合组创面愈合率［(68.42±6.95)%比( 52.78±5.35)%］明、显高于引流组(P<0.05),创面愈合时间［(27.34±2.76)d比( 31.46±3.25)d］明显低于引流组(P<0.05);引流组和联合组治疗后足背动脉血管内径［( 1.95±0.21)mm比( 1.86±0.17)mm,(2.10±0.35)mm比( 1.82±0.15)mm］、 ABI(0.87±0.26比 0.61±0.15,0.75±0.21比 0.64±0.17)均明显升高( P<0.05)且与引流组比较,联合组明显升高( P<0.05);与治疗前比较,引流组和联合组治疗后足背动脉血流速度［( 47.35±4.75)cm/s比(,52.75±5.32)cm/s,(41.78±4.27)cm/s比( 52.14±5.26) cm/s］、 MNSI评分( 4.68±0.51比 5.74±0.61,4.12±0.42比 5.89±0.65)、 CRP［( 35.78±3.62)mg/L比( 41.78±4.25)mg/L,(29.36±3.02) mg/L比( 2.14±4.31)mg/L］、 WBC［( 7.58±0.79)×109/L比( 9.45±1.06)×109/L,(6.42±0.67)×109/L比( 9.12±1.01)×109/L］、 NEU%［( 64.31±6.53)%比( 69.72±7.02)%,(58.14±5.86)%比( 70.54±7.11)%］水平均明显降低( P<0.05)且与引流组比较,联合组明显降低( P<0.05);联合组总有效率( 95.35%)明显高于引流组( 72.09%),联合组临床疗效优于引流,组( Z=2.09,P<0.05)。结论 UCMSCs联合 VSD治疗 DFU可有效减轻炎症反应,改善临床症状,促进创面恢复,效果较好。     

细胞色素P450基因修饰的脐带间充质干细胞联合环磷酰胺对急性B淋巴细胞白血病的治疗作用

目的 研究细胞色素P450（Cyt P450）基因修饰的脐带间充质干细胞（UC-MSCs）联合环磷酰胺（CPA）对急性B淋巴细胞白血病的治疗作用。方法 通过基因工程方法设计Cyt P450 2B6（CYP2B6）基因序列,采用慢病毒载体基因转染UC-MSCs,得到可高表达CYP2B6蛋白的CYP2B6-MSC种子细胞。采用实时荧光定量PCR（qRT-PCR）检测CYP2B6基因的表达评估转染效果;采用流式细胞术检测CYP2B6细胞表面标志物表达;采用体外诱导分化检测CYP2B6细胞分化能力。将CYP2B6-MSC与Nalm-6共培养,CCK8法和Annexin-V FITC/PI试剂盒检测UC-MSCs/CYP2B6-MSC联合CPA对Nalm-6细胞的增殖和凋亡的影响。结果 qRT-PCR检测结果显示,CYP2B6-MSC可高表达CYP2B6蛋白,而hUC-MSC（对照组细胞）不表达CYP2B6蛋白;流式细胞术及诱导分化检测发现,CYP2B6基因修饰后的MSC流式表型及分化能力均未有显著变化;肿瘤杀伤实验发现,CYP2B6-MSC与Nalm-6细胞共培养,同时加入CPA后,与UC-MSC及未加CPA组比较,CYP2B6-MSC+CPA对Nalm-6细胞具有显著的增殖抑制及促凋亡作用（P<0.01）。结论 CYP2B6基因修饰后的MSC联合CPA对Nalm-6具有显著的生长抑制和促凋亡作用。     

Structural and biological investigation of chitosan/hyaluronic acid with silanized-hydroxypropyl methylcellulose as an injectable reinforced interpenetrating network hydrogel for cartilage tissue engineering

Cartilage damage continues to pose a threat to humans, but no treatment is currently available to fully restore cartilage function. In this study, a new class of composite hydrogels derived from water-soluble chitosan (CS)/hyaluronic acid (HA) and silanized-hydroxypropyl methylcellulose (Si-HPMC) (CS/HA/Si-HPMC) has been synthesized and tested as injectable hydrogels for cartilage tissue engineering when combined without the addition of a chemical crosslinking agent. Mechanical studies of CS/HA and CS/HA/Si-HPMC hydrogels showed that as Si-HPMC content increased, swelling rate and rheological properties were higher, compressive strength decreased and degradation was faster. Our results demonstrate that the CS and HA-based hydrogel scaffolds, especially the ones with 3.0% (w/v) Si-HPMC and 2.5/4.0% (w/v) CS/HA, have suitable physical performance and bioactive properties, thus provide a potential opportunity to be used for cartilage tissue engineering. In vitro studies of CS/HA and CS/HA/Si-HPMC hydrogels encapsulated in chondrocytes have shown that the proper amount of Si-HPMC increases the proliferation and deposition of the cartilage extracellular matrix. The regeneration rate of the CS/HA/Si-HPMC (3%) hydrogel reached about 79.5% at 21 days for long retention periods, indicating relatively good in vivo bone regeneration. These CS/HA/Si-HPMC hydrogels are promising candidates for tissue compatibility injectable scaffolds. The data provide proof of the principle that the resulting hydrogel has an excellent ability to repair joint cartilage using a tissue-engineered approach.

RESEARCH HIGHLIGHTS

• An injectable hydrogel based on CS/HA/Si-HPMC composites was developed.
• The CS/HA/Si-HPMC hydrogel displays the tunable rheological with mechanical properties.
• The CS/HA/Si-HPMC hydrogel is highly porous with high swelling and degradation ratio.
• Increasing concentration of Si-HPMC promote an organized network in CS/HA/Si-HPMC hydrogels.
• Injectable CS/HA/Si-HPMC hydrogels have a high potential for cartilage tissue engineering.

     

TRAIL基因修饰脐带间充质干细胞联合5-氟尿嘧啶对U-87MG、A549及HeLa细胞杀伤作用研究

目的 研究TRAIL基因修饰的脐带间充质干细胞（TRAIL-MSCs）联合化疗药5-氟尿嘧啶（5-FU）对人脑胶质瘤细胞U-87MG、人肺癌细胞A549和人宫颈癌细胞HeLa的杀伤作用。方法 通过基因工程手段结合慢病毒转染体系构建高表达十二聚体TRAIL蛋白（dTRAIL）的TRAIL-MSCs,应用流式细胞技术检测TRAIL-MSCs的生物学特性;通过ELISA技术检测TRAIL-MSCs分泌的TRAIL量来评估转染效果;CCK-8法检测低浓度（0、1、2、4、8、16 μg/mL）5-FU对U-87MG、A549、HeLa细胞的增殖抑制率,并计算其对各细胞的半数抑制浓度（IC₅₀）;CCK-8法检测5-FU（IC₅₀）、UC-MSCs培养上清、TRAIL-MSCs培养上清、5-FU（IC₅₀）＋UC-MSCs培养上清和5-FU（1/4 IC₅₀、1/2 IC₅₀和IC₅₀）＋TRAIL-MSCs培养上清对U-87MG、A549、HeLa细胞的增殖抑制率,计算5-FU和TRAIL-MSCs培养上清的相互作用系数（CDI）;Westernblotting法检测5-FU对U-87MG、A549、HeLa细胞死亡受体DR4、DR5蛋白表达的影响;台盼蓝染色法观察5-FU、UC-MSCs、TRAIL-MSCs、5-FU＋UC-MSCs和5-FU＋TRAIL-MSCs对U-87MG、A549、HeLa细胞的杀伤作用。结果 生物学检测结果表明TRAIL-MSCs在维持UC-MSCs生物学特性的同时能够高表达TRAIL蛋白。5-FU对U-87MG、A549、HeLa细胞均有增殖抑制作用,抑制作用由高到底依次为HeLa细胞（IC₅₀为9.15 μg/mL） >A549细胞（IC₅₀为10.62 μg/mL） >U-87MG细胞（IC₅₀为22.37 μg/mL）。与对照组比较,除A549细胞UC-MSCs培养上清组外,各处理组细胞增殖抑制率均显著升高（P<0.01、0.001）;与相应各5-FUIC₅₀及TRAIL-MSCs培养上清组比较,U-87MG、HeLa细胞5-FU （1/4 IC₅₀、1/2 IC₅₀和IC₅₀） ＋TRAIL-MSCs培养上清组细胞增殖抑制率均显著升高（P<0.001）,A549细胞5-FU （IC₅₀） ＋TRAIL-MSCs培养上清组细胞增殖抑制率显著升高（P<0.01、0.001）;5-FU （1/2 IC₅₀） ＋TRAIL-MSCs培养上清组对U-87MG的细胞增殖抑制协同效应最显著（CDI<0.7且P<0.001）; 5-FU （1/4 IC₅₀） ＋TRAIL-MSCs培养上清组对A549的细胞增殖抑制具有协同作用,但其协同效果并不显著; 5-FU （1/4 IC₅₀） ＋TRAIL-MSCs培养上清组对HeLa细胞增殖抑制的协同效果最显著（CDI<0.7且P<0.01）。经5-FU处理后U-87MG、A549、HeLa细胞DR4和DR5的蛋白表达明显升高,其中DR5的表达水平高于DR4,与对照组比较差异均具有统计学意义（P<0.001）。与对照组比较,5-FU、TRAIL-MSCs和5-FU＋TRAIL-MSCs组对于肿瘤细胞U-87MG、A549、HeLa均具显著的杀伤作用（P<0.001）;与5-FU或TRAIL-MSCs组比较,5-FU＋TRAIL-MSCs组的杀伤效果更显著（P<0.001）。结论 TRAIL-MSCs联合低浓度的5-FU对肿瘤细胞U-87MG、A549和HeLa具有显著的细胞杀伤作用,二者联用协同效果显著,机制可能与5-FU提高DR4、DR5蛋白表达、提高肿瘤细胞对TRAIL-MSCs的敏感性相关。     

Novel dual-reverse thermosensitive solid lipid nanoparticle-loaded hydrogel for rectal administration of flurbiprofen with improved bioavailability and reduced initial burst effect

The purpose of this study was to develop novel solid lipid nanoparticle (SLN)-loaded dual-reverse thermosensitive hydrogel (DRTH) for rectal administration of flurbiprofen with improved bioavailability and reduced initial burst effect. The flurbiprofen-loaded SLNs were prepared by hot homogenisation technique, after optimising the amounts of lipid mixture (tricaprin and triethanolamine in 8:2 weight ratio), drug and surfactant. The flurbiprofen-loaded thermosensitive SLN composed of drug, lipid mixture and surfactant at a weight ratio of 10/15/1.3 was a solid at room temperature, and changed to liquid form at physiological temperature due to its melting point of about 32 °C. This SLN gave the mean particle size of about 190 nm and entrapment efficiency of around 90%. The DRTHs were prepared by adding this flurbiprofen-loaded thermosensitive SLN in various poloxamer solutions. Their rheological characterisation, release and stability were investigated while a morphological and pharmacokinetic study was performed after its rectal administration to rats compared with the drug and hydrogel. Poloxamer 188 and SLN decreased the gelation temperature and gelation time, but increased the viscosity at 25 °C, gel strength and mucoadhesive force of DRTHs. In particular, the DRTH composed of [SLN/P 407/P 188 (10%/15%/25%)] with the gelation temperature of about 35 °C existed as liquid at room temperature, but gelled at 30–36 °C, leading to opposite reversible property of SLN. Thus, it was easy to administer rectally, and it gelled rapidly inside the body. This DRTH gave a significantly increased dissolution rate of the drug as compared to the flurbiprofen, but significantly retarded as compared to the hydrogel, including the initial dissolution rate. Moreover, this DRTH gave significantly higher plasma concentration and 7.5-fold AUC values compared to the drug, and lower initial plasma concentration and C_max value compared to the hydrogel due to reduced initial burst effect. No damage in rectal mucosa was observed after the application of DRTH. Thus, this DRTH system with improved bioavailability and reduced initial burst effect would be recommended as an alternative for the flurbiprofen-loaded rectal pharmaceutical products.     

Study of the stability of packaging and storage conditions of human mesenchymal stem cell for intra-arterial clinical application in patient with critical limb ischemia

Critical limb ischemia (CLI) is associated with significant morbidity and mortality. In this study, we developed and characterized an intra-arterial cell suspension containing human mesenchymal stem cells (hMSCs) for the treatment of CLI. Equally, the stability of cells was studied in order to evaluate the optimal conditions of storage that guarantee the viability from cell processing to the administration phase. Effects of various factors, including excipients, storage temperature and time were evaluated to analyze the survival of hMSCs in the finished medicinal product. The viability of hMSCs in different packaging media was studied for 60 h at 4 °C. The best medium to maintain hMSCs viability was then selected to test storage conditions (4, 8, 25 and 37 °C; 60 h). The results showed that at 4 °C the viability was maintained above 80% for 48 h, at 8 °C decreased slightly, whereas at room temperature and 37 °C decreased drastically. Its biocompatibility was assessed by cell morphology and cell viability assays. During stability study, the stored cells did not show any change in their phenotypic or genotypic characteristics and physicochemical properties remained constant, the ability to differentiate into adipocytes and osteocytes and sterility requirements were also unaltered. Finally, our paper proposes a packing media composed of albumin 20%, glucose 5% and Ringer’s lactate at a concentration of 1 × 10⁶ cells/mL, which must be stored at 4 °C as the most suitable to maintain cell viability (>80%) and without altering their characteristics for more than 48 h.     

髓芯减压植骨及自体骨髓间充质干细胞移植治疗早期股骨头缺血性坏死疗效观察

目的 观察在髓芯减压植骨的基础上行自体骨髓间充质干细胞移植治疗早期股骨头缺血性坏死的临床疗效.方法 采用髓芯减压植骨联合自体骨髓间充质干细胞移植治疗41例ARCO分期为Ⅰ、Ⅱ期的股骨头缺血性坏死患者,术后定期复查.结果 所有患者经过11～29个月(平均15个月)的随访,除2例进展为ARCO分期Ⅲ期外,其余患者关节疼痛基本消失,活动范围接近或恢复正常,Harris评分由术前的(55.86±7.12)分提高到术后的(79.54+8.69)分,影像学检查结果显示股骨头囊性变消失,坏死区有新生骨小梁通过.结论 髓芯减压植骨联合自体骨髓间充质干细胞移植治疗早期股骨头缺血性坏死,可改善髋关节活动功能,促进骨坏死修复,具有确切的临床疗效.     

Boosting the anti-inflammatory effect of self-assembled hybrid lecithin–chitosan nanoparticles via hybridization with gold nanoparticles for the treatment of psoriasis: elemental mapping and in vivo modeling

Gold nanoparticles are a promising drug delivery system for treatment of inflammatory skin conditions, including psoriasis, due to their small size and anti-inflammatory properties. The aim of this study was to conjugate gold nanoparticles with anti-psoriatic formulations that previously showed successful results in the treatment of psoriasis (tacrolimus-loaded chitosan nanoparticles and lecithin–chitosan nanoparticles) by virtue of their surface charges, then examine whether the hybridization with gold nanoparticles would enhance the anti-psoriatic efficacy in vivo. Successful formation of gold nanoparticles was examined by elemental mapping and selected area electron diffraction (SAED). Hybrid conjugates were examined in terms of particle size and zeta potential by dynamic light scattering (DLS). Morphological features were captured by transmission electron microscope (TEM) and X-ray diffraction (XRD) analysis was conducted, as well. All characterization was conducted for the conjugated nanoparticles and compared with their bare counterparts. The in vivo results on imiquimod (IMQ)-induced mouse model showed promising anti-psoriatic effects upon application of gold conjugated tacrolimus-loaded lecithin–chitosan hybrid nanoparticles with a significant difference from the bare hybrid nanoparticles in some of the inflammatory markers. The anti-inflammatory effect of the gold conjugate was also evident by a lower spleen to body weight ratio and a better histopathological skin condition compared to other tested formulations.