Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing.

BACKGROUND: Mesenchymal stem cells (MSCs) modulated by various exogenous signals have been applied extensively in regenerative medicine research. Notably, nanosecond pulsed electric fields (nsPEFs), characterized by short duration and high strength, significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways. Consequently, we used transcriptomics to study changes in messenger RNA (mRNA), long noncoding RNA (lncRNA), microRNA (miRNA), and circular RNA expression during nsPEFs application. AIM: To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs. METHODS: The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing. MSCs were pretreated with 5-pulse nsPEFs (100 ns at 10 kV/cm, 1 Hz), followed by total RNA isolation. Each transcript was normalized by fragments per kilobase per million. Fold change and difference significance were applied to screen the differentially expressed genes (DEGs). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions, complemented by quantitative polymerase chain reaction verification. RESULTS: In total, 263 DEGs were discovered, with 92 upregulated and 171 downregulated. DEGs were predominantly enriched in epithelial cell proliferation, osteoblast differentiation, mesenchymal cell differentiation, nuclear division, and wound healing. Regarding cellular components, DEGs are primarily involved in condensed chromosome, chromosomal region, actin cytoskeleton, and kinetochore. From aspect of molecular functions, DEGs are mainly involved in glycosaminoglycan binding, integrin binding, nuclear steroid receptor activity, cytoskeletal motor activity, and steroid binding. Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation. CONCLUSION: Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs, 2 miRNAs, and 65 lncRNAs. Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways, which are involved in vesicular transport, calcium ion transport, cytoskeleton, and cell differentiation.

Bronchogenic Cyst: Skull Base Lesions with Extracranial Extension.

BACKGROUND: Intracranial bronchogenic cysts (BCs) are extremely rare. To our knowledge, this is the first report of a BC in which lesions involve the middle and posterior cranial fossa, as well as the infratemporal fossa. CASE DESCRIPTION: We present the case of a 38-year-old woman who suffered from a cranial nerve dysfunction for 2 years. Magnetic resonance imaging showed that there were skull base communication lesions across the middle and posterior fossa. The patient was operated on through an infratemporal fossa approach. The final diagnosis was BC after histopathologic examination and immunohistochemical verification. The patient's neurologic dysfunction was partially ameliorated at the half-year follow-up. CONCLUSIONS: Intracranial BCs are rare. However, they should be considered in the differential diagnosis for cystic lesions with edge enhancement or extracranial extension.

Rates of periprosthetic infection and surgical revision in Beijing (China) between 2014 and 2016: a retrospective multicenter cross-sectional study.

BACKGROUND: Periprosthetic joint infection (PJI) is a rare but devastating complication after total joint arthroplasty. There is a paucity of data on the incidence and prevalence of periprosthetic infection in mainland China. This study aimed to analyze the rates of surgical revision after arthroplasty due to PJI and the procedures followed in Beijing, China. METHODS: The study involved a retrospective multicenter cross-sectional survey of patients undergoing revisions for periprosthetic infection after hip/knee arthroplasty at nine hospitals in Beijing from 2014 to 2016. Age, gender, body mass index, primary diagnosis, comorbidity, primary surgery, treatment methods, and post-revision complications were analyzed. RESULTS: A total of 38,319 hip/knee arthroplasties and 366 (0.96%) revisions for PJI were identified. Of these, 161 (161/14,110; 1.14%) revisions involved hip arthroplasty, whereas 205 (205/24,209; 0.85%) revisions were due to knee arthroplasty. Procedures for revisions of infected hip included 7 (4.3%) cases of open debridement and prosthesis retention, 32 (19.9%) cases of one-stage exchange, 121 (75.2%) cases of two-stage exchange, and 1 (0.007%) case of hip dissection. As for the infected knee, the procedures included 45 (22.0%) cases of open debridement and prosthesis retention, 13 (6.3%) cases of one-stage exchange, 143 (69.8%) cases of two-stage exchange, and 4 (0.02%) cases of knee fusion. CONCLUSIONS: The study found the rates of revision due to PJI to be low. Nonetheless, the incidence of PJI in mainland China could be higher and calls for more elaborate studies in geographically and socioeconomically diverse health institutions.

Identifying Common Pathogens in Periprosthetic Joint Infection and Testing Drug-resistance Rate for Different Antibiotics: A Prospective, Single Center Study in Beijing.

OBJECTIVE: The aim of this study is to identify the common microorganisms causing PJI as well as the drug-resistant spectrum for each microorganism, to help orthopaedic surgeons to choose appropriate antibiotics. METHOD: One hundred and sixty patients who suffered from failure of primary or revision total hip or knee arthroplasty for different reasons were prospectively recruited. These patients underwent revision or re-revision total hip or knee arthroplasty in our institution between August 2013 to August 2016. The details of patients' medical history and comprehensive physical examination, as well as demographic data were recorded precisely. Routine blood test results, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), high sensitive C-reactive protein (hs-CRP), interleukin-6 (IL-6) levels, and synovial leukocyte counts were collected. Additionally, aspiration was conducted during surgery to avoid pollution unless when PJI was strongly suspected, in which case, joint puncture and aspiration were conducted before surgery. Intraoperatively, the implant-surrounding tissue and the prosthesis were collected under aseptic conditions. Postoperatively, the prosthesis, implant-surrounding tissue and synovium were sent to the laboratory immediately. The sonicate extraction (the prosthesis was sent for ultrasound sonication first), implant surrounding tissue and synovium were sent for microbiologic culture, and the implant-surrounding tissue was also sent for pathological examination. The isolated bacteria strains and drug-resistance rates for each pathogen for different antibiotics were presented. RESULTS: There were 59 PJI cases in the infectious group and 101 cases in the non-infectious group (PJI is diagnosed according to the diagnosing criteria from the Workgroup of the Musculoskeletal Infection Society). Of 69 strains of pathogens isolated, Gram-positive bacterium is the most common pathogenic bacteria causing PJI (60, 86.96%). Staphylococcus epidermidis and Staphylococcus aureus played an important role as well, followed by Gram-negative bacteria (8, 11.59%) and fungus (1, 1.45%). Penicillin (78.57%), erythromycin (66.67%) and clindamycin (44.74%) showed high antibiotic resistance rate. In addition, the second-generation cephalosporin, usually as the prophylactic antibiotic, resistance rate was high (20%) as well. Fortunately, no vancomycin-resistant bacteria were discovered in the current study. CONCLUSION: This study provides some information on the most common pathogens in our institution and the selection of antibiotics in the perioperative period in northern China. Cefuroxime and clindamycin might not be appropriate for use as prophylactic antibiotics in revision total knee or hip arthroplasty. Vancomycin is ideal for empiric antibiotic use in suspected PJI cases because of the low drug-resistance rate.

Verbascoside Inhibits Glioblastoma Cell Proliferation, Migration and Invasion While Promoting Apoptosis Through Upregulation of Protein Tyrosine Phosphatase SHP-1 and Inhibition of STAT3 Phosphorylation.

BACKGROUND/AIMS: As a natural antioxidant, verbascoside (VB) is proved to be a promising method for the treatment of oxidative-stress-related neurodegenerative diseases. Thus, this study aimed to investigate the effects of VB on glioblastoma cell proliferation, apoptosis, migration, and invasion as well as the mechanism involving signal transducer and activator of transcription 3 (STAT3) and Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP-1). METHODS: U87 cells were assigned to different treatments. The MTT assay was used to test cell proliferation, flow cytometry was used to detect cell apoptosis, and a Transwell assay was used for cell migration and invasion. We analyzed the glioblastoma tumor growth in a xenograft mouse model. Western blot analysis was employed to determine the protein expression of related genes. RESULTS: Glioblastoma cells exhibited decreased cell proliferation, migration, invasion, and increased apoptosis when treated with VB or TMZ. Western blot analysis revealed elevated SHP-1 expression and reduced phosphorylated (p)-STAT3 expression in glioblastoma cells treated with VB compared with controls. Correspondingly, in a xenograft mouse model treated with VB, glioblastoma tumor volume and growth were decreased. Glioblastoma xenograft tumors treated with VB showed elevated SHP-1, Bax, cleaved caspase-3, and cleaved PARP expression and reduced p-STAT3, Bcl-2, survivin, MMP-2, and MMP-9 expression. siRNA-SHP-1 inhibited the VB effects on glioblastoma. CONCLUSION: This study demonstrates that VB inhibits glioblastoma cell proliferation, migration, and invasion while promoting apoptosis via SHP-1 activation and inhibition of STAT3 phosphorylation.

Distinct expression of chemokine-like factor 1 in synovium of osteoarthritis, rheumatoid arthritis and ankylosing spondylitis.

Chemokine-like factor 1 (CKLF1) is a newly cloned chemotactic cytokine with CCR4 being its functional receptor. Recent evidence demonstrates a role of CKLF1 in arthritis. The aim of this study was to quantify the expression of CKLF1 as well as assess the correlation between CKLF1 and plasma acute-phase markers. Synovium was obtained from 16 osteoarthritis (OA), 15 rheumatoid arthritis (RA) and 10 ankylosing spondylitis (AS) patients undergoing total joint arthroplasty, with other 11 patients treated for meniscal tears during sport accidents serving as normal controls. Levels of CKLF1 and CCR4 mRNA were detected by qRT-PCR, and the expression of CKLF1 was investigated by immunohistochemistry staining, subsequently analyzed with semiquantitative scores. Plasma acute-phase markers of inflammation were determined by ELISA. CKLF1 was found with a particularly up-regulated expression in synovim from AS and RA patients, and CCR4 mRNA levels increased in RA patients, not in OA or AS patients. Elevated levels of plasma markers of inflammation including CRP, ESR and D-dimer were observed in RA. Further, significantly positive correlations between relative expression levels of CKLF1 and CRP/ESR in RA patients and a positive correlation between CKLF1 and ESR in AS patients were found. There was no detectable correlation between CKLF1 and plasma D-dimer. This study confirms an increased but different level of CKLF1 in RA, OA and AS patients, all significantly higher than that in controls. Additionally, the significant positive correlations between CKLF1 levels and CRP/ESR in RA and between CKLF1 and ESR suggest that CKLF1 might contribute to the inflammation state and clinical symptoms in these rheumatic diseases. Further studies are required to investigate the utility of targeting specific CKLF1 for symptom control or disease modification in RA and AS.

The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a common arthritic disease and multifactorial whole-joint disease. Interactions of chemokines and OA is inadequately documented. RESULTS: In vivo and in vitro studies were conducted to investigate monocyte chemoattractant protein 1 (MCP-1) and receptor chemokine (C-C motif) receptor 2 (CCR2) in chondrocyte degradation and cartilage degeneration. Chondrocytes from 16 OA patients and 6 normal controls were involved in this study. After stimulation of MCP-1, the expression of MCP-1 and CCR2 increased significantly (P < 0.001) and the expression of MMP-13 also increased (P < 0.05). MCP-1 stimulation also induced (or enhanced) the apoptosis of OA chondrocytes (P < 0.05). Additionally, the degradation of cartilage matrix markers (metalloproteinase 3 and 13, MMP3 and MMP13) in the culture medium of normal chondrocytes was also assessed. Furthermore, intra-articular injection of MCP-1 in mouse knees induced cartilage degradation and the CCR2 antagonist did not impede cartilage destroy in rats knees of monosodium iodoacetate (MIA) model. CONCLUSIONS: The results of this study demonstrate that the MCP-1-CCR2 ligand-receptor axis plays a special role in the initiation and progression of OA pathology. Patients with ambiguous etiology can gain some insight from the MCP-1-CCR2 ligand-receptor axis.

Lutein production from biomass: marigold flowers versus microalgae.

Microalgae have faster growth rates and more free lutein than marigold flowers, the current source of lutein. However, no commercial lutein production uses microalgae. This review compares lutein content, cultivation, harvesting, cell disruption, and extraction stages of lutein production using marigold flowers and those using microalgae as feedstock. The lutein production rate of microalgae is 3-6 times higher than that of marigold flowers. To produce 1 kg of pure lutein, marigolds need more land and water, but require less nutrients (N, P, K) and less energy than microalgae. Since lutein is tightly bound in microalgae and microalgae are small, cell disruption and subsequent extraction stages consume a considerable amount of energy. Research and development of affordable lutein production from microalgae are discussed.

The role of MCP-1-CCR2 ligand-receptor axis in chondrocyte degradation and disease progress in knee osteoarthritis

BACKGROUND: Osteoarthritis (OA) is a common arthritic disease and multifactorial whole-joint disease. Interactions of chemokines and OA is inadequately documented. RESULTS: In vivo and in vitro studies were conducted to investigate monocyte chemoattractant protein 1 (MCP-1) and receptor chemokine (C-C motif) receptor 2 (CCR2) in chondrocyte degradation and cartilage degeneration. Chondrocytes from 16 OA patients and 6 normal controls were involved in this study. After stimulation of MCP-1, the expression of MCP-1 and CCR2 increased significantly (P < 0.001) and the expression of MMP-13 also increased (P < 0.05). MCP-1 stimulation also induced (or enhanced) the apoptosis of OA chondrocytes (P < 0.05). Additionally, the degradation of cartilage matrix markers (metalloproteinase 3 and 13, MMP3 and MMP13) in the culture medium of normal chondrocytes was also assessed. Furthermore, intra-articular injection of MCP-1 in mouse knees induced cartilage degradation and the CCR2 antagonist did not impede cartilage destroy in rats knees of monosodium iodoacetate (MIA) model. CONCLUSIONS: The results of this study demonstrate that the MCP-1-CCR2 ligand-receptor axis plays a special role in the initiation and progression of OA pathology. Patients with ambiguous etiology can gain some insight from the MCP-1-CCR2 ligand-receptor axis.

[Clinical significance of arthritis treatment guidelines revealed by outpatients prescriptions].

OBJECTIVE: To investigate the current status of osteoarthritis medications of outpatients for arthritis treatment guidelines, and provide references for the promotion and popularization of traditional Chinese and western medicine in treatment of arthritis. METHODS: The outpatient prescriptions for the treatment of osteoarthritis from all the rheumatology and orthopedics specialists from 2007 February to May in Peking University People's Hospital were chosen and analyzed statistically. RESULTS: There were a total of 2 145 osteoarthritis prescription in this study, including 8 categories: joint lubricants, non-steroid anti-inflammatory drugs (NSAIDs), local anesthetics, cartilage protective agent, adrenal corticosteroids, vitamin AD, analgesic drugs and traditional Chinese medicine. The Chinese medicines were among the drugs with the most species amounted up to 35. The most common route of medication was oral administration (73.2%), which was used more in the department of rheumatology and immunology than in orthopedics. And in oral drugs, the biggest consumption was NSAIDs, accounting for 29.9%. There was no significantly difference between the rheumatology and orthopedic specialists when using non-specific cyclooxygenase (COX) inhibitors. But orthopedic specialists prescribed more COX-1 specific inhibitor than rheumatology specialists. CONCLUSION: Recently the arthritis treatment guidelines have been issued one after another. Many experts have already accepted the treatment of pain. However, in the implementation, the large differences still exist. The use of the Chinese medicine is still very chaotic; there are no clear-cut norms to be followed. Therefore, the implementation of the arthritis treatment guidelines and treatments of arthritis by traditional Chinese medicine are urgent to be standardized.

Clinical attitudes towards pain treatment post-orthopedic surgery: a multicenter study in Beijing.

BACKGROUND: Pain is a common post-operative complication. Incidence of pain directly affects patients' quality of life in terms of patient physiology, psychology, and social characteristics. This study was to understand clinical attitudes with regards to Beijing surgeons, and patients' attitude towards pain treatment after orthopedic surgery. METHODS: A hospital-based cross-sectional and cluster sample survey of 40 hospitals in Beijing was conducted, including 20 level III (tier three) and 20 level II (tier two) general hospitals. Enrolled subjects completed a specifically designed interview-questionnaire. RESULTS: The prevalence of pain 2 weeks post-orthopedic surgery was high in Beijing (96.1%). Meanwhile, collected data indicated most subjects in Beijing suffered moderate to severe pain, 45.1% and 41.4%, respectively, post-surgery. And for the concern of patients before surgery, most subjects chose full recovery from surgery (78.6%), as well as, the pain after operation was 39.2% ranked the third. According to the data from the study, Tramadol use was more common in Level III hospitals, where Somiton was preferred in Level II hospitals. When it came to the education of pain before and after operation, more patients get educated before operation than after it. In our study, case physicians or attending physicians enacted education before and after surgery. Related to the sense of patients, among the surgeons preferring post-operative analgesia, 67.6% considered administration when receiving complaints of moderate level pain, 50.0% indicated they will terminate analgesic treatment once pain degree scale wise decreases to benign pain. CONCLUSIONS: The majority of orthopedic patients experience post-operative pain. Identification of post-operative pain will facilitate future awareness on pain treatment and nursing care in Beijing hospitals, with pain relief through regulated improvements in strategic pain management.

[Experience of using anterior windowing of the femur in revision total hip arthroplasty].

OBJECTIVE: To discuss the clinical results of anterior windowing of the femur to remove the femoral component and cement in revision total hip arthroplasty. METHODS: From September 1999 to May 2011, 31 revision cases received anterior windowing of the femur in operation. There were 12 male and 19 female, with the average age in operation was 61.8 years (from 40 to 83 years). The reason for revision included aseptic loosening in 12 cases, infection in 4 cases, breakage of femoral stem in 11 cases and acetabular liner wear in 4 hemi-arthroplasties. Nine cases were cemented and 22 were non-cemented for the primary stem. The position of the window located in the anterior femur with 6 cases of complete windowing from the proximal to the end of the stem. Another 25 cases received regional windowing just around the tip of the stem. In revisions, non-cemented rectangular revision stem were used for 27 cases and two-stage surgery were used for 4 infection cases. RESULTS: In all revisions, femoral stems and cement fragments were removed successfully and safely without any complications of fracture and perforation of new stems. All femur windows showed successfully union for average 14 weeks (12-18 weeks). There were some postoperative complications. One recurrent dislocation was treated using plaster external fixation for 8 weeks. One case with dislocation and fracture along with the window was revised by cerclage fixation. One periprosthetic fracture due to trauma was treated by cerclage fixation. The average Harris score of the hip improved to 83 points (75 to 90 points) at an average 5.5 years follow-up (0.5 to 12.0 years). Twenty six cases were completely followed up. Revision femoral stems were well-fixed without any subsidence or loosening. CONCLUSIONS: Anterior windowing of the femur is a proven technique which can be used to remove femoral stem and cement conveniently and safely without any fractures in revision total hip arthroplasty.

[Using titanium cerclage band to teat intra- and post-operative femoral fracture in total hip arthroplasty].

OBJECTIVE: To discuss the treatment and clinical result of using titanium cerclage band to teat intra- and post-operative femoral fracture in total hip arthroplasty. METHODS: From August 1999 to September 2007, twenty-two patients who suffered from intra-and post-operative femoral oblique fracture in total hip arthroplasty were reviewed among 2186 consecutive total hip arthroplasty patients performed by one surgeon. There were 12 fractures occurred intraoperative and 10 fractures postoperative. There are 9 primary and 13 revision total hip arthroplasty patients including 10 men and 12 women. All fractures are classified as Vancouver B1 type in 15 cases and B2 type in 7 cases. Two to five titanium cerclage bands had been used to fix the fracture followed by open reduction with 13 cases using 2 cerclage bands and 7 cases using 3 bands and 1 case using 4 bands and 1 case using 5 bands. Whether change the original stem or not depended on the classification of fractures and stability of stems. There were 15 cases of type B1 and 1 case of type B2 without changing stems and 6 cases of type B2 changing to revision stems of the same series of original stems. RESULTS: All fractures showed successfully union for 12 - 20 weeks (average 18 weeks) after operation. The average Harris score of the hip improved to 90 points (80 to 95 points) at an average 6.5 years follow-up (4 to 11 years) and the average range of motion of the hip got to 120° (105° - 135°). Femoral prostheses were well-fixed without any subsidence or lucent lines or loosening. All patients were satisfied with the results with well-fixed and good alignment of femoral stem. CONCLUSIONS: Strong fixation of the fractures and good initial stability of femoral stem can be obtained using titanium cerclage bands to fix the intra- and post-operative femoral oblique fracture. Original stems should be changed to revision stems in case of type B2 and be retained in case of type B1.

[Medium and long-term clinical results of using Zweymüller cup to treat protrusio acetabuli].

OBJECTIVE: To discuss long-term clinical results of using Zweymüller cup to treat protrusio acetabuli. METHODS: From May 1998 to September 2006, 31 patients with 39 hips diagnosed protrusio acetabuli were treated with Zweymüller cup in total hip arthroplasties or revisions. There were 12 men and 19 women with average age of 57.6 years (from 30 to 82 years). The reasons causing protrusio acetabuli were as followed: rheumatoid arthritis 3 cases 6 hips, osteoarthritis followed femoral head necrosis 7 cases 12 hips, acetabular wear after hemi-arthroplasty 11 cases 11 hips and acetabular component loosening 10 cases 10 hips. During all operations, Zweymüller cup was used to fix the acetabular part. RESULTS: All cases were followed up 6 months and one year after operations. Two patients with 3 hips were died for lung cancer and acute myocardial infarction respectively. Twenty-four cases with 31 hips got recent follow-up with average 7.4 years (from 5.0 to 11.5 years). The average Harris score improved from 31.0 (from 14 to 61) preoperatively to 84.7 (from 70 to 95) postoperatively. There was one infection in right hip after bilateral hip arthroplasty treating by removal prosthesis and cement spacer insert. But until now this patient still did not get revision for her internal medicine. One rheumatoid arthritis patient with two-stage bilateral hip arthroplasty was found slight internal migration and loosening line of left acetabular component, but the patient had no pain with good hip function. All other cases had good hip functions and were very satisfied with clinical results. CONCLUSION: Using Zweymüller cup to treat protrusion acetabuli can get strong fixations and perfect medium and long-term clinical results for over 7.4 years.

[Disability and common treatment strategies associated with knee pain in a rural Chinese population].

OBJECTIVE: To estimate the burden of disability and health services use by people with knee pain, with or without radiographic evidence of osteoarthritis (OA), in rural northern China. METHODS: A population-based cross-sectional survey was conducted among 1030 residents of Wuchuan County, Inner Mongolia, aged 50 years old and over. The participants completed an interviewer-based questionnaire and obtained bilateral weight-bearing posterior-anterior semi-flexed knee radiographs. RESULTS: Of 1027 participants with knee radiographs, 513 (50%) participants reported knee pain in most days of at least a month over the past 12 months. Of those with knee pain, 109 (21%) had radiographic OA (Kellgren Lawrence grade > or =2) in symptomatic knees. Adjusting for age, gender, BMI, education and back pain, the presence of knee pain was associated with a significantly greater difficulty in walking, climbing 10 steps, stooping, performing cleaning chores and preparing meals. Among 513 subjects with knee pain, the presence of radiographic disease was significantly associated with the presence of unbearable pain (36% vs. 59%), restricted activity (39% vs. 64%), use of NSAIDs (78% vs. 88%) and consulting a doctor over the last 12 months (33% vs. 59%). CONCLUSION: Knee pain is associated with significant physical disability in rural China The prevalent use of NSAIDs for knee pain and a low use of knee surgery should be of particular concerns. These findings will be useful to guide the distribution of future health care resources and preventive strategies.

Human Wharton's jelly cells can be induced to differentiate into growth factor-secreting oligodendrocyte progenitor-like cells.

Human Wharton's jelly-derived mesenchymal stromal cells (hWJ-MSCs) are capable of differentiating into neural and astroglia-like cell types. However, a reliable means of inducing the selective differentiation of hWJ-MSCs into oligodendrocyte progenitor cells (OPCs) in vitro has not yet been established. In this study, the OPC-like differentiation of hWJ-MSCs was characterized using and immunoblotting. The hWJ-MSC-derived OPC-like cells were able to secrete nerve growth factors and promote neurite outgrowth in vitro. These results show that hWJ-MSCs can be induced to differentiate into cells with the morphologic, phenotypic and functional characteristics of OPC-like cells.

Effects of differentiated versus undifferentiated adipose tissue-derived stromal cell grafts on functional recovery after spinal cord contusion.

Controversies exist concerning the need for mesenchymal stromal cells (MSCs) to be transdifferentiated prior to their transplantation. In the present study, we compared the results of grafting into the rat contused spinal cord undifferentiated, adipose tissue-derived stromal cells (uADSCs) versus ADSCs induced by two different protocols to form differentiated nervous tissue. Using Basso, Beattie, and Bresnahan scores and grid tests, we found that three cell-treated groups, including uADSCs-treated, dADSCs induced by Protocol 1 (dADSC-P1)-treated, and dADSCs induced by Protocol 2 (dADSC-P2)-treated groups, significantly improved locomotor functional recovery in SCI rats, compared with the saline-treated group. Furthermore, functional recovery was better in the uADSC-treated and dADSC-P2-treated groups than in the dADSC-P1-treated group at week 12 postinjury (P < 0.05 for dADSC-P1 group vs. uADSCs or dADSC-P2 groups). Although both protocols could induce high percentages of cells expressing neural markers in vitro, few BrdU-labeled cells survived at the injury sites in the three cell-treated groups, and only a small percentage of BrdU-positive cells expressed neural markers. On the other hand, the number of NF200-positive axons in the uADSC-treated and dADSC-P2-treated groups was significantly larger than those in the dADSC-P1-treated and saline-treated control groups. Our results indicate that ADSCs are able to differentiate into neural-like cells in vitro and in vivo. However, neural differentiated ADSCs did not result in better functional recovery than undifferentiated ones, following SCI. In vitro neural transdifferentiation of ADSCs might therefore not be a necessary pretransplantation step. Furthermore, cellular replacement or integration might not contribute to the functional recovery of the injured spinal cord.

LTB4 can directly stimulate human osteoclast formation from PBMC independent of RANKL.

Leukotriene B4, as a kind of 5-lipoxygenase metabolite of arachidonic acid, is known to influence osteoclast formation and bone resorption. In order to determine whether Leukotriene B4 could directly stimulate human osteoclast differentiation and activation independent of RANKL (ODF), three different concentrations of Leukotriene B4 (10(-9)M, 10(-8)M, 10(-7)M) were added to the culture of CD14+ monocyte fraction of peripheral blood mononuclear cell (PBMC) in the presence of macrophage colony-stimulating factor (M-CSF). Under these conditions, Leukotriene B4 could induce multinucleated cells, which were positive for Tartrate-resistant acidic phosphatase (TRAP) staining and capable of bone resorption. Addition of osteoprotegerin (OPG) to PBMC cultures does not abrogate osteoclast formation induced by LTB4. Osteoclastogenesis induced by Leukotriene B4 were dose-dependently increased and weaker than that of RANKL. These results indicated that Leukotriene B4, elevated in many inflammatory diseases, is directly capable of inducing osteoclast formation by a RANKL-independent mechanism.