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Abstract Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. Objective Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. Methodology The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. Results Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. Conclusions These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.
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Animals , Mice , Mesenchymal Stem Cells , Odontogenesis , Stem Cells , Cell Differentiation , Receptors, Nerve Growth Factor , CD146 Antigen , Mice, Inbred BALB CABSTRACT
BACKGROUND: The development of regenerative medicine and the appearance of tissue engineering technology provide a new solution for cartilage defect reconstruction. In tissue engineering, mesenchymal stem cells are widely used seed cells. However, as a heterogeneous cell group, stem cells play different roles in different subsets. Therefore, the application of key functional subsets of mesenchymal stem cells in cartilage repair has a broad application prospect. OBJECTIVE: To sort CD146 positive subpopulation cells from human adipose-derived mesenchymal stem cells to verify their biological characteristics and potential as seed cells in cartilage tissue engineering. METHODS: Human adipose-derived mesenchymal stem cells were provided by Zhejiang Jinshidai Biotechnology Co., Ltd. Surface markers of human adipose-derived mesenchymal stem cells were identified by flow cytometry. CD146 positive subpopulation cells were sorted from human adipose-derived mesenchymal stem cells using magnetic-activated cell sorting. Molecular characteristics of two kinds of cells were analyzed by gene chip detection technology and bioinformatics analysis technology. Two kinds of cells were induced to chondrocytes in vitro and histologically examined. Cell viability and apoptosis of two kinds of cells were detected before and after cryopreservation. RESULTS AND CONCLUSION: Human adipose-derived mesenchymal stem cells highly expressed stem cell-associated markers CD73 and CD90, but did not express hematopoietic stem cell-associated markers CD34, CD45, and HLA-DR. Bioinformatics analysis results showed that CD146 positive subpopulation had different functions in inflammatory pathways and musculoskeletal diseases compared with human adipose-derived mesenchymal stem cells. CD146 positive subpopulation could differentiate into cartilage, and its chondrogenic differentiation ability was better than that of human adipose-derived mesenchymal stem cells. CD146 positive subpopulation had better apoptosis and activity than human adipose-derived mesenchymal stem cells after resuscitation. These results suggest that CD146 positive subpopulation has good chondrogenic differentiation potential and is a promising seed cell for cartilage tissue engineering.
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Background & objectives: CD9 and CD146 are important adhesion molecules that play a role in the implantation of an embryo. This study was undertaken to correlate the expression of these markers in fertile and infertile women's endometrial stromal cells. Methods: Human endometrial stromal cell culture from endometrial biopsies of fertile (n=50) and infertile females (n=50) was performed and primary cell lines were established. Expression of CD9 and CD146 was studied for all the 100 cell lines with the help of flow cytometry. Gene expression of CD9 and CD146 was performed by real-time polymerase chain reaction. Results: There was a significant difference in endometrial stromal cells of fertile and infertile females. Flow cytometric results revealed significantly lower expression of CD9 (P=0.0126) and CD146 (P=0.0006) in the infertile endometrial stromal cells as compared to fertile endometrial stromal cells. These results were comparable with real-time data. Interpretation & conclusions: This study showed that endometrial stromal cells from infertile females had lower expression of adhesion molecules, CD9 and CD146. Our findings suggest that CD9 and CD146 may have a role in infertility. Infertile female's endometrial stromal cells have decreased expression of CD9 and CD146 which can be the cause of infertility related to implantation failure.
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Objective To investigate the expression and localization of CD146 in corneal neovascularization.Methods Fifty SPF male SD rats with the age of 8 weeks were randomly divided into 1 day group,3 days group,7 days group,14 days group and normal control group by using random number table method,10 rats for each group.Corneal neovascularization models were established by sticking the filter papers with NaOH on the central corneas.The area of corneal neovascularization was observed at different time points by slit lamp microscopy.The expression of CD146 mRNA and protein were detected by reverse transcription PCR and Western blot,respectively.The location of CD146 expression was observed by using immunohistochemical staining method.The animal feeding and use was in accordance with the standards set by the ARVO,and the experiment was approved by the Ethic Committee for Experimental Animal of Affiliated Hospital of Xuzhou Medical University (2016012).Results Corneal neovascularization occurred at 3 days and peaked at 7 days after alkali burn,then gradually subsided.The area of cornealneovascularization was (1.9±0.7),(10.3±1.1),(29.6±2.4) and (11.8±1.0)mm2 in 1 day,3 days,7 days and 14 days after alkali burn,respectively,the overall difference was statistically significant (F =650.976,P =0.000),and the area of corneal neovascularization in 1 day group,3 days group,14 days group was significantly shrinked compared with that in the 7 days group after alkali burn,with significant differences between them (t =-33.293,-20.475,20.744,all at P =0.000),while there was no significant differences between the other groups (all at P＞0.05).The relative expression levels of CD146 mRNA in the 1 day,3 days,7 days and 14 days group after alkali burn was 0.3±0.1,1.1±0.2,3.5±0.4 and 1.3±0.3,and the relative expression levels of CD146 protein was 0.2±0.1,1.4 ± 0.2,4.1 ± 0.5 and 1.3 ± 0.2,respectively,the overall differences between the four groups were statistically significant (F =1 176.920,P =0.000;F =233.127,P =0.000),and the relative expression levels (o)(f) CD146 mRNA and protein in the 1 day,3 days,7 days and 14 days group were lower than those in the 7 days group after alkali burn,with significant differences between them (mRNA:t =-58.109,-33.725,31.006;all at P =0.000.protein:t =-59.873,-38.762,39.153,all at P =0.000).Immunohistochemical results showed that CD146 was highly expressed in corneal neovascular endothelial cells at 7 days after alkali burn,forming lumen structure,while only weak expression of CD146 could be detected in mature corneal neovascularization at 14 days after modeling.Conclusions CD146 is closely related to corneal neovascularization formation,and it is promising as a new target for treatment.
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ABSTRACT:Objective To evaluate the expressions of cell adhesion molecules CD146 and transferrin receptor 1 (TfR1)in ovarian epithelial cancer and investigate their relationship with clinical pathological features of patients with ovarian epithelial cancer (OEC ) so as to further explore the pathogenesis of OEC. Methods Immunohistochemistry was employed to determine the expressions of CD146 and TfR1 in normal,benign and ovarian cancer tissues.Results The immunohistochemical results showed that the positive expressions of CD146 and TfR1 gradually increased with the changes of normal,benign and malignant tissues in OEC.There were significant differences between all the groups (P0.05).There was a significant correlation between expressions of CD146 and TfR1 in OEC (P<0.05,rs=0.532).Combined detection sensitivity and specificity were 82.4% and 75.0%.Conclusion The high expressions of CD146 and TfR1 may play a key role in the occurrance,progression and metastasis of OEC.They may be used as potential markers for diagnosis,postoperative follow-up and targeted therapy.
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Objective To investigate serum soluble CD146 (sCDl46) level in early-stage diabetic nephropathy.Methods Thirty-two SPF-class SD rats were randomly divided into 2 groups as follows:a control group (C group,n =16) and a model group (DN group,n =16).Serum creatinine (Scr),blood urea nitrogen (BUN),sCDl46 level,and renal weight-to-body weight ratio (RW/BW) were estimated at the first day (D 1) and third week (W3) after modelling.Periodic acid-Schiff staining was used for observing the renal cortex and calculating the renal glomerular mesangial membrane area (MMA).Results The serum sCD146 level was significantly higher in the DN group at D1 and W3 (P ＜ 0.05) compared with that in the C group.RW/BW and MMA were significantly higher in the DN group than in the C group (P ＜0.01).Conclusion Serum sCD146 level reacts to increases much sooner and faster in the DN model,which could be used as a sensitive diagnostic index to reflect the severity of early renal damage.
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BACKGROUND: Type 2 diabetes mellitus (T2DM) is a multisystemic, chronic disease accompanied by microvascular complications involving various complicated mechanisms. Intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and cluster of differentiation-146 (CD146) are mainly expressed by endothelial cells, and facilitate the adhesion and transmigration of immune cells, leading to inflammation. In the present study, we evaluated the levels of soluble adhesion molecules in patients with microvascular complications of T2DM. METHODS: Serum and whole blood samples were collected from 58 T2DM patients with microvascular complications and 20 age-matched healthy subjects. Levels of soluble ICAM-1 (sICAM-1) and soluble VCAM-1 (sVCAM-1) were assessed using enzyme-linked immunosorbent assay, while flow cytometry was used to determine CD146 levels. RESULTS: Serum sICAM-1 levels were lower in T2DM patients with microvascular complications than in healthy controls (P<0.05). No significant differences were found in sVCAM-1 and CD146 levels between the study and the control group. Although patients were subdivided into groups according to the type of microvascular complications that they experienced, cell adhesion molecule levels were not correlated with the complication type. CONCLUSION: In the study group, most of the patients were on insulin therapy (76%), and 95% of them were receiving angiotensin-converting enzyme (ACE)-inhibitor agents. Insulin and ACE-inhibitors have been shown to decrease soluble adhesion molecule levels via various mechanisms, so we suggest that the decreased or unchanged levels of soluble forms of cellular adhesion molecules in our study group may have resulted from insulin and ACE-inhibitor therapy, as well as tissue-localized inflammation in patients with T2DM.
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Humans , Cell Adhesion , Cell Adhesion Molecules , Chronic Disease , Diabetes Mellitus , Diabetes Mellitus, Type 2 , Endothelial Cells , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Healthy Volunteers , Inflammation , Insulin , Intercellular Adhesion Molecule-1 , Vascular Cell Adhesion Molecule-1ABSTRACT
Objective To investigate the correlation between the expression of adhesion molecule CD146 and the vulnerability of carotid atherosclerotic plaque.Methods The plaque samples were collected from 40 patients who underwent the carotid endarterectomy and were divided into the stable plaque group and the instable plaque group by ultrasound imaging.Five carotid artery samples were taken from the healthy donors as the control.Immunohistochemistry was applied to test the CD146 expression in all samples.Results Higher expression of CD146 was observed in the atherosclerotic plaques than in the healthy control.Moreover,statistical difference was found in the expression of CD146 in the plaques between the instable plaque group and the stable plaque group (0.31 ± 0.19 vs 0.17 ± 0.07,P ＜ 0.05).The expression of CD146 was positively correlated with the necrotic area (r =0.471 8,P =0.019 9) and the matrix metalloproteinase (MMP)-9 expression in the plaques (r =0.535 6,P =0.000 9).Conclusion The CD146 expression is correlated with the vulnerability of carotid atherosclerotic plaque.
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Objective To explore renal clear cell carcinoma ( CCRCC) expression and significance of CD146 mR-NA in tumor tissue of patients with. Methods ELISA method for quantitative detection was used for CD146 protein and real-time PCR technique for the detection of CCRCC in 102 ( CCRCC) expression in tumor tissue of patients with CD146 mRNA, and 51 cases of renal patients with non tumor tissues as control. Results Found metastasis in patients with CCRCC, the CD146 protein concentration was statistically significant compared with the control group (F=52.1, P<0.01),the average expression of CD146 mRNA value (0.043 8±0.002 4) was significantly high-er than that of in situ CCRCC patients (0.038 2±0.001 1, P= 0.018) and control group (0.034 4±0.001 0, P=0.001 ) . Conclusion Pathological grading and lymph node up regulation of CD146 mRNA expression in renal cell carcinoma and metastasis, is expected to become a new index, evaluation of malignant degree of CCRCC me-tastasis and prognosis, and provide a reliable basis for the intervention of clinical treatment.
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Objective To explore the relationship between CD146(MCAM),microvessel density(MVD)in renal cell carcinoma(RCC) and its clinic-pathology,and to explore their correlation with clinic-pathologic parameter of RCC. Methods Immunohistochemisty was employed to determine the expression of CD146 and MVD in 43 RCC tissues and 20 normal control renal tissues. Results The positive expression of CD146 in RCC(90.7 ％,39/43) was remarkably higher than that in normal renal tissue(30.0 ％,6/20)(x2=27.77,P＜0.05).The expression of CD146 was not correlated with the category of RCC (x2=1.37,all P ＞0.05),but had a significant correlation to(the tumor volume x2=7.57)clinical stage(r=0.62) and metastasis of RCC(x2=19.99,P＜0.05). The MVD of RCC [(78.00±23.10)/200HP]was significantly higher than that of normal renal tissue [(23.05±7.93)/200HP].The MVD of CD146 was not correlated with the tumor volume and category of RCC (t=1.33,t=1.46,au P＞ 0.05),but had a significant correlation to clinical stage and metastasis of RCC (t=2.37,t=2.10,P＜ 0.05). There was a positive correlation between expression of CD146 and MVD in RCC(r=0.74,P＜0.05). Conclusion The overexpression of CD146 in RCC has a significant relation with tumor angiogenesis.The expression of CD146 and angiogenesis might serve as an important indicator of the development, progress and metastasis of RCC.
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In spite of the attention given to the study of mesenchymal stem cells derived periodontal ligament (PDL), there is a lack of information about canine PDL cells. In this study, we characterized canine PDL cells to clarify their stem cell properties, including self renewal, proliferate rate, stem cell markers and multipotency. PDL cells were obtained from extracted premolars of canines, following a colony forming assay and proliferation rate of sub-confluent cultures of cells for self-renewal, immunostaining for STRO-1 and CD146/MUC18 and a differentiation assay for multipotency. Canine PDL cells formed single-cells colonies and 25% of the PDL cells displayed positive staining for BrdU. The cells expressed the mesenchymal stem-cell markers, STRO-1 and CD146/MUC18. Under defined culture conditions, the cells differentiated into osteoblasts and adipocytes, but the cells didn't differentiated into chondrocytes. The findings of this study indicated that the canine PDL cells possess crucial stem cells properties, such as self-renewal and multipotency, and express the mesenchymal stem cell markers on their surface. The isolation and characterization of canine PDL cells makes it feasible to pursue preclinical models of periodontal regeneration in canine.
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Adipocytes , Bicuspid , Bromodeoxyuridine , Chondrocytes , Mesenchymal Stem Cells , Osteoblasts , Periodontal Ligament , Regeneration , Stem CellsABSTRACT
Objective To isolate CESs by dynabeads coated with the specific antibody of CD146 from peripheral blood and its origin was identified.Methods One mL blood with or without admixed HUVECs was diluted(1∶2) in PBS-0.1% BSA.Anti-CD146-coated Dynabeads were added and incubated for 30mins at(4 ℃).Cells bound to anti-CD146-coupled beads were separated from blood in Dynal MPC and then washed and resuspended in(4 mL) buffer.After 4 additional washes with PBS-0.1% BSA using the magnet,the rosetted cells were flushed from the tube wall with (100 ?L) of PBS-BSA with acridine orange or Giemsa,and counted in a hemocytometer in a inverse phase contrast fluorescence microscope.Results The amount of CECs in healthy adult was 10.5(6～16.5)/mL(n=42).The recovery rate was 91%.Isolated CECs were confirmed by positive expression of vWF and CD31.Conclusion Isolation of CECs from blood by immunomagnetic beads coated with antibody of CD146 is characterized by its accuracy,time-saving,high recovery rate,less contaimination of blood and less damage to CECs.It can be used to quantify CECs and to analyze the functional performance of CECs from patients with vascular injury diseases.