Single CD271 marker isolates mesenchymal stem cells from human dental pulp / 国际口腔科学杂志·英文版

Mesenchymal stem cells (MSCs) are a promising tool in regenerative medicine due to their capacity to differentiate into multiple lineages. In addition to MSCs isolated from bone marrow (BMSCs), adult MSCs are isolated from craniofacial tissues including dental pulp tissues (DPs) using various stem cell surface markers. However, there has been a lack of consensus on a set of surface makers that are reproducibly effective at isolating putative multipotent dental mesenchymal stem cells (DMSCs). In this study, we used different combinations of surface markers (CD51/CD140α, CD271, and STRO-1/CD146) to isolate homogeneous populations of DMSCs from heterogeneous dental pulp cells (DPCs) obtained from DP and compared their capacity to undergo multilineage differentiation. Fluorescence-activated cell sorting revealed that 27.3% of DPCs were CD51(+)/CD140α(+), 10.6% were CD271(+), and 0.3% were STRO-1(+)/CD146(+). Under odontogenic conditions, all three subsets of isolated DMSCs exhibited differentiation capacity into odontogenic lineages. Among these isolated subsets of DMSCs, CD271(+) DMSCs demonstrated the greatest odontogenic potential. While all three combinations of surface markers in this study successfully isolated DMSCs from DPCs, the single CD271 marker presents the most effective stem cell surface marker for identification of DMSCs with high odontogenic potential. Isolated CD271(+) DMSCs could potentially be utilized for future clinical applications in dentistry and regenerative medicine.

Characterization of the osteogenic potential of mesenchymal stem cells from human periodontal ligament based on cell surface markers / 国际口腔科学杂志·英文版

Mesenchymal stem cell (MSC)-mediated therapy has been shown to be clinically effective in regenerating tissue defects. For improved regenerative therapy, it is critical to isolate homogenous populations of MSCs with high capacity to differentiate into appropriate tissues. The utilization of stem cell surface antigens provides a means to identify MSCs from various tissues. However, few surface markers that consistently isolate highly regenerative MSCs have been validated, making it challenging for routine clinical applications and making it all the more imperative to identify reliable surface markers. In this study, we used three surface marker combinations: CD51/CD140α, CD271, and STRO-1/CD146 for the isolation of homogenous populations of dental mesenchymal stem cells (DMSCs) from heterogeneous periodontal ligament cells (PDLCs). Fluorescence-activated cell sorting analysis revealed that 24% of PDLCs were CD51(+)/CD140α(+), 0.8% were CD271(+), and 2.4% were STRO-1(+)/CD146(+). Sorted cell populations were further assessed for their multipotent properties by inducing osteogenic and chondrogenic differentiation. All three subsets of isolated DMSCs exhibited differentiation capacity into osteogenic and chondrogenic lineages but with varying degrees. CD271(+) DMSCs demonstrated the greatest osteogenic potential with strong induction of osteogenic markers such as DLX5, RUNX2, and BGLAP. Our study provides evidence that surface marker combinations used in this study are sufficient markers for the isolation of DMSCs from PDLCs. These results provide important insight into using specific surface markers for identifying homogenous populations of DMSCs for their improved utilization in regenerative medicine.

Overexpressions of Vimentin and Integrins in Human Metastatic Spine Tumors

OBJECTIVE: To comparatively investigate the expression of several integrins in specimens of human bone metastases and degenerative bone tissue. METHODS: Degenerative cancellous tissue was obtained from a sample of human degenerative spine. Thirteen human specimens were obtained from metastatic spine tumors, whose primary cancer was colon cancer (n=3), hepatocellular cancer (n=3), lung cancer (n=4), and breast cancer (n=3). The expression of vimentin and integrins alphav, beta1, and beta3 was assessed in metastatic and degenerative specimens by immunohistochemistry and real-time reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS: Immunohistochemical staining showed that vimentin and integrin alphav was broadly expressed in all tissues examined. By contrast, integrin beta1 was weakly expressed only in 38.4% (5/13) of tissues. Integrin beta3 was consistently negative in all cases examined. qRT-PCR analysis showed that vimentin gene expression was higher in all metastatic specimens, as compared to degenerative bone. The gene expression of integrin alphav in breast specimen was significantly higher than others (p=0.045). The gene expression of integrin beta1 was also higher in all metastatic specimens than in degenerative bone tissue. The gene expression of integrin beta3 was variable. CONCLUSION: Spinal metastatic tumors have mesenchymal characteristics such as increased expression of vimentin. The increased expression of integrin alphav and beta1 in spine metastatic tumors suggests that adhesive molecules such as integrin may have implications for the prevention of spine metastasis.

Expression of alpha4, alphav, beta1 and beta3 integrins during the implantation window on blastocyst of a mouse model of polycystic ovarian syndromes

It has been hypothesized that blastocyst integrin expression changes can affect the spontaneous miscarriage in polycystic ovarian syndromes [PCOS]. In this study, the profile of integrin genes and proteins was investigated on blastocyst of the PCOS experimental mouse model. 30 NMRI female mice were equally divided into 3 groups: control, experimental [PCOS that was injected estradiol valerate [40 mg/kg]]. After 8 weeks, each group was hyper stimulated by PMSG and HCG. Vaginal plaque was checked, and mice were investigated 5 days after the test. Progesterone and estradiol levels were determined; alpha4, alpha v, beta 1 and beta 3 integrin genes and protein of blastocysts were examined by real time PCR method and immunohistochemistry, respectively. Estradiol level was significantly increased [p

Effect of zoledronate on the osteoclast adhesion and gene expression of integrin α(v) and β3 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the effect of zoledronate (ZOL) on the osteoclast adhesion and expression of integrin α(v) and β3 in vitro.</p><p><b>METHODS</b>Mice RAW264.7 cells were used for osteoclast differentiation in vitro, and osteoclastogenesis was examined by tartrate-resistant acid phosphatase (TRAP) staining and dentin resorption lacunae examination. The cells were then divided into 2 groups, the control group and ZOL treatment group (treated with 1 x 10(-6) mol · L(-1) ZOL for 2 d). The adhesion ability of osteoclasts and mRNA and the protein expressions of integrin α(v) and β3 were examined by crystal violet staining, real-time fluorescence quantitative polymerase chain reaction, Western blot analysis, and immunofluorescent chemistry.</p><p><b>RESULTS</b>TRAP staining and dentin resorption lacunae examination revealed the formation of multi-nuclear osteoclasts. ZOL treatment significantly decreased the adhesion ability of osteoclasts (P < 0.01). In the ZOL-treated group, the mRNA levels of integrin α(v) and β3 were 0.66 ± 0.05 and 0.59 ± 0.08, respectively. In the control group, the mRNA levels of integrin α(v) and β3, were 1.01 ± 0.01 and 1.01 ± 0.02, respectively; these values were higher than those in the ZOL-treated group (P < 0.01). The protein level of integrin α(v) and β3 in the ZOL-treated group (31,934.84 ± 112.91 and 18,812.79 ± 194.13) was downregulated by approximately 39.19% and 40.17%, respectively, compared with those in the control group (52,517.81 ± 211.72 and 31,441.93 ± 456.87) (P < 0.01). Immunofluorescent examination showed that the fluorescent intensities of integrin α(v) and β3 in the ZOL-treated group (9.491 ± 0.748 and 4.744 ± 0.759) were also significantly decreased compared with those in the control group (15.159 ± 1.143 and 11.418 ± 1.095) (P < 0.01).</p><p><b>CONCLUSION</b>ZOL significantly inhibits osteoclast adhesion and downregulates integrin α(v) and β3, expression, thus contributing to the ZOL-induced inhibition of osteoclast- mediated bone resorption.</p>

Enhanced integrin-mediated human osteoblastic adhesion to porous amorphous calcium phosphate/poly (L-lactic acid) composite / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The initial osteoblastic adhesion to materials characterizes the first phase of cell-material interactions and influences all the events leading to the formation of new bone. In a previous work, we developed a novel amorphous calcium phosphate (ACP)/poly(L-lactic acid) (PLLA) material that demonstrated morphologic variations in its microstructure. The aim of this study was to investigate the initial interaction between this material and osteoblastic cells. Cellular attachment and the corresponding signal transduction pathways were investigated.</p><p><b>METHODS</b>A porous ACP/PLLA composite and PLLA scaffold (as a control) were incubated in fetal bovine serum (FBS) containing phosphate-buffered saline (PBS), and the protein adsorption was determined. Osteoblastic MG63 cells were seeded on the materials and cultured for 1, 4, 8, or 24 hours. Cell attachment was evaluated using the MTS method. Cell morphology was examined using scanning electron microscopy (SEM). The expression levels of the genes encoding integrin subunits α1, α5, αv, β1, focal adhesion kinase (FAK), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined using real-time reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The ACP/PLLA material significantly increased the protein adsorption by 6.4-fold at 1 hour and 2.4-fold at 24 hours, compared with the pure PLLA scaffold. The attachment of osteoblastic cells to the ACP/PLLA was significantly higher than that on the PLLA scaffold. The SEM observation revealed a polygonal spread shape of cells on the ACP/ PLLA, with the filopodia adhered to the scaffold surface. In contrast, the cells on the PLLA scaffold exhibited a spherical or polygonal morphology. Additionally, real-time RT-PCR showed that the genes encoding the integrin subunits α1, αv, β1, and FAK were expressed at higher levels on the ACP/PLLA composite.</p><p><b>CONCLUSIONS</b>The ACP/PLLA composite promoted protein adsorption and osteoblastic adhesion. The enhanced cell adhesion may be mediated by the binding of integrin subunits α1, αv, and β1, and subsequently may be regulated through the FAK signal transduction pathways.</p>

Potential targets for anti-liver fibrosis / 药学学报

Liver fibrosis is a pathological process of the excessive accumulation of extracellular matrix, especially collagen al (I) in liver. Ultimately, hepatic fibrosis leads to cirrhosis or hepatic failure. Liver fibrosis and early cirrhosis can be reversed, thus control of the development of liver fibrosis is very important for preventive treatment of cirrhosis and hepatic failure. This is a review of potential targets for anti-hepatic fibrosis based on plenty of publications, including TGF-β1 and integrin α(v) and so on, aimed at providing novel therapeutic targets in liver fibrosis.

Tanshinone II a protects against lipopolysaccharides-induced endothelial cell injury via Rho/Rho kinase pathway / 中国结合医学杂志

<p><b>OBJECTIVE</b>To test whether tanshinone II A (Tan II A), a highly valued herb derivative to treat vascular diseases in Chinese medicine, could protect endothelial cells from bacterial endotoxin (lipopolysaccharides, LPS)-induced endothelial injury.</p><p><b>METHODS</b>Endothelial cell injury was induced by treating human umbilical vein endothelial cells (HUVECs) with 0.2 μg/mL LPS for 24 h. Y27632 and valsartan were used as positive controls. The effects of tanshinone II A on the LPS-induced cell viability and apoptosis rate of HUVECs were tested by flow cytometry, cell migration by transwell, adhesion by a 96-well plate pre-coated with vitronectin and cytoskeleton reorganization by immunofluorescence assay. Rho/Rho kinase (ROCK) pathway-associated gene and protein expression were examined by microarray assay; quantitative real-time polymerase chain reaction and Western blotting were used to confirm the changes observed by microarray.</p><p><b>RESULTS</b>Tan II A improved cell viability, suppressed apoptosis and protected cells from LPS-induced reductions in cell migration and adhesion at a comparable magnitude to that of Y27632 and valsartan. Tan II A, Y27632 and valsartan also normalized LPS-induced actomyosin contraction and vinculin protein aggregation. A microarray assay revealed increased levels of fibronectin, integrin A5 (ITG A5), Ras homolog gene family member A (RhoA), myosin light chain phosphatase, phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K, or PIP2 in Western blotting), focal adhesion kinase, vascular endothelial growth factor and vascular endothelial growth factor receptor 2 in the damaged HUVECs, which were attenuated to different degrees by Tan II A, Y27632 and valsartan.</p><p><b>CONCLUSION</b>Tan II A exerted a strong protective effect on HUVECs, and the mechanism was caused, at least in part, by a blockade in the Rho/ROCK pathway, presumably through the down-regulation of ITG A5.</p>

Expression and significance of integrins subunits in laryngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#This study was to investigate the expression and significance of Integrins subunits in laryngeal squamous cell carcinoma (LSCC).@*METHOD@#The expression of Integrins subunits was detected by cDNA microarray in 4 cases of primary LSCC tissues and corresponding adjacent normal tissues. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) were used to identify the different expression of Integrins subunits in 24 cases of primary LSCC tissues and corresponding adjacent normal tissues.@*RESULT@#A cDNA microarray analysis revealed significant changes in the expression of Integrins subunits, with IntegrinalphaV, Integrinbeta8 being up-regulated and Integrinalpha8 being down-regulated. The result of RT-PCR was consistent with that of cDNA microarray. The mRNA levels of IntegrinalphaV and Integrinbeta8 were significantly higher in LSCC tissues than that in corresponding adjacent normal tissues (1.0131 +/- 0.4780 vs 0.7591 +/- 0.4678 for IntegrinalphaV, P<0.05, 1.7362 +/- 1.3849 vs 1.2267 +/- 0.9363 for Integrinbeta8, P<0.05). The mRNA levels of Integrinalpha8 were significantly lower in LSCC tissues than that in corresponding adjacent normal tissues (0.2646 +/- 0.2622 vs 0.5457 +/- 0.3827, P<0.05).@*CONCLUSION@#The expression of IntegrinalphaV, Integrinbeta8, Integrinalpha8 were significantly up-regulated or down-regulated in laryngeal squamous cell carcinoma, which may relate to tumorigenesis and development of laryngeal squamous cell carcinoma.

Effects of bushen zhuyun recipe on protein expressions of estrogen receptor, progesterone receptor and integrin alpha5 and beta3 in endometrium of rats at the implantation stage / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of Bushen Zhuyun Recipe (BZR) on protein expressions of estrogen receptor (ER), progesterone receptor (PR) and integrin alpha5 and beta3 in endometrium of rats at the implantation stage, for exploring the possible mechanism of the recipe in treating luteal phase defect (LPD) infertility.</p><p><b>METHODS</b>Female SD rats were randomly divided into 6 groups, the blank group, the model group, the WM group treated by Western medicine, and the three BZR groups treated by low-, middle- and high-dose BZR respectively. Rats were made to pregnancy and sacrificed at the implantation stage, their middle segment of uterus, about 1 cm in length was gotten for detecting the protein expressions by Western blot. Results The protein expressions of endometrial ER and PR were significantly higher, while those of integrin alpha5 and beta3 were significantly lower than those of the control group (P < 0.05). The protein expressions of endometrial ER and PR were significantly lower, but those of integrin alpha5 and integrin beta3 were higher in rats treated by middle- and high- dose BZR than those in model rats (P < 0.05).</p><p><b>CONCLUSION</b>BZR can raise the receptivity of rats' endometrium through down-regulating the expressions of ER, PR and increasing the protein expression of integrin alpha5 and beta3 in endometrium and thus to enhance the pregnant rate.</p>

Netrin-1 Specifically Enhances Cell Spreading on Fibronectin in Human Glioblastoma Cells

Netrins are secreted molecules and involved in axon guidance, cell migration and tumor development. Recent studies revealed that netrins perform novel functions in such processes as epithelial development and angiogenesis without operating through the classical netrin receptors, DCC (Deleted in Colorectal Cancer) and Unc5h. In the present study, we investigated the roles of netrin-1 and its receptors in cell spreading of human glioblastoma cells, and found that netrin-1 haptotactically enhanced fibronectin-induced cell spreading and focal adhesion formation in U373 glioblastoma cells. Netrin-1 binding to the U373 cell membrane was blocked by an antibody against alpha v integrin subunit, but not by an anti-DCC or anti-Unc5h antibody. In addition, enhancement of the fibronectin response by netrin-1 was abrogated by a function blocking antibody against integrin alpha v beta 3. Since the alpha v subunit of the integrin family plays an important role in the pathophysiological aspects of cell migration, including tumor angiogenesis and metastasis, our data provide important insight into the molecular mechanism of netrin function.

Expression of alpha-v integrin subunit in the laryngeal and hypopharyngeal squamous cell carcinomas and its relationship with tumor angiogenesis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of integrin alpha-v subunit in laryngeal and hypopharyngeal squamous cell carcinomas (LHSCC) and to correlate the expression ratio with clinic and pathologic features of LHSCC, meanwhile, to investigate the relationship between the expression of integrin alpha-v subunit and tumor angiogenesis.</p><p><b>METHODS</b>A tissue microarray of LHSCC was designed and made. Using this microarray, the expression of integrin alpha-v subunit in LHSCC was studied by immunohistochemistry, and the expression disparity in different clinic and pathologic staging of LHSCC was analyzed. The immunohistochemical staining of CD105 was done in the same microarray, the intratumor microvessel density (IMVD) was calculated by CD105 staining. The relationship between integrin alpha-v subunit expression and the IMVD was analyzed.</p><p><b>RESULTS</b>In primary cancer tissue, the expression ratio of integrin alpha-v subunit was 68.0% (51/75), significantly higher than normal tissue beside cancer (10.3%, 3/29, chi2 = 28.68, P < 0.001); the expression ratio of integrin alpha-v subunit in lymph node metastatic carcinoma was 100.00% (20/20), significantly higher than normal tissue (chi2 = 38.77, P < 0.001) and primary cancer tissue (chi2 = 12.69, P < 0.05); in group with lymph node metastasis, the expression ratio of integrin alpha-v subunit was significantly higher than group without lymph node metastasis (chi2 = 10.87, P < 0.001); the IMVD of the group with integrin alpha-v subunit positive expression was significantly higher than the group with integrin alpha-v subunit negative expression (P < 0.001).</p><p><b>CONCLUSIONS</b>There was significant relationship between integrin alpha-v subunit expression and lymphatic metastasis of LHSCC. Overexpression of the integrin alpha-v subunit may have contributed to the tumor angiogenesis and lead to lymphatic metastasis. Integrin alpha-v subunit may become a novel lymphatic metastasis marker of LHSCC.</p>

The Expression of HOX D3 and Vascular Endothelial Growth Factor-C in Skin Tumors / 대한피부과학회지

BACKGROUND: The anatomical relation between a malignant tumor and its vascular and lymphatic bed is an important influencing metastasis. Hox D3 is required for these expressions of integrin alpha v beta3 and urokinase plasminogen activator (uPA), which contribute to endothelial cell adhesion, invasion, and migration during angiogenesis. Recent studies in different tumor types have shown that vascular endothelial growth factor-C (VEGF-C), which displays a high specificity for lymphatic endothelium, is involved in tumor-induced lymphagiogenesis and lymphatic metastatic spread. OBJECTIVE: This study was designed to measure the expression of HOX D3 and VEGF-C in different skin cancers. METHODS: The expression of HOX D3 and VEGF-C was examined by immunohistochemical staining of 40 skin cancer tissue samples, including 8 keratoacanthomas, 8 extramammary paget's disease, 8 basal cell carcinomas, 8 squamous cell carcinomas and 8 malignant melanomas. RESULTS: Immunohistochemical analysis of 40 skin cancer tissue samples revealed a high expression of HOX D3 and VEGF-C in the more aggressive and invasive skin tumors, including squamous cell carcinomas and malignant melanomas. On the other hand, low expression was seen in the less-invasive skin tumors, including keratoacanthomas, extramammary paget's disease and basal cell carcinomas. Also the degree of expression of HOX D3 and VEGF-C showed a statistically-significant correlation with each skin tumor (p<0.05). CONCLUSION: These findings provide evidence that the upregulation of HOX D3 and VEGF-C might be involved in the promotion of angiogenesis and lymphagiogenesis in skin tumors and play an important role in metastasis.

Cloning and sequence analysis of cDNA encoding porcine alphav subunit for FMDV receptor / 生物工程学报

Receptors play a crucial role in determining the pathogenesis and tissue tropism of virus. Foot-and-mouth disease virus (FMDV) has been showed to use four integrins, alphavbeta1, alphavbeta3, alphavbeta6 and alphavbeta8 as receptors to initiate infection. In this study, the porcine integrin alphav gene was cloned by RT-PCR from the lung tissue of healed pig infected experimently with FMDV, and compared its nucleotide and deduced amino acid sequence with the av gene of other animals. The 3141bp cDNA of bovine integrin alphav encodes a polypeptide of 1046 amino acids consisting of a 30-residue putative signal peptide, a 955-residue ectodomain, a 29-residue transmembrane domain, and a 32-residue cytoplasmic domain. The ectodomain contains 11 potential N-linked glycosylation sites (NXT/NXS), 2 calcium binding domains (DX[D/N] XDGXXD) and 18 cysteine residues. The nucleotide sequence similarities of integrin alphav between pig and cattle, human, rheses monkey, house mouse, chicken, dog are 93.3%, 91.5%, 91.4%, 85.6%, 73.2% and 89.9% respectively; and the amino acid sequence similarities are 96.3%, 94.6%, 94.1%, 90.8%, 81.6% and 93.8%, respectively. The alphav gene of cattle and pig exhibited the highest sequence homology. It is possible that host tropism of FMDV may related to divergence in receptors among different species.

Statins Inhibited the ADP-Stimulated Activation of Integrins alpha v beta5 and alpha v beta3 of Vascular Smooth Muscle Cells

BACKGROUND AND OBJECTIVES: Integrins mediate the migration, adhesion and proliferation of vascular smooth muscle cells. Adenosine diphosphate (ADP) can activate vascular integrins. We assessed the hypothesis that 'statins inhibit the ADP-stimulated activation of integrins alpha v beta5 and alpha v beta3 in human aortic smooth muscle cells (HASMC)'. MATERIALS AND METHODS: The expressions of integrins were analyzed using flow cytometry. The activations of integrins were evaluated using the adhesion assay, with prothrombin as an activation-dependent ligand. The MTT assay was used to evaluate the proliferation. RESULTS: Statins did not suppress the expressions of the integrins, alpha v beta5 and alpha v beta3. The ADP-stimulated adhesion was partially prevented by LM609, which blocked integrin alpha v beta3 (13% inhibition), and markedly prevented by P1F5, which blocked integrin alpha v beta5 (76% inhibition; n=5, p<0.05). However, the proliferation was inhibited by c7E3 and LM609, but not by P1F5. Statins inhibited the ADP-stimulated adhesions in a dose-dependent manner after 15 min of pretreatment. After incubating HASMC with statins for 1 day, simvastatin and fluvastatin inhibited the adhesion by 70 and 66%, respectively (n=5, p<0.05 vs. no statin). Statins also inhibited the ADP-stimulated proliferation of HASMC. CONCLUSION: Statins did not suppress the expressions of the integrins, alpha v beta5 and alpha v beta3, but did inhibit the ADP-stimulated activation of the integrins of HASMC.

Effects of Ovariectomy on Bone Mineral Density and Integrin Expression in Maxilla of Rats / 대한해부학회지

In postmenopausal osteoporosis, estrogen deficiency leads to unbalance of bone metabolism, decreased bone formation and increased bone resorption, and the result is reduced bone mineral density (BMD) and bone stiffness. The processes of bone formation and resorption involves the expression of integrins in anchoragedependent cells, such as osteoblast and osteoclast. The osteoporosis-induced rats frequently demonstrated the loss of trabecular bone volume in the tibia, vertebra and mandible due to estrogen depletion. However, in maxilla, study has been rare because of its anatomical limits. So the objective of this study was to investigate bony change and property of integrin expression in maxilla of osteoporosis-induced rats. 12-week-old female Sprague-Dawley rats were bilaterally ovariectomized (OVX). At 1, 2, 3, 4, 8 and 12 weeks, control and OVX group rats were sacrificed respectively. BMD of maxilla of the rats was measured using dual-energy X-ray absorptiometry (DEXA). And then the histopathologic observation, histomorphometric analysis and immunohistochemistry with CD44, alpha2 integrin, alpha5 integrin, alpha6 integrin, alphav integrin and beta3 integrin were done. BMD of alveolar bone in maxilla was decreased with significance statistically after OVX 4 weeks and was decreased 18.15% at OVX 12 weeks group compared to control group. From OVX 4 to 12 weeks, the thickness of periodontal ligament space was decreased, the number of osteoclast and the size of marrow stroma were increased than control group. By histomorphometric analysis, the size of marrow stroma of alveolar bone in maxilla was increased 86.42% at OVX 12 weeks group compared to control group. CD44 was widely expressed throughout the odontoblast, cementoblast, dental pulp, preiodontal ligament, osteocyte, osteoclast and perivascular tissue at control group, and CD44 immunoreactivity was increased the odontoblast, osteoblast and osteoclast at OVX groups. alpha2 integrin was expressed the odontoblast and osteoblast at control group, but alpha2 integrin immunoreactivity was decreased the osteoblast at OVX 12 weeks group. alpha5 integrin was expressed the cementoblast, osteoblast and osteoclast at control group, and alpha5 integrin immunoreactivity was decreased the osteoblast and was increased the osteoclast from OVX 4 weeks group. alpha6 integrin was weakly expressed the odontoblast, cementoblast, osteoblast and osteoclast at control group, and alpha6 integrin immunoreactivity was weakly increased the osteoclast from OVX 4 weeks. alphav integrin was expressed the odontoblast and osteoclast at control group, and alphav integrin immunoreactivity was strongly increased the osteoclast from OVX 4 weeks. beta3 integrin was expressed the osteocyte and osteoclast at control group, and beta3 integrin immunoreactivity was strongly increased the osteoclast from OVX 4 weeks. From these results, alveolar bone in maxilla of OVX rats was decreased BMD gradually. Moreover, alpha2 and alpha5 integrin expression of osteoblast was decreased, and alpha5, alphav and beta3 integrin expression of osteoclast was increased in OVX rats. Thus, this study indicates that consideration of reduced BMD is necessary in dental procedure of postmenopausal women.

Study of the expression of integrin alphaV beta3 in mandibular distraction osteogenesis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and distribution pattern of Integrin alphaV beta3 (ItgalphaV beta3) in course of distraction 44 adults New Zealand white osteogenesis and to quest for the function of ItgalphaV beta3 in distraction osteogenesis (DO).</p><p><b>METHODS</b>rabbits were selected and divided randomly into 3 groups: DO, bone fracture and normal group. There were 20, 20 and 4 rabbits in each group separately. Animal models were established and animals were killed at different time points. Sections were stained by IHC method. Distribution and expression pattern of ItgalphaV beta3 were observed by semi-quantitative technique, and the results were managed with statistic methods.</p><p><b>RESULTS</b>The expression of ItgalphaV beta3 was found both in the DO and bone fracture groups. The positive staining was seen mainly in vascular endothelial cells on cytomembrane and in cytoplasm. The staining intensity of group of DO was higher than that of the bone fracture group.</p><p><b>CONCLUSION</b>ItgalphaV beta3 plays an important role in promoting the process of DO.</p>

In vivo characterization of the integrin beta3 as a receptor for Hantaan virus cellular entry

Binding of viruses to cell surface molecules is an essential step in viral infection. In vitro studies suggested that the alpha v beta3 integrin receptor is the epithelial cell receptor for Hantaan virus (HTNV). Whether beta3 is in vivo the only or central cellular receptor for HTNV infection is not known. To investigate the role of beta3 integrin for cellular entry of HTNV, we established an HTNV infection model in newborn murine pups. Infected pups died at an average age of 14.2 +/- 1.1 days with high levels of viral antigen detected in their brain, lung, and kidney. Pre-injection of blocking monoclonal antibodies (mAb) specific for either beta3 or av prolonged survival significantly to a maximal average survival of 19.7 +/- 1.5 days (P<0.01) and 18.4 +/- 0.9 days (P<0.01), respectively. XT-199, a chemical blocker of the alpha v beta3 receptor also prolonged survival to 19.5 +/- 1.3 days (P<0.01). In contrast to these receptor blockades, anti-HTNV antibody was not only able to prolong survival, but 20% of infected pups achieved long-term survival. An anti-murine beta1 antibody comparatively prolonged survival (19.0 +/- 1.2 days), suggesting that HTNV infection is partly mediated through integrin beta1 receptors as well as through beta3 receptors in vivo. Our data demonstrate that the beta3 receptor is important for HTNV infection in vivo, but also suggest that HTNV may utilize additional receptors beyond beta3 for cellular entry within an organism.

The Patterns of Integrin Subunit Distribution in Glomerular Disease / 대한신장학회잡지

BACKGROUND: It has been known that integrins are not only simple glue which mediate cell-to-cell or cell to extra-cellular matrix but also function as signaling molecules at the surface of cell by conformational change. Because of the diversity of subunits and versatility of ligand recognition, integrins have been recognized as important molecules in kidney and the expression of integrin in normal human kidney has already been reported. METHODS: To know the significance of integrin expression in the pathogenesis of various renal diseases, we stained kidney specimens from the patients with membrano- proliferative glomerulonephritis(MPGN), minimal change nephrotic syndrome(MCNS) and membraneous nephropathy(MN) of lupus nephritis with 13 monoclonal antibodies of integrins. RESULTS: There was a significant change in the distribution of integrin subunits according to the types of glomerular diseases. Integrin subunit alpha2 was helpful in confirming the mesangial interposition as there was intense staining of subendotherial area in MPGN, while only the mesangium was stained with this unit in normal. In addition, an abundant staining of alpha4 and beta2 was observed and this finding indicated non-glomerular resident cell is participating in the MPGN, while in MN type of lupus nephritis, alpha2 staining was restricted to mesangium and alpha4 or beta2 integrin staining was negative. In the MN type of lupus nephritis, the remarkable finding was a ragged appearance in the subepithelial area, just below the glomerular basement membrane, which was visible in alpha3 and alphaV integrin staining. In MCNS, there was no difference in the staining pattern of integrin. CONCLUSION: Integrin subunits can be useful in evaluating the specific cell types which is actively involved in pathogenesis and these results suggest that alteration of integrin distribution can play an important role in the pathogenesis of glomerular diseases.

Expression of Adhesion Molecules in Placenta and Correlation with Uterine Artery Doppler Velocimetry in Pregnancy-induced Hypertension / 대한산부인과학회잡지

INTRODUCTION: The pathophysiology of PIH remains unclear. Recently, placental abnormalitiesare stressed as a possible cause of PIH. Abnormal shallow invasion of trophoblasts, confinedto decidua, without involving myometrium is believed to result in reduced uteroplacentalperfusion, endothelial injury, and activation of coagulation cascade system. Integrin, one of theadhesive membrane proteins, is expected to be related to the regulation of trophoblasts invasion. PURPOSE: The purpose of this study is to investigate the expression of adhesion moleculesin placenta and the correlation between uterine artery Doppler findings and integrinexpressions in the placentas of PIH patients. SUBJECTS: Thirty-six cases of severe PIH patients were enrolled in the study with 10number of normal control pregnant women. The integrin subunit expressions withimmunohistochemical staining were observed in floating villi, maternal-side cytotropholbasts, andfetal-side cytotrophoblasts. Uterine artery Doppler study was also performed, and the S/Dratio was evaluated. Abnormal Doppler findings was defined as S/D ratio>or=2.6. RESULTS: Cytoplasmic staining of villi and placental bed cytotrophoblast for theintegrin alpha1 subunit in PIH specimen was weaker than those in normal controls. Theexpression of integrin beta1 subunit was negative for both controls and PIH group. Thepositive cytoplasmic stain was observed among PIH placenta in contrast to normal control inwhich the expression of integrin beta4 subunit was not detected. The expression of alpha v beta3 introphoblast with PIH was positive staining, but not in control group. Uterine artery Dopplervelocimetry was performed in 25 cases with PIH. Trace(+/-) or - staining of integrin alpha1 subunit were observed in 60.0% of abnormal S/D(>or=2.6) group, 20.0% of normal S/Dratio group patients, respectively. Trace or + staining of integrin beta4 subunit were observedin 50.0% of abnormal S/D group and 6.7% of normal S/D group and this is in statisticallysignificant. Trace or + staining of integrin alpha v beta3 subunit were observed 70.0% ofabnormal S/D group and 26.7% of normal S/D group, and this statistically significant. CONCLUSION: In PIH the abnormality in the invasion of cytotrophoblats results inabnormal integrin subunit expression, but it is also correlated to the abnormal uterine arteryDoppler velocimetry which shows a S/D ratio of greater than 2.6. Thus, the uterine arteryDoppler velocimetry reflects abnormal placentation.