Progress on the mechanism of PDGF-BB signaling pathway in the pulmonary vascular remodeling of neonatal hypoxic pulmonary hypertension / 国际儿科学杂志

Neonatal hypoxic pulmonary hypertension(HPH)is a common acute critical disease in NICU, and is one of the diseases leading to neonatal death.At present, the specific pathogenesis is still unclear.Current studies have shown that pulmonary vascular remodeling is an important pathological feature of pulmonary hypertension, and the excessive proliferation and migration of pulmonary artery smooth muscle cell is the main cause of pulmonary vascular remodeling.Platelet-derived growth factor(PDGF-BB)is a powerful mitogenic factor which involved in cell proliferation and migration.Currently, plenty of studies have found that PDGF-BB plays an important role in multiple diseases, including tumor, atherosclerosis, pulmonary hypertension and pulmonary fibrosis.In view of the mechanism of PDGF-BB, this article reviews the possible mechanism of PDGF-BB in pulmonary vascular remodeling with neonatal HPH, aiming to provide a new direction for the therapies of reversing pulmonary vascular remodeling with neonatal HPH.

PDGF-BB promotes angiogenesis of human osteosarcoma through exosomes / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore whether PDGF-BB can be transmitted through exosome and verify its angiogenic function in human osteosarcoma. Methods: Exosomes from a variety of human osteosarcoma cells were isolated. The expression of PDGF-BB in cells and exosomes was detected by WB. Exosomes derived from osteosarcoma SJSA-1 cells were co-incubated with HUVEC, and the pattern of exosomal PDGF-BB entering HUVEC was observed using Immunofluorescence and confocal scanning microscope. SJSA-1 cell lines with PDGF-BB over-expression or knockdown were constructed by lentiviral infection, and the exosomes derived from transfected SJSA-1 cells were isolated and incubated with HUVEC. Microtubule formation experiment was conducted to detect their effects on angiogenesis; SJSA-1 cell transplanted xenograft model was established in nude mice, and the exosomes derived from SJSA-1 cells with PDGF-BB over-expression or knockdown were infused into nude mice to observe their effects on tumor growth. Results: The exosomes derived from osteosarcoma cells were successfully isolated, in which a large amount of PDGF-BB was confirmed. The exosomes entered HUVEC by endocytosis. The SJSA-1 cell lines with PDGF-BB over-expression or knockdown were successfully constructed, and the corresponding exosomes were isolated. Compared with the control group, exosomes with high PDGF-BB content significantly promoted HUVEC angiogenesis (P < 0.01 , t=13.51) and tumor growth (P < 0.01 ), while exosomes with low PDGF-BB content reduced the angiogenesis ability of HUVEC (P < 0.01 , t=8.226) and inhibited tumor growth (P < 0.01 ). Conclusion: The exosomal PDGF-BB secreted by osteosarcoma cells can be directly absorbed by HUVEC and induce tumor angiogenesis, further promoting the growth of osteosarcoma.

Efeitos do PDGF-BB na taxa de proliferação e na adesão de células derivadas da granulação óssea a fragmentos radiculares / Effects of PDGF-BB on the rate of proliferation and on the adhesion of human cells derived from bone granulation tissue to root fragments

O objetivo deste estudo foi investigar o papel do fator de crescimento derivado de plaquetas-BB (PDGF-BB) na concentração de 300ng/ml na taxa de proliferação e adesão de células derivadas da granulação óssea humana a fragmentos radiculares periodontalmente comprometidos. Na primeira etapa do estudo, foi estabelecida cultura primária de células da granulação óssea de dois pacientes adultos, sistemicamente saudáveis, não fumantes. Após a expansão celular, as células foram caracterizadas para determinação do fenótipo por meio de ensaios de viabilidade celular, MTT, ensaio de atividade de fosfatase alcalina, ensaio de mineralização e caracterização imunohistoquímica por meio de citometria de fluxo (segunda etapa). Na terceira etapa do estudo, os efeitos da adição de PDGF-BB recombinante humano na concentração de 300ng/ml na taxa de proliferação e adesão de células derivadas da granulação óssea a superfícies radiculares periodontalmente comprometidas foram investigados. A taxa de proliferação celular estimulada pelo PDGF-BB (grupo teste) ou pelo meio de cultura (grupo controle) foi investigada por meio de contagem de células viáveis nos frascos de cultura após 1, 3, 5 e 7 dias do cultivo celular. Foram obtidos 30 fragmentos dentários a partir de dentes extraídos por razões periodontais. Os fragmentos foram raspados com curetas Gracey e condicionados com solução em gel de EDTA a 24% durante 3 minutos, lavados com solução de soro fisiológico, secos e posicionados em placas de 24 poços. Foram incubadas sobre os fragmentos tratados 1x104 células GO por 24 horas, seguido por fixação e preparo para análise por microscopia eletrônica de varredura (MEV). O número de células aderidas sobre os fragmentos foi analisado nas fotomicrografias. O padrão de crescimento das células GO foi compatível com células ósseas, com modificação do padrão do crescimento com o aumento do número de passagens. Houve atividade de fosfatase alcalina em meio osteogênico e convencional, com pico máximo aos 7 dias e atividade de mineralização estimulada ou não por meio osteogênico, com pico máximo aos 21 dias. A análise por meio de citometria de fluxo demonstrou que as células GO não expressaram CD105 e CD166 na 14a passagem, indicando sua diferenciação celular avançada nesse período. A adição de rhPDGF-BB resultou em mudança na taxa de proliferação celular, observando-se pico máximo de crescimento aos 7 dias, com diferenças estatisticamente significantes (p < 0.005; ANOVA post hoc Tukey) em relação aos períodos de 1, 3 e 5 dias. O ensaio de MTT demonstrou maior viabilidade celular no período de 48 hs, comparativamente aos períodos de 24 e 72 horas, quando a densidade óptica celular diminuiu de forma significativa (p< 0.05; Friedmann pósteste Dunn). No ensaio de adesão celular, pode-se observar que a adição de rhPDGFBB aumentou significativamente o número de células aderidas aos fragmentos dentários (p< 0.05; teste t não pareado com correção Welch), com alteração da morfologia celular. Esses resultados sugerem que as células GO tem características compatíveis com linhagem de células osteoblásticas, de fenótipo mais diferenciado após a 12a passagem. A adição de rhPDGF-BB (300ng/ml) resulta em aumento da taxa de proliferação das células GO e do número de células aderidas a fragmentos radiculares, indicando que, nesta concentração, o fator de crescimento é citocompatível, favorecendo a proliferação e adesão celular.(AU)

The goal of this study was to investigate the effects of recombinant human platelet derived growth factor (rhPDGF-BB) at the concentration of 300ng/ml in the proliferation and adhesion of human bone granulation cells to periodontally diseased root fragments. At the first stage of the study, the granulation tissue existent in healing sockets (21 days after its creation) was collected from two systemically healthy nonsmoking adults to the establishment of primary culture. The in vitro properties of bone granulation (BG) cell lineage were characterized by cell viability, MTT, alkaline phosphatase activity and mineralization assays. The effects of culture medium (control) and rhPGDF-BB 300ng/ml (test) in the proliferation and adhesion of BG cells were investigated. The rate of BG cells proliferation was investigated by the number of viable cells present at 1, 3, 5 and 7 days after platting. Thirty root fragments were obtained from teeth extracted for periodontal reasons. Root fragments were scaled and root planed, conditioned with EDTA 24% for 3 minutes, rinsed in saline solution, air-dryed and positioned in 24-well plates. Each fragment was seeded with 104 BG cells, fixated after 24 hours and prepared for analysis in SEM. The number of cells adhered to the fragments was analysed in photomicrographies. BG cells growth pattern was compatible with osteogenic cell lineage, showing modification with the increasing number of cell passage. GO cells expressed alkaline phosphatase activity in conventional and osteogenic culture medium, with maximum peak at 7 days, as well as mineralization activity stimulated or not by osteogenic or non-osteogenic culture medium, with maximum peak at 21 days. The analysis by flow cytometer showed that BG cells have not expressed CD105 and CD106 at the 14th passage, indicating its advanced cell differentiation. The addition of rhPDGF-BB resulted in modification of proliferation rate, with maximum peak observed at 7 days, significantly different from 1-, 3- and 5-day periods (p< 0.005; ANOVA post hoc Tukey). MTT assay showed greater cell viability after 48 hours than after 24 and 72 hours, when optical density has significantly diminished (p< 0.05; Friedmann post hoc Dunn). At cell adhesion assay, it could be observed that the adhesion of rhPDGF-BB has significantly increased the number of cells adhered to root fragments (p< 0.05; unpaired t test with Welchs correction), and alterations in cell morphology. These results suggest that BG cells present in vitro characteristics compatible with osteoblastic cell lineages, with a more differentiated phenotype after the 12th passage. The addition of rhPDGF-BB (300 ng/ml) results in increase of the rate of BG cell proliferation and in the number of cells adhered to root fragments, indicating that, at this concentration, the growth factor is compatible with BG cells and favors cells proliferation and adhesion.(AU)

In vitro culture and migration assay of mouse smooth muscle progenitor cells / 中国药理学通报

Aim To establish a reliable method for the culture of mouse smooth muscle progenitor cells and in-vestigate their migration ability .Methods Mesenchy-mal stem cells from compact bones were obtained from C57 BL/6 mice and stimulated with PDGF-BB to in-duce these cells to differentiate into smooth muscle pro-genitor cells. Morphological analysis , immunocyto-chemical and flow cytometric analysis were used to i-dentify the cell type and the migration ability was in-vestigated by the transwell system and flow cytometry . Result After PDGF-BB stimulation for 7 days, the cells showed spindle shape and started to express α-SMA as demonstrated by immunocytochemistry .After 21 days induction , Flow cytometric analysis revealed that over 70%of the cells expressed both CD 34 andα-SMA and 58.5%of the cells expressed SM-MHC.Mi-gration assay showed that the smooth muscle progenitor cells from culture could migrate in vivo and in vitro. Conclusions The culture of smooth muscle progenitor cells from compact bone-derived mesenchymal stem cells is easily operated with high yield rate and shorten culture period . Obtained smooth muscle progenitor cells from culture could migrate in vivo and in vitro, which is suitable for the mechanism studies .

Estimation of the use of fibrin and collagen membranes as carriers for platelet-derived growth factor-BB (PDGF-BB) in the presence of amoxicillin.

The effect of homogeneous fibrin (Fb), collagen (Coll) and composite fibrin-heparin (Fb-Hp), fibrin-collagen (Fb-Coll) membranes on in vitro release of platelet-derived growth factor (PDGF-BB) was evaluated in the presence or absence of amoxicillin using of the ELISA immunoassay test. Amoxicillin concentration was determined spectrophotometrically at 272 nm. The process of the PDGF-BB growth factor and amoxicillin release from the studied membranes was of a two-phase nature in the majority of the systems analysed. The PDGF-BB was released in the highest amount from the Coll membrane (M7) without the presence of amoxicillin – 546.2 ±7.47 pg, t0.5 = 0.88 h and 202.5 ± 6.83 pg, t0.5 = 26.65 h during the first phase and second phase, respectively. The lowest PDGF-BB release was observed from composite M4 (Fb-Hp) membrane – 5.88 ± 0.81 pg, t0.5 = 1.69 h; and 110.2 ± 6.48 pg, t0.5 = 855.6 h during first and second phase respectively. An optimal release of amoxicillin was observed in the case of the composite M6 (Fb-Coll) membrane – only in the second phase: 64.2 ± 7.8 mg, t0.5 = 83.5 h. The lowest and delayed amoxicillin release was achieved for M4 membrane (approx. 17.1 ± 1.12 mg, t0.5 = 46.5 h). The results of the PDGF-BB release and amoxicillin from membranes indicated a correlation between the level of release and composition of the film. Our results suggested that fibrin and collagen membranes may be beneficial to enhance periodontal bone regeneration.

AICAR Blocks the Proliferation of Pulmonary Artery Smooth Muscle Cells by Inhibition of Cell Cycle in G0/G1 / 医学研究杂志

Objective To establish the PDGF-BB-induced proliferation of rat pulmonary artery smooth muscle cells(PASMCs), and investigate the effects of the AMPK agonist AICAR on the cell cycle of PASMCs, in order to search new drugs for prevention of pulmo-nary vascular remodeling. Methods 20ng/ml PDGF-BB was used to induced the proliferation of PASMCs, and the effect of 0. 5mmol/L AICAR on the proliferation of PASMCs was observed. Western blot was used to detect the total and phosphorylated AMPK. The prolifer-ation of PASMCs was determined by CCK-8. The mRNA expression of cyclinD1, cyclinE and CDK2/4/6 were detected by flow cytometry analysis cell cycle,quantitative real-time PCR. Results Western blot Results indicated AICAR could promote the activation of AMPK. CCK-8 test Results showed that AICAR blocked the proliferation of PASMCs induced by PDGF-BB. Flow cytometry analysis indicated that AICAR arrested the cell cycle in G0/G1 to S phase. RT-PCR Results demonstrated that AICAR inhibited the mRNA expression of cy-clinD1, cyclinE and CDK2/4/6. Conclusion The AMPK agonist AICAR can block the proliferation of PASMCs induced through arrest cell cycle in G0/G1-S phase by regulation the mRNA expression of cyclin D1, cyclinE, CDK2/4/6, and AICAR has a potential applica-tion in preventing pulmonary vascular remodeling.

Influence of extracellular matrix proteins and PDGF-BB on airway smooth muscle cells / 医用生物力学

Objective To investigate the influence of three extracellular matrix (ECM) proteins, namely, laminin (LN), collagen type I (Col I), fibronectin (FN) on the morphology and contractility changes in airway smooth muscle cells (ASMCs) induced by platelet-derived growth factor-BB (PDGF-BB). Methods ASMCs were seeded on the culture dish coated with LN, Col I, or FN, respectively, and divided into two groups to be cultured either in the absence or presence of PDGF-BB (10 mg/L) for 0~5 d. Subsequently, cell morphology was examined by the optical microscopy and quantified as the ratio of cell width to length, and the KCl/histamine-induced contractile responses of the cell were measured by optical magnetic twisting cytometry (OMTC). Results ASMCs cultured in the presence of PDGF-BB generally appeared in longer and thinner cell shapes, namely, a smaller ratio of cell width to length, but the cell width/length ratio for ASMCs adhered on LN was relatively bigger than that on Col I or FN. In the absence of PDGF-BB, contractility of ASMCs to KCl increased with the duration of culture, which was independent of the ECM proteins. In contrast, in the presence of PDGF-BB, contractility of ASMCs to KCl or histamine decreased in all situations, but degree of the decrease was smaller for ASMCs adhered on LN than those on Col I or FN. Conclusions The morphology and contractility changes in ASMCs induced by PDGF-BB are influenced by ECM proteins on which cells are grown. For ASMCs adhered on LN, the morphology and contractility changes are relatively smaller than those on Col I and FN. The differential effect of ECM proteins on PDGF-BB induced changes in morphology and contractility of ASMCs is important to fully understand the interactions between ECM proteins, inflammatory factors, ASMCs, and their relation to the pathophysiological mechanism of asthma.

Effect from different pore sizes of co-culture inserts on the permeability of platelet derived growth factor / 医用生物力学

Objective To investigate the effect from different pore sizes of co culture inserts on the permeability of biomacromolecules through polyethylene terephthalate (PET) membrane so as to solve the key technology problem in mechanobiology experiment on vascular cells. Methods Inserts with 0.4 μm and 1.0 μm pores on the PET membrane were studied using flow chamber system. Low shear stress was subjected to the co-cultured system of endothelial cell (EC)/vascular smooth muscle cell (VSMC) and the concentration of platelet-derived growth factor BB (PDGF-BB) was detected by ELISA. Under the static condition, vascular cells were cultured on the plate (with no cell on PET membrane), on the outer side of PET membrane, and on the both sides of PET membrane, respectively. Then the recombinants PDGF-BB (rPDGF-BB) were added on the different sides of PET membrane. Western blotting was used to detect the change in expressions of p-ERK1/2, p-Akt and Lamin after cells were stimulated by rPGDF BB. Results After low shear stress subjection for 12 h, the concentration of PDGF-BB in the medium from VSMC side was significantly higher than that from EC-side. rPDGF-BB passed through 0.4 μm and 1.0 μm pores on the PET membrane and modulated expressions of p-ERK1/2, p-Akt and Lamin A in cells cultured on the opposite side of PET membrane and cells cultured on the plate separately. When cells were cultured on the both sides of PET membrane, rPDGF-BB only stimulated cells cultured on the same side of 0.4 μm pores on PET membrane, but had no specific effect on cells cultured on the opposite side. Conclusions PET membrane with both 0.4 μm and 1.0 μm pores was permeable to PDGF-BB, and cells cultured on the membrane could affect the permeability. The efficiency of PDGF BB passing through 0.4 μm pores was significantly repressed with cells cultured on the both sides, which was more similar to that in vivo.

Increased Growth Factors in Children with Mycoplasma pneumoniae Pneumonia / 소아알레르기및호흡기학회지

PURPOSE: Mycoplasma pneumoniae is a common causative agent of community acquired pneumonia in children and is well known to cause various respiratory and extrapulmonary diseases. We determined whether growth factors, including transforming growth factor (TGF)-beta1 and platelet-derived growth factor (PDGF-BB) that regulate airway fibrosis and remodeling, are increased in young children with M. pneumoniae pneumonia and also investigated if there was any difference in relation to the clinical status of the patients. METHODS: Fifty-two patients (3 to 6 years of age) who were admitted with M. pneumoniae pneumonia were enrolled and divided into 2 groups: the patients with frequent wheezing episodes (group A, n=28) and the patients with no previous history of wheeze (group B, n=24). The former group included the patients who had recurrent wheeze more than 3 times before admission. Fifteen children admitted with minor surgical problems were also studied as controls. TGF-beta1 and PDGF-BB were measured in the plasma samples collected on admission using ELISA in both patient groups and controls. RESULTS: Plasma TGF-beta1 and PDGF-BB levels were increased significantly in the patients with M. pneumoniae pneumonia as compared to the controls (P<0.01, respectively). TGF-beta1 and PDGF-BB were higher in group A than in group B, but the difference was not statistically significant (P=0.08 vs. P=0.05). In group A, TGF-beta1 was significantly higher in atopic patients than in non-atopic patients (P<0.05). CONCLUSION: Our study showed significantly increased TGF-beta1 and PDGF-BB in patients with M. pneumoniae pneumonia. It is suggested that these growth factors may play an important role in the pathogenesis of lower airway infection by M. pneumoniae.

Transcriptional profiling and Wnt signaling activation in proliferation of human hepatic stellate cells induced by PDGF-BB / 대한간학회지

BACKGROUND/AIMS: This study aimed to better understand gene expression profiles of human hepatic stellate cell (HSC) activation and the relationship with the Wnt signaling pathway. METHODS: The global transcript levels in platelet derived growth factor-BB (PDGF-BB)-stimulated hTERT HSCs were analyzed using oligonucleotide microarrays. Oligonucleotide microarrays with 19K human oligo chips were performed to obtain gene expression profiles associated with proliferation in human hTERT HSCs. The microarray data was verified by real time quantitative PCR and expression of the components of Wnt signaling was analyzed by Western blot. RESULTS: Microarray data showed 243 up-regulated and 265 down-regulated genes in PDGF-BB-treated HSCs. The changes in expression of glypican3 and BH3 interacting domain death agonist (BID) mRNA in real time quantitative PCR, especially among the highly up- or down-regulated genes, were statistically consistent with the microarray data. The Wnt signaling pathway components, frizzled10 (FZD10) and calcium/calmodulin-dependent protein kinase II alpha (CAMK2A), showed increased expression in the short time course microarray and the up-regulation of FZD10 also occurred at the protein level. Our data showed various gene expression profiles during activation of human HSC. CONCLUSIONS: The up-regulated expression of FZD10 and CAMK2A suggests that the Wnt/Ca2+ signaling pathway is active in hTERT HSCs and may participate in HSC activation and proliferation

Effect of Qingfei Oral Liquid on Transforming Growth Factor-?_1 and Platelet-Derived Growth Factor-BB mRNA Gene Expression of Human Embryonic Lung Fibroblast Cells Infected by Adenovirus Type 3I and 7b / 中国中医药信息杂志

Objective To study the effect of Qingfei oral liquid contained serum on the transforming growth factor-?1 (TGF-?1) and platelet-derived growth factor-BB (PDGF-BB) mRNA gene expression of human embryonic lung fibroblast cells infected by adenovirus (ADV) type 3I and 7b. Method TGF-?1 and PDGF-BB mRNA expression of human embryonic lung fibroblast cells infected by ADV type 3I and 7b were determined by in situ hybridization before and after treated with Qingfei oral liquid contained serum. Results ADV could up-regulate TGF-?1 and PDGF-BB mRNA of human embryonic lung fibroblast cells (P

Effects of bFGF and PDGF-BB on osteoblast differentiation of bone marrow-derived mesenchymal stem cell in rat

In this study we evaluate the effects of bFGF-BB and PDGF on in vitro proliferation, differentiation and mineralization of mesenchymal stem cells (MSCs) from rat. MSCs were prepared from the bone marrow of 6 or 7-week-old male rats with a technique previously described by Maniatopoulos et al. in 1988. Lineage differentiation to osteogenesis, chondrogenesis and adipogenesis were performed. At first, we characterized the cultured cell on passage 1, 3, 5, 7 with immunocytochemical staining using CD29, 44, 34, 45, alpha-SMA and type I collagen. And to study the effects of bFGF and PDGF-BB on proliferation, differentiation and mineralization, we seeded the expanded cell at a density of 6 x 10(3) cells/cm2 to 100-mm dish for evaluation of cell proliferation and MTT assay was carried out on day 2, 4, 7, 9. We also resuspended the cells with same density (6 x 10(3) cells/cm2) to 24 well plates for subculture. On the following day, the attached cells were exposed to 2.5ng/ml bFGF and/or 25ng/ml PDGF-BB daily during 5 days. The osteocalcin (OC) level was assessed and mineral contents were evaluated with alizarin red S staining on subculture day 2, 7, 14, 21. We identified the mesenchymal stem cell from the bone marrow derived cells of rat through their successful multi-differentiation and stable display of its phenotype. And bFGF and PDGF-BB showed the effect that inhibited osteoblastic differentiation and mineralization mildly in above concentration at in vitro culture.

PD98059 Inhibited the Activation of Pancreatic Stellate Cells Mediated by Platelet-Derived Growth Factor BB in Rats / 华中科技大学学报（医学）（英德文版）

Summary: To determine the biological effects of extracelluar signal regulated kinase (ERK) specific inhibitor PD98059 on pancreatic stellate cells (PSCs) activated by platelet-derived factor-BB (PDGF-BB), cultured rat PSCs were co-incubated at 37 ℃ for 24 h with 25 ng/ml PDGF-BB and different doses of PD98059 (ranging from 5 ng/ml to 40 ng/ml). Expression of pERK1 protein was detected by Western blot and collagen α1 (Ⅰ) mRNA was tested by reverse transcription-polymerase chain reaction (RT-PCR). Our results showed that there were statistical differences in the expression of pERK1 protein in all groups. Expression of pERK1 protein was up-regulated in the group treated by PDGF-BB, and gradually down-regulated in the other groups treated by PD98059 of different doses. An excellent positive correlation was revealed between the inhibitory effect and PD98059 concentrations. It was also observed that the expression of collagen α1 (Ⅰ) mRNA had similar response to pERK1. The level of collagen α1 (Ⅰ) mRNA was the highest in the PDGF-BB group, and gradually reduced in the other groups treated by PD98059 of different doses. It is concluded that expression of pERK1 protein and collagen α1 (Ⅰ) mRNA was up-regulated in rat PSCs activated by PDGF-BB. Meanwhile, PD98059 could inhibit PSCs activation mediated by PDGF. It is suggested that ERK1 protein plays an important role on PSCs activation mediated by PDGF signal pathway.

Inhibitory effect of panaxynol on the proliferation of rat aortic smooth muscle cell and its mechanisms / 中国药理学通报

Aim To evaluate the inhibitory effect of panaxynol(PNN) on the proliferation of rat aortic smooth muscle cell(RASMC) and its mechanisms.Methods Cell proliferation was determined using cell count and -TdR incorporation test.Fura-3/AM and confocal were used to measure intracelluar free Ca~(2+) concentration.Expression of mitochondrial transcription factor 1(mtTF1) mRNA was tested by using RT-PR.Results PNN inhibited the RASMC proliferation and DNA synthesis induced by serum and PDGF-BB in a dose-dependent manner.9 ?mol?L~(-1) of PNN inhibited the increase of intracelluar free Ca~(2+) concentration induced by PDGF-BB.PDGF-BB upregulated the expression of mtTF1 mRNA,which could be suppressed by 3,9 ?mol?L~(-1) of PNN significantly.Conclusions PNN can inhibit RASMC proliferation significantly,which might be related to the decrease of intracellular free Ca~(2+) concentration and mtTF1 mRNA expression.

Clinical Significance of Serum PDGF-BB and IGF-1 in Coal Workers' Pneumoconiosis / 결핵

BACKGROUND: Coal workers' pneumoconiosis(CWP) is a fibrotic lung disease resulting from the chronic inhalation of coal dust. Various cytokines and growth factors secreted from macrophages and monocytes play a key r ole in the pathogenesis of penumoconiosis. The platelet-derived growth factor (PDGF)-BB and the insulin-like growth factor(IGF)-1 secreted from the macrophages and monocytes are believed to stimulate the accumulation of mesenchymal cells and fibrosis of the lower respiratory tract that is observed in fibrotic lung disease. The serum concentration of PDGF-BB and IGF-1 in 30 CWP patients and 10 healthy controls were measured in order to determine if PDGF-BB and IGF-1 can be used as sensitive biomarkers in CWP. METHODS: Serum was collected from 30 patients with CWP (13 with simple CWP and 17 with complicated CWP) and 10 healthy controls. The serum concentrations of PDGF-BB and IGF-1 were measured using ELISA (RandD system, Minneapolis, MN). RESULTS: The serum PDGF-BB concentration in patients with complicated CWP (10083.76+/-639.07 pg/ml) was significantly higher than in the patients with simple CWP(8493.88+/-848.51 pg/ml) and the healthy controls (3726.17+/-292.20pg/ml)(p0.05). CONCLUSIONS: These results show the important role of the PDGF-BB mediated pathways in the pathogenesis of CWP. These data suggests that the PDGF-BB serum concentration is a useful biomarkers of the fibrotic extent in CWP patients.

The effect of PDGF-BB loaded TCP/ chitosan microgranules on new bone formation / 대한치주과학회지

The purpose of this study was to evaluate newly fabricated tricalcium phosphate(TCP)/chitosan microgranuls as bone substitutes. TCP/chitosan microgranules were fabricated by dropping TCP-chitosan suspension into the NaOH/ethanol solution. The size of microgranules could be controllable via airflow rate. PDGF-BB was loaded into the fabricated granules via freeze-drying methods(300 ng/20 mg). To evaluate cell proliferation, cultured osteoblasts cell lines(MC3T3-E1) was dropped on the BioOss(R), chitosan microgranules, TCP/chitosan microgranules and cultured for 1, 7, 14, and 28 days. Scanning electron microscopic observation was done after 7 days of culture and light microscopic examination was done after 28 days of culture. PDGF-BB release from the microgranules was tested. Rabbit calvarial defects(8 mm in diameter) were formed and chitosan, TCP/chitosan, PDGF-TCP/chitosan microgranules, and BioGran(R) were grafted to test the ability of new bone formation. At SEM view, the size of prepared microgranules was 250-1000 um and TCP powders were observed at the surface of TCP/chitosan microgranules. TCP powders gave roughness to the granules and this might help the attachment of osteoblasts. The pores formed between microgranules might be able to allow new bone ingrowth and vascularization. There were no significant differences in cell number among BioOss(R) and two microgranules at 28 day. Light and scanning electron microscopic examination showed that seeded osteoblastic cells were well attached to TCP/chitosan microgranules and proliferated in a multi-layer. PDGF-BB released from TCP/chitosan microgranules was at therapeutic concentration for at least 1 week. In rabbit calvarial defect models, PDGF-TCP/chitosan microgranules grafted sites showed thicker bone trabeculae pattern and faster bone maturation than others. These results suggested that the TCP/chitosan microgranules showed the potential as bone substitutes.

The Effect of Interleukin 1-beta, Platelet Derived Growth Factor-BB and Transforming Growth Factor-betaon the expression of PDLs17 mRNA in the Cultured Human Periodontal Ligament Fibroblasts / 대한치주과학회지

The molecular mechanisms control the function of PDL(periodontal ligament) cells and/or fibroblasts remain unclear. PDLs17, PDL-specific gene, had previousely identified the cDNA for a novel protein from cultured PDL fibroblasts using subtraction hybridization between gingival fibroblasts and PDL fibroblasts. The purpose of this study was to determine the regulation by growth factors and cytokines on PDLs17 gene expression in cultured human periodontal ligament cells and observe the immunohistochemical localization of PDLs17 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant IL-1beta, PDGF-BB and TGF betafor 48h and 2 weeks. At each time point total RNA was extracted and the levels of transcription were assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). Polyclonal antiserum raised against PDLs17 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLs17 mRNA levels were increased in response to PDGF (10ng/ml) and TGF beta(20ng/ml) after treatment of the IL-1beta, PDGF-BB and TGFbetafor 48 h. 2. PDLs17 was up-regulated only by TGFbeta(20 ng/ml) after treatment of the IL-1beta, PDGF-BB and TGF betafor 2 weeks and unchanged by the other stimulants. 3. PDLs17 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLs17 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLs17 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLs17 might upregulated by PDGF-BB or TGFbetaand acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLs17 during the differentiation of bone marrow mesenchymal and PDL cells.

The Effect of PDGF-Loaded Biodegradable Membrane on Early Healing Stage in Guided Tissue Regeneration / 대한치주과학회지

The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect of a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/cm2 platelet derived growth factor. 2 beagle dogs were used in the experiment. 5mmx6mm alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test group or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new bone formation could be seen. Small amount of bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at 3 weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane and the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.

Effects of platelet-derived growth factor loaded bioresorbable membrane on periodontal regeneration / 대한치주과학회지

PDGF-BB has been recognized as a highly potential growth factor for guided tissue regeneration in periodontal defect. This study carried out histologic and histometric evaluation of 200ng/cm2 PDGF-BB loaded bioresorbable membrane made from polyglycolic and polylactic acid. It was tested for its biocompatibility, ability to prevent epithelial downgrowth and amount of periodontal regeneration. Without membrane and PDGF-BB unloaded bioresorbable membrane were used as control. Healthy six beagle dogs were used. Each dog was anesthetized and buccal flaps were reflected in the mandibular and maxillary premolar areas. Buccal alveolar bone between the mesiobuccal and distobuccal line angles was surgically removed on the lower 2nd and 4th premolar in mandible, 2nd premolar in maxilla, to a level 4mm apical to the cementoenamel junction with creating a Class II buccal furcation defect for available space. Care was taken not to remove the root cementum layer and rubber impression materials were placed over each surgically created defect. Flaps were repositioned and sutured. Reconstructive surgery was performed 1 month after defect preparation. PDGF-BB loaded membranes and controls were randomly placed on maxillary 2nd premolars and mandibular 2nd and 4th premolars. Plaque control regimen was instituted with daily brushing with a 0.1% chlorhexidine digluconate during experimental periods. The animals were sacrificed 2 and 5 weeks after surgery and undecalcified specimens were prepared for histologic evaluation. The degree of coronal regrowth of new bone, new cementum and the amonut of new bone areas formed on the defected area of the PDGF-BB loaded membrnae turned superior to without membrane and drug unloaded membrane. Experimental membrane could prevent the epithelial downgrowth irrespective of drug loaded or not and showed good biocompatiblity. These results implicated that PDGF-BB loaded bioresorbable membrane could be highly useful tool for guided tissue regeneration of periodontal defects.

Cellular activity and guided bone regenerative effect of drug-loaded biodegradable membranes / 대한치주과학회지

The purpose of this study was to evaluate the effects of tetracycline(TC), flurbiprofen, and PDGF-BB loaded biodegradable membranes on the cell-attachment, the activity of loaded PDGF-BB, in vivo release kinetics, and guided bone regenerative potentials. To evaluate the cell attachment to membranes, the number of gingival fibroblasts attached to each membrane(10% TC, 10% flurbiprofen, 200ng/cm2 PDGF-BB loaded membranes, drug-unloaded membrane) was counted by coulter counter and the morphologic pattern of attached cells was examined under SEM. To determine whether the activity of loaded PDGF-BB is sustained, the cellular growth and survival rate of gingival fibroblasts was used for both standard PDGF-BB and loaded PDGF-BB. For evaluation of in vivo release kinetics, drug-loaded membranes were implanted on the dorsal skin of the rats. On 1, 3, 7, 10, 14, 21, and 28 days after implantation, the amount of remaining drugs were measured by HPLC assay for TC and flurbiprofen, and by gamma-scintillation counter for PDGF-BB(I125). For evaluation of guided regenerative potential, the amount of new bone in the calvarial defects(5mm in diameter) of the rat was measured by histomorphometry 1 and 2 weeks after implantation of membranes. The number of cells attached to the PDGF-BB loaded membrane was largest as compared with the other membranes.(p<0.05) The activity of loaded PDGF-BB was not significantly different from the activity of standard PDGF-BB.(p< 0.05) After initial burst release of drug during the first 24 hours, drugs were gradually released for 4 weeks. Especially the release rate of PDGF-BB was nearly constant during 4 weeks. PDGF-BB loaded membranes(200, 400ng/cm2) were effective in guided bone regeneration as compared with drug-unloaded membrane. These results implicate that drug-loaded biodegradable membranes might be a useful for guided bone regeneration.