Effects of total flavonoids from Nymphaea candida on hepatic fibrosis induced by carbon tetrachloride in rats / 中草药

Objective: To investigate the preventive effect of total flavonoids from Nymphaea candida (NCTF) on carbon tetrachloride (CCl4)-induced hepatic fibrosis in rats. Methods: Wistar rats were randomly divided into control group, model group, NCTF groups (50, 100, 200 mg/kg) and colchicine group (0.2 mg/kg). Rats were subcutaneously injected with 50% CCl4 peanut oil solution (0.1 mL/100g, twice a week for 12 weeks) to induce hepatic fibrosis. In addition to control and model group given with 0.5% CMC-Na, the other groups were intragastrically administered with drugs (1.0 mL/100 g, once a day for 12 weeks), all rats were put to death. Blood, hepatic and splenic tissue were collected to detect liver indexes. Pathological histology observation by Masson and HE staining were performed for other hepatic tissues. The expression of α-smooth muscle actin (α-SMA) in hepatic tissue was analyzed by immunohistochemical technique. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein (TP) and albumin (ALB), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and nitric oxide (NO) were determined by automatic biochemical analyzer. The levels of serum laminin (LN), type III procollagen (PCIII), type IV collagen (CIV), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured by enzyme-linked immunoassay assay (ELISA). Serum hyaluronic acid (HA) was detected by radioimmunoassay. Hepatic tissue hydroxyproline (Hyp) was measured by spectrophotometric method. Result Compared with the control group, the rats showed energielos, liver and spleen index of those were increased, as well as inflammatory cell infiltration and obvious fibrosis of hepatic tissue were appeared in the model group; The levels of ALT, AST, ALP, HA, LN, PCIII, CIV, MDA, NO, TNF-α, IL-1β and IL-6 in serum in model group were increased significantly (P < 0.05, 0.01); The levels of TP, ALB, GSH and SOD in serum were decreased significantly (P < 0.01); The expressions of α-SMA and Hyp in hepatic tissue were significantly improved (P < 0.01). Compared with the model group, the levels of ALT, AST, ALP, MDA, NO in serum in NCTF group were significantly reduced (P < 0.05, 0.01); The levels of HA, LN, PC III, CIV, TNF-α, IL-1β and IL-6 were significantly reduced (P < 0.05, 0.01); The levels of TP, ALB, GSH and SOD were significantly increased (P < 0.05, 0.01); The expressions of α-SMA and Hyp in hepatic tissue were significantly reduced (P < 0.05, 0.01). Compared with the model group, the pathological damage of hepatic tissue was significantly improved, the degeneration and necrosis of liver cells in rat were significantly reduced (P < 0.05). Conclusion: NCTF has a better anti-hepatic fibrosis effect, its mechanism is related to antioxidant, regulation of collagen synthesis and inhibition of the proinflammatory factors expression.

Study on the Effects and Mechanism of Matrine on Proliferation and Collagen Synthesis of Hepatic Stellate Cells CFSC-8B Activated by Acetaldehyde / 中国药房

OBJECTIVE：To study the effects of matrine on proliferation and collagen synthesis of rat hepatic stellate cells CFSC-8B activated by acetaldehyde ，and to investigate its possible mechanism. METHODS ：CFSC-8B cells cultured in vitro were divided into blank control group ，model group ，positive control group （2.5 μmol/L colchicine）and matrine low ，medium and high concentration groups （30，60，120 μmol/L）. Except for blank control group ，other groups were activated with 200 μmol/L acetaldehyde for 24 h；medicine groups were intervened with relevant medicine for 24 h（blank control group and model group were intervened with equal volume blank medium ）. Survival rate of cell was detected by CCK- 8 assay. C ells were divided into blank control group ，model group ，positive control group （2.5 μmol/L colchicine），matrine medium and high concentration groups （60，120 μmol/L），then activated and treated with same method. Hydroxyprolin （Hyp）content in cell culture solution was tested by enzyme digestion. The contents of Col- Ⅰ and Col- Ⅲ in cell culture solution were determined by ELISA. mRNA expressionss of α-SMA，TGF-β1，TβR-І，TβR-Ⅱ，Smad3，Smad4 and Smad 7 in cells were detected by RT-PCR. The protein expressions of α-SMA，TGF-β1，TβR-Ⅰ，TβR- Ⅱ，Smad3，Smad4 and Samd 7 in cells were detected by Western blotting. RESULTS ：Compared with blank control group ，survival rate of cells in model group was increased significantly （P＜0.05）；the contents of Hyp ，Col-Ⅰ and Col- Ⅲ in cell culture solution ，mRNA and its protein expressions of α-SMA，TGF-β1，TβR-Ⅰ，TβR-Ⅱ，Smad3，Smad4 in cells were increased significantly in model group （P＜0.05），while the mRNA and protein expression of Smad 7 was decreased significantly（P＜0.05）. Compared with model group ，survival rate of cells ，the contents of Hyp ，Col-Ⅰ and Col- Ⅲ in cell culture solution，the mRNA and protein expressions of α-SMA and Smad 4 were decreased significantly in positive control group and matrine medium and high concentration groups （P＜0.05）， while the mRNA and protein expression of Smad 7 was WF-0099） increased significantly （P＜0.05）；the mRNA and proteinexpressions of TGF-β1，TβR-Ⅰ，TβR-Ⅱ and Smad 3 were decreased significantly in positive control group and matrine high concentration group （P＜0.05）. Compared with matrine medium concentration group ，all above indexes were improved significantly in matrine high concentration group （P＜0.05）. CONCLUSIONS ：Matrine can suppress the proliferation and collagen synthesis of CFSC- 8B cells activated by acetaldehyde ，with a centain concentrlation dependence ，the mechanism of which may be associated with regulating the conduction of TGFβ/Smad signal pathway.

Comparative Study Of Conventional Local Steroid Injections Only With Multi-Drugs Therapy And Steroid Injections In Oral Submucosal Fibrosis (OSMF)

Oral submucous fiosis (OSMF) is defined as the chronic inflammatory disease and progressive fiosis with localized collagen disorder involving oral mucosa. There are multiple pathophysiological changes that occur in this disease. The aim of this study is to establish that multidrugs needed for effective conservative treatment in oral submucous fiosis. A total number of 35 cases of OSMF managed in our ENT out patient department (OPD) over one year period ( March 2010 to Feuary 2011) were followed up until July 2011 and the results were analyzed retrospectively to find out any incidence of recurrence of this disease. All the cases received injection steroid and hyaluronidase with multivitamins (A,C and E in higher doses) for 10 weeks. At the end of treatment all the patients were relieved of burning sensation of mouth and got nearnormal mouth opening. The oral mucosa also changed from pale to pink. During follow-up no recurrence was detected. The study suggested that Vitamins A, C and E in higher doses are required to ensure best possible result apart from intralesional steroid and hyaluronidase as a standard conservative treatment protocol in oral submucous fiosis.

Effects of Chelidonine on Proliferation ，Collagen Synthesis and TGF-β1 Receptor of Activated Hepatic Stellate Cells CFSC- 8B in Rats / 中国药房

OBJECTIVE： To investigate the effects of chelidonine on proliferation， collagen synthesis and TGF-β1 receptor of activated hepatic stellate cells CFSC-8B. METHODS： CFSC-8B cells in logarithmic phase were collected and then divided into normal control group， model group， solvent group （ethanol）， positive control group （1 μg/mL colchicine ethanol solution）， chelidonine low， medium and high concentration groups （2.1， 4.2， 8.4 μg/mL chelidonine ethanol solution）. Except for normal control group， other groups were activated with 20 μg/L TGF-β1 for 24 h； the latter 5 groups were intervened with relevant medicine for 24 h. Cell proliferation of activated cells was assayed by CCK-8 assay. Hydroxyprolin （Hyp） content was assayed by enzyme digestion； the levels of typeⅠ collagen （Col-Ⅰ） and type Ⅲ collagen （Col-Ⅲ） were assayed by ELISA； the expressions of TβR-Ⅰ and TβR-Ⅱ protein were assessed by Western blot； mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ in hepatic stellate cells were assessed by RT-PCR. RESULTS： Compared with normal control group， cell proliferation rate， Hyp content， the levels of Col-Ⅰ and Col-Ⅲ， the protein expressions of TβR-Ⅰ and TβR-Ⅱ as well as mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ were increased significantly （P＜0.05）. Compared with model group， there were no significant difference in above indexes of hepatic stellate cells in solvent group （P＞0.05）； there were no significant difference in the proliferation rate of hepatic stellate cells in chelidonine low concentration group （P＞0.05）， above indexes of hepatic stellate cells were decreased significantly in positive control group and chelidonine high concentration group （P＜0.05）. The decrease of Hyp and Col-Ⅲ levels were not significant in chelidonine medium concentration， but other above indexes were decreased significantly （P＜0.05）. Compared with chelidonine medium concentration group， the rate of cell proliferation， Col-Ⅰ level， protein and mRNA expressions of TβR-Ⅰ and TβR-Ⅱ were decreased significantly in chelidonine high concentration group （P＜0.05）. CONCLUSIONS： Chelido- nine can inhibit the proliferation， collagen synthesis as well as the protein and mRNA expressions of TβR-Ⅰand TβR-Ⅱ in activated CFSC-8B cells.

Effects of thrombopoietin on TGFβ1-induced myofibroblast transdifferentiation in hu-man lung fibroblasts / 中国肿瘤临床

Objective: To investigate the effects of thrombopoietin (TPO) on proliferation and collagen synthesis in pulmonary fibro-blasts induced by TGFβ1. Methods: Cultured human embryonic lung fibroblasts (HFLs) were treated with recombinant human TGF-β1 to induce myofibroblast differentiation. Different concentrations of recombinant human TPO were applied individually or in combina-tion. Cell proliferation rate was determined using the CCK8 assay. Q-PCR and immunofluorescence assay were employed to examine the mRNA and protein expression of α-smooth muscle actin (αSMA) and type I collagen (COL1)A2. Results: TGFβ1 treatment induced HFL transdifferentiation to myofibroblasts was determined by the expression of αSMA, a myofibroblast-specific marker. Cell prolifera-tion increased during the induction. COL1 gene and protein expression were upregulated by TGFβ1 induction (P<0.05). The TGFβ1-in-duced mRNA and protein expression of αSMA and COL1A2 was decreased by TPO treatment (P<0.05), as determined by reverse tran-scription quantitative polymerase chain reaction and immunofluorescence analysis, respectively. The inhibitory rate showed a dose de-pendent effect within a certain TPO concentration range. The CCK8 assay demonstrated that TPO downregulated the TGFβ1-induced proliferation (P<0.05). Furthermore, the expression of heme oxygenase-1 (HO-1) was downregulated in TGFβ1-induced lung fibro-blasts, and these effects were attenuated by TPO administration (P<0.05). Conclusions: TPO can inhibit the TGFβ1-induced prolifera-tion and differentiation of human lung fibroblasts. These effects may be mediated in part by HO-1-related signaling pathways.

Effects of total saponins of Schizocapsa plantaginea Hance on hepatic fibrosis in rats and its mechanism / 中国药理学通报

Aim To study the effect of total saponins of Schizocapsa plantaginea Hance ( SFSP) on hepatic fibrosis in rats induced by CC14 and to investigate the molecular mechanisms of these effects. Methods 5 SD rats were randomly divided into five groups: Normal control group, model group, colchicine (Col) as positive control group, low-dose SFSP group (SFSP-L), and high-dose SFSP group ( SFSP-H ). After four weeks of continuous administration, serum AST, ALT and hydroxyproline (Hyp) were measured by biochemical detection method. The four indicators of liver fibrosis ( CIV, PC HJ , LN, HA) in serum were measured by enzyme linked immunosorbent assay. The location of liver fibrous tissues was observed by Masson staining. Liver sections were stained with HE, and observed for pathologic changes. Liver ultrastructural changes were observed using a transmission electron microscope. The expressions of mRNA of α-smooth muscle actin(α-SMA) , transforming growth factor pi (TGF-β1) , Smad4 and Smad7 in liver were detected by qPCR. Results In the liver fibrosis model group, the contents of AST, ALT and CIV, PCI, LN, HA, Hyp reflecting liver injury and fibrosis significantly increased ( P < 0. 05 ) , while in SFSP-H group and Col group the contents of the above indicators were significantly reduced, and the pathological results of liver tissues were consistent with it. SFSP significantly inhibited the expression of TGF-β1, Smad4 and α-SMA mR-NA ( P < 0. 01 ) , and increased the expression of Smad7 as well (P < 0. 01). Conclusion SFSP has an antifibrogenic effect through down-regulating the TGF-(β1/Smad signaling pathway in rats.

Inhibitory effect and mechanism of paeoniflorin on proliferation of hypertrophic scar fibroblasts / 中国比较医学杂志

Objective To explore the inhibitory effect and related mechanism of paeoniflorin on proliferation of hypertrophic scar(HS)fibroblasts. Methods HS fibroblasts were cultured with 0(control),200,400,800 μmol/L of paeoniflorin for 24,48,and 72 h. Cell viability was detected by MTT assay. Cell apoptosis was detected using Hoechst staining. Cell cycle was measured by flow cytometry. The levels of type I collagen(COL I)and type III collagen(COL III)were detected by enzyme linked immunosorbent assay(ELISA). The expressions of transforming growth factor-β (TGF-β)/Smad signaling pathway related proteins, as well as matrix metalloproteinase 1(MMP1)and MMP13 were detected by Western blot. Results 200, 400, 800 μmol/L of paeoniflorin reduced the cell viability of HS fibroblasts significantly(P < 0.01), with the nuclei turning pale and shrunken, and caused cell cycle arrest at G1 phase(P <0.01). Moreover, the levels of COL I and COL III in the cells were decreased significantly(P < 0.01), and the expressions of MMP1, MMP13, TGF-β1, p-Smad2 and p-Smad3 were down-regulated significantly(P < 0.01). Conclusions Paeoniflorin can obviously inhibit the proliferation and collagen synthesis of hypertrophic scar fibroblasts,probably through inhibition of the TGF-β1/Smad signaling pathway.

Pulmonary Toxicity in Rats Caused by Exposure to Intratracheal Instillation of SiO2 Nanoparticles / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>The effect of the silica nanoparticles (SNs) on lungs injury in rats was investigated to evaluate the toxicity and possible mechanisms for SNs.</p><p><b>METHODS</b>Male Wistar rats were instilled intratracheally with 1 mL of saline containing 6.25, 12.5, and 25.0 mg of SNs or 25.0 mg of microscale SiO2 particles suspensions for 30 d, were then sacrificed. Histopathological and ultrastructural change in lungs, and chemical components in the urine excretions were investigated by light microscope, TEM and EDS. MDA, NO and hydroxyproline (Hyp) in lung homogenates were quantified by spectrophotometry. Contents of TNF-α, TGF-β1, IL-1β, and MMP-2 in lung tissue were determined by immunohistochemistry staining.</p><p><b>RESULTS</b>There is massive excretion of Si substance in urine. The SNs lead pulmonary lesions of rise in lung/body coefficients, lung inflammation, damaged alveoli, granuloma nodules formation, and collagen metabolized perturbation, and lung tissue damage is milder than those of microscale SiO2 particles. The SNs also cause increase lipid peroxidation and high expression of cytokines.</p><p><b>CONCLUSION</b>The SNs result into pulmonary fibrosis by means of increase lipid peroxidation and high expression of cytokines. Milder effect of the SNs on pulmonary fibrosis comparing to microscale SiO2 particles is contributed to its elimination from urine due to their ultrafine particle size.</p>

Clinical Assessment of Periodontal Status in Patients on Long Term Corticosteroid Therapy.

Corticosteroids (Cs) are used widely for their anti-inflammatory and immunosuppressive properties. However their long-term administration may lead to impaired periodontal health. The aim of the present study was to clinically assess the periodontal status in patients on long-term corticosteroid therapy. Periodontal health of 100 patients under long-term corticosteroid therapy for a minimum of 6 months duration was compared with sex- and age-matched 100 healthy controls. The periodontal examination included measuring oral hygiene index-simplified (OHI-S), gingival index (GI), sulcus bleeding index (SBI), probing pocket depth (PPD) and clinical attachment loss (CAL). The results showed that mean values of OHI-S, GI and SBI did not differ significantly (p>0.05) between cases and controls. Mean PPD and CAL was significantly higher in cases when compared to the controls (p = 0.0003). Within the limitations of the study, it can be concluded that there is a positive correlation between periodontal status and long term steroid therapy.

Expressão gênica do colágeno em ferida cutânea de equinos tratada com plasma rico em plaquetas / Collagen gene expression in skin wound of horses treated with platelet-rich plasma

[...]The objective of this study was to evaluate type I and III collagen gene expression during different phases of the healing process of PRP-treated skin. Eight healthy crossbred geldings, aged 16 and 17 years (16.37±0.52) were used. Three quadrangular-shaped lesions (6.25cm²) were surgically induced in the right and left gluteal regions of all the animals. Twelve hours after induction of the lesions, 0.5mL of PRP was administered in each of the four edges of the wounds (T=treated group) in one of the gluteal regions, randomly chosen. The contralateral region was used as control (NT=non-treated group). The wounds were submitted to daily cleaning with Milli-Q water, and the samples were obtained with a 6mm diameter biopsy Punch. Six skin biopsies were obtained, with the first being performed on the day the lesions were induced (T0), and the others 1 (T1), 2 (T2), 7 (T3), and 14 (T4) days, after the wound was induced. The sixth biopsy (T5) was performed after fully healed of the skin. Evaluation of type I and III collagen gene expression was carried out by the qRT-PCR technique. The data were analyzed by the Bonferroni test, Student t-test, paired t-test, and regression analysis (p<0,05). Difference (p<0.05) between groups were observed for both collagen gene expressions from T1 to T4, being higher in the animals of group T. The peak for type I and III collagen gene expressions occurred in T5 for both groups, but the highest expression was different (p<0.05) from zero time, starting in T3. In the animals of treated group, collagen expression started to establish at T5, while in the horses of NT group, the values remained increased. Local administration of a single PRP dose in cutaneous wound of the gluteal region of horses results in a higher local gene expression of type I and III collagens. However, this expression does not alter the maximum time of macroscopic healing of the wound.

[...] Objetivou-se avaliar a expressão dos genes dos colágenos tipos I e III durante diferentes fases do processo de cicatrização da pele tratada com PRP. Foram utilizados oito equinos machos castrados, mestiços, hígidos, com idade entre 16 e 17 (16,37±0,52) anos. Três feridas em formato quadrangular (6,25cm²) foram confeccionadas nas regiões glúteas direita e esquerda de todos os animais. Doze horas após indução das lesões, 0,5mL do PRP foi administrado em cada uma das quatro extremidades das feridas (T=grupo tratado), de uma das regiões glúteas, escolhida aleatoriamente. A região contralateral foi utilizada como controle (NT=grupo não tratado). As feridas foram submetidas à limpeza diária com água Milli Q, e amostras foram obtidas com biópsias utilizando-se Punch de 6mm de diâmetro. Seis biópsias de pele foram obtidas a primeira no dia de indução das lesões (T0), e as demais com 1 (T1) 2 (T2) 7 (T3) e 14 (T4) dias após a realização das feridas. A sexta biópsia (T5) foi realizada após o completo fechamento da pele. A avaliação da expressão dos genes dos colágenos tipos I e III foi realizada pela técnica qRT-PCR e os dados analisados pelo teste de Bonferroni, t de Student, t pareado e análise de regressão (p<0,05). Diferenças (p<0,05), entre grupos, foram observadas para a expressão de ambos os colágenos nos T1 a T4, sendo maior nos animais do grupo T. O pico de expressão dos colágenos tipos I e III ocorreu no T5 para ambos os grupos, mas a maior expressão foi diferente (p<0,05) do tempo zero a partir do T3. Nos animais do grupo tratado a expressão dos colágenos começou a estabilizar no T5, enquanto que nos equinos do NT os valores permaneceram elevados. A administração local de uma única dose do PRP em ferida cutânea na região glútea de equinos, resulta em maior expressão gênica local dos colágenos tipos I e III. Entretanto, essa expressão não altera o tempo máximo de fechamento macroscópico da ferida.

Protection of peimisine on hepatic fibrosis of rats induced by CCl4 / 中草药

Objective: To investigate the protective effect of peimisine on carbon tetrachloride (CCl4)-induced hepatic fibrosis in rats. Methods: The rats were divided into control, model, low-, mid-, and high-dose (2.5, 5, and 10 mg/kg) peimisine groups. Hepatic fibrosis models were induced by ip injection of CCl4 in rats once every 3 d for 8 weeks. The rats in the treatment groups were administered four weeks after the model establishment, once daily until the end of the week 4 after the model establishment. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glutamyl transpeptidasecc (GGT), hyaluronie acid (HA), laminin (LN), type III procollagenc (PC-III), and collegen type IV (IV-C) were assayed, and hepatic tissue contents of hydroxyprolinc (Hyp), superoxide dismutase (SOD), and malondialdehyde (MDA) were determined. The effect of peimisine on hepatic fibrosis in rats was observed. Results: Compared with the model control group, the hepatic fibrosis of rats in peimisine groups was improved obviously, the levels of ALT, AST, ALP, and GGT in serum were lowered obviously (P < 0.05, 0.01), also the serum levels of HA, LN, PC-III, IV-C, and the contents of Hyp and MDA in liver tissue were decreased (P < 0.01), while the level of SOD was increased (P < 0.01). Conclusion: Peimisine has the protective effect on the experimental hepatic fibrosis formation. The possible mechanisms are associated with inhibiting fibrogenesis and fibrosis accumulation, and decreasing lipid peroxidation.

Effect of Xilei Powder on primary cultured fibroblast functions related to wound healing / 中成药

AIM:Xilei Powder,a traditional Chinese prescription,has been used to treat wounds for hundreds of years,but the mechanism has not been fully understood.METHODS:The effects of Xilei Powder on fibroblast proliferation,collagen accumulation,matrix metalloproteinases-2,9 ( MMP-2,9 ) activities and tissue inhibitor of metalloproteinase 1 (TIMP-1) production were investigated by MTT,chloramine T method,gelatin zymography and enzyme-linked immunosorbent assays ( ELISA),respectively.RESULTS: The aqueous extract of Xilei Powder significantly promoted fibroblasts proliferation in a time and concentration manner,the population doubling time (125 μg/mL) was 33.8 h,it also significantly (P ＜0.05 ) promoted collagen production.Both of the aqueous and alcoholic extracts could significantly ( P ＜ 0.05 ) increase MMPo2 activity,and also very significantly ( P ＜ 0.01 )promote TIMP-1 production.CONCLUSION: Xilei Powder could promote fibroblasts proliferation,collagen and TIMP-1 production,this might be parts of mechanism to promote wound healing.

Zinc may increase bone formation through stimulating cell proliferation, alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells

Zinc is an essential trace element required for bone formation, however not much has been clarified yet for its role in osteoblast. We hypothesized that zinc would increase osteogenetic function in osteoblasts. To test this, we investigated whether zinc treatment enhances bone formation by stimulating osteoblast proliferation, bone marker protein alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells. MC3T3-E1 cells were cultured and treated with various concentrations of zinc (0, 1, 3, 15, 25 uM) along with a normal osteogenic medium (OSM) as control for 1, 5, 10 days. As measured by MTT assay for mitochondrial metabolic activity, cell proliferation was stimulated even at low zinc treatment (1-3 micrometer) compared to OSM, and it was stimulated in a zinc concentration-dependent manner during 5 and 10 days, with the most pronounced effect at 15 and 25 uM Zn. Cellular (synthesized) alkaline phosphatase (ALP) activity was increased in a zinc concentration-dependent manner, so did medium (secreted) ALP activity. Cellular collagen concentration was increased by zinc as time went by, therefore with the maximum zinc stimulatory effect in 10 days, and medium collagen concentration showed the same pattern even on 1 and 5 day. This zinc stimulatory effect of collagen synthesis was observed in cell matrix collagen staining. The study results imply that zinc can increase osteogenic effect by stimulating cell proliferation, ALP activity and collagen synthesis in osteoblastic cells.

The comparison of gene expression from human dental pulp cells and periodontal ligament cells / 대한치과보존학회지

The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA microarray assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well. According to this study, the results were as follows: 1. From the microarray assay, 51 genes were more expressed (2 fold) from PC than PDLC. 2. RT-PCR confirmed that ITGA4 and TGF beta2 were more expressed in PC than in PDLC 3. From the microarray assay, 19 genes were more expressed (2 fold) from PDLC than PC. 4. RT-PCR confirmed that LUM, WISP1, and MMP1 were more expressed in PDLC than in PC. From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.

Inhibitory effect of cyclosporin A on growth and collagen synthesis of rat cardiac fibroblasts induced by arginine vasopressin / 药学学报

Aim To investigate the effects of cyclosporin A (CsA) on growth and collagen synthesis of cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods CFs of neonatal Sprague-Dawley rats were isolated by trypsinization and cultured; growth-arrested CFs were stimulated with 1×10-7 mol·L-1 AVP in the presence or absence of CsA (0.05, 0.5 and 5 μmol·L-1). MTT and flow cytometry techniques were adopted to measure cell number and analyze cell cycle respectively. Collagen synthesis was determined by measurement of hydroxyproline content in culture supernatant with colorimetry. Calcineurin activity was estimated by chemiluminescence. Trypan blue staining to test the viability of CFs. Results 0.05, 0.5 and 5 μmol·L-1 CsA inhibited the increase of CFs number induced by 1×10-7 mol·L-1 AVP in a dose-dependent manner, with the inhibitory rates by 12%, 24% and 29%, respectively (P＜0.05). Furthermore, cell cycle analysis showed 0.5 μmol·L-1 CsA decreased the S stage percentage and proliferation index of CFs stimulated by AVP (P＜0.05). In culture medium, the hydroxyproline content induced by AVP decreased by 0.5 and 5 μmol·L-1 CsA (P＜0.05), with the inhibitory rates of 29% and 33%, respectively. CsA completely inhibited the increment of calcineurin activity induced by AVP (P＜0.01), but CsA itself had no effect on the baseline of calcineurin activity and CFs viability. Conclusion CsA inhibits proliferation and collagen synthesis of CFs by virtue of blocking calcineurin signaling pathway and might provide a novel target for prevention and treatment to cardiac fibrosis.

Effect of vitamin C deficiency on the rate of orthodontic tooth movement and alveolar bone remodeling / 대한치과교정학회지

This study was undertaken to investigate the effect of vitamin C deficiency on the orthodontic tooth movement and bony remodeling processes. Thirty six male guinea pigs were divided on the basis of the given amount of vitamin C (normal group: 5 mg/day, deficient group: 0.2 mg/day) and 75 gm of force was applied to the maxillary incisors. Experimental animals were sacrificed at day 0, day 1, day 3, day 5, day 7 and day 14 after force application and the amount of tooth movement was measured and tissues were studied histologically. The results showed that the amount of collagen fiber in the periodontal ligament and alveolar bone of the deficient group was less than that of the normal group. In the stretched side, the osteoblastic activity and alveolar bone formation of the normal group increased in a time dependent manner during experimental periods, but the deficient group showed less activity and formation. The amount of tooth movement in the deficiency group was more than in the normal group at day 0, day 1, day 3, day 5, and day 7. According to the above results, a deficiency of vitamin C resulted in a defect of collagen synthesis of the periodontium and inhibition of bone formation and stimulation of bone resorption with rapid tooth movement in early periods of force application.

The Effect of an AP-1 decoy on Collagen Synthesis in Wound Healing on Rat Skin: The use of Gene Therapy to Minimise Scarring / 대한피부과학회지

BACKGROUND: The wound healing process is composed of inflammation, cellular growth, migration, angiogenesis and an extracellular matrix composition. In this process, fibroblasts proliferate but leave scarring due to their excessive growth. The process is controlled by platelet derived growth factor (PDGF) and basic fibroblast growth factor (bFGF), while PDGF and transforming growth factor beta(TGF-beta) play a major role in controlling extracellular matrix composition. Numerous modalities have been tried to treat this abnormal response, but the results were unsatisfactory. TGF-beta is activated by AP-1 (Activator protein). Theregone, AP-1 decoy oligodeoxynucleotides(ODN) had recently been used for regulation of TGF-beta transcription as a gene treatment. OBJECTIVE: The purpose of this study was to examine the effect of blocking TGF-beta transcription by an AP-1 decoy ODN on collagen synthesis in wound healing on rat skin. METHODS: In this study, the effect of AP-1 decoy ODN on collagen synthesis was examined by Hematoxylin and eosin (H&E) staining, Masson's trichrome staining, and immunohistochemical staining for TGF-beta on damaged rat skin. RESULTS: In the H&E stain and Masson's trichrome stain of the damaged rat skin, the number of collagen fibers of AP-1 decoy ODN treated group had decreased in compared to the control group, especially on the 15th day after incision. With immunohistochemical stain, the expression of TGF-beta in fibroblasts, inflammatory cells and the endothelium of vessel walls in the dermis had also decreased, compared to the control group. TGF-beta was expressed in the dermis from the 3rd day, and predominantly in the fibroblasts on the 15th day after incision. CONCLUSION: These results indicate that AP-1 decoy ODN is a powerful down-regulator of collagen synthesis in wound healing through significant suppression of TGF-beta expression in damaged skin. Therefore, AP-1 decoy ODN can be used effectively to treat or minimize scarring on damaged skin.

Variable Effect of Estrogen on Fibroblast Proliferation and Collagen Synthesis by Gender and Age

It was assumed that the effect of estrogen on wound healing would be variable according to patient's gender and age since estrogen is a sex steroid. This study was designed to determine the variability of the effect of estrogen on proliferation of human dermal fibroblasts and collagen synthesis which are most important in wound healing considering patient's gender and age. Fibroblasts were isolated from the dermis of female patients in premenstrual, menstrual, or postmenopausal age group and that of male patients. The isolated fibroblasts were cultivated in the presence of estrogen(1.0microgram/ml). The cells were seeded at 5.0x103cell/ well in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 5% fetal bovine serum in 96-well plates. The cells were incubated for 3 days. For fibroblast proliferation MTT assay method was used. To measure the production of collagen, the collagen type I carboxy- terminal propeptide enzyme immunoassay was carried out. Estrogen stimulated the proliferation of fibroblasts in female patients, but not in male patients. The greatest cell proliferation and collagen synthesis was seen at women in menstrual and postmenopausal age. These results demonstrated that effects of estrogen on dermal fibroblast proliferation and collagen synthesis were variable with gender and age.

Effect of Estrogen on Fibroblast Proliferation and Collagen Synthesis

Estrogen is a sex steroid hormone which is known to be helpful in preventing aging process in various ways. It is frequently used in hormone replacement therapy (HRT). Basically wound healing is likely to have same process on the view of cell proliferation and extracellular matrix formation. However, it is not determined yet whether estrogen up-regulates or down-regulates wound healing. This study was designed to determine the effect of estrogen on proliferation of human dermal fibroblasts and collagen synthesis in vitro which are most important in wound healing. Fibroblasts were isolated from the dermis of adults and cultivated in the presence of either one of 5 concentrations of estrogen(0, 0.01, 0.1, 1.0, 10microgram/ml). The fibroblasts were seeded at 5.0x103cell/well in Dulbecco's Modified Eagle's Medium/Ham's F-12 nutrient including 10% fetal bovine serum with either one of 5 different concentration of estrogen in 96-well plates. The cells were incubated for 3 days. For fibroblast proliferation MTT assay method was used. To measure the production of collagen, the collagen type I carboxy- terminal propeptide enzyme immunoassay was carried out. All concentrations of estrogen stimulated the proliferation of fibroblasts. The best proliferation and maximum stimulation of collagen synthesis was seen at supplementation of 1.0microgram/ml of estrogen. The collagen synthesis per cell was also maximal at concentration of 1.0microgram/ml estrogen. These results demonstrate that estrogen influences human dermal fibroblast proliferation and the estrogen concentration is critically important factor in vitro.

The Effect of 1320 nm Nd: YAG Laser and Long-pulsed 1064 nm Nd: YAG Laser Irradiation on Hairless Mouse Skin / 대한피부과학회지

BACKGROUND: The treatment of facial rhytides has traditionally centered around methods that involve removal of the epidermis and superficial dermis, encouraging the production of a new epidermis with collagenesis and remodelling. But all of the resurfacing techniques lead to postoperative complications such as oozing, bleeding, infections, "downtime" as the skin begins to reepithelialize, and the occasional incidence of posttreatment, postinflammatory pigmentary changes. 1320nm and 1064nm wavelengths are nonspecifically absorbed in the human dermis and are unique for its significant horizontal scattering. These wavelengths, when used for localized facial areas, have been shown to produce new collagen formation and improvement in the quality of treated skin. OBJECTIVE: The purpose of this study was to examine the effect of 1320nm Nd: YAG laser and long-pulsed 1064nm Nd: YAG laser irradiation on hairless mouse skin and rat skin. METHODS: In this study, the effect of 1320nm Nd: YAG laser and long-pulsed 1064nm Nd: YAG laser irradiation were examined by Hematoxylin and eosin (H&E) stain, Masson's trichrome stain, immunohistochemical stain for type I collagen and dot-blot hybridization for alpha1(I) procollagen mRNA. RESULTS: In the H&E stain, Masson's trichrome stain, and immunohistochemical stain of the hairless mouse skin, the number of collagen fibers with a greater density of fibers increased, compared to the non-irradiated controls on both the 1320nm Nd: YAG laser and the long-pulsed 1064nm Nd: YAG laser. In the dot-blot hybridization in the hairless mouse, levels of alpha1(I) procollagen mRNA were increased 3.0-fold, 4.8-fold and 5.2-fold at each 1 week, 4 weeks and 12 weeks after irradiation in 1320nm Nd: YAG laser and 1.8-fold and 2.1-fold at each 4 weeks and 12 weeks after irradiation in long-pulsed 1064nm Nd: YAG laser, compared to the non-irradiated controls. But in the 1320nm Nd: YAG laser irradiation on the rat, there is no significant change in the number and density of collagen fibers, compared to the non- irradiated controls. CONCLUSION: These results indicate that the 1320nm Nd: YAG laser and the long-pulsed 1064nm Nd: YAG laser may be a powerful up-regulator of collagen synthesis through significant dermal damage and the 1320nm Nd: YAG laser is better than the long-pulsed 1064nm Nd: YAG laser for collagen synthesis. Therefore, the 1320nm Nd: YAG laser can be more effective clinically than the long-pulsed 1064nm Nd: YAG laser for the treatment of photodamaged skin.