OBJECTIVEThe aim of this study was to investigate the expression of collagen in the process of masseter muscle reattachment to the cortical and cancellous bones of mandible.

METHODSA total of nine adult goats were used in the study. One was the control. The other eight were treated with bilateral detachment of the masseter muscles. The biopsies of bone and muscle were taken at 2, 4, 8 and 12 weeks after the operation. The characteristics of the healing muscle-bone interfaces were examined using immunohistochemical techniques.

RESULTSImmunohistochemical analysis illustrated that the locations of collagen type I, II and III were different during the healing process, but similar in the cortical and cancellous bones.

CONCLUSIONThis study demonstrates that the distribution of the three types of collagens at the muscle-bone interfaces is associated with time, but not related with their locations.

Animals ; Collagen ; biosynthesis ; genetics ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type II ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Female ; Goats ; Male ; Mandible ; metabolism ; pathology ; surgery ; Masseter Muscle ; metabolism ; pathology ; surgery ; Wound Healing ; physiology

OBJECTIVETo investigate the role of recombinant human transforming growth factor beta3 (rhTGFbeta3) on fibroblast and its possible mechanism.

METHODSNormal skin fibroblast (NSFb) and hypertrophic scar fibroblast (HSFb) were cultured in vitro, and were processed by different concentrations of rhTGFbeta3. NSFb and HSFb in DMEM solution without rhTGFbeta3 were employed as control. The changes in the protein and mRNA expression of type I and III collagen in NSFb and HSFb were observed.

RESULTS(1) The expression of type I and III procollagen in NSFb was evidently different from that of HSFb (2) The synthesis of type I and III procollagen in all test groups was increased obviously after rhTGFbeta3 process (P < 0.001) while the ratio of type I to III procollagen was decreased when compared with that in control group. (3) The effects of rhTGFbeta3 on the biological behavior exhibited an obvious dose- effects relationship. The contents of type I to III procollagen in HSFb were higher than those in NSFb when the dose of rhTGFbeta3 was same.

CONCLUSIONrhTGFbeta3 could effectively promote the synthesis of type I and III procollagen, especially type III procollagen in fibroblasts. This might be beneficial to the accelerate of wound healing and to inhibit or prevent scar formation.

Cicatrix, Hypertrophic ; metabolism ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Fibroblasts ; drug effects ; metabolism ; Humans ; In Vitro Techniques ; Recombinant Proteins ; pharmacology ; Transforming Growth Factor beta3 ; pharmacology

OBJECTIVETo evaluate the effects of taurine in diet on the expression of type I and III collagen and collagen ratio at different time points in rats lung by image process technology.

METHODSWistar rats were randomly divided into three groups: the saline instilled with a control diet (the saline treated group); silica instilled with a control diet (the silica treated group); and silica instilled with a diet containing 2.5% taurine (the taurine treated group). Animal models were established by the direct tracheal instillation of silica into rat lungs exposed surgically. The taurine concentration of serum was analyzed by means of HPLC. Paraffin embedded lung sections were stained with Sirius red. Polarization microscopy and Image Pro Plus Version 4.5 for windows were used for detecting type I and III collagen.

RESULTSThe concentration of taurine in serum of the taurine treated group was significantly elevated compared to the saline treated and silica treated group (P < 0.05 or P < 0.01). Sirius red polarization microscopy showed that type I and III collagen positive area percentage were elevated in the silica treated rats compared with the saline treated group. On the 7th, 14th, 21st, 28th day after silica instillation type I collagen positive area percentage was increased by 3.84, 3.77, 3.73, 9.83 respectively (P < 0.01), and type III collagen positive area percentage were elevated by a little in the silica treated rats compared with saline treated group. The taurine treatment significantly decreased elevation of silica type I collagen positive area percentage of lung by 2.39, 1.62, 7.13 at the 7th, 21st, 28th day respectively (P < 0.05 or P < 0.01), and type III collagen positive area percentage of lung by 2.62 at the 28th day (P < 0.05) compared with the silica treated group. The ratio of type I to III collagen was increased from the 7th day to 28th day after silica instillation, and reached 1.87 at the 28th day with the maximal ratio in the silica-treated group.

CONCLUSIONTreatment with taurine can effectively attenuate type I and III collagen expression in the rat lung induced by silica particles at different time points in our study.

Animals ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Female ; Lung ; drug effects ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Taurine ; pharmacology

AIMTo observe the effects of hypoxia on DNA synthesis and the expression of collagen type I and III mRNA in cultured adult rat cardiac fibroblasts.

METHODSCardiac fibroblasts(CFs) were isolated from adult Wistar rat ventricule and cultured in vitro either in normoxic or hypoxic condition. Studies were conducted with the second passage of CFs. The changes of DNA synthesis was determined by measuring the incorporation of 3H-TdR into DNA and the changes of expression of pro-alpha1 (I) collagen, pro-alpha1(III) mRNA were measured by in situ hybridization respectively.

RESULTSThe 3H-TdR incorporation of CFs was increased by 34% (P < 0.05) and 36% (P < 0.01) after 6 h, 12 h hypoxia (2% O2) exposure respectively. The level of pro-alpha1(I) collagen mRNA expression was significantly elevated in the cells under hypoxia for 4 h, 8 h, and 12 h. The expression of pro-alpha1(III) mRNA increased when cells were cultured under hypoxia for 2 h.

CONCLUSIONThese results suggest that hypoxia alone can upregulate DNA synthesis and expression of collagen type I and III mRNA in adult rat cardiac fibroblasts. It may be one of the important mechanisms by which hypoxic myocardial fibrosis occur.

Animals ; Cell Hypoxia ; Cells, Cultured ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; DNA ; biosynthesis ; Fibroblasts ; metabolism ; Male ; Myocytes, Cardiac ; cytology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

OBJECTIVETo observe the in vitro effects of gamma-interferon (IFNgamma) on gene expression of collagen I (Col I), III (Col III) and tissue inhibitor of metalloprotenase 1 (TIMP1) of HSC-T6 cells.

METHODSCultured HSC-T6 cells were exposed to IFNgamma at concentrations of 0.1, 1, 10, 10(2), 10(3), 10(4), 2.5 x 10(5), 5 x10(5) U/ml for 48 hours. 4,5-simethylthiazaoly colormetric assay was used to evaluate the effect of IFNgamma on HSC-T6 cell proliferation. After incubating with IFNgamma (1 U/ml, 10(2) U/ml and 10(4) U/ml) for 48 hours, HSC-T6 cells were harvested to detect Col I, Col III and TIMP1 steady state mRNA levels by quantitative reverse-transcription polymerase chain reaction (RT-PCR).

RESULTSThe Col I, Col III and TIMP1 mRNA levels of the control group were 2.86+/-0.21, 2.00+/-0.23 and 3.90+/-0.81, respectively. Col I and Col III mRNA levels in HSC-T6 cells treated by different concentrations of IFNgamma were lower than that of the controls (P < 0.01). There was no significant difference in TIMP1 mRNA levels between IFNgamma groups and controls.

CONCLUSIONIFNgamma suppresses expression of Col I and Col III whereas it has no effect on TIMP1 mRNA expression. The antifibrotic mechanism of IFNgamma may be partly due to its down-regulation of Col I and Col III mRNA levels in HSC-T6 cells.

Animals ; Cell Line ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Hepatocytes ; cytology ; metabolism ; Interferon-gamma ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of PDGF receptor-beta and its correlation with extracellular matrix in hepatic tissue during hepatic fibrosis.

METHODSThe model of hepatic fibrosis in rats was induced by carbon tetrachloride. PDGF receptor-beta subunit, collagen I, collagen III and a-SMA in hepatic tissues of these rats were examined using immunohistochemistry. The correlation between PDGF receptor-beta subunit and collagen I, III was analyzed using SAS software after the results of immunohistochemistry were semi-quantified.

RESULTSPDGF receptor-beta subunit and a-SMA were not detected in normal controls. Collagen I and III were distributed in the portal tracts and beneath the endothelia of the central veins and of the Disse spaces. Two weeks after CCl4 injection, the PDGF receptor-beta and a-SMA were detected, and the expression of collagen I and III increased. At the end of 4 and 6 weeks, the above four proteins were further increased. Two weeks after CCl4 injection, PDGF receptor-beta had no apparent correlation with collagen I and III. However, PDGF receptor-beta had a significant correlation with collagen I and III 2 weeks later, and the correlation coefficient was 0.74 and 0.60 respectively at 4 weeks, and 0.83 and 0.67 respectively at 6 weeks. PDGF receptor-beta had a significant correlation with a-SMA during the whole process of hepatic fibrosis and the correlation coefficient was 0.62, 0.69 and 0.81, respectively at the time of 2, 4 and 6 weeks after CCl4 injection.

CONCLUSIONThe PDGF receptor-beta was overexpressed during the process of hepatic fibrosis development, and it significantly correlated with collagen I and collagen III.

Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Extracellular Matrix ; metabolism ; Liver ; metabolism ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; biosynthesis ; genetics

To investigate the effect of different frequencies of cyclic tensile strain on extracellular matrix (ECM) of vascular smooth muscle cells (VSMCs) and to research the relationship between tensile strain and vascular remodeling, the aortic vascular smooth muscle cells of rats grown on dishes coated with collagen I were subjected to 10% elongation and various frequencies of mechanical strain using the Flexercell 4000 Strain Unit. The expression of extracellular matrix including fibronectin, collagen I and collagen III was detected by Real-time RT-PCR, and p38 activity by western blot. The result showed that the expression of extracellular matrix was induced by mechanical strain in a nonlinear frequency-dependent manner, which was mediated by p38 pathway. These results demonstrate that the variety of frequencies of cyclic tensile strain could modulate the expression of ECM. It may have important influence on vascular remodeling.
Animals ; Aorta ; cytology ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Extracellular Matrix ; metabolism ; Fibronectins ; biosynthesis ; Male ; Mechanotransduction, Cellular ; physiology ; Muscle, Smooth, Vascular ; cytology ; physiology ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical

Aldosterone ; pharmacology ; Animals ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Dose-Response Relationship, Drug ; Liver ; cytology ; drug effects ; metabolism ; Liver Cirrhosis ; etiology ; RNA, Messenger ; analysis ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; genetics

OBJECTIVETo investigate the effects of Bushen Huoxue Fang on the proliferation of rat cardiac fibroblasts and collagen production in the cells.

METHODSRat cardiac fibroblasts were isolated and cultured in DMEM containing 10% (group A) or 20% (group B) or no (group C) serum from rats treated with Bushen Huoxue Fang, with cells cultured in DMEM containing 10% FBS as the control (group D). After 72 h of cell culture, the proliferation of the fibroblasts was detected using CCK-8 kit, and collagen mRNA and protein expressions were examined using RT-PCR and Western blotting, respectively.

RESULTSCompared with that in groups C and D, the cell proliferation decreased significantly in groups A and B, and especially in the latter (P<0.05). RT-PCR demonstrated significant reductions of the mRNAs of type 1 and 3 collagens in groups A and B (P<0.05), and their protein levels were also significantly lowered (P<0.05).

CONCLUSIONBushen Huoxue Fang can effectively inhibit the proliferation of rat cardiac fibroblasts and reduced collagen type 1 and 3 productions in the cells in vitro.

Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Fibrosis ; prevention & control ; Myocardium ; cytology ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the mechanism of Shengji Huayu Recipe (SJHYR) and its 3 disassembled formulae in accelerating wound healing toward skin restoration.

METHODSUsing in vitro cultured fibroblasts from granulation tissue of wound and compared with the normal skin fibroblast of suckling rat, who were treated separately by drug serum containing high and low dose SJHYR and its disassembled prescriptions (Shengji formula and Huayu formula). The type I and III collagen contents in the fibroblasts were determined with immunocytochemical ABC method.

RESULTSShengji formula could increase the levels of type I and III collagens in fibroblasts, which was significantly higher than those in model cells and control (P < 0.01). Huayu formula lowered them to the levels below those in model cells (P < 0.01), while in comparing the levels in the cells treated by high or low dose SJHYR with those in control, no significant difference was shown(P > 0.05).

CONCLUSIONSJHYR might accelerate wound healing toward skin restoration through regulating the ratio of collagen type I and III, to adjust their metabolism.

Animals ; Animals, Newborn ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; metabolism ; pathology ; Male ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Wound Healing ; drug effects ; Wounds and Injuries ; pathology