BACKGROUND: Changes in the composition of the extracellular matrix (ECM) occur between the proliferating and involuted phases of infantile hemangiomas (IH), and are associated with angiogenic growth. We examined the composition of the ECM in proliferating and involuted IHs and assessed correlations between the composition of the ECM and whether the IH was in the proliferating or the involuted phase. METHODS: We evaluated IH samples from a cohort of patients who had five proliferating IHs and five involuted IHs. The following ECM molecules were analyzed using enzyme-linked immunosorbent assays and immunohistochemistry: laminin, fibronectin, collagen type I, collagen type II, and collagen type III. RESULTS: The involuted IHs had higher levels of deposition of collagen type III than the proliferating IHs. The median values (interquartile ranges) were 1.135 (0.946-1.486) and 1.008 (0.780-1.166) (P=0.019), respectively. The level of laminin was higher in involuted IHs than in proliferating IHs, with median values (interquartile ranges) of 3.191 (2.945-3.191) and 2.479 (1.699-3.284) (P=0.047), respectively. Abundant collagen type III staining was found in involuted IHs. Laminin alpha4 chain staining was clearly present within the basement membrane adjacent to the blood vessels, and was significantly more intense in involuted IHs than in proliferative IHs. CONCLUSIONS: Involuted hemangiomas showed extensive deposition of collagen III and laminin, suggesting that differences in the composition of the ECM reflect stages of the development of IHs. This pattern may be due to the rapid senescence of IHs.
Aging ; Basement Membrane ; Blood Vessels ; Cohort Studies ; Collagen ; Collagen Type I ; Collagen Type II ; Collagen Type III ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix* ; Fibronectins ; Hemangioma* ; Humans ; Immunohistochemistry ; Laminin

OBJECTIVETo study the changes in sciatic nerve blood flow and the expression of collagen type I after electric injury of rabbit nerve with different voltages.

METHODSThirty-six healty rabbits were randomized into 3 groups before receiving injury with electricity in voltages, i.e. 50 v, 75 v, and 100 v groups. The changes in blood flow of sciatic nerve were observed with Laser Doppler Flowmeter immediately after injury and 1, 4, 8 weeks after injury. The changes in the expression of collagen type I was observed by immunohistochemical method, and the positive expression rate was calculated.

RESULTSThe sciatic nerve blood flow increased in all groups immediately after electric injury. In the 75 v and 100v groups, the nerve blood flow [(53 +/- 3 ), (48 +/- 5) PU] was obviously lower than that of normal value [(62 +/- 4) PU, P < 0.05]. There was little collagen type I deposition in 50 v group, while brown collagenous fibers in epineurium and perineurium were observed in 75 v and 100v groups 4 and 8 weeks after injury. The expression of collagen type I in all groups were obviously higher than that of normal value, and that in 75v and 100 v groups were higher than that in 50 v group at bachl time-point (P < 0.01).

CONCLUSIONThe restoration of sciatic nerve blood flow is postponed following by the injury with increase of the electrical voltage. The collagen deposition after electrical injury may be one of the reasons for nerve blood flow decrease.

Animals ; Collagen Type I ; biosynthesis ; Electric Injuries ; blood ; physiopathology ; Nerve Regeneration ; Rabbits ; Random Allocation ; Sciatic Nerve ; blood supply ; injuries

Osteoporosis is a major health problem worldwide, and is projected to increase exponentially due to the aging of the population. The absolute fracture risk in individual subjects is calculated by the use of algorithms which include bone mineral density (BMD), age, gender, history of prior fracture and other risk factors. This review describes the laboratory investigations into osteoporosis which include serum calcium, phosphate, creatinine, alkaline phosphatase and 25-hydroxyvitamin D and, additionally in men, testosterone. Parathyroid hormone (PTH) is measured in patients with abnormal serum calcium to determine its cause. Other laboratory investigations such as thyroid function testing, screening for multiple myeloma, and screening for Cushing's syndrome, are performed if indicated. Measurement of bone turnover markers (BTMs) is currently not included in algorithms for fracture risk calculations due to the lack of data. However, BTMs may be useful for monitoring osteoporosis treatment. Further studies of the reference BTMs serum carboxy terminal telopeptide of collagen type I (s-CTX) and serum procollagen type I N-terminal propeptide (s-PINP) in fracture risk prediction and in monitoring various treatments for osteoporosis may help expedite their inclusion in routine clinical practice.
Algorithms ; Biological Markers/*blood ; Clinical Laboratory Techniques ; Collagen Type I/blood ; Fractures, Bone/prevention & control ; Humans ; Osteoporosis/*diagnosis ; Peptide Fragments/blood ; Peptides/blood ; Procollagen/blood

The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 (1 x 10(5) cells/ml/well) were cultured at 12-well plate at 37degrees C for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin. According to this study, the results were as follows: 1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05). 2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days. 3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively). 4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05). 5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels. This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.
Blood Vessels ; Collagen ; Collagen Type I* ; Dental Pulp* ; Eosine Yellowish-(YS) ; Fibroblasts ; Gingiva ; Hematoxylin ; Humans ; NIH 3T3 Cells ; Skin ; Tissue Engineering*

PURPOSE: We evaluated the differentiation potential of CD34-cells expanded from human cord blood into several differentiated cells, namely, osteoblasts, chondrocytes and adipocytes. MATERIALS AND METHODS: CD34- cells, extracted from cord blood and isolated using a MiniMACS system, were cultured. Cells labeled with appropriate antibodies were analyzed by FACScan. The phenotypes of adipogenic and osteogenic differentiation were evaluated by Oil Red O, alkaline phosphatase, and von Kossa staining. Chondrogenic differentiation was evaluated by RT-PCR using primers for aggrecan, collagen types I, II and X. RESULTS: CD34- cells showed a fibroblast-like morphology and expressed CD105, CD29, and CD44 antigens. These cells showed the deposition of a mineralized matrix and the expression of alkaline phosphatase in the osteogenic medium, and stained with Oil Red O in the adipogenic medium. In terms of chondrogenic differentiation, the expressions of aggrecan, and collagen types II and X showed a gradual increase, whereas the expression of type I collagen gradually decreased. CONCLUSION: Based on their differentiation potentials in at least three different tissue types, i.e., fat, bone, and cartilage, cord blood-derived CD34- cells can be visualized as attractive target cells for cellular or gene transfer therapeutic options.
Adipocytes ; Aggrecans ; Alkaline Phosphatase ; Antibodies ; Antigens, CD44 ; Cartilage ; Chondrocytes ; Collagen ; Collagen Type I ; Fetal Blood* ; Humans* ; Mesenchymal Stromal Cells ; Osteoblasts ; Phenotype

The purpose of the present study is to explore the effect of aliskiren on the proliferation of cardiac fibroblasts (CFs) in AGT-REN double transgenic hypertensive (dTH) mice. The cultured CFs from AGT-REN dTH mice were divided into AGT-REN group (dTH) and aliskiren group (ALIS). Cultured CFs from C57B6 mice were served as control (WT). The effect of different concentration of aliskiren (1 × 10, 1 × 10, 1 × 10, 1 × 10mol/L) on CFs proliferation was determined by MTT assay. After treatment with 1 × 10mol/L aliskiren for 24 h, α-SMA, collagen I, III and NADPH oxidase (NOX) protein expression in CFs of AGT-REN dTH mice were detected by Western blot. The collagen synthesis in CFs was assessed by hydroxyproline kit. The expression of ROS was determined by DHE. Results showed that the blood pressure and plasma Ang II levels were significantly increased and CFs proliferation was significantly increased as well in AGT-REN dTH mice compared with WT group. However, aliskiren intervention decreased CFs proliferation, myofibroblast transformation, as well as the collagen I and III synthesis in CFs of AGT-REN dTH mice. Meanwhile, aliskiren inhibited ROS content and NOX2/NOX4 protein expression in CFs of AGT-REN dTH mice. These results suggest that aliskiren decreases the cell proliferation, myofibroblast transformation and collagen production in CFs of AGT-REN dTH mice, which might be through inhibition of oxidative stress response.
Amides ; Animals ; Blood Pressure ; Cell Proliferation ; Cells, Cultured ; Collagen ; Collagen Type I ; Fumarates ; Heart ; Hydroxyproline ; Hypertension ; Mice ; Mice, Transgenic ; Myocardium ; Myofibroblasts ; NADPH Oxidases

OBJECTIVESome research has shown that resveratrol can ameliorate myocardial injury and improve cardiac function in mice with acute viral myocarditis (VMC), and can inhibit cardiac fibroblast proliferation and myofibroblast differentiation in vitro. This study was designed to investigate whether resveratrol has similar effects in the mouse model of chronic VMC.

METHODSOne hundred mice were inoculated with 0.3 mL of Coxsackievirus B3 1*106 TCID50. Thirty days later, the survivors (n=62) were used as a model of chronic VMC, and were randomly assigned to 4 groups: untreated VMC, and low- (10 mg/kg), middle- (100 mg/kg) and high-dose (1 000 mg/kg) resveratrol-treated VMC (once daily, for 30 days). Ten mice which received neither Coxsackievirus B3 nor resveratrol treatment served as the control group. After 30 days of resveratrol treatment, the mice were sacrificed. Serum concentrations of collagenous pre-peptides (PINP, PICP and PIIINP) were assessed using ELISA. Hematoxylin-eosin staining, picrosirius red staining and circularly polarized light were used to examine the histochemistry of myocardial collagen.

RESULTSThe myocardial collagen volume fraction in the high-dose (0.74+/-0.19) and the middle-dose (1.07+/-0.12) resveratrol-treated VMC groups was significantly lower than that in the untreated VMC (2.33+/-0.18) and the low-dose resveratrol-treated VMC (2.17+/-0.19) groups (P<0.05). Compared with the untreated VMC group, serum concentrations of PICP and PIIINP in the high-dose and the middle-dose resveratrol-treated VMC groups were significantly reduced (P<0.05), while PINP concentrations increased significantly (P<0.05).

CONCLUSIONSResveratrol can inhibit hyperplasia of myocardial collagen in the mouse model of chronic VMC, acting as an effective anti-fibrotic agent in the myocardium.

Animals ; Chronic Disease ; Collagen Type I ; analysis ; Collagen Type II ; analysis ; Coxsackievirus Infections ; drug therapy ; Enterovirus B, Human ; Fibrosis ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; drug therapy ; Myocardium ; pathology ; Peptide Fragments ; blood ; Procollagen ; blood ; Stilbenes ; therapeutic use

BACKGROUND: Aldosterone induces renal fibrosis and administration of spironolactone showed beneficial effect in various animal models of renal injury such as unilateral ureteral obstruction model and hypertensive renal injury models. The aim of this study is to demonstrate that inhibition of aldosterone may have additional beneficial effect independent of angiotensin blockade in diabetic nephropathy. METHODS: We investigated the effect of aldosterone blockade on renal function in animal model of type II diabetic nephropathy. Diabetic rats were treated with spironolactone (20 mg/kg/day), starting at 20 weeks of age. RESULTS: In diabetic rats, proteinuria was increased since 24 weeks, and presented five-fold increment at 52 weeks comparing to that at 24 weeks. Aldosterone blockade significantly reduced proteinuria without affecting blood glucose concentration and blood pressure. In the diabetic kidney, profibrogenic molecules such as CTGF and type I collagen expression was enhanced and spironolactone treatment decreased their expression. It is of interest that plasma aldosterone level showed a weak but significant relationship with blood glucose levels. CONCLUSION: Aldosterone blockade prevents fibrotic process in the kidney associated with decrement in profibrotic molecule such as CTGF and type I collagen. Furthermore, aldosterone blockade resulted in decrease in urinary protein excretion and glomerulosclerosis. These results suggest that aldosterone blockade may be a new therapeutic target for retarding the progression of diabetic nephropathy.
Aldosterone ; Angiotensins ; Animals ; Blood Glucose ; Blood Pressure ; Collagen Type I ; Diabetic Nephropathies ; Fibrosis ; Kidney ; Models, Animal* ; Plasma ; Proteinuria ; Rats* ; Spironolactone* ; Ureteral Obstruction

OBJECTIVETo study the bone mineral density (BMD) of the third lumbar vertebrae and the Ward's triangle in the femoral neck and serum type I collagen C-telopeptide (CTx) level in female adults and provide scientific evidence for diagnosis and prevention of osteoporosis.

METHODSAccording to the inclusion criteria, 653 female adults were examined for BMD of the third lumbar vertebrae and the Ward's triangle in the femoral neck and the level of serum CTx.

RESULTSThe peak of BMD in the third lumbar vertebrae occurred in 30-39 years of age, and that of the Ward's triangle in the femoral neck occurred in 20-29 years of age, showing significant differences between the age groups (P<0.01). The BMD of the Ward's triangle in the femoral neck decreased earlier than that of the third lumbar vertebrae by one age period. The level of serum CTx increased with age, showing significant differences between the age groups (P<0.01) but not between the 20-29 years and the 30-39 groups, or between 30-39 and the 40-49 years groups. The prevalence of osteoporosis increased also with age, and osteoporosis in the third lumbar vertebrae increased rapidly in the 50-59 years and the 60-69 years groups, maintaining a high level in older ages. The prevalence of osteoporosis in the Ward's triangle in the femoral neck increased obviously after the peak of BMD, maintaining a significantly higher level than that of the third lumbar vertebrae in the same age group.

CONCLUSIONSThe BMD in the third lumbar vertebrae and the level of serum CTx undergo obvious changes with age, especially in menopause. There is no obvious relation between decreased BMD in the Ward's triangle in the femoral neck and menopause.

Adult ; Aged ; Bone Density ; China ; Collagen Type I ; blood ; Female ; Femur Neck ; metabolism ; Humans ; Lumbar Vertebrae ; metabolism ; Middle Aged ; Peptides ; blood ; Young Adult

OBJECTIVE: To determine the relation between serum concentration of retinol binding protein (RBP) 4 and markers of bone metabolism, bone mineral density (BMD) in patients with type 2 diabetes mellitus (T2DM). METHODS: A total of 82 patients newly diagnosed with T2DM and 46 subjects with normal glucose tolerance (NGT) enrolled in the cross-sectional study. Subset analyses were performed, dividing subjects on the basis of gender into M-T2DM, F-T2DM, M-NGT, and F-NGT. The serum concentrions of RBP4, osteocalcin (OC) and C-terminal telopeptide of collagen type I (CTX) were measured with ELISA. The BMD was measured by dual-energy X-ray absorptiometry (DXA) with a Hologic QDR4500A device. RESULTS: In both the T2DM groups, lnRBP4 showed a positive relationship with lnCTX (M-T2DM, r=0.564, P<0.01; F-T2DM, r=0.386, P=0.018), but no association with lnOC. After adjusting for age, smoking, creatinine clearance rate (CCr), and waist-to-hip ratio (WHR), lnRBP4 still showed a strong association with lnCTX in the M-T2DM group (r'=0.536, P<0.01), but not in F-T2DM (r'=0.317, P=0.072). In the NGT group, there was no relation between lnRBP4 and lnCTX or lnOC. LnRBP4 showed no association with BMD in all groups. CONCLUSION The level of serum RBP4 may be correlated with the bone metabolism in patients with T2DM.
Adult ; Aged ; Bone Density ; Bone and Bones ; metabolism ; Collagen Type I ; blood ; Cross-Sectional Studies ; Diabetes Mellitus, Type 2 ; blood ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Osteocalcin ; blood ; Peptides ; blood ; Retinol-Binding Proteins, Plasma ; metabolism