OBJECTIVETo study the inhibitory effect of Typhonium gigantewm Engl. (AEoTGE) on the proliferation and apoptosis of KFB in vitro and to survey the death rate.

METHODSSamples of hypertrophic scars were collected and cultured. Only 4-8 passage cells were selected for experiment. Inverted microscope and transmission electron microscope were used to observe the morphogenesis and ultrastructure of KFB. The KFB cells were treated with AEoTGE in different concentrations(3. 125,6.250, 12.500, 25.000, 50. 000,100.000 g/L) for 24 hours. The effect of AEoTGE on the proliferation and the IC50 of KFB was observed with MTT assay and EdU. The effect of AEoTGE on apoptosis of KFB was detected by flow cytometry.

RESULTSIt showed that AEoTGE could inhibit the proliferation of KFB in an concentration-dependent style within the range of 3. 125-100.000 g/L. The AEoTGE could obviously increase the apoptosis rate of the KFB compared with blank control group(P <0.05). The IC50 of AEoTGE was 35 g/L. FITC-Annexin V/PI showed that apoptosis rate of KFB in the AEoTGE group was (72. 07 +/- 0. 70)% , while it was 23. 5% in blank control group (P < 0. 05).

CONCLUSIONSAEoTGE could significantly inhibit the proliferating activity and induce apoptosis of KFB after co-culture for 24 hours. The IC50 is 35 g/L and the rate of apoptosis is (72.07 +/- 0.70)%.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; drug effects ; pathology ; Humans ; Keloid

PURPOSE: Long-term use of topical medication is needed for glaucoma treatment. One of the most commonly prescribed classes of hypotensive agents are prostaglandin analogs (PGs) used as both first-line monotherapy; as well as in combination therapy with other hypotensive agents. Several side effects of eye drops can be caused by preservatives. The purpose of this study was to evaluate the effects of PGs with varying concentrations of benzalkonium chloride (BAC), alternative preservatives, or no preservatives on human conjunctival fibroblast cells. METHODS: Primary human conjunctival fibroblast cells were used in these experiments. Cells were exposed to the following drugs: BAC at different concentrations, bimatoprost 0.01% (with BAC 0.02%), latanoprost 0.005% (with BAC 0.02%), tafluprost 0.0015% with/without 0.001% BAC and travoprost 0.004% (with 0.001% Polyquad) for 15 and 30 minutes. Cell cytotoxicity was evaluated by phase-contrast microscopy to monitor morphological changes of cells, Counting Kit-8 (CCK-8) assay to cell viability, and fluorescent activated cell sorting (FACS) analysis to measure apoptosis. RESULTS: BAC caused cell shrinkage and detachment from the plate in a dose-dependent manner. Morphological changes were observed in cells treated with bimatoprost 0.01% and latanoprost 0.005%. However, mild cell shrinkage was noted in cells treated with tafluprost 0.0015%, while a non-toxic effect was noted with travoprost 0.004% and preservative-free tafluprost 0.0015%. CCK-8 assay and FACS analysis showed all groups had a significantly decreased cell viability and higher apoptosis rate compared with the control group. However, travoprost 0.004% and preservative-free tafluprost 0.0015% showed lower cytotoxicity and apoptosis rate than other drugs. CONCLUSIONS: This in vitro study revealed that BAC-induced cytotoxicity is dose-dependent, although it is important to emphasize that the clinical significance of toxicity differences observed among the different PGs formulations has not yet been firmly established. Alternatively preserved or preservative-free glaucoma medications seem to be a reasonable and viable alternative to those preserved with BAC.
Apoptosis ; Cell Line ; Cell Survival/drug effects ; Conjunctiva/drug effects/*pathology ; Fibroblasts/drug effects/pathology ; Glaucoma/drug therapy/pathology ; Humans ; Preservatives, Pharmaceutical/*pharmacology ; Prostaglandins, Synthetic/*pharmacology

OBJECTIVETo investigate the effect of HMME-PDT on the proliferation and cell distribution of hypertrophic scar fibroblast (HSF).

METHODSHSF were cultured and only 4-6th passages were used in this study. Argyrophilic protein in nucleolar organizer regions(AgNORs) were calculated by I. S% after argyrophilic staining. Flow cytometry was applied to analyze the cell cycle and proliferation index (PI).

RESULTS1) I. S% of HSF after HMME-PDT was reduced markedly. 2) HMME-PDT inhibited HSF entering S stage from G, stage, cell percentage in S stage was decreased to (11.2 +/- 2.3)%. 3) PI in HMME-PDT group was less than that in control group [(35.0 +/- 3.4)% vs (27.2 +/- 3.1)%, P < 0.05].

CONCLUSIONSHMME-PDT can inhibit proliferation of HSF, and chang cell distribution.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; drug therapy ; pathology ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans ; Photochemotherapy

OBJECTIVETo investigate the effects of sinomenine (SIN) and methotrexate (MTX) on the proliferation and apoptosis of in vitro cultured fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) patients, as well as the expression of osteoclast differentiation factor in FLS.

METHODSFLS were isolated from the synovium of RA patients and cultured in vitro. FLS were incubated with different concentrations of SIN and MTX respectively or combined: 0.001, 0.010, 0.100, 1.000 mg/mL SIN; 0.001, 0.010, 0.100, 1.000 mg/mL MTX; 0.001 mg/mL SIN + 0.001 mg/mL MTX, 0.010 mg/mL SIN + 0.010 mg/mL MTX, 0.100 mg/mL SIN + 0.100 mg/mL MTX, 1.000 mg/mL SIN + 1.000 mg/mL MTX, namely SIN1, 2, 3, 4 groups; MTX1, 2, 3, 4 groups and the combination 1, 2, 3, 4 groups. The medium without drugs was used as a control group. There was a total of 13 groups, each group with 3 complex holes. MTT was applied to detect the growth of FLS. The flow cytometry was applied to detect the apoptosis of FLS. The expressions of FLS receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and osteoprotegerin (OPG) mRNA were observed by semi-quantitative RT-PCR.

RESULTSCompared with the control group, RA FLS proliferation OD values of all the drug groups were lower (P < 0.05). The RA FLS apoptosis OD value of the combination 3 group increased, the OPG mRNA expression increased, the expression of RANKL mRNA decreased with statistical difference (P < 0.05). The RA proliferation OD values of the SIN3 group and the MTX3 group increased when compared with the combination 3 group (P < 0.05).

CONCLUSIONSSIN and MTX had synergistic effects in inhibiting FLS. This might be one of the mechanisms for inhibiting RA bone damage.

Apoptosis ; drug effects ; Arthritis, Rheumatoid ; pathology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fibroblasts ; drug effects ; Humans ; Methotrexate ; pharmacology ; Morphinans ; pharmacology ; Synovial Membrane ; cytology ; drug effects

The wound healing time, tension of wound edge, proliferation of fibroblast, and extracellular matrix deposition are the important factors of scar formation, and botulinum toxin type A can regulate the above. Prevention and treatment of scar with botulinum toxin type A is one of the hot topics of clinical research in recent years. This paper briefly reviews researches by scholars at home and abroad on the mechanism, clinical application, complications, and adverse effects of botulinum toxin type A in scar prevention and treatment.
Botulinum Toxins, Type A/therapeutic use* ; Cicatrix/prevention & control* ; Extracellular Matrix/pathology* ; Fibroblasts/drug effects* ; Humans ; Wound Healing/drug effects*

Failure of a glaucoma filtering operation mainly results from scarring at the filtering wound, and postoperative proliferation and migration of fibroblasts play an important role histologically in the formation of scar tissue. As an inhibitory agent for fibroblast proliferation, gamma-interferon has been introduced, and the application of gamma-interferon following filtering surgery is now being made on a trial basis. We studied the effect of gamma-interferon histologically on the fibroblast proliferation and collagen synthesis occurring at the filtering site by comparing the effect of gamma-interferon on the experimental group with that of 5-fluorouracil on the control group, using 10 rabbits (20 eyes) after posterior lip sclerectomy. Both groups showed similar flat and diffused bleb grossly and also showed a similar inhibitory effect on fibroblast proliferation and collagen fiber synthesis histologically. Our findings seem to justify the clinical use of gamma-interferon. Further studies on adequate dosage, method of administration, and local and systemic complications would be desired.
Animals ; Anterior Chamber/drug effects ; Cell Division/drug effects ; Collagen/*biosynthesis ; Fibroblasts/drug effects ; Fluorouracil/pharmacology ; Glaucoma/pathology/*surgery ; Interferon-gamma/*pharmacology ; Rabbits ; Sclera/pathology ; *Sclerostomy

OBJECTIVETo investigate the effects of lanthanum chloride on the apoptosis of fibroblasts in trauma tissue.

METHODSFifty adult female SD rats were used and linear incisions were made on the back near the joints of extremities of the rats. One of the cuts receiving no treatment was designated as blank control (C). 0.25 ml of distilled water, lanthanum chloride (50 mmol/L) and the antibody of (TGFbeta(1)) transforming growth factor beta(1) (0.2 mg/ml) were respectively injected into the both sides of the other three wounds subcutaneously and the wounds were divided into simulating control (SC), lanthanum chloride (LC) and antibody (A) groups. The fibroblast apoptosis in the wound tissue samples and the change in intracellular calcium concentration (Ca(2+)) were determined by flow cytometry (FCM) and TUNEL methods on the 14th and 28th day after the injection.

RESULTSApoptosis of fibroblasts was enhanced significantly after 14 days of injection in LC and A groups compared with that in C and SC groups (P < 0.05 approximately 0.01). Furthermore, intracellular Ca(2+) was increased evidently in LC group (P < 0.01).

CONCLUSIONIt is indicated that lanthanum chloride might be effective in preventing scar development.

Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cicatrix ; prevention & control ; Female ; Fibroblasts ; drug effects ; pathology ; Flow Cytometry ; Lanthanum ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Wound Healing ; drug effects

OBJECTIVETo evaluate the effects of nicotine and tobacco extract (ST) on PDLFs morphology, structure, proliferation and attachment.

METHODSPDLFs were cultured in the presence of nicotine and ST at various concentration. The cell changes in the morphology and structure were examined by histological and transmission electrical microscope (TEM). The growth and attachment of cell were measured by MTT method.

RESULTSThe size of the cells became smaller gradually and their shapes changed from shuttle type to oval or round when the concentration of nicotine and ST increased, the polarity of the cells was in disorder, ultrastructure showed that the organelles, especially rough-surfaced endoplasmic reticulum and golgi complex decreased in number, microtubule and microfilaments were disassembled, the nuclei became fewer or shrunk, the growth and attachment were dose-dependently inhibited.

CONCLUSIONSNicotine and ST can change PDLFs' morphology and structure, they may inhibit the growth and attachment through disruption of the cytoskeleton, suggesting nicotine and ST may have pathological role on human periodontitis.

Cell Adhesion ; drug effects ; Cell Division ; drug effects ; Fibroblasts ; cytology ; Humans ; Nicotine ; toxicity ; Periodontal Ligament ; drug effects ; pathology ; ultrastructure ; Plant Extracts ; toxicity ; Tobacco

OBJECTIVETo study the interaction between transformation of human pulmonary epithelial cells and activation of fibroblasts induced by Yunnan tin mine dust.

METHODS(1) The immortalized human bronchial epithelial cell line BEAS-2B and human embryo lung fibroblast cell line WI-38 were grown in MEM medium containing 5% and 10% FBS, respectively, at 37 degrees C and 5% CO2 with saturated humidity. The cells were subcultured every 6 days. BEAS-2B cells and WI-38 cells were induced with Yunnan tin mine dust on every other generation at the concentration of 100 microg/ml. From the 11th generation, the cells were co-cultured. Epithelial cell transformation was tested using concanavalin A (ConA) agglutination and anchorage-independent growth assays. The cell cycles were analyzed through flow cytometry. The expressions of alpha-SMA in fibroblasts were determined with immunocytochemistry.

RESULTS(1) Cell morphology of mine dust-exposed epithelial cells began to transform at the 28th generation. Similar transformations were observed with mine dust-induced epithelial cells co-cultured with fibroblasts from the 20th generation and mine dust-induce epithelial cells co-cultured with mine dust-induced fibroblasts from the 16th generation. ConA agglutination assay and anchorage-independent growth assays were negative in normal BEAS-2B cells. At the 26 th generation, the agglutination test result of the mine dust-exposed epithelial cells was positive. Co-cultured with fibroblasts and mine dust-exposed fibroblasts, the agglutination time of the mine dust-exposed epithelial cells became short. Epithelial cell anchorage-independent growth assay was positive for mine dust-exposed epithelial cells co-cultured with fibroblasts at the 36th generations and for mine dust-exposed epithelial cells co-cultured with mine dust-exposed fibroblasts at the 26th generations. The clone formation rate of the 26th generation was 6.00 per thousand +/- 1.00 per thousand and 15.33 per thousand +/- 2.52 per thousand respectively, with the significant differences (P < 0.05). With generation adding, the portion of S phase increased for mine dust-exposed epithelial cells. (2) At the 26th generations, fibroblasts expressed alpha-SMA. Co-cultured with epithelial cell, the alpha-SMA expression of fibroblasts increased. Especially, positive cell numbers and intensity of staining dramatically increased with generation adding.

CONCLUSIONS(1) The tin mine dust can induce malignant transformation of human pulmonary epithelial cells BEAS-2B and activation of fibroblasts WI-38. (2) The epithelial cells are major target in carcinogenesis induced by Yunnan tin mine dust. (3) Transformation of epithelia and activation of fibroblasts co-evolve in the developing process of induced lung cancer by Yunnan tin mine dust.

Cell Cycle ; drug effects ; Cell Line ; Cell Transformation, Neoplastic ; drug effects ; Coculture Techniques ; Dust ; Epithelial Cells ; pathology ; Fibroblasts ; metabolism ; pathology ; Humans ; Tin ; toxicity

OBJECTIVETo seek an effective drug for treatment of human hyperplastic scar through studying the effects of curcumin on fibroblast growth and collagen synthesis.

METHODSFibroblasts derived from scar tissue and from normal epidermal tissue were isolated and cultured separately with tissue-block method, their morphology were observed under invert phase contrast microscope, their growth curve was drawn respectively to determine the speed of growth. Then, fibroblasts from scar were stimulated with curcumin in different concentrations (0, 12.5, 25, 50 and 100 micromol/L) for detecting the inhibitory effect of curcumin on growth of fibroblasts using MTT methods and that on activity of procollagen alpha-1 gene transcription in fibroblast was detected by RT-PCR.

RESULTSThe cell growth curve showed that double-multiplying time was 5 days in fibroblasts from scar and 4 days in those from normal dermis, showing significant difference between them (P < 0.05). MTT showed that curcumin in 12.5 micromol/L showed a cell proliferation enhancing trend, and its absorbance value was significantly higher than that in the normal group, but the effect turned to inhibition when concentration increased to over 25-100 micromol/L, and became significant inhibition at concentration of 50 and 100 micromol/L. Besides, curcumin also showed markedly inhibition on collagen type I synthesis in fibroblasts (P < 0.01).

CONCLUSIONHigh concentration curcumin can inhibit effectively the fibroblast proliferation and collagen I synthesis in hyperplastic scar, therefore, may has therapeutic effect on the disease in human being.

Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen Type I ; biosynthesis ; Curcumin ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans