Objective To study the effects of dermal template on the biological behaviors of fibroblasts during wound healing. Methods A total of 120 rats were made fullthickness wound modes on the dorsum and divided into 4 groups,in group 1, the wounds were allowed to heal by contraction（ConT）；in group2, the wounds covered with fullthickness skin grafts( FTSG); in group 3, the wounds were with split thickness skin grafts (STSG); and ingroup 4, the wounds were covered by dermal regeneration template with overlying thin splitthickness autograft (ADMT).The specimens were obtained at one week, two weeks, four weeks, six weeks,and twelve weeks respectively. The expressions of α smooth muscle actin(αSMA，characteristic of MFB）,fibronectin(FN),integrin α2,β1 and transforming growth factor β1(TGF-β1) were examined by immunohistochemical analysis. Results Positive expression of α-SMA、FN、integrin α2β1 and TGF-β1 in ADMT groups was significantly lower than that in STSG group and ConT group, but higher than that in FTSG group（P＜0.05）. Conclusion Dermal regeneration template can inhibit the transformation of FB to MFB and restrain the expressionof FN,integrin α2,β1,and TGF-β1 in fibroblasts which might reduce thepossibility of hypertrophyic scaring, and improve wound healing.
OBJECTIVE To review the recent advances in fibroblast study and its role in wound repair. METHODS Recent original articles related to wound repair were retrieved extensively, and the effect of fibroblast on every stages of repair were summed up and comprehended. RESULTS Fibroblast plays important roles in granulation formation, wound contraction, matrix synthesis, wound repair, scar formation and scarless repair by means of growth factors modulation. CONCLUSION The understanding of fibroblast in the wound repair can promote the progress of biological therapy of wound repair and scar prevention.
Objective To const ruct art ificial derm is on co llagen2chondront in sulfate (CS) scaffo ld. Methods Co llagen w as compounded from CS and 1-ethyl-3-(13-dimethyl am inop ropyl) carbodiim ide (EDC) used as a cro sslinker. Physical and chem ical p ropert ies of the scaffo ld w ere characterized by elect ron spect ro scopy fo r chem ical analysis (ESCA ) , scanning elect ron m icrograph (SEM ) , HE staining, and mechanical p roperty test. Derm is fibroblasts w ere iso lated from human embryo and w ere cultured on the scaffo lds. Th rough h isto logical test ing, immunoh istochem ical test ing and biochem ical p roperty test ing, the p roperty of co llagen-CS art ificial derm is w as compared w ith that of colla gen spongy art ificial derm is. Results Co llagen-CS had th ree2dimension st ructure w ith po rous. Compared w ith co llagen scaffo ld, themechanical p roperty of co llagen2CS scaffo ld imp roved. There w eremo re po lar group s on the surface of co llagen-CS scaffo ld. The fibroblasts on the co llagen-CS scaffo ld grew w ell, and art ificial derm is w as const ructed. Conclus ion Co llagen-CS art ificial derm is has mo re excellent bio logical and mechanical p ropert ies. F ibroblasts at tach and p ro liferate bet ter on co llagen2CS scaffo ld than on co llagen scaffo lds.
OBJECTIVE: To study the stimulating effects of basic fibroblast growth factor(bFGF) on fibroblast function and its ability to expression of c-fos gene. Furthermore, to explore the possible network action between bFGF and oncogene in modulating wound healing. METHODS: Cultured rat fibroblasts were divided into bFGF stimulating group and control group. Fibroblasts in bFGF stimulating group were treated with bFGF in a dosage of 40 ng/culture hole, while the control fibroblasts were treated with the same vehicle without bFGF. The morphology, cell vitality and their ability to express c-fos gene in the fibroblasts in both groups were studied with MTT and immunohistochemical methods. RESULTS: All fibroblasts in bFGF treated groups were enlarged and showed increased vitality with MTT method. C-fos gene expression in bFGF stimulating group was increased, especially in nucleus when compared with those in control group. CONCLUSION: The results show that the function and the ability to express c-fos gene in bFGF treated fibroblasts are enhanced. Combined with our previous studies, it may make a conclusion that there is a network regulation mechanism between growth factors and some oncogenes.
ObjectiveTo investigate the histological changes and vascularization of the porcine acellular dermal matrix (P-ADM) processed with matrix metalloproteinase 7 (MMP-7) (P-ADM-pm) after implanted into rats.
MethodsSixty-two pieces of porcine reticular layer dermis which were from the pig abdominal skin and obtained by using a mechanical method, were randomly divided into group A (n=31) and group B (n=31). The porcine reticular layer dermis in 2 groups were treated with decellularization (P-ADM), then the P-ADM in group B were treated with processing by MMP-7 (P-ADM-pm). Thirty adult male Wistar rats were selected. P-ADM (group A) and P-ADM-pm (group B) were subcutaneously transplanted into the left and right fascia lacuna, respectively. The implants were harvested from 6 rats at 3, 7, 14, 21, and 28 days after implantation, respectively. Gross, histochemical, and immunohistochemical observations, and scanning electron microscopy (SEM) examination were performed to observe host cells, microvessels infiltration and histological changes in the implants.
ResultsNo rat died in the experiment, incision healed well and no obvious inflammatory reaction was seen in all rats. Gross observation suggested that the implants of 2 groups were encapsulated by a thin layer of connective tissue at 7 days after implantation. With the time of implantation, the microvessels increased and coarsened, and the changes of group B were more obvious than those of group A. At 21 days, the microvessels of 2 groups decreased, and the implants of group B showed complete vascularization. The histochemical and immunohistochemical observations showed that group A had more severe inflammatory response than group B. Fibroblasts and microvessels in group B appeared in the superficial zone of implant at 3 and 7 days after implantation and they could be observed in the center zone of implant at 14 and 21 days. However, fibroblasts and microvessels in group A appeared in the superficial zone of implant at 3 and 14 days and they could not be observed in the center zone of implant at 28 days. Fibroblasts and microvessels of group B were significantly more than those of group A (P < 0.05). SEM examination showed that more fibroblasts and new collagen fibrils were observed in group B at 14 days.
ConclusionThe host response to P-ADM-pm is similar to normal wound healing, and P-ADM-pm as implantable scaffold material plays a good template conduction role.
Objective To investigate the influence of enamel matrix proteins (EMPs) on the attachment, prol iferation and pre-mRNA of type I collagen synthesis of cultured human dermal fibroblast cells. Methods Human dermal fibroblast cells were obtained from human acrobystia and cultured in DMEM medium with 10% FBS. The 3rd to 6th passage cells were used. Ninety-six-well plates and 6-well plates were pre-coated with different concentrations of EMPs (50, 100, 150 and200 μg/ mL). ① The cell attachment experiment: 0.2 mL cells suspension at the concentration of 1 × 106/mL was added to the pre-coated 96-well plates as the experimental groups (groups A, B, C and D based on different concentrations of EMPs). At 1.5, 3.0, and 4.5 hours after inoculation, the attached cells were measured by MTT method. ② The cell prol iferation experiment: 0.2 mL cells suspension at the concentration of 5 × 104/mL was added to the pre-coated 96-well plates as the experimental groups (groups A1, B1, C1 and D1 based on the different concentrations of EMPs). At 2, 4, 6 and 8 days after inoculation, the cells were measured by MTT method. ③ The synthesis experiment of pre-mRNA: 2 mL cells at the concentration of 1 × 106/mL was added to the pre-coated 6-well plates as the experimental groups (groups A2, B2, C2 and D2 based on different concentrations of EMPs). At 5 days after inoculation, the synthesis of pre-mRNA was measured by RT-PCR method. Human dermal fibroblast cells were added to the un-coated plates as the control groups. Results ① The cell attachment experiment: There were significant differences in attachment cells between the control group, group A and the groups B, C and D (P lt; 0.05). There were no significant difference between group A and control group (P lt; 0.05). ② The cell prol iferation experiment: At 2 days, there were no significant differences in absorbance between the control group and the experimental groups (P gt; 0.05); at 4 days and 6 days, the absorbance of groups B1 (0.598 ± 0.020 and 0.639 ± 0.016 ), C1 (0.582 ± 0.017 and 0.641 ± 0.020) and D1 (0.574 ± 0.021and 0.635 ± 0.021) was significantly higher than that of the control group (0.548 ± 0.021 and 0.605 ± 0.019, P lt; 0.05); at 8 days, the absorbance of group B1 (0.629 ± 0.012) and group C1 (0.631 ± 0.014) was significantly higher than that of the control group (0.606 ± 0.031, P lt; 0.05). ③ The synthesis experiment of pre-mRNA: The synthesis of type I collage pre-mRNA of groups B2, C2 and D2 was significantly higher than that of the control group. Conclusion EMPs stimulate human dermal fibroblast cell attachment, prol iferation and synthesis of type I collage pre-mRNA, and its maximal effect can be achieved at the concentration of 100 μg /mL.
Objective
To observe the affection of optic nerve under acute ocular hypertension and the effect of protection of bFGF on optic nerve.
Methods
BSS was perfused into anterior chamber of rabbits to increase the intraocular pressure to cause retinal ischemia. A computer image analysis system was used to count the optic nerve axons.Eyes were intravitreally injected with bFGF and then the number of optic nerve axons of the normal rabbits,and hypertension with and without bFGE treatment groups were counted respectively.
Results
The number of optic nerve axons in ocular hypertension eyes was less than the normal eyes(P=0.00003).The bFGF treated eyes had more optic nerve axons than the controls(P=0.0078).
Conclusions
The acute ocular hypertension may cause the loss of the nerve axons,and bFGF may be effective in protecting optic nerve in acute ocular hypertension.
(Chin J Ocul Fundus Dis,2000,16:94-96)
ObjectiveTo investigate the effects of liver X receptor agonist, T0901317, on the proliferation, migration and hydroxyproline production of human embryonic lung fibroblasts (HELF).
MethodsHELF cells were devided into a control group, a growth factor group, a T0901317 group and three growth factor+T0901317 groups. The cells of the control group were treated with Dulbecco's modified Eagle medium. The cells of T0901317 group were treated with 1.00 μmol/L T0901317. The growth factor+T0901317 groups were incubated with different doses of T0901317 (0.25 μmol/L, 0.50 μmol/L and 1.00 μmol/L) for 2 h. Then the cells of the growth factor+T0901317 groups and the growth factor group were incubated with basic fibroblast growth factor and transforming growth factor-β1 for 24 h. The proliferation, migration and collagen production of HELF were determined by cell counting kit-8 method, transwell chamber, and hydroxyproline method.
ResultsCompared with the control group, T0901317 had no effect on the proliferation, migration and hydroxyproline production of HELF. Growth factors could promote the proliferation, migration and hydroxyproline production of HELF significantly. T0901317 could inhibit those effects of growth factors with a dosage-dependent manner.
ConclusionT0901317 may inhibit the proliferation, migration and hydroxyproline production of HELF induced by growth factors.
OBJECTIVE :To investigale effect of subretinal fluld(SRF)on proliferalion of the cellular elements of PVR.
METHOD:The effect of SRF of 28 patients with rhegmatogenous retinal detachment proliferation of the cultured human retinal pigment epithelial cells(RPE),retinal glial cells (RG),and fibroblast (FB)was observed and detected by the methods of cell-counting and 3H-TdR in DNA synthesis.
RESULTS:The range of proliferatinn-stimulating activity was 52.5%~233.3%,
36.4% ~ 177.8%，55.4% ~277.8% above the baseline in 1:10 dilution of these 3 kinds ,of cellular elements,and there was no significant difference among them.
CONCLUSION ;The stimulating effect of SRF on the cellular proliferation was thougt to be due to the actions from certain growth factors.
（Chin J Ocul Fundus Dis,1996,12: 233-235）
Porpose
To investigate the optimal concentration of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on DNA synthesis and their synergism indensity arrested human retinal pigment epithelial (RPE) cells. Methods
Growth factor effects in cultured human RPE of the 6th generation were assessed by [3 H]-thymidine incorporation and radioautography. Results
EGF and bFGF were potent stimulators when used alone,and their optimal concentrations were 10ng/ml in DMEM and 1ng/ml in 2% serum DMEM.When used in combination (10ng/ml EGF and 10ng/ml bFGF),they caused a significant enhancement of [3 H]-thymidine incorporation about 2.96 times. Conclusion
EGF and bFGF were potent stimulators in RPE cells,and demonstrated synergism in their action. (Chin J Ocul Fundus Dis,1998,14:98-100)
