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Objective To investigate the osteogenic differentiation of rabbit dental pulp stem cells (DPSCs) induced by transforming growth factor-β3 (TGF-β3) in vitro. Methods DPSCs and osteoblasts (OBs) were respectively obtained from rabbit dental pulp and skull by enzymetic digestion method. The morphology of the cells was observed by a light microscopy. Immunohistochemical staining and alizarin red staining were carried out to identify OBs. The third generation of DPSCs and OBs were divided into three groups, including DPSCs group (blank control), OBs group (positive control) and DPSCs+TGF-β3 group (experimental group). The expression of Runt-related transcription factor 2 (Runx-2) in each group was detected by immunohistochemical staining on the 5th day of culture. The activity of alkaline phosphatase (ALP) in each group was detected by assay kit on the 7th day of culture. Western Blot was used to detect the expression of the bone-specific markers Runx-2 and TGF-β3 proteins on the 1st, 3rd, 5th and 7th days of culture. Results The rabbit DPSCs were mostly long spindle-shaped with many synapses. The OBs were mostly short spindle-shaped or fibroblast-like, and plump with few synapses. The identification result showed that the DPSCs and OBs were positive. On the 5th day of culture, the expression of Runx-2 protein in the OBs group and DPSCs+TGF-β3 group showed strong positive. On the 7th day of culture, there was no significant difference in ALP activity between the above two groups (P>0.05). The results of Western Blot showed that the relative expressions of Runx-2 and TGF-β3 protein in the DPSCs group were significantly different from those in the other two groups, and the differences were statistically significant (all P<0.05). On the 7th day of culture, the relative expression of Runx-2 and TGF-β3 protein in the DPSCs+TGF-β3 group was higher than that in the other two groups, and the differences were statistically significant (all P<0.01). Conclusions TGF-β3 can promote the expression of early osteogenic specific proteins in DPSCs.
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OBJECTIVE:To study the preventive and therapeutic effect and its mechanism of medicine pair decoction liquid of Curcuma phaeocaulis-Sparganium stoloniferum on rats with uterine myoma. METHODS:Rats were randomly divided into normal group,model group,positive group(mifepristone,2.25 mg/kg)and medicine pair decoction liquid group(C. phaeocaulis-S. stolon-iferum decoction liquid,6.0 g/kg),10 in each group. Except for the normal group,rats in other groups were injected Estradiol ben-zoate injection (0.5 mg/kg) intramuscularly every Monday,Wednesday and Friday,for 12 weeks. From the 13th week,rats in modeling group were added Progesterone injection(5 mg/kg)intramuscularly as well as relevant medicines intragastrically,once a day,until the 16th week. After administration,uterine coefficient of rats was detected. HE staining was used to observe the patho-logical changes of uterus and determine the thickness of smooth muscle;immunohistochemical staining was adopted to detect the transforming growth factor β3 (TGF-β3),matrix metalloproteinase 11 (MMP-11) protein expressions in uterus tissue of rats. RE-SULTS:Compared with normal group,uterine coefficient was increased in model group,pathological changes were obvious in uterus,thickness of smooth muscle was increased,TGF-β3 and MMP-11 protein expressions were enhanced(P<0.05 or P<0.01). Compared with model group,above-mentioned changes were improved significantly in positive group and medicine pair decoction liquid groups(P<0.05 or P<0.01). CONCLUSIONS:Medicine pair decoction liquid of C. phaeocaulis-S. stoloniferum shows cer-tain preventive and therapeutic effect on rats with uterine myoma. The mechanism may be associated with downregulating the TGF-β3,MMP-11 protein expressions in uterus tissue.
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Objective To investigate the feasibility of combing the application of transforming growth factor-β3 (TGF-β3) with concentration of 100 ng/μl and dental pulp stem cells (DPSCs) for recovering rabbit facial nerve transverse trauma.Methods Thirty-six healthy adult Zelanian rabbits of clean-grade were selected and randomly divided into group DPSCs+TGF-β3 (experimental group), group TGF-β3 (control group 1) and group PBS (control group 2) with 12 rabbits in each group.The operations for all three groups were applied at rabbit's left cheek.A model of traumatic transection was set on upper buccal branch, then 100 ng/μl TGF-β3 solution and 0.1 ml of 1 ×108/L DPSCs suspension were added into regeneration chamber for the experimental group, while the same amount of 100 ng/μl TGF-β3 solution was added for group TGF-β3 and the same amount of PBS for group PBS.The recovery of facial nerve regeneration with the prepared animal's specimen was evaluated in the 1st, 4th and 12th week after the operation on sacrificed rabbits.Results The effects on nerve regeneration recovery for the experimental group was superior to that of control groups 1 and 2 with all the 36 models included in the result analysis, and that of control group 1 was superior to that of control group 2, which was getting better with the extension of time.Conclusions The combined application of TGF-β3 and DPSCs can effectively promote the facial nerve regeneration, which is better than that of single application of TGF-β3.Meanwhile, the effect with TGFβ3 application is better than that with application of PBS.
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Objective To investigate the ability of human exfoliated deciduous teeth-derived stem cells (SHED) to differentiate into odontoblast-like cells. Methods SHEDs were isolated by enzyme digestion method. The 3nd passage SHEDs were incubated with 25 ng/mL recombinant human TGF-β3 , or with TGF-β3 in combination with heparin. The DSPP expression was detected by Q-PCR and Western-blotting assay. Alizarin red staining, immunhistochemistry assay and alkaline phosphatase(AKP) activity assay were performed, respectively. Result The AKP activity was enhance by TGF-β3 in combination with heparin. Alizarin red staining was positive in TGF-β3-heparin groups, with the increase of DSPP expression at both mRNA and protein level. Conclusion TGF-β3 in combination with heparin can enhance the differentiation of human exfoliated deciduous teeth-derived stem cells into odontoblast-like cells.
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ObjectiveTo investigate the effects of using transforming growth factor-β3 (TGF-β3) combined with dental pulp stem cells (DPSCs) on the regeneration of rabbit facial nerve.MethodsSixteen New Zealand adult rabbits were selected randomly.These rabbits were divided into three groups (TGF-β3 and DPSCs treated group,TGF-β3 control group and PBS control group.treatment group (silicone guidance channel with collagen protein sponge were filled with TGF-β3 and DPSCs),TGF-β3 control group (silicone guidance channel with collagen protein sponge were filled with TGF-β3) and PBS control group(silicone guidance channel with collagen protein sponge were filled with PBS).After 3 months,a series of examinations were performed,including gross morphology,histological staining,neuroelectrophysiological tset.ResultsIn the experimental group,the diameter of the regenerating nerve and near distal nerve stem were almost the same.There were no formation of neuroma.The adventitial angiogenesis was rich and with tough texture.3 months after operation,in the experimental group,the facial nerve membrane integrity,nerve fibers arranged in neat rows,form a more complete,myelin swelling.The total number of regeneration of nerve fibers in the experimental group were more than of control groups,statistical analysis was significant (P＜0.05).Diameter of regenerating nerve fibers in the experimental group were greater than that of control groups,statistical analysis was significant (P＜0.05).The ultra-thin section showed that the regenerated fibers in the treatment group were mainly myelinated never fiber.The layer structure of myelin sheath was clear,and there were rich organells axoplasma.The neuroelectrophysiological examinations revealed that the latency of nerve and muscle action conduction in the treatment group was shorter than that of the control groups,the treatment group:(1.96±0.32) ms,the TGF-β3 control group:(2.35±0.41) ms,the PBS control group:(3.42±0.55) ms.The wave amplitude of nerve and muscle action conduction in the treatment group was obviously higher than that of control groups,the treatment group:(11.06±3.25) mV,the TGF-β3 control group:(8.40± 1.68) mV,the PBS control group:(4.62±0.77) mV.ConclusionThe combination of TGF-β3 and DPSCs can improve the effects on the repair of facial nerve injury.
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Objective To investigate effect of transforming growth factor β3(TGFβ3)on expression of MMP-9,MMP-2,TIMP-1 and collagen I in rats with liver fibrosis through transduction by recombinant adeno-associated virus 2(rAAV2).Methods Rats were randomly divided into 4 groups:normal control group,model group,negative control group and TGFβ3 group.Liver fibrosis model was induced by hypodermic injection of 40% CCl4.rAAV2-TGFβ3,was injected via vena caudalis of the rats one week before CCl4 Was given.All rats were sacrificed and the liver tissues were taken 8 weeks afler injection of CCl4.The histopathological changes were observed on HE sections;the expressions of MMP-9,MMP-2,TIMP-1 and collagen I were examined by histochemistry and the positive area rates were semi-quantitatively analyzed.Results Compared with the model group and negative control group,the decreases of inflammatory infiltration and collagen fibers hyperplasia were observed in the TGFβ3 group,and the expression of MMP-9 increased(q=23.664,27.746,P<0.01),the expression of collagen I(q=5.503,5.251,P<0.01)and TIMP-1(q=5.800,8.608,P<0.01)decreased,but that of MMP-2 was not changed(q=2.1 08,0.996,P>0.05).Conclusion rAAV2-TGFβ3 can reduce the histological damage and degree of liver fibrosis in rats by inhibiting expression of TIMP-1 and collagen I,and upregulating expression of MMP-9.