Objective To study the effects of drug-containing serum of Ficus Hirta on oxidative damage of spleen lymphocyte due to aging in aged mice; To discuss its mechanism of action.Methods Forty aged mice were randomly divided into control group and high-, medium- and low-dose Ficus Hirta groups. Control group was given 0.9% sodium chloride solution for gavage, while high-, medium- and low-dose Ficus Hirta groups were given 6.6, 4.4, and 2.2 g/kg aqueous extract of Ficus Hirta for gavage. The spleen index was observed for optimum dose in aged mice. The optimum time and dilution of drug-containing serum of Ficus Hirta were confirmed by MTT method in lymphocyte proliferation test. The positive rate of senescent cells, the activity of T-SOD and the contents of MDA and ROS were determined in cellular antioxidant experiment after treated by optimal drug-containing serum for 48 h. Results Compared with the control group, the spleen index was significantly improved in high-, medium- and low-dose Ficus Hirta groups (P<0.05,P<0.01). 20% drug-containing serum of Ficus Hirta cultivated for 48 h had the best effects on lymphocyte proliferation in aged mice. 20% drug-containing serum of Ficus Hirta could significantly decrease the positive rate of senescent cells (P<0.01), improve T-SOD activity and decrease the contents of MDA and ROS (P<0.05,P<0.01).Conclusion The drug-containing serum of Ficus Hirta can improve the proliferative activity of spleen lymphocyte in aged mice and the mechanism of action may be involved in decreasing the positive rate of senescent cells and increasing antioxidant ability of lymphocyte.

OBJECTIVETo evaluate the effects of Icariin (ICA) on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in vitro and in vivo.

METHODSAn enzymatic digestion block was used in vitro to culture hPDLSCs, which were separated and purified by limited dilution cloning. The hPDLSCs were identified using cell-surface markers and cocultured with 1 x 10(-7) mol.L-1 ICA solution. The proliferation ability of these cells was determined by thiazolyl blue tetrazolium bromide (MTT) assay. After staining with alkaline phosphatase (ALP), osteogenesis was detected by enzyme-linked immunosorbent assay. Osteoblast-related genes were analyzed by reverse transcription-polymerase chain reaction. Alizarin red staining was performed to measure the level of calcium deposition. The hPDLSCs were cocultured with 1 x 10(-7) mol.L-1 ICA and nano-hydroxyapatite scaffolds in vivo before transplantation into subcutaneous tissues of nude mice. Osteogenic abilities were histochemically analyzed after 30 days of induction.

RESULTSThe hPDLSCs were affected by 1 x 10(-7) mol.L-1 ICA, and MTT assay showed that the proliferation of the groups treated with ICA in vitro was better than that of the control groups on the second day. The ALP activity of the treated hPDLSCs was significantly enhanced after cell culture for 3, 5, and 7 days. The gene expression of osteoblastic markers was also significantly enhanced after 7 days. The deposition of mineralization after incubation with 1 x 10(-7) mol.L-1 ICA increased compared with the control after cell culture for 14, 21, and 28 days. Furthermore, the bone expression of the treatment groups in vivo was significantly enhanced compared with that of the control groups.

CONCLUSIONTreatment with 1 x 10(-7) mol.L-1 ICA can significantly promote proliferation and differentiation of hPDLSCs in vitro and in vivo. ICA can effectively function as a bioactive growth factor in periodontal tissue engineering to replace traditional growth factors.

Alkaline Phosphatase ; Animals ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Coculture Techniques ; Enzyme-Linked Immunosorbent Assay ; Flavonoids ; Humans ; Mice ; Mice, Nude ; Osteoblasts ; Osteogenesis ; Periodontal Ligament ; Stem Cells

Objective??To?evaluate?the?effects?of?Icariin?(ICA)?on?the?proliferation?and?osteogenic?differentiation?of?human?periodontal?ligament?stem?cells?(hPDLSCs)?in vitro and?in vivo. Methods??An?enzymatic?digestion?block?was?used?in vitro?to?culture?hPDLSCs,?which?were?separated?and?purified?by?limited?dilution?cloning.?The?hPDLSCs?were?identified?using?cell-surface?markers?and?cocultured?with?1×10?7 mol·L?1 ICA?solution.?The?proliferation?ability?of?these?cells?was?determined?by?thiazolyl?blue?tetrazolium?bromide?(MTT)?assay.?After?staining?with?alkaline?phosphatase?(ALP),?osteogenesis?was?detected?by?enzyme-linked?immunosorbent?assay.?Osteoblast-related?genes?were?analyzed?by?reverse?transcription-polymerase?chain?reaction.?Alizarin?red?staining?was?performed?to?measure?the?level?of?calcium?deposition.?The?hPDLSCs?were?cocultured?with?1×10?7 mol·L?1 ICA?and?nano-hydroxyapatite?scaffolds?in vivo?before?transplantation?into?subcutaneous?tissues?of?nude?mice.?Osteogenic?abilities?were?histochemically?analyzed?after?30?days?of?induction.?Results??The?hPDLSCs?were?affected?by?1×10?7 mol·L?1 ICA,?and?MTT?assay?showed?that?the?proliferation?of?the?groups?treated?with?ICA?in vitro?was?better?than?that?of?the?control?groups?on?the?second?day.?The?ALP?activity?of?the?treated?hPDLSCs?was?significantly?enhanced?after?cell?culture?for?3,?5,?and?7?days.?The?gene?expression?of?osteoblastic?markers?was?also?significantly?enhanced?after?7?days.?The?deposition?of?mineralization?after?incubation?with?1×10?7 mol·L?1?ICA?increased?compared?with?the?control?after?cell?culture?for?14,?21,?and?28?days.?Furthermore,?the?bone?expression?of?the?treatment?groups?in vivo?was?significantly?enhanced?com-pared?with?that?of?the?control?groups.?Conclusion??Treatment?with?1×10?7 mol·L?1 ICA?can?significantly?promote?proliferation?and?differentiation?of?hPDLSCs?in vitro?and?in vivo.?ICA?can?effectively?function?as?a?bioactive?growth?factor?in?periodontal?tissue?engineering?to?replace?traditional?growth?factors.