ABSTRACT
Objective To investigate the the multi-directional differentiation potential between pluripotent of human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLCs). Methods HPDLCs and HGFs were obtained from the primary culture. HPDLCs and HGFs at 3rd-4th passage were cultured in osteogenic, adipogenic or chondrogenic me-dium. Cells without differentiation were taken as control. Alizarin red, Alcian blue and oil red O staining were performed to detect osteogenic differentiation, chondrogenic and adipogenic differentiation in vitro, respectively. Reverse transcription polymerase chain reaction (RT-PCR) was applied to examine the expression of osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and collagen 1 (Col 1), peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and collagen 10 (Col 10). Results HPDLCs and HGFs cultured in osteogenic medium showed massive calicium nodulus at day 28, but HP-DLCs formed more calicium nodulus than those of HGFs. The expressions of OCN, RUNX2 and Col 1 were significantly high-er in HPDLCs than those in HGFs (P<0.05). In chondrogenic medium both cells were found blue deposit at day 14, and the expression of Col 10 was significantly higher in HGFs than that of HPDLCs (P<0.01). Furthermore, in adipogenic medium HGFs showed more lipid-filled droplets stained with oil red O than HPDLCs at day 21. The expression of PPARγ2 was sig-nificantly higher in HGFs than that of HPDLCs (P<0.01). Conclusion HPDLCs has the better potency of osteogenic differ-etiation than HGFs, however, HGFs has the better potency of adipogenic and chondrogenic differentiation.
ABSTRACT
Marrow stromal cells ( MSCs) , component of the mar-row stroma, play an important role in the growth of bone and metabolic balance. This paper mainly summarizes and reviews the molecular regulaory effects of vitamin D and its active form 1, 25 dihydroxy vitamin D3[1,25(OH)2D3] on the osteogenet-ic differentiation of MSCs by Wnt signaling pathway, Wnt5a/ROR2 axis, BMP/TGF-β/Samd signaling pathway and ROS/ERK signaling pathway, so as to clarify the molecular signaling pathway through which vitamin D regulates MSCs metabolically.
ABSTRACT
Objective To investigate the pluripotency of human gingival fibroblasts (hGFs), and provide a novel cell source for tissue engineering. Methods With informed consent from volunteers, fresh and healthy gingiva were collected. The hGFs were obtained from the gingiva by tissue culture. The third passage of hGFs was cultured in osteogenic medium, chondrogenic medium and adipogenic medium. Cells without differentiation were taken as control. Cells were examined by al?kaline phosphatase (ALP) staining, Alizarin red staining, Alcian blue staining and oil red O staining for detecting of the abili?ty of differentiation pluripotency. Real-time polymerase chain reaction was applied to examine the expression of osteogenic marker genes ALP, runt-related transcript factor 2 (Runx2), chondrogenic marker aggrecan (AGR) and adipogenic marker peroxisome proliferator-activated receptor gamma 2 (PPARγ2). Results The hGFs cultured in osteogenic medium showed massive violet deposit at day 7 and calcium nodulus at day 28, meanwhile, the expressions of ALP and Runx2 were higher than those of control (P<0.01). In chondrogenic group cells were found blue deposit at day 14. In adipogenic group lipid-filled droplets stained with oil red O were found in cells at day 14. However, hGFs in control group had no any positive stain?ing. Furthermore, expressions of AGR and PPARγ2 were significantly higher than those of control (P<0.01). Conclusion Human gingival fibroblasts have the pluripotency of osteogenic, adipogenic and chondrogenic differentiation.
ABSTRACT
Aim To investigate the role of Wnt/β-cate-nin signaling pathway on the baicalin-induced osteo-genic differentiation in rat bone marrow derived mesen-chymal stem cells ( rBMSC ) . Methods rBMSC was isolated and cultured by adherence screening method. Alkaline phosphatase ( ALP) amount, CFU-FALP and mineralized nodules were compared between each ba-icalin group and vehicle control group at different time points. Real time q-PCR was employed to evaluate the mRNA level of Wnt signaling-related marker ( Wnt10a, GSK-3β,β-catenin and LEF1) after baica-lin treatment. Protein expression of β-catenin and Runx2 was measured by Western blot. Results Ba-icalin significantly increased ALP activities from day 3 to day 7 . The formation of CFU-FALP and mineralized nodules remarkably increased after rBMSC was treated with1, 10, 50 μmol · L-1 baicalin. mRNA levels of Wnt10a, β-catenin, GSK-3β, LEF1and osteocalcin were enhanced significantly in baicalin-treated group compared to control group. Protein expression of β-catenin and Runx2 was also elevated. Conclusion Baicalin ( 0. 1 to 50 μmol · L-1 ) promotes the osteo-genic differentiation and maturation of rBMSC, in which Wnt/β-catenin signaling pathway might be in-volved.