Table 1.
Oligonucleotide primers used in RT-PCR.
Figure 1.
SFMSCs obtained from TMD patients.
(A) Dissociated single adherent cells were observed after culture of synovial fluid samples for 48(B) Cells were observed growing from small tissue masses after culture of synovial fluid samples for 5 days. (C) Microscopic image showing the typical morphology of synovial fluid-derived cells. (D) Immunofluorescent staining of synovial fluid-derived cells demonstrated positive expression of VCAM-1. Scale bars = 100 µm.
Figure 2.
SFs and SFCs obtained from the synovial fluid of TMD patients.
(A) Gross morphology of a SF under a stereomicroscope (M205A, Leica, Germany). (B) An HE-stained section of a SF. (C) Typical morphology of SFCs. (D) Immunofluorescent staining showed that almost all SFCs were positive for VCAM-1. Scale bar = 1 mm in (A) and 100 µm in (B-D).
Figure 3.
Cell growth curve of SFCs and SFMSCs.
Figure 4.
Flow cytometric analysis of SFCs and SFMSCs.
Both SFCs and SFMSCs expressed similar surface markers, including CD90, CD44, CD105, and CD73. Stro-1, CD146 CD45, CD34, CD11b, CD19, and HLA-DR were not detected on the surfaces of these cells. Black lines represent negative controls, and red lines are the results for the experimental groups.
Figure 5.
Osteogenic differentiation of SFCs and SFMSCs after culture in induction medium for 28 days.
Alizarin red staining identified calcium deposits in cultured (A) SFCs and (C) SFMSCs. von Kossa staining identified calcium deposits in cultured (B) SFCs and (D) SFMSCs. Scale bars = 100 µm. (E) Upregulated expression of runx2 and (F) ALP activity in SFCs and SFMSCs after osteogenic induction compared to that in control cells. Significant differences between the two groups are indicated by single and double asterisks (*p<0.05, **p<0.01).
Figure 6.
Adipogenic differentiation of SFCs and SFMSCs after culture in induction medium for 28 days.
Lipid droplets were observed in SFCs after the induction period by (A) oil red O staining and (B) Sudan black B staining. Lipid droplets were also observed in SFMSCs after the induction period by (C) oil red O staining and (D) Sudan black B staining. Scale bars = 100 µm. (E) After culture in induction medium for 28 days, lpl expression was upregulated in both SFCs and SFMSCs compared to the respective control cells. Significant differences between the two groups are indicated by single and double asterisks (*p<0.05, **p<0.01).
Figure 7.
Chondrogenic differentiation of SFCs and SFMSCs after culture in induction medium for 21 days.
(A) Cartilage nodule formed by SFCs after the induction period. (B) Safranin O staining and (C) collagen type II immunohistochemical staining of cartilage nodules formed by SFCs. (D) Cartilage nodule formed by SFMSCs after the induction period. (E) Safranin O staining and (F) collagen type II immunohistochemical staining of cartilage nodules formed by SFMSCs. Scale bars = 1 mm in (A, D) and 100 µm in (B, C, E, F).
Figure 8.
Neurogenic differentiation of SFCs and SFMSCs after culture in induction medium for 12 hours.
(A) SFCs acquired a bipolar and stellate morphology by the end of the induction period and stained positively for (B) nestin expression and (C) glial fibrillary acidic protein expression. (D) SFMSCs also acquired a bipolar and stellate morphology by the end of the induction period and stained positively for (E) nestin expression and (F) glial fibrillary acidic protein expression. Scale bars = 100 µm.