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Fibroblast growth factor 18 increases the trophic effects of bone marrow mesenchymal stem cells on chondrocytes isolated from late stage osteoarthritic patients.

Zhang Z, Wang Y, Li M, Li J, Wu J - Stem Cells Int (2014)

Bottom Line: Finally, coimplantation of MSCs with OA chondrocytes produces more matrix than chondrocytes only.In conclusion, FGF18 can restore the responsiveness of OA chondrocytes to the trophic effects of MSCs.Coimplantation of MSCs and OA chondrocytes treated with FGF18 may be a good alternative cell source for regenerating cartilage tissue that is degraded during OA pathological changes.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, The First Affiliated Hospital of Harbin Medical University, 23 You Zheng Street, Nangang District, Harbin 150001, China.

ABSTRACT
Coculture of mesenchymal stem cells with chondrocytes increases production of cartilaginous matrix. Chondrocytes isolated from late stage osteoarthritic patients usually lost their phenotype of producing cartilaginous matrix. Fibroblast growth factor 18 is believed to redifferentiate OA chondrocyte into functionally active chondrocytes. The aim of this study is to investigate the supportive effects of MSCs on OA chondrocytes and test if FGF18 could enhance the responsiveness of OA chondrocytes to the support of MSCs in a coculture system. Both pellet and transwell co-cultures were used. GAG quantification, hydroxyproline assay, and qPCR were performed. An ectopic models of cartilage formation was also applied. Our data indicated that, in pellets coculture of MSCs and OA chondrocytes, matrix production was increased in the presence of FGF18, comparing to the monoculture of chondrocytes. Results from transwell coculture study showed that expression of matrix producing genes in OA chondrocytes increased when cocultured with MSCs with FGF18 in culture medium, while hypertrophic genes were not changed by coculture. Finally, coimplantation of MSCs with OA chondrocytes produces more matrix than chondrocytes only. In conclusion, FGF18 can restore the responsiveness of OA chondrocytes to the trophic effects of MSCs. Coimplantation of MSCs and OA chondrocytes treated with FGF18 may be a good alternative cell source for regenerating cartilage tissue that is degraded during OA pathological changes.

No MeSH data available.


Related in: MedlinePlus

Coimplantation of MSCs and chondrocyte in alginate hydrogel subcutaneously in nude mice. (a) Overview of tissue engineering constructs containing chondrocytes only or mixture of MSC and chondrocyte. Hydrogel was loaded with or without FGF18 (10 ng/mL). Bar = 5 mm. (b) Tissue engineering constructs were sectioned and stained with Safranin O. Lower panel is the enlarged pictures of the inserts in upper panel. Bar = 50 μm. (c) GAGs were quantified in tissue engineering constructs GAG assay (n = 3). Total GAGs (μg) were normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (d) Collagen contents were measured in tissue engineering constructs by hydroxyproline assay (n = 3). Total collagen (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (e) Collagen type II deposition was tested in tissue engineering constructs by ELISA (n = 3). Total COL II (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance.
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fig3: Coimplantation of MSCs and chondrocyte in alginate hydrogel subcutaneously in nude mice. (a) Overview of tissue engineering constructs containing chondrocytes only or mixture of MSC and chondrocyte. Hydrogel was loaded with or without FGF18 (10 ng/mL). Bar = 5 mm. (b) Tissue engineering constructs were sectioned and stained with Safranin O. Lower panel is the enlarged pictures of the inserts in upper panel. Bar = 50 μm. (c) GAGs were quantified in tissue engineering constructs GAG assay (n = 3). Total GAGs (μg) were normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (d) Collagen contents were measured in tissue engineering constructs by hydroxyproline assay (n = 3). Total collagen (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (e) Collagen type II deposition was tested in tissue engineering constructs by ELISA (n = 3). Total COL II (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance.

Mentions: To evaluate previous findings in vivo, chondrocytes and MSCs (ratio 1 : 1) were coimplanted into subcutaneously immune-deficient mice with alginate as carrying vehicle. Before implantation, TE constructs were cultured in serum-free medium with or without FGF18. Then, eight weeks after implantation, histological examination as well as GAG and collagen quantification was performed. Figure 1(a) shows the gross-looking of TE constructs under stereomicroscope. In general, TE constructs of all groups look similar. Those ones not cultured in FGF18 seem to have more angiogenesis. As shown in Figure 3(b), there was some accumulation of ECM components in the pericellular regions in all experimental groups. However, TE constructs cultured in FGF18 obviously deposited more GAGs than constructs cultured without FG18. Quantification of GAG and collagen confirmed that coimplantation after FGF18 culture group produced most GAGs and collagens when comparing with other experimental groups (Figures 3(c) and 3(d)). Then, ELISA was used to quantify cartilage specific COL II in TE constructs. Comparing the groups cultured with or without FGF18, at least threefold increase was observed in COL II production between two groups (Figure 3(e)). Again, coimplantation TE constructs contain significantly more COL II than chondrocyte TE constructs.


Fibroblast growth factor 18 increases the trophic effects of bone marrow mesenchymal stem cells on chondrocytes isolated from late stage osteoarthritic patients.

Zhang Z, Wang Y, Li M, Li J, Wu J - Stem Cells Int (2014)

Coimplantation of MSCs and chondrocyte in alginate hydrogel subcutaneously in nude mice. (a) Overview of tissue engineering constructs containing chondrocytes only or mixture of MSC and chondrocyte. Hydrogel was loaded with or without FGF18 (10 ng/mL). Bar = 5 mm. (b) Tissue engineering constructs were sectioned and stained with Safranin O. Lower panel is the enlarged pictures of the inserts in upper panel. Bar = 50 μm. (c) GAGs were quantified in tissue engineering constructs GAG assay (n = 3). Total GAGs (μg) were normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (d) Collagen contents were measured in tissue engineering constructs by hydroxyproline assay (n = 3). Total collagen (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (e) Collagen type II deposition was tested in tissue engineering constructs by ELISA (n = 3). Total COL II (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Coimplantation of MSCs and chondrocyte in alginate hydrogel subcutaneously in nude mice. (a) Overview of tissue engineering constructs containing chondrocytes only or mixture of MSC and chondrocyte. Hydrogel was loaded with or without FGF18 (10 ng/mL). Bar = 5 mm. (b) Tissue engineering constructs were sectioned and stained with Safranin O. Lower panel is the enlarged pictures of the inserts in upper panel. Bar = 50 μm. (c) GAGs were quantified in tissue engineering constructs GAG assay (n = 3). Total GAGs (μg) were normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (d) Collagen contents were measured in tissue engineering constructs by hydroxyproline assay (n = 3). Total collagen (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance. (e) Collagen type II deposition was tested in tissue engineering constructs by ELISA (n = 3). Total COL II (μg) was normalized to wet weight (mg). Data was presented as mean ± standard deviation. P values were calculated with Student's t-test. NS: nonsignificance.
Mentions: To evaluate previous findings in vivo, chondrocytes and MSCs (ratio 1 : 1) were coimplanted into subcutaneously immune-deficient mice with alginate as carrying vehicle. Before implantation, TE constructs were cultured in serum-free medium with or without FGF18. Then, eight weeks after implantation, histological examination as well as GAG and collagen quantification was performed. Figure 1(a) shows the gross-looking of TE constructs under stereomicroscope. In general, TE constructs of all groups look similar. Those ones not cultured in FGF18 seem to have more angiogenesis. As shown in Figure 3(b), there was some accumulation of ECM components in the pericellular regions in all experimental groups. However, TE constructs cultured in FGF18 obviously deposited more GAGs than constructs cultured without FG18. Quantification of GAG and collagen confirmed that coimplantation after FGF18 culture group produced most GAGs and collagens when comparing with other experimental groups (Figures 3(c) and 3(d)). Then, ELISA was used to quantify cartilage specific COL II in TE constructs. Comparing the groups cultured with or without FGF18, at least threefold increase was observed in COL II production between two groups (Figure 3(e)). Again, coimplantation TE constructs contain significantly more COL II than chondrocyte TE constructs.

Bottom Line: Finally, coimplantation of MSCs with OA chondrocytes produces more matrix than chondrocytes only.In conclusion, FGF18 can restore the responsiveness of OA chondrocytes to the trophic effects of MSCs.Coimplantation of MSCs and OA chondrocytes treated with FGF18 may be a good alternative cell source for regenerating cartilage tissue that is degraded during OA pathological changes.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, The First Affiliated Hospital of Harbin Medical University, 23 You Zheng Street, Nangang District, Harbin 150001, China.

ABSTRACT
Coculture of mesenchymal stem cells with chondrocytes increases production of cartilaginous matrix. Chondrocytes isolated from late stage osteoarthritic patients usually lost their phenotype of producing cartilaginous matrix. Fibroblast growth factor 18 is believed to redifferentiate OA chondrocyte into functionally active chondrocytes. The aim of this study is to investigate the supportive effects of MSCs on OA chondrocytes and test if FGF18 could enhance the responsiveness of OA chondrocytes to the support of MSCs in a coculture system. Both pellet and transwell co-cultures were used. GAG quantification, hydroxyproline assay, and qPCR were performed. An ectopic models of cartilage formation was also applied. Our data indicated that, in pellets coculture of MSCs and OA chondrocytes, matrix production was increased in the presence of FGF18, comparing to the monoculture of chondrocytes. Results from transwell coculture study showed that expression of matrix producing genes in OA chondrocytes increased when cocultured with MSCs with FGF18 in culture medium, while hypertrophic genes were not changed by coculture. Finally, coimplantation of MSCs with OA chondrocytes produces more matrix than chondrocytes only. In conclusion, FGF18 can restore the responsiveness of OA chondrocytes to the trophic effects of MSCs. Coimplantation of MSCs and OA chondrocytes treated with FGF18 may be a good alternative cell source for regenerating cartilage tissue that is degraded during OA pathological changes.

No MeSH data available.


Related in: MedlinePlus