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Platelet-rich plasma releasate differently stimulates cellular commitment toward the chondrogenic lineage according to concentration.

do Amaral RJ, Matsiko A, Tomazette MR, Rocha WK, Cordeiro-Spinetti E, Levingstone TJ, Farina M, O'Brien FJ, El-Cheikh MC, Balduino A - J Tissue Eng (2015)

Bottom Line: In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation.Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum.Our data support the hypothesis of platelet-rich plasma chondrogenic potential, allowing fetal bovine serum substitution for platelet-rich plasma releasate at specific concentrations in culture medium when chondrogenic commitment is desired on specific cell types and moments of culture.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brasil ; Excellion Serviços Biomédicos, Amil/UnitedHealth Group, Petrópolis, Brasil.

ABSTRACT
Platelet-rich plasma has been used to treat articular cartilage defects, with the expectations of anabolic and anti-inflammatory effects. However, its role on cellular chondrogenic or fibrogenic commitment is still a controversy. Herein, the role of platelet-rich plasma releasate, the product obtained following platelet-rich plasma activation, on cellular commitment toward the chondrogenic lineage was evaluated in vitro. Human nasoseptal chondrogenic cells and human bone marrow mesenchymal stromal cells were used as cell types already committed to the chondrogenic lineage and undifferentiated cells, respectively, as different concentrations of platelet-rich plasma releasate were tested in comparison to commonly used fetal bovine serum. Low concentration of platelet-rich plasma releasate (2.5%) presented similar effects on cellular growth compared to 10% fetal bovine serum, for both cell types. In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation. Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum. Histological analyses and measurements of pellet area demonstrated that even low concentrations of platelet-rich plasma releasate were enough to prevent nasoseptal chondrogenic cells from losing their chondrogenic potential due to in vitro expansion thereby promoting their recommitment. Low concentration of platelet-rich plasma releasate supplemented in chondrogenic medium also increased the chondrogenic potential of mesenchymal stromal cells seeded on collagen-hyaluronic acid scaffolds, as observed by an increase in chondrogenic-related gene expression, sulfated glycosaminoglycan production, and compressive modulus following in vitro culture. On the contrary, higher concentration of platelet-rich plasma releasate (10%) hampered some of these features. In conclusion, platelet-rich plasma releasate was able to prevent cellular chondrogenic capacity loss, inducing regain of their phenotype, and modulate cell commitment. Our data support the hypothesis of platelet-rich plasma chondrogenic potential, allowing fetal bovine serum substitution for platelet-rich plasma releasate at specific concentrations in culture medium when chondrogenic commitment is desired on specific cell types and moments of culture.

No MeSH data available.


Related in: MedlinePlus

Quantification of NCCs pellet areas. Areas (mm2) of (a) passage 3 pellets cultured for 21 days, (b) passage 3 pellets cultured for 35 days, (c) passage 5 pellets cultured for 21 days, (d) and passage 5 pellets cultured for 35 days, in medium without (w/o) or with (w/) TGF-β. Asterisks represent statistical significance compared to 10% FBS pre-treated pellets (p < 0.05).NCCs: nasoseptal chondrogenic cells; FBS: fetal bovine serum.
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fig4-2041731415594127: Quantification of NCCs pellet areas. Areas (mm2) of (a) passage 3 pellets cultured for 21 days, (b) passage 3 pellets cultured for 35 days, (c) passage 5 pellets cultured for 21 days, (d) and passage 5 pellets cultured for 35 days, in medium without (w/o) or with (w/) TGF-β. Asterisks represent statistical significance compared to 10% FBS pre-treated pellets (p < 0.05).NCCs: nasoseptal chondrogenic cells; FBS: fetal bovine serum.

Mentions: Hereafter, NCCs were cultured in two-dimensions (2D) in 2.5% PRPr, 10% PRPr, and 10% FBS in passage 2 and then transferred to 3D pellet culture in passage 3 with or without TGF-β, to evaluate cartilage-like formation (Figure 2). Similar cartilage-like pellets were formed in all three conditions. However, it can be noticed that, after 21 days of culture, 2.5% PRPr treatment resulted in pellets that best resemble a hyaline cartilage–like structure, with intense ECM staining and more cells with chondrocyte-like morphology. Although no statistical significance was observed, PRPr-treated pellets, in special 10% PRPr, were also larger than 10% FBS–treated pellets (Figure 4(a)). Only after 35 days, 10% FBS treatment resulted in pellets with intense ECM staining, high number of cells with chondrocyte morphology and as large as PRPr-treated ones (Figures 2 and 4(b)).


Platelet-rich plasma releasate differently stimulates cellular commitment toward the chondrogenic lineage according to concentration.

do Amaral RJ, Matsiko A, Tomazette MR, Rocha WK, Cordeiro-Spinetti E, Levingstone TJ, Farina M, O'Brien FJ, El-Cheikh MC, Balduino A - J Tissue Eng (2015)

Quantification of NCCs pellet areas. Areas (mm2) of (a) passage 3 pellets cultured for 21 days, (b) passage 3 pellets cultured for 35 days, (c) passage 5 pellets cultured for 21 days, (d) and passage 5 pellets cultured for 35 days, in medium without (w/o) or with (w/) TGF-β. Asterisks represent statistical significance compared to 10% FBS pre-treated pellets (p < 0.05).NCCs: nasoseptal chondrogenic cells; FBS: fetal bovine serum.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4555349&req=5

fig4-2041731415594127: Quantification of NCCs pellet areas. Areas (mm2) of (a) passage 3 pellets cultured for 21 days, (b) passage 3 pellets cultured for 35 days, (c) passage 5 pellets cultured for 21 days, (d) and passage 5 pellets cultured for 35 days, in medium without (w/o) or with (w/) TGF-β. Asterisks represent statistical significance compared to 10% FBS pre-treated pellets (p < 0.05).NCCs: nasoseptal chondrogenic cells; FBS: fetal bovine serum.
Mentions: Hereafter, NCCs were cultured in two-dimensions (2D) in 2.5% PRPr, 10% PRPr, and 10% FBS in passage 2 and then transferred to 3D pellet culture in passage 3 with or without TGF-β, to evaluate cartilage-like formation (Figure 2). Similar cartilage-like pellets were formed in all three conditions. However, it can be noticed that, after 21 days of culture, 2.5% PRPr treatment resulted in pellets that best resemble a hyaline cartilage–like structure, with intense ECM staining and more cells with chondrocyte-like morphology. Although no statistical significance was observed, PRPr-treated pellets, in special 10% PRPr, were also larger than 10% FBS–treated pellets (Figure 4(a)). Only after 35 days, 10% FBS treatment resulted in pellets with intense ECM staining, high number of cells with chondrocyte morphology and as large as PRPr-treated ones (Figures 2 and 4(b)).

Bottom Line: In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation.Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum.Our data support the hypothesis of platelet-rich plasma chondrogenic potential, allowing fetal bovine serum substitution for platelet-rich plasma releasate at specific concentrations in culture medium when chondrogenic commitment is desired on specific cell types and moments of culture.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brasil ; Excellion Serviços Biomédicos, Amil/UnitedHealth Group, Petrópolis, Brasil.

ABSTRACT
Platelet-rich plasma has been used to treat articular cartilage defects, with the expectations of anabolic and anti-inflammatory effects. However, its role on cellular chondrogenic or fibrogenic commitment is still a controversy. Herein, the role of platelet-rich plasma releasate, the product obtained following platelet-rich plasma activation, on cellular commitment toward the chondrogenic lineage was evaluated in vitro. Human nasoseptal chondrogenic cells and human bone marrow mesenchymal stromal cells were used as cell types already committed to the chondrogenic lineage and undifferentiated cells, respectively, as different concentrations of platelet-rich plasma releasate were tested in comparison to commonly used fetal bovine serum. Low concentration of platelet-rich plasma releasate (2.5%) presented similar effects on cellular growth compared to 10% fetal bovine serum, for both cell types. In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation. Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum. Histological analyses and measurements of pellet area demonstrated that even low concentrations of platelet-rich plasma releasate were enough to prevent nasoseptal chondrogenic cells from losing their chondrogenic potential due to in vitro expansion thereby promoting their recommitment. Low concentration of platelet-rich plasma releasate supplemented in chondrogenic medium also increased the chondrogenic potential of mesenchymal stromal cells seeded on collagen-hyaluronic acid scaffolds, as observed by an increase in chondrogenic-related gene expression, sulfated glycosaminoglycan production, and compressive modulus following in vitro culture. On the contrary, higher concentration of platelet-rich plasma releasate (10%) hampered some of these features. In conclusion, platelet-rich plasma releasate was able to prevent cellular chondrogenic capacity loss, inducing regain of their phenotype, and modulate cell commitment. Our data support the hypothesis of platelet-rich plasma chondrogenic potential, allowing fetal bovine serum substitution for platelet-rich plasma releasate at specific concentrations in culture medium when chondrogenic commitment is desired on specific cell types and moments of culture.

No MeSH data available.


Related in: MedlinePlus