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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

Biochemical and biomechanical effects of PRP on CHyA seeded scaffolds following 14 days of culture. sGAG synthesis was evaluated by biochemical analysis and the values were normalized by DNA content. Cultures were supplemented with 2.5% PRPr, 10% PRPr, chondrogenic medium with 2.5% PRPr (CM + 2.5% PRPr), chondrogenic medium with 10% PRPr (CM + 10% PRPr), chondrogenic medium with 10% FBS in cells expanded in 2.5% PRPr (CM + 10% FBS_PRP exp), and chondrogenic medium with 10% FBS in cells expanded in 10% FBS (CM + 10% FBS_FBS exp), which was considered the control condition, by which all results were normalized. (a) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). Compressive modulus of CHyA seeded scaffolds. (b) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). The data are presented as a mean and the error bars correspond to standard deviation (SD).PRP: platelet-rich plasma; CHyA: collagen-hyaluronic acid; sGAG: sulfated glycosaminoglycan; PRPr: platelet-rich plasma releasate; CM: chondrogenic medium; FBS: fetal bovine serum.
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fig6-2041731415594127: Biochemical and biomechanical effects of PRP on CHyA seeded scaffolds following 14 days of culture. sGAG synthesis was evaluated by biochemical analysis and the values were normalized by DNA content. Cultures were supplemented with 2.5% PRPr, 10% PRPr, chondrogenic medium with 2.5% PRPr (CM + 2.5% PRPr), chondrogenic medium with 10% PRPr (CM + 10% PRPr), chondrogenic medium with 10% FBS in cells expanded in 2.5% PRPr (CM + 10% FBS_PRP exp), and chondrogenic medium with 10% FBS in cells expanded in 10% FBS (CM + 10% FBS_FBS exp), which was considered the control condition, by which all results were normalized. (a) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). Compressive modulus of CHyA seeded scaffolds. (b) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). The data are presented as a mean and the error bars correspond to standard deviation (SD).PRP: platelet-rich plasma; CHyA: collagen-hyaluronic acid; sGAG: sulfated glycosaminoglycan; PRPr: platelet-rich plasma releasate; CM: chondrogenic medium; FBS: fetal bovine serum.

Mentions: To verify whether higher chondrogenic gene expression on PRPr groups resulted in higher biochemical and biomechanical improvement, sGAG production and compressive modulus evaluation were performed. Biochemical analyses evidenced that higher sGAG production was observed in the CM supplemented with 2.5% PRPr. On the other hand, other conditions showed lower amount of sGAG production than the control group (Figure 6(a)). The groups in which PRPr replaced FBS in the CM presented higher mechanical properties than those maintained in FBS. In contrast, PRPr alone, that is, without the addition of CM, was not enough to induce higher levels of compressive modulus, with values remaining even lower than in CM groups supplemented with 10% FBS (Figure 6(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)

Biochemical and biomechanical effects of PRP on CHyA seeded scaffolds following 14 days of culture. sGAG synthesis was evaluated by biochemical analysis and the values were normalized by DNA content. Cultures were supplemented with 2.5% PRPr, 10% PRPr, chondrogenic medium with 2.5% PRPr (CM + 2.5% PRPr), chondrogenic medium with 10% PRPr (CM + 10% PRPr), chondrogenic medium with 10% FBS in cells expanded in 2.5% PRPr (CM + 10% FBS_PRP exp), and chondrogenic medium with 10% FBS in cells expanded in 10% FBS (CM + 10% FBS_FBS exp), which was considered the control condition, by which all results were normalized. (a) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). Compressive modulus of CHyA seeded scaffolds. (b) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). The data are presented as a mean and the error bars correspond to standard deviation (SD).PRP: platelet-rich plasma; CHyA: collagen-hyaluronic acid; sGAG: sulfated glycosaminoglycan; PRPr: platelet-rich plasma releasate; CM: chondrogenic medium; FBS: fetal bovine serum.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

fig6-2041731415594127: Biochemical and biomechanical effects of PRP on CHyA seeded scaffolds following 14 days of culture. sGAG synthesis was evaluated by biochemical analysis and the values were normalized by DNA content. Cultures were supplemented with 2.5% PRPr, 10% PRPr, chondrogenic medium with 2.5% PRPr (CM + 2.5% PRPr), chondrogenic medium with 10% PRPr (CM + 10% PRPr), chondrogenic medium with 10% FBS in cells expanded in 2.5% PRPr (CM + 10% FBS_PRP exp), and chondrogenic medium with 10% FBS in cells expanded in 10% FBS (CM + 10% FBS_FBS exp), which was considered the control condition, by which all results were normalized. (a) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). Compressive modulus of CHyA seeded scaffolds. (b) Asterisks and cross represent p < 0.05 compared to 10% FBS, and similar symbols between groups represent statistic similarity among them (p > 0.05). The data are presented as a mean and the error bars correspond to standard deviation (SD).PRP: platelet-rich plasma; CHyA: collagen-hyaluronic acid; sGAG: sulfated glycosaminoglycan; PRPr: platelet-rich plasma releasate; CM: chondrogenic medium; FBS: fetal bovine serum.
Mentions: To verify whether higher chondrogenic gene expression on PRPr groups resulted in higher biochemical and biomechanical improvement, sGAG production and compressive modulus evaluation were performed. Biochemical analyses evidenced that higher sGAG production was observed in the CM supplemented with 2.5% PRPr. On the other hand, other conditions showed lower amount of sGAG production than the control group (Figure 6(a)). The groups in which PRPr replaced FBS in the CM presented higher mechanical properties than those maintained in FBS. In contrast, PRPr alone, that is, without the addition of CM, was not enough to induce higher levels of compressive modulus, with values remaining even lower than in CM groups supplemented with 10% FBS (Figure 6(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