Ann Dermatol.  2011 Nov;23(4):424-431.

Can Platelet-rich Plasma Be Used for Skin Rejuvenation? Evaluation of Effects of Platelet-rich Plasma on Human Dermal Fibroblast

• Kim DH
• Je YJ
• Kim CD
• Lee YH
• Seo YJ
• Lee JH
• Lee Y

Abstract

BACKGROUND
Autologous platelet-rich plasma has attracted attention in various medical fields recently, including orthopedic, plastic, and dental surgeries and dermatology for its wound healing ability. Further, it has been used clinically in mesotherapy for skin rejuvenation.
OBJECTIVE
In this study, the effects of activated platelet-rich plasma (aPRP) and activated platelet-poor plasma (aPPP) have been investigated on the remodelling of the extracellular matrix, a process that requires activation of dermal fibroblasts, which is essential for rejuvenation of aged skin.
METHODS
Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared using a double-spin method and then activated with thrombin and calcium chloride. The proliferative effects of aPRP and aPPP were measured by [3H]thymidine incorporation assay, and their effects on matrix protein synthesis were assessed by quantifying levels of procollagen type I carboxy-terminal peptide (PIP) by enzyme-linked immunosorbent assay (ELISA). The production of collagen and matrix metalloproteinases (MMP) was studied by Western blotting and reverse transcriptase-polymerase chain reaction.
RESULTS
Platelet numbers in PRP increased to 9.4-fold over baseline values. aPRP and aPPP both stimulated cell proliferation, with peak proliferation occurring in cells grown in 5% aPRP. Levels of PIP were highest in cells grown in the presence of 5% aPRP. Additionally, aPRP and aPPP increased the expression of type I collagen, MMP-1 protein, and mRNA in human dermal fibroblasts.
CONCLUSION
aPRP and aPPP promote tissue remodelling in aged skin and may be used as adjuvant treatment to lasers for skin rejuvenation in cosmetic dermatology.

Keyword

Fibroblast; Plasma; Platelet; Rejuvenation

MeSH Terms

Aged
Blood Platelets
Blotting, Western
Calcium Chloride
Cell Proliferation
Collagen
Collagen Type I
Dermatology
Enzyme-Linked Immunosorbent Assay
Extracellular Matrix
Fibroblasts
Humans
Matrix Metalloproteinases
Mesotherapy
Orthopedics
Plasma
Plastics
Platelet Count
Platelet-Rich Plasma
Rejuvenation
RNA, Messenger
Skin
Thrombin
Wound Healing
Calcium Chloride
Collagen
Collagen Type I
Matrix Metalloproteinases
Plastics
RNA, Messenger
Thrombin

Figure

• 

  Fig. 1 Platelet concentrations after double-spin method (n=5). (A) Microscopic examination of stained platelet smears (×40). (B) Quantified platelet counts indicate increased number of platelets in platelet-rich plasma. *p<0.05 vs. Control. nPPP: non-activated platelet-poor plasma, nPRP: non-activated platelet-rich plasma.

• 

  Fig. 2 aPPP and aPRP treatment increased the proliferation of cultured fibroblast. After serum-starvation for 24 h, fibroblasts were cultured with FBS, aPPP, or aPRP at concentrations as indicated. [3H]thymidine incorporation was measured at 3 and 5 days after the addition of various concentrations of aPRP and aPPP to the growth medium. (A) HDF proliferation on day 3 after control (serum-free medium), 5% FBS, aPPP, and aPRP additions. (B) HDF proliferation on day 5 after control (serum-free medium), 5% FBS, aPPP, and aPRP additions. *p<0.05 vs. control, †p<0.05 vs. 5% FBS (positive control). Con: control (serum-free medium), aPPP: activated platelet-poor plasma, aPRP: activated platelet-rich plasma, HDF: human dermal fibroblasts, FBS: fetal bovine serum.

• 

  Fig. 3 Procollagen type I carboxy-terminal peptide (PIP) production in aPRP and aPPP treated human dermal fibroblasts. After 24 h of serum-starvation, cells were cultured in serum-free (Con), 5% FBS or 1, 5, or 10% aPRP or aPPP treated culture medium for 48 h. PIP was quantified using an ELISA kit. *p <0.05 vs. control, Con: control (serum-free medium), aPPP: activated platelet-poor plasma, aPRP: activated platelet-rich plasma, FBS: fetal bovine serum.

• 

  Fig. 4 aPRP and aPPP treatment in culture medium increased the expression of collagen type I, alpha1 and collagen type I, alpha2 mRNA and protein expression in cultured human dermal fibroblasts. After 24 h of serum-starvation, cells were cultured in serum-free (Con), 5% FBS, 5% aPRP, or aPPP treated culture medium for 48 h. α1 chains of type I collagen mRNA and protein expression were determined by (A) RT-PCR, (B) western blotting, respectively. (C) α2 chains of type I collagen mRNA expression was determined by RT-PCR, (D) western blotting. *p<0.05 vs. control, †p<0.05 vs. 5% FBS (positive control). Con: control (serum-free medium), aPPP: activated platelet-poor plasma, aPRP: activated platelet-rich plasma, RT-PCR: reverse transcriptase-polymerase chain reaction, FBS: fetal bovine serum.

• 

  Fig. 5 aPRP and aPPP treatment in culture medium increased the expression of MMP-1 and MMP-3 protein expression in cultured HDF. (A) After 24 h of serum-starvation, cells were cultured in serum-free (Con), 5% FBS, 5% aPRP, or aPPP treated culture medium for 48 h. MMP-1 protein expression was detected by western blotting. (B) After 24 h of serum-starvation, cells were cultured in serum-free (Con), 5% FBS, 5% aPRP, or aPPP treated culture medium for 48 h. MMP-3 protein expression was detected by western blotting. *p<0.05 vs. control, †p<0.05 vs. 5% FBS (positive control). Con: control (serum-free medium), aPPP: activated platelet-poor plasma, aPRP: activated platelet-rich plasma, MMP: matrix metalloproteinases, HDF: human dermal fibroblasts, FBS: fetal bovine serum.

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