Yonsei Med J.  2017 Nov;58(6):1195-1203. 10.3349/ymj.2017.58.6.1195.

Effects of Autologous Platelet-Rich Plasma on Regeneration of Damaged Endometrium in Female Rats

• Jang HY
• Myoung SM
• Choe JM
• Kim T
• Cheon YP
• Kim YM
• Park H

• KMID: 2418902
• DOI: http://doi.org/10.3349/ymj.2017.58.6.1195

Abstract

PURPOSE
To investigate whether autologous platelet-rich plasma (PRP) treatment can improve regeneration of the endometrium in an experimental model of ethanol-induced damage.
MATERIALS AND METHODS
Sixty female Sprague-Dawley rats were randomly assigned into three groups: control group, ethanol group, and PRP-treated group (administration of 0.25 mL of PRP into both uterine cavities 72 hours after ethanol injection). After 15 days of endometrial damage, all the animals were sacrificed during the estrous cycle, and samples were taken from the mid-uterine horn. Functional and structural recovery of the endometrium was analyzed by hematoxylin-eosin (H&E) and Masson trichrome (MT) staining, real-time polymerase chain reaction (PCR) assay, and immuno-histochemical (IHC) analyses.
RESULTS
H&E and MT staining confirmed significantly decreased fibrosis and increased cellular proliferation in the PRP-treated group, compared to the ethanol group. The endometrial areas in the ethanol and PRP-treated groups were 212.83±15.84 µm² and 262.34±12.33 µm² (p=0.065). Significantly stronger IHC expression of cytokeratin, homeobox A10 (HOXA10), vascular endothelial growth factor (VEGF), and Ki-67 was found in the PRP-treated group, compared to the ethanol group. In real-time PCR analyses, interleukin-1β mRNA was down-regulated, while c-Kit mRNA was up-regulated, in the PRP-treated group, compared to the ethanol group.
CONCLUSION
Intrauterine administration of autologous PRP stimulated and accelerated regeneration of the endometrium and also decreased fibrosis in a murine model of damaged endometrium.

Keyword

Platelet-rich plasma (PRP); fibrosis; regeneration; thin endometrium

MeSH Terms

Animals
Cell Proliferation
Endometrium/*metabolism
Female
Interleukin-1beta
Platelet-Rich Plasma/*metabolism
Random Allocation
Rats
Rats, Sprague-Dawley
Real-Time Polymerase Chain Reaction
Regeneration/*physiology
Vascular Endothelial Growth Factor A/*metabolism
Interleukin-1beta
Vascular Endothelial Growth Factor A

Figure

• 

  Fig. 1 Outline of the experimental schedule. PRP, platelet-rich plasma; PMSG, pregnant mare serum gonadotropin.

• 

  Fig. 2 Comparison of the endometrial morphology, area, and extent of fibrosis. (A) On H&E staining, distinctive morphological changes in the endometrial characteristics were found in the glands, epithelial lining, and stroma between the ethanol group and the PRP-treated group. (B) On MT staining, the collagen-deposited fibrotic regions were shown in blue, and the extent of these regions was increased in the ethanol group compared to the other groups (left, control group; middle, ethanol group; right, PRP-treated group). (C) Comparison of the area of the endometrium. (D) Comparison of the extent of endometrial fibrosis (blue area on MT staining). Statistical comparisons were performed among the groups using the Kruskal-Wallis test and Mann-Whitney test. All images were captured at magnifications of ×40. H&E, hematoxylin-eosin; PRP, platelet-rich plasma; MT, Masson Trichrome.

• 

  Fig. 3 Expression of (A) CK, (B) HOXA10, (C) VEGF, and (D) Ki-67 with IHC staining (left, control group; middle, ethanol group; right, PRP-treated group). (E) Comparisons of the expression of CK, HOXA10, and VEGF. Quantitative assessment of five random sections of endometrial tissue slides are shown as four zones with total/high/medium/low positive intensity. (F) Quantitative analysis of nuclear markers for Ki-67 IHC staining. Statistical comparisons of groups are based on the chi-square (χ2) tests, with statistical significance defined as p<0.05. *Significant when compared to the control, †Significant when compared between the ethanol group and the PRP-treated group. All images were captured at magnifications of ×400. CK, cytokeration; HOXA10, homeobox A10; VEGF, vascular endothelial growth factor; IHC, immuno-histochemical; PRP, platelet-rich plasma; DAB, diaminobenzidene tetrahydrochloride.

• 

  Fig. 4 The quantitative fold changes in the expression of the target gene (HOXA10, VEGF-A, c-Kit, Oct-4, IL-1β, IL-10, and NF-κB) relative to the internal control gene (β-actin) were analyzed using real-time PCR. (A) Significantly increased expression of HOXA10 in the PRP-treated group compared to the ethanol group, but no significant change in the expression of VEGF-A. (B) Significantly increased expression of the c-Kit gene in the PRP-treated group compared to the ethanol group, but no significant change in the expression of the Oct-4 gene. (C) Significantly increased expression of IL-1β in the ethanol group when compared to the other groups, but no significant change in the expressions of IL-10 and NF-κB on comparison of the ethanol group and the PRP-treated group. Statistical comparisons of groups are based on the Mann-Whitney test, with statistical significance defined as p<0.05. *Significant when compared to the control, †Significant when compared between the ethanol group and the PRP-treated group. HOXA10, homeobox A10; VEGF-A, vascular endothelial growth factor-A; Oct-4, octamer-binding transcription factor 4; IL, interleukin; NF-κB, nuclear factor-κB; PCR, polymerase chain reaction; PRP, platelet-rich plasma.

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