Effect of platelet-rich plasma on bone regeneration in ovariectomized osteoporotic rats

Jong-Moon Cho; Jeong-Kyung Kang; Kyu-Won Suh; Jae-Jun Ryu

doi:10.4047/jkap.2010.48.1.16

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Cho, Kang, Suh, and Ryu: Effect of platelet-rich plasma on bone regeneration in ovariectomized osteoporotic rats

ORIGINAL ARTICLE

J Korean Acad Prosthodont 2010;48(1):16-27.

Published online: 18 December 2010

DOI: https://doi.org/10.4047/jkap.2010.48.1.16

Effect of platelet-rich plasma on bone regeneration in ovariectomized osteoporotic rats

Jong-Moon Cho, DDS, MSD¹, Jeong-Kyung Kang, DDS, MSD², Kyu-Won Suh, MSD, DDS, PhD², Jae-Jun Ryu, DDS, MSD, PhD^2,^∗

^¹Department of Dentistry, Graduate School of Medical Science

^²Graduate School of Clinical Dentistry, Korea University, Seoul, Korea

∗Corresponding Author: Jae-Jun Ryu Korea University Ansan Hospital, Gojan 1-dong, Danwon-Gu, Ansan, Kyunggi-Do, 425-707, Korea +82 31 412 5878: e-mail, koprosth@unitel.co.kr

Received 4 November 20099 December 2009 Accepted 4 January 2010

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

The aim of this experimental study is to observe the effect of platelet-rich plasma (PRP) on early bone regeneration of rats both in normal condition and in osteoporosis induced by ovariectomy.

Material and methods

Total 40 Sprague-Dawley female rats were divided into 4 groups. A 8-mm-diameter calvarial critical-sized defect (CSD) was made by drilling with trephine at the center of calvaria in cranium of every rat. Every CSD was augmented with an osteoconductive synthetic alloplastic substitute (MBCP^TM) and PRP as follows. Group A; 10 non-ovariectomized rats grafted with only MBCP^TM. Group B; 10 non-ovariectomized rats grafted with MBCP^TM and PRP. Group C; 10 ovariectomized rats grafted with only MBCP^TM. Group D; 10 ovariectomized rats grafted with MBCP^TM and PRP. At 4 weeks after graft, every rat was sacrificed. And histomorphometric analysis was performed by measuring calcified area of new bone formation within the CSD.

Results

Comparing four groups, results were obtained as follows. 1. In non-ovariectomized groups, PRP showed a positive effect somewhat but not significant (P > .05). 2. In ovariectomized groups, PRP showed a positive effect significantly (P < .05). 3. In PRP untreated groups, ovariectomy diminished bone regeneration significantly (P < .05). 4. In PRP treated groups, ovariectomy diminished bone regeneration somewhat but not significant (P > .05).

Conclusion

Within the limitation of this study, it can be concluded that PRP in combination with an osteoconductive synthetic alloplastic substitute has an effect on bone regeneration more significantly in ovariectomized osteoporotic rats than in normal rats. (J Korean Acad Prosthodont 2010;48:16-27)

Keywords: Bone regeneration, Critical-sized defect (CSD), Osteoporosis, Ovariectomy, Platelet-rich plasma (PRP)

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Fig. 1.

Surgical procedure of ovariec-Etomy (OVX). A, Detecting the small flower-like shaped ovary and taking it out after abdominal incision; B, Tight-suturing gently at the blood vessel to ovary; C, Excision of ovary and around tissue with scissors at the point between ovary and tight-sutured point; D, OVX accomplished in both right and left side; E, Surgically removed ovary and around fatty tissue.

Fig. 2.

Surgical procedure of bone graft. A, Trephine drilling under saline irrigation; B, Careful removal of 8-mm-diameter calvarial bone; C, A prepared calvarial critical-sized defect (CSD); D, A CSD filled with synthetic alloplastic substitute.

Fig. 3.

Whole blood sample 5 ml in A and centrifuging separator (PLACON^TM: Platelet Concentrator, Oscotec Inc., Cheonan, Korea) in B.

Fig. 4.

Bone sampling after sacrifice. Cutting carefully the calvarial bone more widely than outline (arrowheads) of a critical-sized defect.

Fig. 5.

Measuring the calcified area of new bone formation. Yellow lines represent outlines of newly calcified bone after bone graft which was stained in reddish color between the non-structural synthetic alloplastic substitute (MBCP^TM) particles. Green words represent serial numbers of yellow lines.

Fig. 6.

Comparison of serum alkaline phosphatase (ALP) level between normal and ovariectomized groups (OVX). Thick solid lines indicate standard deviation. ∗∗ means significant difference at P < .05 statistically.

Fig. 7.

Light-microscopic view. A, Calcified area of new bone (arrowheads) mainly around outlines of synthetic alloplastic substitute particles and host calvarial bone (asterisk) at 100× magnification; B, Osteoblasts (arrowheads) lining in a row at 200× magnification; C, Multinucleated cells (arrowheads) on the surface of a synthetic alloplastic substitute particle and newly forming blood vessel (asterisk) at 400× magnification.

Fig. 8.

Light-microscopic view of calcified area of new bone formation in each group at 100× magnification. In group C, calcified area which was stained in reddish color is relatively smaller than that of other three groups.

Fig. 9.

Comparison of new bone formation area. OVX; Ovariectomized group, PRP; PRP treated group. Thick solid lines indicate standard deviation. ∗∗means significant difference at P < .05 statistically.

Fig. 10.

Comparison of new bone formation area between group C and group D. In ovariectomized (osteoporotic) groups (OVX), platelet-rich plasma showed a positive effect on bone regeneration significantly. PRP; PRP treated group. Thick solid lines indicate standard deviation. ∗∗means significant difference at P < .05 statistically.

Fig. 11.

Comparison of new bone formation area between group B and group D. In PRP treated groups (PRP), ovariectomy diminished bone regeneration somewhat. However, statistically it is not significant (P > .05). OVX; Ovariectomized group. Thick solid lines indicate standard deviation.

Table I.

Experimental groups according to OVX and PRP treatments

Group	OVX	PRP	Graft material	N
A	No	No	MBCP^TM	10
B	No	Yes	MBCP^TM	10
C	Yes	No	MBCP^TM	10
D	Yes	Yes	MBCP^TM	10

Rats in group C and D were ovariectomized. In combination with bone graft material, PRP was treated in group B and D. OVX, Ovariectomy; PRP, Platelet-rich plasma; MBCP^TM , Micro- & macro-porous biphasic calcium phosphate; N, Number of specimen.

Table II.

Results of serum ALP level (IU/L)

Group	N	Mean (IU/L)	SD (IU/L)
A & B (non-OVX groups)	20	50.45	13.65
C & D (OVX groups)	20	78.95	13.64

Mean value of OVX groups was higher than that of non - OVX groups. ALP; Alkaline phosphatase. OVX; Ovariectomized. N; Number of specimen. SD; Standard deviation.

Table III.

Results of measuring calcified area of new bone formation (μ m²)

Group	OVX	PRP	Mean (μ m²)	SD (μ m²)
A	non - OVX (normal)	No	49335.85	20739.41
B	non - OVX (normal)	Yes	66919.09	30418.69
C	OVX (osteporosis)	No	13824.49	8298.45
D	OVX (osteporosis)	Yes	49988.67	24042.91

Mean value of group C was lower than those of other three groups. Mean value of group D (rats which were PRP treated after OVX) was as high as that of group A (rats which were normal and PRP untreated). OVX; Ovariectomized, PRP; Platelet-rich plasma, SD; Standard deviation.

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