Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jan 26, 2020; 12(1): 55-69
Published online Jan 26, 2020. doi: 10.4252/wjsc.v12.i1.55
Early therapeutic effect of platelet-rich fibrin combined with allogeneic bone marrow-derived stem cells on rats' critical-sized mandibular defects
Muhammad A Awadeen, Fouad A Al-Belasy, Laila E Ameen, Mohamad E Helal, Mohammed E Grawish
Muhammad A Awadeen, Mohammed E Grawish, Department of Oral Biology, Faculty of Oral and Dental Medicine, Delta University for Science and Technology, Mansoura 11152, Egypt
Fouad A Al-Belasy, Department of Oral Surgery and Anesthesia, Faculty of Oral and Dental Medicine, Delta University for Science and Technology, Mansoura 11152, Egypt
Laila E Ameen, Mohamad E Helal, Mohammed E Grawish, Department of Oral Biology, Faculty of Dentistry, Mansoura University, Mansoura 35516, Egypt
Author contributions: Awadeen MA generated the figures, tables and wrote the manuscript; Al-Belasy FA, Grawish ME, Ameen LE and Helal ME provided an intellectual contribution to the content, contributed to writing of the manuscript, and made all critical revisions; all authors read, reviewed and approved the final version of the manuscript.
Institutional animal care and use committee statement: All experimental procedures were performed under a protocol approved by the Ethics Committee of the Faculty of Dentistry, Mansoura University, Egypt. The rats were cared for and housed in standard cages by the guidelines of the Faculty of Medicine, Medical Research Centre, Mansoura University, Egypt.
Conflict-of-interest statement: Authors declare no conflict of interests for this article.
Data sharing statement: The datasets supporting the conclusions of this article are included in the article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Mohammed E Grawish, BDS, MSc, PhD, Professor, Department of Oral Biology, Faculty of Dentistry, Mansoura University, Dakahlia Governorate, Mansoura 35516, Egypt. grawish2005@yahoo.com
Received: March 7, 2019
Peer-review started: March 8, 2019
First decision: April 16, 2019
Revised: May 13, 2019
Accepted: November 29, 2019
Article in press: November 29, 2019
Published online: January 26, 2020
Abstract
BACKGROUND

Critically sized bone defects represent a significant challenge to orthopaedic surgeons worldwide. These defects generally result from severe trauma or resection of a whole large tumour. Autologous bone grafts are the current gold standard for the reconstruction of such defects. However, due to increased patient morbidity and the need for a second operative site, other lines of treatment should be introduced. To find alternative unconventional therapies to manage such defects, bone tissue engineering using a combination of suitable bioactive factors, cells, and biocompatible scaffolds offers a promising new approach for bone regeneration.

AIM

To evaluate the healing capacity of platelet-rich fibrin (PRF) membranes seeded with allogeneic mesenchymal bone marrow-derived stem cells (BMSCs) on critically sized mandibular defects in a rat model.

METHODS

Sixty-three Sprague Dawley rats were subjected to bilateral bone defects of critical size in the mandibles created by a 5-mm diameter trephine bur. Rats were allocated to three equal groups of 21 rats each. Group I bone defects were irrigated with normal saline and designed as negative controls. Defects of group II were grafted with PRF membranes and served as positive controls, while defects of group III were grafted with PRF membranes seeded with allogeneic BMSCs. Seven rats from each group were killed at 1, 2 and 4 wk. The mandibles were dissected and prepared for routine haematoxylin and eosin (HE) staining, Masson's trichrome staining and CD68 immunohistochemical staining.

RESULTS

Four weeks postoperatively, the percentage area of newly formed bone was significantly higher in group III (0.88 ± 0.02) than in groups I (0.02 ± 0.00) and II (0.60 ± 0.02). The amount of granulation tissue formation was lower in group III (0.12 ± 0.02) than in groups I (0.20 ± 0.02) and II (0.40 ± 0.02). The number of inflammatory cells was lower in group III (0.29 ± 0.03) than in groups I (4.82 ± 0.08) and II (3.09 ± 0.07).

CONCLUSION

Bone regenerative quality of critically sized mandibular bone defects in rats was better promoted by PRF membranes seeded with BMSCs than with PRF membranes alone.

Keywords: Platelet-rich fibrin membrane, Bone marrow-derived stem cells, Critical-sized mandibular defects, Rats, Histological and immunohistochemical staining

Core tip: Our findings are derived from a rat model for treating critical-sized mandibular bone defects. Defects were grafted with platelet-rich fibrin (PRF) membranes seeded with allogeneic bone marrow-derived stem cells (BMSCs). Our findings confirm the in vivo anti-inflammatory effects of allogenic BMSCs. In addition, BMSCs seeded on the PRF membranes exhibited beneficial syngeneic effects in promoting and accelerating the healing of critically sized mandibular defects. Routine and specific histological and immunohistochemical staining demonstrated for the first time that experimentally treated critically sized mandibular defects with PRF membrane and BMSC combined therapy increased the amount and the rate of the newly formed bone and decreased the amount of granulation tissue with a reduction in the number of inflammatory cell infiltrates.