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Submitted: 2021-05-31
Accepted: 2021-07-09
Published online: 2021-08-03
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Evaluation of the effects of Ankaferd hemostat application on bone regeneration in rats with calvarial defects: histochemical, immunohistochemical and scintigraphic study

M. Turgut12, S. Karademir2, H. K. Başaoğlu23, C. Tomruk4, E. O. Cetin5, Y. Uyanikgil467, A. Cengiz8
DOI: 10.5603/FM.a2021.0074
·
Pubmed: 34355787
Affiliations
  1. Department of Neurosurgery, Aydın Adnan Menderes University Faculty of Medicine, 09010 Aydın,, Turkey
  2. Department of Histology and Embryology, Aydın Adnan Menderes University Health Sciences Institute, 09010 Aydın, Turkey
  3. Department of Histology and Embryology, Aydın Adnan Menderes University Faculty of Medicine, 09010 Efeler, Aydın, Turkey.
  4. Department of Histology and Embryology, Ege University Faculty of Medicine, 35100 Bornova, İzmir, Turkey.
  5. Department of Pharmaceutical Technology, Department of Biopharmaceutics and Pharmacokinetics, EgeUniversity Faculty of Pharmacy, 35100 İzmir, Turkey
  6. Department of Stem Cell, Ege University, Health Science Institue, 35100 Bornova, Izmir, Turkey.
  7. Cord Blood, Cell and Tissue Research and Application Centre, Ege University, 35100 Bornova, Izmir, Turkey
  8. Department of Nuclear Medicine, Aydın Adnan Menderes University Faculty of Medicine, 09010 Efeler, Aydın, Turkey.

open access

Ahead of Print
ORIGINAL ARTICLES
Submitted: 2021-05-31
Accepted: 2021-07-09
Published online: 2021-08-03

Abstract

Background: Bone wax, a hemostatic agent, is widely used in craniospinal surgical procedures for a long time, inspite of controversial results regarding its negative influence upon bone regeneration. In this experimental study, the effects of Ankaferd Blood Stopper (ABS), as an alternative hemostatic agent, were evaluated through histochemical, immunohistochemical and scintigraphic studies.

Materials and methods: The total of 30 adult female Wistar-Albino rats was randomly divided into three groups: intact control group (n=10), bone wax group (n=10), and ABS group (n=10). Surgically, a 3.0 mm hole in diameter was drilled on the right side of calvarium of the rats using a Class Mini Grinder set in all three groups, as described previously. At the end of 8 weeks, bone healing and connective tissue alterations surrounding drilled calvarial defect areas of the rats were determined via Hematoxylin and Eosin (H&E) and the Mallory's trichrome staining and anti-bone sialoprotein (BSP) immunohistochemistry. Image Pro Express 4.5 program was used for histomorphometric calculation of new bone and fibrotic tissue areas. All statistical analyzes were made with SPSS 25.0 and analysis of variance (one-way ANOVA) followed by Bonferroni post hoc test was performed, p<0.001 was considered as significance level.

Results: Histomorphometrically, it was found that he had the largest hole diameter and the least fibrotic scar area in the bone-wax group. In the bone wax group, it was observed that the material closed the hole and there was only a fibrotic scar tissue in the area between the bone tissue at the edge of the hole and bone wax, and a fibrotic tissue was formed in the bone wax area. During the histological procedure, this bone-wax material was poured and the sections were seen as a gap in this area. In the ABS hemostat group, the smallest hole diameter and the least fibrotic scar tissue were observed. Fibrotic scar tissue close to each other was found in the ABS hemostat and bone wax groups. Histological analysis of samples also showed a statistical significance for fibrotic connective tissue area between groups (p <0.05). Scintigraphically, osteoblastic activity related to blood flow in the animal taken from the group with application of ABS hemostat was more pronounced compared to the other two groups.

Conclusions: In our study, it has been concluded that the ABS yields affirmative effects on the bone healing, while bone wax leads to negative impact on the bone regeneration. Scintigraphic, histochemical and immunohistochemical data support the affirmative impact of the ABS hemostat application upon the bone regeneration apart from the quick stop of hemorrhage.

Abstract

Background: Bone wax, a hemostatic agent, is widely used in craniospinal surgical procedures for a long time, inspite of controversial results regarding its negative influence upon bone regeneration. In this experimental study, the effects of Ankaferd Blood Stopper (ABS), as an alternative hemostatic agent, were evaluated through histochemical, immunohistochemical and scintigraphic studies.

Materials and methods: The total of 30 adult female Wistar-Albino rats was randomly divided into three groups: intact control group (n=10), bone wax group (n=10), and ABS group (n=10). Surgically, a 3.0 mm hole in diameter was drilled on the right side of calvarium of the rats using a Class Mini Grinder set in all three groups, as described previously. At the end of 8 weeks, bone healing and connective tissue alterations surrounding drilled calvarial defect areas of the rats were determined via Hematoxylin and Eosin (H&E) and the Mallory's trichrome staining and anti-bone sialoprotein (BSP) immunohistochemistry. Image Pro Express 4.5 program was used for histomorphometric calculation of new bone and fibrotic tissue areas. All statistical analyzes were made with SPSS 25.0 and analysis of variance (one-way ANOVA) followed by Bonferroni post hoc test was performed, p<0.001 was considered as significance level.

Results: Histomorphometrically, it was found that he had the largest hole diameter and the least fibrotic scar area in the bone-wax group. In the bone wax group, it was observed that the material closed the hole and there was only a fibrotic scar tissue in the area between the bone tissue at the edge of the hole and bone wax, and a fibrotic tissue was formed in the bone wax area. During the histological procedure, this bone-wax material was poured and the sections were seen as a gap in this area. In the ABS hemostat group, the smallest hole diameter and the least fibrotic scar tissue were observed. Fibrotic scar tissue close to each other was found in the ABS hemostat and bone wax groups. Histological analysis of samples also showed a statistical significance for fibrotic connective tissue area between groups (p <0.05). Scintigraphically, osteoblastic activity related to blood flow in the animal taken from the group with application of ABS hemostat was more pronounced compared to the other two groups.

Conclusions: In our study, it has been concluded that the ABS yields affirmative effects on the bone healing, while bone wax leads to negative impact on the bone regeneration. Scintigraphic, histochemical and immunohistochemical data support the affirmative impact of the ABS hemostat application upon the bone regeneration apart from the quick stop of hemorrhage.

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Keywords

Ankaferd hemostat, bone wax, calvarium, osteogenesis, rat

About this article
Title

Evaluation of the effects of Ankaferd hemostat application on bone regeneration in rats with calvarial defects: histochemical, immunohistochemical and scintigraphic study

Journal

Folia Morphologica

Issue

Ahead of Print

Article type

Original article

Published online

2021-08-03

DOI

10.5603/FM.a2021.0074

Pubmed

34355787

Keywords

Ankaferd hemostat
bone wax
calvarium
osteogenesis
rat

Authors

M. Turgut
S. Karademir
H. K. Başaoğlu
C. Tomruk
E. O. Cetin
Y. Uyanikgil
A. Cengiz

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