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. 2020 Jul 31;13(15):3393.
doi: 10.3390/ma13153393.

Native Bovine Hydroxyapatite Powder, Demineralised Bone Matrix Powder, and Purified Bone Collagen Membranes Are Efficient in Repair of Critical-Sized Rat Calvarial Defects

Alexey Veremeev  1   2 , Roman Bolgarin  1 , Vladimir Nesterenko  2 , Alexander Andreev-Andrievskiy  3 , Anton Kutikhin  4
Affiliations

Native Bovine Hydroxyapatite Powder, Demineralised Bone Matrix Powder, and Purified Bone Collagen Membranes Are Efficient in Repair of Critical-Sized Rat Calvarial Defects

Alexey Veremeev et al. Materials (Basel). .

Abstract

Here we evaluated the efficacy of bone repair using various native bovine biomaterials (refined hydroxyapatite (HA), demineralised bone matrix (DBM), and purified bone collagen (COLL)) as compared with commercially available bone mineral and bone autografts. We employed a conventional critical-sized (8 mm diameter) rat calvarial defect model (6-month-old male Sprague-Dawley rats, n = 72 in total). The artificial defect was repaired using HA, DBM, COLL, commercially available bone mineral powder, bone calvarial autograft, or remained unfilled (n = 12 animals per group). Rats were euthanised 4 or 12 weeks postimplantation (n = 6 per time point) with the subsequent examination to assess the extent, volume, area, and mineral density of the repaired tissue by means of microcomputed tomography and hematoxylin and eosin staining. Bovine HA and DBM powder exhibited excellent repair capability similar to the autografts and commercially available bone mineral powder while COLL showed higher bone repair rate. We suggest that HA and DBM powder obtained from bovine bone tissue can be equally applied for the repair of bone defects and demonstrate sufficient potential to be implemented into clinical studies.

Keywords: bone autografts; bone collagen; bone defects; bone repair; clinical translation; critical-sized rat calvarial defect; demineralised bone matrix; hydroxyapatite; microcomputed tomography; xenogeneic implants.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, and in the decision to publish the results.

Figures

Figure 1
Figure 1
Representative microcomputed tomography images demonstrating filling of the critical-sized (8 mm diameter) rat calvarial defect upon the implantation of bovine-derived HA, DBM, and COLL at the baseline (A) and in a (B) short- and (C) long-term. Unfilled defect, autograft, and Geistlich Bio-Oss® represent a negative control, positive control, and reference product, respectively. HA—refined hydroxyapatite powder, DBM—demineralised bone matrix powder, COLL—purified bone collagen membranes.
Figure 2
Figure 2
Representative hematoxylin and eosin-stained sections of rat calvarial bone tissues demonstrating filling of the critical-sized (8 mm diameter) rat calvarial defect upon the implantation of bovine-derived HA, DBM, and COLL in a (A) short- and (B) long-term. Close-ups (red squares demarcating the inserts) show areas with ongoing repair. Unfilled defect, autograft, and Geistlich Bio-Oss® represent a negative control, positive control, and reference product, respectively. HA—refined hydroxyapatite powder, DBM—demineralised bone matrix powder, COLL—purified bone collagen membranes.
Figure 3
Figure 3
Evaluation of bone repair features upon the implantation of bovine-derived HA, DBM, and COLL into a critical-sized (8 mm diameter) rat calvarial defect in a short- and long-term. Microcomputed tomography measurements of (A) bone tissue volume; (B) bone tissue area; (C) tissue mineral density; and (D) bone mineral density. Unfilled defect, autograft, and Geistlich Bio-Oss® represent a negative control, positive control, and reference product, respectively. N = 12 animals per group, one-way analysis of variance with Tukey’s multiple comparisons test. UD—unfilled defect, AU—autograft, Ref—reference product (Geistlich Bio-Oss®), HA—refined hydroxyapatite powder, DBM—demineralised bone matrix powder, COLL—purified bone collagen membranes. $ means statistically significant differences (p ≤ 0.05) compared with UD, # means statistically significant differences compared with AU; % means statistically significant differences compared with Ref; ^ means statistically significant differences compared with HA.
Figure 4
Figure 4
Representative images demonstrating the implantation of HA into the critical-sized (8 mm diameter) rat calvarial defect. (A) Access to the calvarial bones; (B) excision of the calvarial bones using a trephine; (C) calvarial bones ready to be excised; (D) calvarial defect upon the excision of the respective bone tissue; (E) filling of the calvarial defect with HA; and (F) closure of the periosteum over the implant. HA—hydroxyapatite.
See this image and copyright information in PMC

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