J Periodontal Implant Sci.  2012 Aug;42(4):136-143. 10.5051/jpis.2012.42.4.136.

Novel analysis model for implant osseointegration using ectopic bone formation via the recombinant human bone morphogenetic protein-2/macroporous biphasic calcium phosphate block system in rats: a proof-of-concept study

• Park JC
• Lee JB
• Daculsi G
• Oh SY
• Cho KS
• Im GI
• Kim BS
• Kim CS

• KMID: 1783649
• DOI: http://doi.org/10.5051/jpis.2012.42.4.136

Abstract

PURPOSE
The osseointegration around titanium mini-implants installed in macroporous biphasic calcium phosphate (MBCP) blocks was evaluated after incubation with recombinant human bone morphogenetic protein-2 (rhBMP-2) in an ectopic subcutaneous rat model.
METHODS
Mini-implants (phi1.8x12 mm) were installed in MBCP blocks (bMBCPs, 4x5x15 mm) loaded with rhBMP-2 at 0.1 mg/mL, and then implanted for 8 weeks into subcutaneous pockets of male Sprague-Dawley rats (n=10). A histomorphometric analysis was performed, and the bone-to-implant contact (BIC) and bone density were evaluated.
RESULTS
Significant osteoinductive activity was induced in the rhBMP-2/bMBCP group. The percentage of BIC was 41.23+/-4.13% (mean+/-standard deviation), while bone density was 33.47+/-5.73%. In contrast, no bone formation was observed in the bMBCP only group.
CONCLUSIONS
This model represents a more standardized tool for analyzing osseointegration and bone healing along the implant surface and in bMBCPs that excludes various healing factors derived from selected animals and defect models.

Keyword

Animals; Dental implants; Osseointegration

MeSH Terms

Animals
Bone Density
Bone Morphogenetic Protein 2
Calcium
Dental Implants
Humans
Hydroxyapatites
Male
Nitrogen Mustard Compounds
Osseointegration
Osteogenesis
Rats
Rats, Sprague-Dawley
Recombinant Proteins
Titanium
Transforming Growth Factor beta
Bone Morphogenetic Protein 2
Calcium
Dental Implants
Hydroxyapatites
Nitrogen Mustard Compounds
Recombinant Proteins
Titanium
Transforming Growth Factor beta

Figure

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  Figure 1 Kinetics of recombinant human bone morphogenetic protein-2 (rhBMP-2) release from macroporous biphasic calcium phosphate block observed in vitro. Sustained release of rhBMP-2 was observed for up to 2 weeks.

• 

  Figure 2 Clinical photograph of a macroporous biphasic calcium phosphate block (bMBCP) and titanium mini-implant installation. One vertical hole and three horizontal holes are present in the bMBCP. The titanium mini-implant was installed manually.

• 

  Figure 3 Clinical photograph of a macroporous biphasic calcium phosphate block (bMBCP) retrieved after necropsy. (A) High-density connective tissue formation was observed with infiltration of microvessels in the test group (recombinant human bone morphogenetic protein-2/bMBCP). (B) Loose connective tissue, which was easily wiped off, dominantly formed over the bMBCP in the control group (phosphate buffer solution/bMBCP).

• 

  Figure 4 Histological overview of the retrieved titanium mini-implants and recombinant human bone morphogenetic protein-2/macroporous biphasic calcium phosphate blocks (rhBMP-2/bMBCPs) or phosphate buffer solution/bMBCPs (H&E staining). (A, B) The rhBMP-2-treated test group presenting significant new bone formation along the implant surface and within the bMBCP (A, ×40; B, ×200). (C) The newly formed bone exhibited a lamellated pattern under polarized-light microscopy, suggesting the occurrence of waves of differentiated bone formation over time (×200). (D, E) The untreated control group demonstrated no mineralized tissue formation along the implant surface or within the bMBCP (D, ×40; E, ×200). (F) No mineralized tissue was observed under polarized-light microscopy in the control tissue (×200). Ti: titanium mini-implant, M: bMBCP, NB: new bone.

• 

  Figure 5 (A) Scanning electron microscopy (SEM) images of the test group (recombinant human bone morphogenetic protein-2/macroporous biphasic calcium phosphate blocks [bMBCPs]) 8 weeks after subcutaneous implantation in rats (×50). A higher magnification of the boxed area is shown in B. New bone formation is evident along the implant surface. The bMBCP is revealed as dark-gray areas, while MBCP and the titanium mini-implant appear as light-gray and white, respectively. (B) A higher magnification view of the boxed area clearly shows newly formed bone tissue (×100). The yellow arrowheads indicate the obvious formation of NB. (C) The SEM images of the control group (phosphate buffer solution/bMBCP) (×50). (D) Polarized-light microscopy showing the absence of mineralized tissue (×100). Ti: titanium mini-implant, M: bMBCP, NB: new bone.

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