J Periodontal Implant Sci.  2016 Jun;46(3):176-196. 10.5051/jpis.2016.46.3.176.

Evaluation of bone substitutes for treatment of peri-implant bone defects: biomechanical, histological, and immunohistochemical analyses in the rabbit tibia

• dos Santos PL
• de Molon R
• Queiroz TP
• Okamoto R
• de Souza Faloni AP
• Gulinelli
• Luvizuto E
• Garcia IR

• KMID: 2382982
• DOI: http://doi.org/10.5051/jpis.2016.46.3.176

Abstract

PURPOSE
We sought to evaluate the effectiveness of bone substitutes in circumferential peri-implant defects created in the rabbit tibia.
METHODS
Thirty rabbits received 45 implants in their left and right tibia. A circumferential bone defect (6.1 mm in diameter/4 mm depth) was created in each rabbit tibia using a trephine bur. A dental implant (4.1 mm × 8.5 mm) was installed after the creation of the defect, providing a 2-mm gap. The bone defect gaps between the implant and the bone were randomly filled according to the following groups: blood clot (CO), particulate Bio-Oss® (BI), and Bio-Oss® Collagen (BC). Ten animals were euthanized after periods of 15, 30, and 60 days. Biomechanical analysis by means of the removal torque of the implants, as well as histologic and immunohistochemical analyses for protein expression of osteocalcin (OC), Runx2, OPG, RANKL, and TRAP were evaluated.
RESULTS
For biomechanics, BC showed a better biological response (61.00±15.28 Ncm) than CO (31.60±14.38 Ncm) at 30 days. Immunohistochemical analysis showed significantly different OC expression in CO and BC at 15 days, and also between the CO and BI groups, and between the CO and BC groups at 60 days. After 15 days, Runx2 expression was significantly different in the BI group compared to the CO and BC groups. RANKL expression was significantly different in the BI and CO groups and between the BI and BC groups at 15 days, and also between the BI and CO groups at 60 days. OPG expression was significantly higher at 60 days postoperatively in the BI group than the CO group.
CONCLUSIONS
Collectively, our data indicate that, compared to CO and BI, BC offered better bone healing, which was characterized by greater RUNX2, OC, and OPG immunolabeling, and required greater reversal torque for implant removal. Indeed, along with BI, BC presents promising biomechanical and biological properties supporting its possible use in osteoconductive grafts for filling peri-implant gaps.

Keyword

Bone substitutes; Bone transplantation; Dental implantation; Osseointegration; Rabbits

MeSH Terms

Animals
Bone Substitutes*
Bone Transplantation
Collagen
Dental Implantation
Dental Implants
Osseointegration
Osteocalcin
Rabbits
Tibia*
Torque
Transplants
Bone Substitutes
Collagen
Dental Implants
Osteocalcin

Figure

• 

  Figure 1 Flowchart of the classification of the study groups.

• 

  Figure 2 Bone defects were filled with: I - blood clot (CO), II - Bio-Oss® (BI), and III - Bio-Oss® Collagen (BC).

• 

  Figure 3 Removal torque analysis in all groups and periods. a)Statistically significant between indicated groups, P<0.05.

• 

  Figure 4 Light micrographs of bone portions from implant threads in the CO (4A, 4D, and 4G), BI (4B, 4E, and 4H), and BC (4C, 4F, and 4I) groups at 15 (4A-4C), 30 (4D-4F), and 60 days (4G-4H). Hematoxylin and eosin staining. B, bone trabeculae; C, connective tissue; IT, implant thread; BM, biomaterial.

• 

  Figure 5 Light micrographs from upper cortical region and bone defects in the CO (5A, 5D and 5G), BI (5B, 5E and 5H), and BC (5C, 5F and 5I) groups at 15 (5A-5C), 30 (5D-5F), and 60 days (5G-5I). Hematoxylin and eosin staining. B, bone trabeculae; C, connective tissue; UC, upper cortical region; BM, biomaterial; MB, medullary bone.

• 

  Figure 6 Light micrographs of sections submitted to immunohistochemical analysis for detection of osteocalcin (OC). Samples from implant thread tissues in CO (6A, 6D and 6G), BI (6B, 6E and 6H), and BC (6C, 6F and 6I) groups at 15 (6A-6C), 30 (6D-6F) and 60 days (6G-6H). Graphic representation of balanced OC labeling over time(6J). Blue arrows: labeling in cytoplasm; White arrows: background labeling. a)Statistically significant between indicated groups, P<0.05.

• 

  Figure 7 Light micrographs of immunohistochemical sections representing RUNX2 labeling in samples from implant threads in the CO (7A, 7D, and 7G), BI (7B, 7E, and 7H), and BC (7C, 7F, and 7I) groups at 15 (7A-7C), 30 (7D-7F), and 60 (7G-7I) days. Graphic representation of balanced RUNX2 labeling over time(7J). Blue arrows: labeling in cytoplasm. a)Statistically significant between indicated groups, P<0.05.

• 

  Figure 8 Light micrographs of immunohistochemical sections representing OPG labeling in implant thread tissue in CO (8A, 8D and 8G), BI (8B, 8E and 8H), and BC (8C, 8F and 8I) groups at 15 (8A-8C), 30 (8D-8F), and 60 days (8G-8I). Graphic representation of balanced OPG labeling over time (8J). Blue arrows: labeling in cytoplasm. a)Statistically significant between indicated groups, P<0.05.

• 

  Figure 9 Light micrographs of immunohistochemical slices representing RANKL labeling in implant thread tissue in CO (9A, 9D and 9G), BI (9B, 9E and 9H), and BC (9C, 9F and 9I) groups at 15 (9A-9C), 30 (9D-9F), and 60 days (9G-9I). Graphic representation of balanced RANKL labeling over time (9J). Blue arrows: labeling in cytoplasm. a)Statistically significant between indicated groups, P<0.05.

• 

  Figure 10 Light micrograph immunohistochemical sections representing TRAP labeling containing samples from implant threads in CO (10A, 10D and 10G), BI (10B, 10E and 10H), and BC (10C, 10F and 10I) groups at 15 (10A-10C), 30 (10D-10F), and 60 (10G-10I) days. Graphic representation of balanced TRAP labeling over time(10J). Blue arrows: labeling in cytoplasm.

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