Quantitative Analysis of New Bone Formation in Various Bone Grafts by In - Vivo Tetracycline Labeling

The aim of this study was to investigate the behavior of dental-derived human mesenchymal stem cells (d-hMSCs) in response to differently surface-treated implants and to evaluate the effect of d-hMSCs on local osteogenesis around an implant in vivo. d-hMSCs derived from alveolar bone were established and cultured on machined, sandblasted and acid-etched (SLA)-treated titanium discs with and without osteogenic induction medium. Their morphological and osteogenic potential was assessed by scanning electron microscopy (SEM) and real-time polymerase chain reaction (RT-PCR) via mixing of 5 × 106 of d-hMSCs with 1 mL of Metrigel and 20 μL of gel-cell mixture, which was dispensed into the defect followed by the placement of customized mini-implants (machined, SLA-treated implants) in New Zealand white rabbits. Following healing periods of 2 weeks and 12 weeks, the obtained samples in each group were analyzed radiographically, histomorphometrically and immunohistochemically. The quantitative change in osteogenic differentiation of d-hMSCs was identified according to the type of surface treatment. Radiographic analysis revealed that an increase in new bone formation was statistically significant in the d-hMSCs group. Histomorphometric analysis was in accordance with radiographic analysis, showing the significantly increased new bone formation in the d-hMSCs group regardless of time of sacrifice. Human nuclei A was identified near the area where d-hMSCs were implanted but the level of expression was found to be decreased as time passed. Within the limitations of the present study, in this animal model, the transplantation of d-hMSCs enhanced the new bone formation around an implant and the survival and function of the stem cells was experimentally proven up to 12 weeks post-sacrifice.

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In Vivo and In Vitro Evaluation of Coated HAp Films with Different Surface Morphology

Materials Science Forum ◽

10.4028/www.scientific.net/msf.539-543.710 ◽

2007 ◽

Vol 539-543 ◽

pp. 710-715

Author(s):

Kotaro Kuroda ◽

Ryoichi Ichino ◽

Masazumi Okido

Keyword(s):

Bone Formation ◽

Surface Morphology ◽

New Bone Formation ◽

Contact Ratio ◽

Bone Implant ◽

Ft Ir ◽

Mouse Osteoblast ◽

Surface Morphologies

Hydroxyapatite (HAp) coatings were formed on cp titanium plates and rods by the thermal substrate method in an aqueous solution that included 0.3 mM Ca(H2PO4)2 and 0.7 mM CaCl2. The coating experiments were conducted at 40-140 oC and pH = 8 for 15 or 30 min. The properties for the coated samples were studied using XRD, EDX, FT-IR, and SEM. All the specimens were covered with HAp, which had different surface morphologies such as net-like, plate-like and needle-like. After cleaning and sterilization, all the coated specimens were subjected to in vivo and vitro testing. In the in vitro testing, the mouse osteoblast-like cells (MC3T3-E1) were cultured on the coated and non-coated specimens for up to 30 days. Moreover, the specimens (φ2 x 5 mm) were implanted in rats femoral for up to 8 weeks, the osseoinductivity on them were evaluated. In in vitro evaluations, there were not significant differences between the different surface morphologies. In in vivo evaluations, however, two weeks postimplantation, new bone formed on both the HAp coated and non-coated titanium rods in the cancellous and cortical bone. The bone-implant contact ratio, which was used for the evaluation of new bone formation, was significantly dependent on the surface morphology of the HAp, and the results demonstrated that the needle-like coating appears to promote rapid bone formation.

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The genesis of membrane bone in the embryonic chick maxilla: epithelial-mesenchymal tissue recombination studies

Development ◽

10.1242/dev.56.1.269 ◽

1980 ◽

Vol 56 (1) ◽

pp. 269-281

Author(s):

Mary S. Tyler ◽

David P. McCobb

Keyword(s):

Bone Formation ◽

Hind Limb ◽

Bone Grafts ◽

Chick Embryos ◽

Embryonic Chick ◽

Limb Buds ◽

Mesenchymal Tissue ◽

Membrane Bone ◽

Tissue Recombination

In the present study, the question of whether a relatively non-specific epithelial requirement exists for membrane bone formation within the maxillary mesenchyme was investigated. Organ rudiments from embryonic chicks of three to five days of incubation (HH 18–25) were enzymatically separated into the epithelial and mesenchymal components. Maxillarymesenchyme (from embryos HH 18–19) which in the absence of epithelium will not form bone was recombined with epithelium from maxillae of similarly aged embryos (homotypichomochronic recombination) and of older embryos (HH 25) (homotypic-heterochronicrecombination). Heterotypic recombinations were made between maxillary mesenchyme (HH 18–19) and the epithelium from wing and hind-limb buds (HH 19–22). Recombinants were grown as grafts on thechorioallantoic membranes of host chick embryos. Grafts of intact maxillae, isolated maxillary mesenchyme, and isolated epithelia from the maxilla, wing-, and hind-limb buds weregrown as controls. The histodifferentiation of grafted intact maxillae was similar to that in vivo; both cartilage and membrane bone differentiated within the mesenchyme. Grafts of maxillary mesenchyme (from embryos HH 18–19) grown in the absence of epithelium formed cartilage but did not form membrane bone. Grafts of maxillary mesenchyme (from embryos HH 18–19) recombined with epithelium in homotypichomochronic, homotypic-heterochronic, and heterotypic tissue combinations formed membrane bone in addition to cartilage. These results indicate that maxillary mesenchyme requires the presence of epithelium to promote osteogenesis and that this epithelial requirement is relatively non-specific in terms of type and age of epithelium.

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Effects of condensation and compressive strain on implant primary stability

Bone and Joint Research ◽

10.1302/2046-3758.92.bjr-2019-0161 ◽

2020 ◽

Vol 9 (2) ◽

pp. 60-70 ◽

Cited By ~ 3

Author(s):

Zhijun Li ◽

Masaki Arioka ◽

Yindong Liu ◽

Maziar Aghvami ◽

Serdar Tulu ◽

...

Keyword(s):

Bone Density ◽

Bone Formation ◽

Compressive Strain ◽

Primary Stability ◽

New Bone Formation ◽

Implant Stability ◽

Bone Microdamage ◽

A Site ◽

Femoral Implants

Aims Surgeons and most engineers believe that bone compaction improves implant primary stability without causing undue damage to the bone itself. In this study, we developed a murine distal femoral implant model and tested this dogma. Methods Each mouse received two femoral implants, one placed into a site prepared by drilling and the other into the contralateral site prepared by drilling followed by stepwise condensation. Results Condensation significantly increased peri-implant bone density but it also produced higher strains at the interface between the bone and implant, which led to significantly more bone microdamage. Despite increased peri-implant bone density, condensation did not improve implant primary stability as measured by an in vivo lateral stability test. Ultimately, the condensed bone underwent resorption, which delayed the onset of new bone formation around the implant. Conclusion Collectively, these multiscale analyses demonstrate that condensation does not positively contribute to implant stability or to new peri-implant bone formation. Cite this article: Bone Joint Res. 2020;9(2):60–70.

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In Vitro and In Vivo Evaluation Of Calcium Phosphate Bone Graft Substitutes

Materials Science Forum ◽

10.4028/www.scientific.net/msf.654-656.2065 ◽

2010 ◽

Vol 654-656 ◽

pp. 2065-2070

Author(s):

Ho Yeon Song ◽

Young Hee Kim ◽

Jyoti M. Anirban ◽

In Seon Byun ◽

Kyung A Kwak ◽

...

Keyword(s):

Calcium Phosphate ◽

Bone Formation ◽

Bone Graft ◽

Cellular Proliferation ◽

New Bone Formation ◽

Hydroxy Apatite ◽

Defect Sites ◽

Bone Graft Substitutes

Calcium phosphate ceramics such as hydroxy apatite (HA), β-tricalcium phosphate (β-TCP) and bicalcium phosphate (BCP) have been used as a bone graft biomaterial because of their good biocompatibility and similarity of chemical composition to natural bones. To increase the mechanical and osteoconductive properties, the granules and spongy type porous bone graft substitutes were prepared by fibrous monolithic process and polyurethane foam replica methods, respectively. The pore sizes obtained using these approaches ranged between 100-600 µm. The cytotoxicity, cellular proliferation, differentiation and ECM deposition on the bone graft substitutes were observed by SEM and confocal microscopy. Moreover, the scaffolds were implanted in the rabbit femur. New bone formation and biodegradation of bone graft were observed through follow-up X-ray, micro-CT analysis and histological findings. After several months (2, 3, 6, 12 and 24 months) of implantation, new bone formation and ingrowths were observed in defect sites of the animal by CaP ceramics and 2 to 3 times higher bone ingrowths were confirmed than that of the normal trabecular bones in terms of total bone volume (BV).

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Effect of Particle Size on New Bone Formation In Vivo Using Calcium Phopshate Glass

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.330-332.165 ◽

2007 ◽

Vol 330-332 ◽

pp. 165-168

Author(s):

Hyun Ju Moon ◽

Racquel Z. LeGeros ◽

Kyoung Nam Kim ◽

Kwang Mahn Kim ◽

Seong Ho Choi ◽

...

Keyword(s):

Particle Size ◽

Calcium Phosphate ◽

Bone Formation ◽

Phosphate Glass ◽

Glass Powder ◽

New Bone Formation ◽

Sprague Dawley ◽

Constant Weight ◽

Calvarial Defects

The purpose of this study was to compare the bone regenerative effect of calcium phosphate glass according to the particle size in vivo. We prepared two different sizes, that is 400 μm and 40 μm, of calcium phosphate glass powder using the system CaO-CaF2-P2O5-MgO-ZnO. Critical-sized calvarial defects were created in 60 male Sprague-Dawley rats. The animals were divided into 3 groups of 20 animals each. Each defect was filled with a constant weight of 0.5 g calcium phosphate glass powder mixed with saline. As controls, the defect was left empty. The rats were sacrificed 2 or 8 weeks after postsurgery, and the results were evaluated using histological as well as histomorphometrical studies. The particle size of the calcium phosphate was crucial; 400 μm particles promoted new bone formation, while 40 μm particles inhibited it because of severe inflammation.

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RhBMP-2 and -7 combined with absorbable collagen sponge carrier enhance ectopic bone formation: An in vivo bioassay

Asian Biomedicine ◽

10.5372/1905-7415.0501.010 ◽

2011 ◽

Vol 5 (1) ◽

pp. 85-92

Author(s):

Kanok Preativatanyou ◽

Sittisak Honsawek

Keyword(s):

Bone Formation ◽

Synergistic Effect ◽

Expression System ◽

Collagen Sponge ◽

In Vivo Studies ◽

Blood Vessel Invasion ◽

New Bone Formation ◽

Absorbable Collagen Sponge ◽

In Vivo Bioassay

Abstract Background: Recombinant human bone morphogenetic proteins (rhBMPs) have been characterized especially chondrogenic and osteogenic activity both in vitro and in vivo studies. However, delivery of more than one growth factor by sustained release carrier to orthopedic site has yet been questionable in terms of efficacy and synergism. Objective: Evaluate osteoinductivity and synergistic effect of rhBMP-2 and -7 using absorbable collagen sponge (ACS) carrier system in vivo. Methods: cDNA of BMP-2 and -7 active domains were cloned and expressed in Escherichia coli BL21 StarTM (DE3) using pRSETc expression system. Then, the purified rhBMPs were loaded onto ACS and evaluated by in vivo rat subcutaneous bioassay. Two and eight weeks postoperatively, all treated groups were histologically verified for evidence of new bone formation and neovascularization by hematoxylin-eosin staining and light microscopy. Results: The Wistar rat treated with rhBMP-2 or -7/ACS exhibited new bone formation, compared to ACS control. The group treated with ACS supplemented with both rhBMP-2 and -7 significantly showed the osteoid matrix very well-organized into trabeculae-like structure with significant blood vessel invasion. Conclusion: The osteogenic induction of rhBMPs was combined with ACS carrier in the in vivo bioassay. In addition, the combination of both two potent recombinant osteoinductive cytokines, rhBMP-2 and -7, with ACS carrier demonstrated synergistic effect and might be a more promising and effective choice for therapeutic applications.

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Evaluation of Gamma Irradiated Human Bone in Nude Rats

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.288-289.269 ◽

2005 ◽

Vol 288-289 ◽

pp. 269-272

Author(s):

Y. Yu ◽

Jin Biao Chen ◽

J.L. Yang ◽

D.A.F. Morgan ◽

W.R. Walsh

Keyword(s):

Bone Formation ◽

Gamma Irradiation ◽

Soft Tissues ◽

Bone Grafts ◽

Human Bone ◽

New Bone Formation ◽

Nude Rats ◽

Defect Area ◽

Post Operation ◽

Gamma Irradiated

Deep-frozen morselized human bone grafts showed osteoconductivity and osteoinductivity when implanted into tibial window defects of nude rats. The osteoconductivity was assessed by measuring the total area of newly formed bone bridged by the implanted bone grafts in the entire defect area. The osteoinductivity was evidenced by the presence of active osteoblast-like cells and new bone formation around the implanted bone grafts, which were surrounded by soft tissues distant from the host cortex. Gamma irradiation at the doses of 15 or 25 kGray reduced the osteoconductivity (ANOVA and LSD tests, p<0.05) at 3 weeks post operation. The 25 kGray group had a significantly lower level of new bone formation compared with the 0 and 15 kGray groups. The evidences of osteoinductivity were only noted in the 0 and 15 kGray groups. Our data indicate that 25 kGray gamma irradiation reduces the osteoconductive and osteoinductive properties of the morselized human bone graft.

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Biological Reaction to Electrically Polarized Hydroxyapatite

MRS Proceedings ◽

10.1557/proc-662-ll3.4 ◽

2000 ◽

Vol 662 ◽

Cited By ~ 1

Author(s):

Tomoko Sakai ◽

Masato Ueshima ◽

Sadao Morita ◽

Satoshi Nakamura ◽

Kimihiro Yamashita

Keyword(s):

Bone Formation ◽

Body Fluid ◽

Bone Defects ◽

New Bone Formation ◽

In Vitro Experiment ◽

In Vivo Experiments ◽

Biological Reaction ◽

Positively Charged

AbstractWe have studied the polarized hydroxyapatite (HAp) whose surface was negatively or positively charged. In this study, we assessed the interfaces in vitro and in vivo periodically. As in vitro experiment, samples were immersed in simulated body fluid for 7 days and the surface was examined by scanning electron microscope (SEM). As in vivo experiments, cortical bone defects were created on the femoral trochanters and the condyles of the six Japanese white rabbits and the samples were implanted. The rabbits were sacrificed at 1, 2 and 4 W after the operation to analyze the surfaces by the SEM and the optical microscopy. In this study, a new thick apatite layer was formed on the negatively charged surface (N-surface) after 1week immersion in SBF in vitro. Besides, significant new bone formation was found at 2 weeks after the operation on N-surface in vivo, which was earlier than positively charged or non-polarized HAp surface. From this study negatively charged HAp surface by polarization accelerated the HAp crystal growth or the new bone formation. Thus, this N-surface will be promising for earlier fixation of the prosthesis or better recovery of the bone defect.

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