In vivo 3D analysis with micro-computed tomography of rat calvaria bone regeneration using periosteal cell sheets fabricated on temperature-responsive culture dishes

2010 ◽  
Vol 5 (6) ◽  
pp. 483-490 ◽  
Author(s):  
Hiroto Uchiyama ◽  
Masayuki Yamato ◽  
Ryo Sasaki ◽  
Hidekazu Sekine ◽  
Joseph Yang ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Bone Regeneration ◽  
3D Analysis ◽  
Micro Computed Tomography ◽  
Cell Sheets ◽  
Temperature Responsive ◽  
Rat Calvaria ◽  
Periosteal Cell

Demineralized bone matrix-based microcarrier scaffold favors vascularized large bone regeneration in vivo in a rat model

2018 ◽  
Vol 33 (2) ◽  
pp. 182-195 ◽  
Author(s):  
Qiannan Li ◽  
Wenjie Zhang ◽  
Guangdong Zhou ◽  
Yilin Cao ◽  
Wei Liu ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Stem Cells ◽  
Bone Marrow ◽  
Bone Regeneration ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Matrix Group ◽  
Micro Computed Tomography ◽  
Demineralized Bone

Insufficient neo-vascularization of in vivo implanted cell-seeded scaffold remains a major bottleneck for clinical translation of engineered bone formation. Demineralized bone matrix is an ideal bone scaffold for bone engineering due to its structural and biochemical components similar to those of native bone. We hypothesized that the microcarrier form of demineralized bone matrix favors ingrowth of vessels and bone regeneration upon in vivo implantation. In this study, a rat model of femoral vessel pedicle-based bone engineering was employed by filling the demineralized bone matrix scaffolds inside a silicone chamber that surrounded the vessel pedicles, and to compare the efficiency of vascularized bone regeneration between microcarrier demineralized bone matrix and block demineralized bone matrix. The results showed that bone marrow stem cells better adhered to microcarrier demineralized bone matrix and produced more extracellular matrices during in vitro culture. After in vivo implantation, microcarrier demineralized bone matrix seeded with bone marrow stem cells formed relatively more bone tissue than block demineralized bone matrix counterpart at three months upon histological examination. Furthermore, micro-computed tomography three-dimensional reconstruction showed that microcarrier demineralized bone matrix group regenerate significantly better and more bone tissues than block demineralized bone matrix both qualitatively and quantitatively (p < 0.05). Moreover, micro-computed tomography reconstructed angiographic images also demonstrated significantly enhanced tissue vascularization in microcarrier demineralized bone matrix group than in block demineralized bone matrix group both qualitatively and quantitatively (p < 0.05). Anti-CD31 immunohistochemical staining of (micro-) vessels and semi-quantitative analysis also evidenced enhanced vascularization of regenerated bone in microcarrier demineralized bone matrix group than in block demineralized bone matrix group (p < 0.05). In conclusion, the microcarrier form of demineralized bone matrix is an ideal bone regenerative scaffold due to its advantages of osteoinductivity and vascular induction, two essentials for in vivo bone regeneration.

scholarly journals Spatio-temporal characterization of fracture healing patterns and assessment of biomaterials by time-lapsed in vivo micro-computed tomography

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Esther Wehrle ◽  
Duncan C. Tourolle né Betts ◽  
Gisela A. Kuhn ◽  
Erica Floreani ◽  
Malavika H. Nambiar ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Bone Regeneration ◽  
Healing Process ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Collagen Scaffolds ◽  
Non Union ◽  
Spatio Temporal

AbstractThorough preclinical evaluation of functionalized biomaterials for treatment of large bone defects is essential prior to clinical application. Using in vivo micro-computed tomography (micro-CT) and mouse femoral defect models with different defect sizes, we were able to detect spatio-temporal healing patterns indicative of physiological and impaired healing in three defect sub-volumes and the adjacent cortex. The time-lapsed in vivo micro-CT-based approach was then applied to evaluate the bone regeneration potential of functionalized biomaterials using collagen and bone morphogenetic protein (BMP-2). Both collagen and BMP-2 treatment led to distinct changes in bone turnover in the different healing phases. Despite increased periosteal bone formation, 87.5% of the defects treated with collagen scaffolds resulted in non-unions. Additional BMP-2 application significantly accelerated the healing process and increased the union rate to 100%. This study further shows potential of time-lapsed in vivo micro-CT for capturing spatio-temporal deviations preceding non-union formation and how this can be prevented by application of functionalized biomaterials. This study therefore supports the application of longitudinal in vivo micro-CT for discrimination of normal and disturbed healing patterns and for the spatio-temporal characterization of the bone regeneration capacity of functionalized biomaterials.

scholarly journals Micro-Computed Tomography Analysis of Subchondral Bone Regeneration Using Osteochondral Scaffolds in an Ovine Condyle Model

2021 ◽  
Vol 11 (3) ◽  
pp. 891
Author(s):  
Taylor Flaherty ◽  
Maryam Tamaddon ◽  
Chaozong Liu
Keyword(s):  
Computed Tomography ◽  
Bone Regeneration ◽  
Subchondral Bone ◽  
Volume Ratio ◽  
Bone Volume ◽  
Osteochondral Defects ◽  
Micro Ct ◽  
Micro Computed Tomography ◽  
Trabecular Thickness ◽  
Osteochondral Scaffold

Osteochondral scaffold technology has emerged as a promising therapy for repairing osteochondral defects. Recent research suggests that seeding osteochondral scaffolds with bone marrow concentrate (BMC) may enhance tissue regeneration. To examine this hypothesis, this study examined subchondral bone regeneration in scaffolds with and without BMC. Ovine stifle condyle models were used for the in vivo study. Two scaffold systems (8 mm diameter and 10 mm thick) with and without BMC were implanted into the femoral condyle, and the tissues were retrieved after six months. The retrieved femoral condyles (with scaffold in) were examined using micro-computed tomography scans (micro-CT), and the micro-CT data were further analysed by ImageJ with respect to trabecular thickness, bone volume to total volume ratio (BV/TV) ratio, and degree of anisotropy of bone. Statistical analysis compared bone regeneration between scaffold groups and sub-set regions. These results were mostly insignificant (p < 0.05), with the exception of bone volume to total volume ratio when comparing scaffold composition and sub-set region. Additional trends in the data were observed. These results suggest that the scaffold composition and addition of BMC did not significantly affect bone regeneration in osteochondral defects after six months. However, this research provides data which may guide the development of future treatments.

scholarly journals Longitudinal in vivo evaluation of bone regeneration by combined measurement of multi-pinhole SPECT and micro-CT for tissue engineering

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Philipp S. Lienemann ◽  
Stéphanie Metzger ◽  
Anna-Sofia Kiveliö ◽  
Alain Blanc ◽  
Panagiota Papageorgiou ◽  
...  
Keyword(s):  
Computed Tomography ◽  
High Resolution ◽  
Bone Formation ◽  
Bone Regeneration ◽  
Bone Defects ◽  
Histological Evaluation ◽  
Biological Processes ◽  
Micro Ct ◽  
Pinhole Spect

Abstract Over the last decades, great strides were made in the development of novel implants for the treatment of bone defects. The increasing versatility and complexity of these implant designs request for concurrent advances in means to assess in vivo the course of induced bone formation in preclinical models. Since its discovery, micro-computed tomography (micro-CT) has excelled as powerful high-resolution technique for non-invasive assessment of newly formed bone tissue. However, micro-CT fails to provide spatiotemporal information on biological processes ongoing during bone regeneration. Conversely, due to the versatile applicability and cost-effectiveness, single photon emission computed tomography (SPECT) would be an ideal technique for assessing such biological processes with high sensitivity and for nuclear imaging comparably high resolution (<1 mm). Herein, we employ modular designed poly(ethylene glycol)-based hydrogels that release bone morphogenetic protein to guide the healing of critical sized calvarial bone defects. By combined in vivo longitudinal multi-pinhole SPECT and micro-CT evaluations we determine the spatiotemporal course of bone formation and remodeling within this synthetic hydrogel implant. End point evaluations by high resolution micro-CT and histological evaluation confirm the value of this approach to follow and optimize bone-inducing biomaterials.

scholarly journals Bone Regeneration Capacity of Newly Developed Uncalcined/Unsintered Hydroxyapatite and Poly-l-lactide-co-glycolide Sheet in Maxillofacial Surgery: An In Vivo Study

Nanomaterials ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 22
Author(s):  
Huy Xuan Ngo ◽  
Quang Ngoc Dong ◽  
Yunpeng Bai ◽  
Jingjing Sha ◽  
Shinji Ishizuka ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Reconstructive Surgery ◽  
Early Stage ◽  
Maxillofacial Surgery ◽  
Male Rats ◽  
Regeneration Ability ◽  
Sprague Dawley ◽  
Micro Computed Tomography ◽  
Rat Mandible

Uncalcined/unsintered hydroxyapatite and poly-l-lactide-co-glycolide (u-HA/PLLA/PGA) is a new bioresorbable nanomaterial with superior characteristics compared with current bioresorbable materials, including appropriate mechanical properties, outstanding bioactive/osteoconductive features, and remarkably shorter resorption time. Nevertheless, the bone regeneration characteristics of this nanomaterial have not been evaluated in maxillofacial reconstructive surgery. In this study, we used a rat mandible model to assess the bone regeneration ability of u-HA/PLLA/PGA material, compared with uncalcined/unsintered hydroxyapatite and poly-l-lactide acid (u-HA/PLLA) material, which has demonstrated excellent bone regenerative ability. A 4-mm-diameter defect was created at the mandibular angle area in 28 Sprague Dawley male rats. The rats were divided into three groups: u-HA/PLLA/PGA (u-HA/PLLA/PGA graft + defect), u-HA/PLLA (u-HA/PLLA graft + defect), and sham control (defect alone). At 1, 3, 8, and 16 weeks after surgeries, the rats were sacrificed and assessed by micro-computed tomography, histological analysis with hematoxylin and eosin staining, and immunohistochemical analyses. The results confirmed that the accelerated bone bioactive/regenerative osteoconduction of u-HA/PLLA/PGA was comparable with that of u-HA/PLLA in the rat mandible model. Furthermore, this new regenerative nanomaterial was able to more rapidly induce bone formation in the early stage and had great potential for further clinical applications in maxillofacial reconstructive surgery.

3D-printed Poly-Lactic Co-Glycolic Acid (PLGA) scaffolds in non-critical bone defects impede bone regeneration in rabbit tibia bone

2021 ◽  
pp. 1-7
Author(s):  
Jin Xi Lim ◽  
Min He ◽  
Alphonsus Khin Sze Chong
Keyword(s):  
Computed Tomography ◽  
Bone Regeneration ◽  
Bone Defect ◽  
Volume Ratio ◽  
Bone Defects ◽  
Control Group ◽  
Micro Computed Tomography ◽  
Rabbit Tibia ◽  
3D Printed ◽  
Critical Bone Defects

BACKGROUND: An increasing number of bone graft materials are commercially available and vary in their composition, mechanism of action, costs, and indications. OBJECTIVE: A commercially available PLGA scaffold produced using 3D printing technology has been used to promote the preservation of the alveolar socket after tooth extraction. We examined its influence on bone regeneration in long bones of New Zealand White rabbits. METHODS: 5.0-mm-diameter circular defects were created on the tibia bones of eight rabbits. Two groups were studied: (1) control group, in which the bone defects were left empty; (2) scaffold group, in which the PLGA scaffolds were implanted into the bone defect. Radiography was performed every two weeks postoperatively. After sacrifice, bone specimens were isolated and examined by micro-computed tomography and histology. RESULTS: Scaffolds were not degraded by eight weeks after surgery. Micro-computed tomography and histology showed that in the region of bone defects that was occupied by scaffolds, bone regeneration was compromised and the total bone volume/total volume ratio (BV/TV) was significantly lower. CONCLUSION: The implantation of this scaffold impedes bone regeneration in a non-critical bone defect. Implantation of bone scaffolds, if unnecessary, lead to a slower rate of fracture healing.

Micro-Computed Tomography Analysis on Administration of Mesenchymal Stem Cells - Bovine Teeth Scaffold Composites for Alveolar Bone Tissue Engineering

2021 ◽  
Vol 52 ◽  
pp. 86-96
Author(s):  
Desi Sandra Sari ◽  
Fourier Dzar Eljabbar Latief ◽  
Ferdiansyah ◽  
Ketut Sudiana ◽  
Fedik Abdul Rantam
Keyword(s):  
Computed Tomography ◽  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Regeneration ◽  
Bone Growth ◽  
Alveolar Bone ◽  
Micro Computed Tomography ◽  
Freeze Dried ◽  
Significant Difference

The tissue engineering approach for periodontal tissue regeneration using a combination of stem cells and scaffold has been vastly developed. Mesenchymal Stem Cells (MSCs) seeded with Bovine Teeth Scaffold (BTSc) can repair alveolar bone damage in periodontitis cases. The alveolar bone regeneration process was analyzed by micro-computed tomography (µ-CT) to observe the structure of bone growth and to visualize the scaffold in 3-Dimensional (3D). The purpose of this study is to analyze alveolar bone regeneration by µ-CT following the combination of MSCs and bovine teeth scaffold (MSCs-BTSc) implantation in the Wistar rat periodontitis model. Methods. MSCs were cultured from adipose-derived mesenchymal stem cells of rats. BTSc was taken from bovine teeth and freeze-dried with a particle size of 150-355 µm. MSCs were seeded on BTSc for 24 hours and transplanted in a rat model of periodontitis. Thirty-five Wistar rats were made as periodontitis models with LPS induction from P. gingivalis injected to the buccal section of interproximal gingiva between the first and the second mandibular right-molar teeth for six weeks. There were seven groups (control group, BTSc group on day 7, BTSc group on day 14, BTSc group on day 28, MSCs-BTSc group on day 7, MSCs-BTSc group on day 14, MSCs-BTSc group on day 28). The mandibular alveolar bone was analyzed and visualized in 3D with µ-CT to observe any new bone growth. Statistical Analysis. Group data were subjected to the Kruskal Wallis test followed by the Mann-Whitney (p <0.05). The µ-CT qualitative analysis shows a fibrous structure, which indicates the existence of new bone regeneration. Quantitative analysis of the periodontitis model showed a significant difference between the control model and the model with the alveolar bone resorption (p <0.05). The bone volume and density measurements revealed that the MSCs-BTSc group on day 28 formed new bone compared to other groups (p <0.05). Administration of MSCs-BTSc combination has the potential to form new alveolar bone.

scholarly journals In vivo bone regeneration assessment of offset and gradient melt electrowritten (MEW) PCL scaffolds

2020 ◽  
Vol 24 (1) ◽  
Author(s):  
Naghmeh Abbasi ◽  
Ryan S. B. Lee ◽  
Saso Ivanovski ◽  
Robert M. Love ◽  
Stephen Hamlet
Keyword(s):  
Bone Regeneration ◽  
Size Structure ◽  
Three Dimensional ◽  
Immunohistochemical Analysis ◽  
Repair Process ◽  
Micro Computed Tomography ◽  
Calvarial Defects ◽  
Promising Solution ◽  
Poly Caprolactone

Abstract Background Biomaterial-based bone tissue engineering represents a promising solution to overcome reduced residual bone volume. It has been previously demonstrated that gradient and offset architectures of three-dimensional melt electrowritten poly-caprolactone (PCL) scaffolds could successfully direct osteoblast cells differentiation toward an osteogenic lineage, resulting in mineralization. The aim of this study was therefore to evaluate the in vivo osteoconductive capacity of PCL scaffolds with these different architectures. Methods Five different calcium phosphate (CaP) coated melt electrowritten PCL pore sized scaffolds: 250 μm and 500 μm, 500 μm with 50% fibre offset (offset.50.50), tri layer gradient 250–500-750 μm (grad.250top) and 750–500-250 μm (grad.750top) were implanted into rodent critical-sized calvarial defects. Empty defects were used as a control. After 4 and 8 weeks of healing, the new bone was assessed by micro-computed tomography and immunohistochemistry. Results Significantly more newly formed bone was shown in the grad.250top scaffold 8 weeks post-implantation. Histological investigation also showed that soft tissue was replaced with newly formed bone and fully covered the grad.250top scaffold. While, the bone healing did not happen completely in the 250 μm, offset.50.50 scaffolds and blank calvaria defects following 8 weeks of implantation. Immunohistochemical analysis showed the expression of osteogenic markers was present in all scaffold groups at both time points. The mineralization marker Osteocalcin was detected with the highest intensity in the grad.250top and 500 μm scaffolds. Moreover, the expression of the endothelial markers showed that robust angiogenesis was involved in the repair process. Conclusions These results suggest that the gradient pore size structure provides superior conditions for bone regeneration.

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