scholarly journals Synergic effects of decellularized bone matrix, hydroxyapatite, and extracellular vesicles on repairing of the rabbit mandibular bone defect model

2020 ◽  
Author(s):  
Asrin Emami ◽  
Tahereh Talaei-Khozani ◽  
Saeid Tavanafar ◽  
Nehleh Zareifard ◽  
Negar Azarpira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Bone Defect ◽  
Extracellular Vesicles ◽  
Bone Cells ◽  
Bone Matrix ◽  
Bone Defects ◽  
Dna Quantification ◽  
Mg63 Cells

Abstract Background Extracellular vesicles (ECV) have beneficial effects on the treatment of some pathological cases such as bone defects. Besides, the bone extracellular matrix (ECM) has been used to restore bone damage. Since major bone defects showed low spontaneous repairing ability, the purpose of this study was to find the synergic effect of decellularized bone (DB) ECM and ECVs on the repair of rabbit z. Methods The quality of decellularized sheep bones was confirmed by H&E, Hoechst, and DNA quantification. Also, immunohistochemistry, histochemical staining, and scanning electron microscopy (SEM) were done to evaluate the retaining of ECM constitutions. Osteoblast-derived ECVs were evaluated by internalization test, Transmission electron microscopy, Dynamic light scattering, and flow cytometry for CD9, CD63, CD81 markers. The hydrogel containing DB and hydroxyapatite (HA) with or without ECVs was evaluated for osteoblast functions and bone repair both in vitro and in vivo. Results SEM, histochemistry, DNA quantification, and immunohistochemistry indicated that the ECM was preserved after decellularization, while the cell and nuclear debris were removed sufficiently. In vitro assessments revealed that mineralization and alkaline phosphatase activity did not improve after treatment of MG63 cells by ECVs, while in vivo assessments showed synergic effects of ECVs and DB+HA hydrogels compared to the control, DB+HA and ECV-treated groups. Morphometric estimations revealed that the number of the bone-specific cells and vessel and bone area increased in bone defect treated with DB+HA/ECVs. Conclusion The DB enriched with ECVs can be an ideal scaffold for bone tissue engineering and may provide a suitable niche for migration and differentiation of the bone cells.

scholarly journals Synergic effects of decellularized bone matrix, hydroxyapatite, and extracellular vesicles on repairing of the rabbit mandibular bone defect model

2020 ◽  
Author(s):  
Asrin Emami ◽  
Tahereh Talaei-Khozani ◽  
Saeid Tavanafar ◽  
Nehleh Zareifard ◽  
Negar Azarpira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Bone Repair ◽  
Bone Matrix ◽  
Bone Cell ◽  
Histochemical Staining ◽  
Beneficial Effects ◽  
Mg63 Cells

Abstract Background: Extracellular vesicles (ECV) and bone extracellular matrix (ECM) have beneficial effects on the treatment of some pathological conditions. The purpose of this study was to find the synergic effects of decellularized bone (DB) ECM and ECVs on the repair of rabbit. Methods: The quality of decellularized sheep bones was confirmed by H&E, Hoechst, DNA quantification, immunohistochemistry, histochemical staining, and scanning electron microscopy (SEM). Osteoblast-derived ECVs were evaluated by internalization test, Transmission electron microscopy, Dynamic light scattering, and flow cytometry for CD9, CD63, CD81 markers. The hydrogel containing DB and hydroxyapatite (HA) with or without ECVs was evaluated for osteoblast functions and bone repair both in vitro and in vivo. Results: The data indicated ECM preservation after decellularization as well as cell depletion. In vitro assessments revealed that mineralization and alkaline phosphatase activity did not improve after treatment of MG63 cells by ECVs, while in vivo morphomatrical estimations showed synergic effects of ECVs and DB+HA hydrogels on increasing the number of bone-specific cells and vessel and bone area compared to the control, DB+HA and ECV-treated groups. Conclusions: The DB enriched with ECVs can be an ideal scaffold for bone tissue engineering and may provide a suitable niche for bone cell migration and differentiation.

scholarly journals Synergic effects of decellularized bone matrix, hydroxyapatite, and extracellular vesicles on repairing of the rabbit mandibular bone defect model

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Asrin Emami ◽  
Tahereh Talaei-Khozani ◽  
Saeid Tavanafar ◽  
Nehleh Zareifard ◽  
Negar Azarpira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Bone Repair ◽  
Bone Matrix ◽  
Bone Cell ◽  
Histochemical Staining ◽  
Beneficial Effects ◽  
Mg63 Cells

Abstract Background Extracellular vesicles (ECV) and bone extracellular matrix (ECM) have beneficial effects on the treatment of some pathological conditions. The purpose of this study was to find the synergic effects of decellularized bone (DB) ECM and ECVs on the repair of rabbit. Methods The quality of decellularized sheep bones was confirmed by H&E, Hoechst, DNA quantification, immunohistochemistry, histochemical staining, and scanning electron microscopy (SEM). Osteoblast-derived ECVs were evaluated by internalization test, Transmission electron microscopy, Dynamic light scattering, and flow cytometry for CD9, CD63, CD81 markers. The hydrogel containing DB and hydroxyapatite (HA) with or without ECVs was evaluated for osteoblast functions and bone repair both in vitro and in vivo. Results The data indicated ECM preservation after decellularization as well as cell depletion. In vitro assessments revealed that mineralization and alkaline phosphatase activity did not improve after treatment of MG63 cells by ECVs, while in vivo morphomatrical estimations showed synergic effects of ECVs and DB + HA hydrogels on increasing the number of bone-specific cells and vessel and bone area compared to the control, DB + HA and ECV-treated groups. Conclusions The DB enriched with ECVs can be an ideal scaffold for bone tissue engineering and may provide a suitable niche for bone cell migration and differentiation.

scholarly journals Synergic effects of decellularized bone matrix, hydroxyapatite, and extracellular vesicles on repairing of the rabbit mandibular bone defect model

2020 ◽  
Author(s):  
Asrin Emami ◽  
Tahereh Talaei-Khozani ◽  
Saeid Tavanafar ◽  
Nehleh Zareifard ◽  
Negar Azarpira ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Bone Repair ◽  
Bone Matrix ◽  
Bone Cell ◽  
Histochemical Staining ◽  
Beneficial Effects ◽  
Mg63 Cells

Abstract Background: Extracellular vesicles (ECV) and bone extracellular matrix (ECM) have beneficial effects on the treatment of some pathological conditions. The purpose of this study was to find the synergic effects of decellularized bone (DB) ECM and ECVs on the repair of rabbit. Methods: The quality of decellularized sheep bones was confirmed by H&E, Hoechst, DNA quantification, immunohistochemistry, histochemical staining, and scanning electron microscopy (SEM). Osteoblast-derived ECVs were evaluated by internalization test, Transmission electron microscopy, Dynamic light scattering, and flow cytometry for CD9, CD63, CD81 markers. The hydrogel containing DB and hydroxyapatite (HA) with or without ECVs was evaluated for osteoblast functions and bone repair both in vitro and in vivo. Results: The data indicated ECM preservation after decellularization as well as cell depletion. In vitro assessments revealed that mineralization and alkaline phosphatase activity did not improve after treatment of MG63 cells by ECVs, while in vivo morphomatrical estimations showed synergic effects of ECVs and DB+HA hydrogels on increasing the number of bone-specific cells and vessel and bone area compared to the control, DB+HA and ECV-treated groups. Conclusion: The DB enriched with ECVs can be an ideal scaffold for bone tissue engineering and may provide a suitable niche for bone cell migration and differentiation.

scholarly journals Increased serum and bone matrix levels of transforming growth factor β1 in patients with GH deficiency in response to GH treatment

2011 ◽  
Vol 165 (3) ◽  
pp. 393-400 ◽  
Author(s):  
Thor Ueland ◽  
Tove Lekva ◽  
Kari Otterdal ◽  
Tuva B Dahl ◽  
Nicoleta Cristina Olarescu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cortical Bone ◽  
Transforming Growth Factor ◽  
Bone Cells ◽  
Gh Deficiency ◽  
Bone Matrix ◽  
Transforming Growth Factor Β1 ◽  
Gh Treatment

ObjectivePatients with adult onset GH deficiency (aoGHD) have secondary osteoporosis, which is reversed by long-term GH substitution. Transforming growth factor β1 (TGFβ1 or TGFB1) is abundant in bone tissue and could mediate some effects of GH/IGFs on bone. We investigated its regulation by GH/IGF1in vivoandin vitro.Design and methodsThe effects of GH substitution (9–12 months, placebo controlled) on circulating and cortical bone matrix contents of TGFβ1 were investigated in patients with aoGHD. The effects of GH/IGF1 on TGFβ1 secretion in osteoblasts (hFOB), adipocytes, and THP-1 macrophages as well as the effects on release from platelets were investigatedin vitro.ResultsIn vivoGH substitution increased TGFβ1 protein levels in cortical bone and serum.In vitro, GH/IGF1 stimulation induced a significant increase in TGFβ1 secretion in hFOB. In contrast, no major effect of GH/IGF1 on TGFβ1 was found in adipocytes and THP-1 macrophages. Finally, a minor modifying effect on SFLLRN-stimulated platelet release of TGFβ1 was observed in the presence of IGF1.ConclusionGH substitution increases TGFβ1in vivoandin vitro, and this effect could contribute to improved bone metabolism during such therapy, potentially reflecting direct effect of GH/IGF1 on bone cells.

scholarly journals FORMATION OF BONE TISSUE IN CULTURE FROM ISOLATED BONE CELLS

1974 ◽  
Vol 61 (2) ◽  
pp. 427-439 ◽  
Author(s):  
Itzhak Binderman ◽  
Dan Duksin ◽  
Arieh Harell ◽  
Ephraim Katzir (Katchalski) ◽  
Leo Sachs
Keyword(s):  
Electron Microscopy ◽  
Bone Tissue ◽  
Bone Cells ◽  
Light And Electron Microscopy ◽  
Bone Collagen ◽  
The Matrix ◽  
Three Stages ◽  
And Function

A system is described for the formation of bone tissue in culture from isolated rat bone cells. The isolated bone cells were obtained from embryonic rat calvarium and periosteum or from traumatized, lifted periosteum of young rats. The cells were cultured for a period of up to 8 wk, during which time the morphological, biochemical, and functional properties of the cultures were studied. Formation of bone tissue by these isolated bone cells was shown, in that the cells demonstrated osteoblastic morphology in light and electron microscopy, the collagen formed was similar to bone collagen, there was mineralization specific for bone, and the cells reacted to the hormone calcitonin by increased calcium ion uptake. Calcification of the fine structure of the cells and the matrix is described. Three stages in the calcification process were observed by electron microscopy. It is concluded that these bone cells growing in vitro are able to function in a way similar to such cells in vivo. This tissue culture system starting from isolated bone cells is therefore suitable for studies on the structure and function of bone.

scholarly journals Bone Defect Repair Using a Bone Substitute Supported by Mesenchymal Stem Cells Derived from the Umbilical Cord

2020 ◽  
Vol 2020 ◽  
pp. 1-15 ◽  
Author(s):  
Michal Kosinski ◽  
Anna Figiel-Dabrowska ◽  
Wioletta Lech ◽  
Lukasz Wieprzowski ◽  
Ryszard Strzalkowski ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Bone Defect ◽  
Bone Substitute ◽  
Bone Defects ◽  
Bone Transplantation ◽  
Congenital Defects

Objective. Bone defects or atrophy may arise as a consequence of injury, inflammation of various etiologies, and neoplastic or traumatic processes or as a result of surgical procedures. Sometimes the regeneration process of bone loss is impaired, significantly slowed down, or does not occur, e.g., in congenital defects. For the bone defect reconstruction, a piece of the removed bone from ala of ilium or bone transplantation from a decedent is used. Replacement of the autologous or allogenic source of the bone-by-bone substitute could reduce the number of surgeries and time in the pharmacological coma during the reconstruction of the bone defect. Application of mesenchymal stem cells in the reconstruction surgery may have positive influence on tissue regeneration by secretion of angiogenic factors, recruitment of other MSCs, or differentiation into osteoblasts. Materials and Methods. Mesenchymal stem cells derived from the umbilical cord (Wharton’s jelly (WJ-MSC)) were cultured in GMP-grade DMEM low glucose supplemented with heparin, 10% platelet lysate, glucose, and antibiotics. In vitro WJ-MSCs were seeded on the bone substitute Bio-Oss Collagen® and cultured in the StemPro® Osteogenesis Differentiation Kit. During the culture on the 1st, 7th, 14th, and 21st day (day in vitro (DIV)), we analyzed viability (confocal microscopy) and adhesion capability (electron microscopy) of WJ-MSC on Bio-Oss scaffolds, gene expression (qPCR), and secretion of proteins (Luminex). In vivo Bio-Oss® scaffolds with WJ-MSC were transplanted to trepanation holes in the cranium to obtain their overgrowth. The computed tomography was performed 7, 14, and 21 days after surgery to assess the regeneration. Results. The Bio-Oss® scaffold provides a favourable environment for WJ-MSC survival. WJ-MSCs in osteodifferentiation medium are able to attach and proliferate on Bio-Oss® scaffolds. Results obtained from qPCR and Luminex® indicate that WJ-MSCs possess the ability to differentiate into osteoblast-like cells and may induce osteoclastogenesis, angiogenesis, and mobilization of host MSCs. In animal studies, WJ-MSCs seeded on Bio-Oss® increased the scaffold integration with host bone and changed their morphology to osteoblast-like cells. Conclusions. The presented construct consisted of Bio-Oss®, the scaffold with high flexibility and plasticity, approved for clinical use with seeded immunologically privileged WJ-MSC which may be considered reconstructive therapy in bone defects.

scholarly journals Evaluation of Mesenchymal Stem Cell Sheets Overexpressing BMP-7 in Canine Critical-Sized Bone Defects

2018 ◽  
Vol 19 (7) ◽  
pp. 2073 ◽  
Author(s):  
Yongsun Kim ◽  
Byung-Jae Kang ◽  
Wan Kim ◽  
Hui-suk Yun ◽  
Oh-kyeong Kweon
Keyword(s):  
Bone Defect ◽  
Therapeutic Potential ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Bone Defects ◽  
Trophic Factors ◽  
Gene Expressions ◽  
Cell Sheets ◽  
Demineralized Bone

The aim of this study was to investigate the in vitro osteogenic capacity of bone morphogenetic protein 7 (BMP-7) overexpressing adipose-derived (Ad-) mesenchymal stem cells (MSCs) sheets (BMP-7-CS). In addition, BMP-7-CS were transplanted into critical-sized bone defects and osteogenesis was assessed. BMP-7 gene expressing lentivirus particles were transduced into Ad-MSCs. BMP-7, at the mRNA and protein level, was up-regulated in BMP-7-MSCs compared to expression in Ad-MSCs. Osteogenic and vascular-related gene expressions were up-regulated in BMP-7-CS compared to Ad-MSCs and Ad-MSC sheets. In a segmental bone-defect model, newly formed bone and neovascularization were enhanced with BMP-7-CS, or with a combination of BMP-7-CS and demineralized bone matrix (DBM), compared to those in control groups. These results demonstrate that lentiviral-mediated gene transfer of BMP-7 into Ad-MSCs allows for stable BMP-7 production. BMP-7-CS displayed higher osteogenic capacity than Ad-MSCs and Ad-MSC sheets. In addition, BMP-7-CS combined with demineralized bone matrix (DBM) stimulated new bone and blood vessel formation in a canine critical-sized bone defect. The BMP-7-CS not only provides BMP-7 producing MSCs but also produce osteogenic and vascular trophic factors. Thus, BMP-7-CS and DBM have therapeutic potential for the treatment of critical-sized bone defects and could be used to further enhance clinical outcomes during bone-defect treatment.

scholarly journals Reconstructing Bone with Natural Bone Graft: A Review of In Vivo Studies in Bone Defect Animal Model

Nanomaterials ◽  
2018 ◽  
Vol 8 (12) ◽  
pp. 999 ◽  
Author(s):  
Mengying Liu ◽  
Yonggang Lv
Keyword(s):  
Bone Graft ◽  
Bone Defect ◽  
Bone Structure ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Bone Defects ◽  
Autologous Bone Graft ◽  
In Vivo Studies ◽  
Natural Bone

Bone defects caused by fracture, disease or congenital defect remains a medically important problem to be solved. Bone tissue engineering (BTE) is a promising approach by providing scaffolds to guide and support the treatment of bone defects. However, the autologous bone graft has many defects such as limited sources and long surgical procedures. Therefore, xenograft bone graft is considered as one of the best substitutions and has been effectively used in clinical practice. Due to better preserved natural bone structure, suitable mechanical properties, low immunogenicity, good osteoinductivity and osteoconductivity in natural bone graft, decellularized and demineralized bone matrix (DBM) scaffolds were selected and discussed in the present review. In vivo animal models provide a complex physiological environment for understanding and evaluating material properties and provide important reference data for clinical trials. The purpose of this review is to outline the in vivo bone regeneration and remodeling capabilities of decellularized and DBM scaffolds in bone defect models to better evaluate the potential of these two types of scaffolds in BTE. Taking into account the limitations of the state-of-the-art technology, the results of the animal bone defect model also provide important information for future design of natural bone composite scaffolds.

scholarly journals Implant removal using thermal necrosis—an in vitro pilot study

2020 ◽  
Vol 25 (1) ◽  
pp. 265-273
Author(s):  
Kristian Kniha ◽  
Eva Miriam Buhl ◽  
Benita Hermanns-Sachweh ◽  
Faruk Al-Sibai ◽  
Anna Bock ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Bone Matrix ◽  
Implant Removal ◽  
Threshold Temperature ◽  
Thermal Necrosis ◽  
Cold Temperatures ◽  
Bone Necrosis ◽  
Temperature Thresholds

Abstract Objectives The purpose of this pilot porcine cadaver study was to evaluate the feasible temperature thresholds, which affect osteocyte viability and bone matrix in a preclinical setup, assessing the potential of thermal necrosis for implant removal for further in vivo investigations. Materials and methods After implant bed preparation in the upper and lower jaw, temperature effects on the bone were determined, using two tempering pistons with integrated thermocouples. To evaluate threshold temperature and time intervals leading to bone necrosis, one piston generated warm temperatures at 49 to 56 °C for 10 s and the other generated cold temperatures at 5 to 1 °C for 30 s. Effects were assessed by a semi-quantitative, histomorphometrical scoring system, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and transmission electron microscopy (TEM). Results The bone matrix was significantly degenerated starting at 51 °C for 10 s and 5 °C for 30 s. The osteocyte condition indicated significant bone damage beginning at cold temperatures of 2 °C. Temperature inputs starting at 53 °C led to decalcification and swollen mitochondria, which lost the structure of their inner cristae. Conclusions This study identified temperatures and durations, in both heat and cold, so that the number of samples may be kept low in further studies regarding temperature-induced bone necrosis. Levels of 51 °C for 10 s and 5 °C for 30 s have presented significant matrix degeneration. Clinical relevance Temperature thresholds, potentially leading to thermo-explantation of dental implants and other osseointegrated devices, were identified.

Fabrication of piezoelectric porous BaTiO3 scaffold to repair large segmental bone defect in sheep

2020 ◽  
Vol 35 (4-5) ◽  
pp. 544-552 ◽  
Author(s):  
Wenwen Liu ◽  
Di Yang ◽  
Xinghui Wei ◽  
Shuo Guo ◽  
Ning Wang ◽  
...  
Keyword(s):  
Electric Field ◽  
Bone Defect ◽  
Piezoelectric Effect ◽  
Bone Defects ◽  
Segmental Bone Defect ◽  
Segmental Bone ◽  
Large Bone ◽  
Large Bone Defects

Porous titanium scaffolds can provide sufficient mechanical support and bone growth space for large segmental bone defect repair. However, they fail to restore the physiological environment of bone tissue. Barium titanate (BaTiO3) is considered a smart material that can produce an electric field in response to dynamic force. Low-intensity pulsed ultrasound stimulation (LIPUS), as a kind of micromechanical wave, can not only promote bone repair but also induce BaTiO3 to generate an electric field. In our studies, BaTiO3 was coated on porous Ti6Al4V and LIPUS was externally applied to observe the influence of the piezoelectric effect on the repair of large bone defects in vitro and in vivo. The results show that the piezoelectric effect can effectively promote the osteogenic differentiation of bone marrow stromal cells (BMSCs) in vitro as well as bone formation and growth into implants in vivo. This study provides an optional alternative to the conventional porous Ti6Al4V scaffold with enhanced osteogenesis and osseointegration for the repair of large bone defects.

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