scholarly journals Biohybrid Bovine Bone Matrix for Controlled Release of Mesenchymal Stem/Stromal Cell Lyosecretome: A Device for Bone Regeneration

2021 ◽  
Vol 22 (8) ◽  
pp. 4064
Author(s):  
Elia Bari ◽  
Ilaria Roato ◽  
Giuseppe Perale ◽  
Filippo Rossi ◽  
Tullio Genova ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Bone Matrix ◽  
Stromal Vascular Fraction ◽  
In Vitro Tests ◽  
Burst Release ◽  
Bovine Bone ◽  
Class Iii ◽  
Freeze Dried ◽  
Quantification Analysis

SmartBone® (SB) is a biohybrid bone substitute advantageously proposed as a class III medical device for bone regeneration in reconstructive surgeries (oral, maxillofacial, orthopedic, and oncology). In the present study, a new strategy to improve SB osteoinductivity was developed. SB scaffolds were loaded with lyosecretome, a freeze-dried formulation of mesenchymal stem cell (MSC)-secretome, containing proteins and extracellular vesicles (EVs). Lyosecretome-loaded SB scaffolds (SBlyo) were prepared using an absorption method. A burst release of proteins and EVs (38% and 50% after 30 min, respectively) was observed, and then proteins were released more slowly with respect to EVs, most likely because they more strongly adsorbed onto the SB surface. In vitro tests were conducted using adipose tissue-derived stromal vascular fraction (SVF) plated on SB or SBlyo. After 14 days, significant cell proliferation improvement was observed on SBlyo with respect to SB, where cells filled the cavities between the native trabeculae. On SB, on the other hand, the process was still present, but tissue formation was less organized at 60 days. On both scaffolds, cells differentiated into osteoblasts and were able to mineralize after 60 days. Nonetheless, SBlyo showed a higher expression of osteoblast markers and a higher quantity of newly formed trabeculae than SB alone. The quantification analysis of the newly formed mineralized tissue and the immunohistochemical studies demonstrated that SBlyo induces bone formation more effectively. This osteoinductive effect is likely due to the osteogenic factors present in the lyosecretome, such as fibronectin, alpha-2-macroglobulin, apolipoprotein A, and TGF-β.

scholarly journals Freeze-Dried Secretome (Lyosecretome) from Mesenchymal Stem/Stromal Cells Promotes the Osteoinductive and Osteoconductive Properties of Titanium Cages

2021 ◽  
Vol 22 (16) ◽  
pp. 8445
Author(s):  
Elia Bari ◽  
Fulvio Tartara ◽  
Fabio Cofano ◽  
Giuseppe di Perna ◽  
Diego Garbossa ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Bone Matrix ◽  
In Vitro Tests ◽  
Alizarin Red ◽  
Cellular Processes ◽  
Freeze Dried ◽  
Good Biocompatibility ◽  
Titanium Cages ◽  
Bone Matrix Protein

Titanium is one of the most frequently used materials in bone regeneration due to its good biocompatibility, excellent mechanical properties, and great osteogenic performance. However, osseointegration with host tissue is often not definite, which may cause implant failure at times. The present study investigates the capacity of the mesenchymal stem cell (MSC)-secretome, formulated as a ready-to-use and freeze-dried medicinal product (the Lyosecretome), to promote the osteoinductive and osteoconductive properties of titanium cages. In vitro tests were conducted using adipose tissue-derived MSCs seeded on titanium cages with or without Lyosecretome. After 14 days, in the presence of Lyosecretome, significant cell proliferation improvement was observed. Scanning electron microscopy revealed the cytocompatibility of titanium cages: the seeded MSCs showed a spread morphology and an initial formation of filopodia. After 7 days, in the presence of Lyosecretome, more frequent and complex cellular processes forming bridges across the porous surface of the scaffold were revealed. Also, after 14 and 28 days of culturing in osteogenic medium, the amount of mineralized matrix detected by alizarin red was significantly higher when Lyosecretome was used. Finally, improved osteogenesis with Lyosecretome was confirmed by confocal analysis after 28 and 56 days of treatment, and demonstrating the production by osteoblast-differentiated MSCs of osteocalcin, a specific bone matrix protein.

scholarly journals Dynamics of reparative histogenesis of bone tissue in presence of some osteoplastic materials in vitro

2020 ◽  
Vol 4 (34) ◽  
pp. 46-50
Author(s):  
S. Yu. Ivanov ◽  
A. V. Volkov ◽  
D. A. De
Keyword(s):  
Bone Regeneration ◽  
Bone Tissue ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Bone Defects ◽  
Control Group ◽  
Bovine Bone ◽  
Reparative Osteogenesis ◽  
The Impact

Currently, to solve the bone deficiency problem in the maxillofacial region, osteoplastic materials based on allogeneic and xenogenic collagen bone matrix are used, both in pure and in activated forms, by adding growth factors. It is impossible to determine the effectiveness and mechanisms of the osteoplastic materials effect on bone regeneration without a comprehensive study, including not only histological, but also morphometric studies of the structural components and cellular reactions in the impact area. Such studies provide reliable and objective information on the main processes taking place in bone regeneration.Purpose. To determine the spatial distribution of reparative osteogenesis in the presence of some osteoplastic materials in vitro.Materials and methods. Svetlogorsk breed pigs were used as a biomodel. Depending on the osteoplastic preparations used, the animals were divided into four groups of the two in each: 1st — a preparation based on a natural bovine bone graft was injected into bone defects. 2nd — a preparation based on collagenized porcine transplant was injected into bone defects. 3rd — a preparation consisting of 60 % hydroxyapatite (HA) and 40 % beta-tri-calcium phosphate; 4th — control group — the bone defect healed under a blood clot. Animals were removed from the experiment on the 45th day. We examined sections with a thickness of 20 μm using the method of light and fluorescence microscopy.Results. The results indicate different dynamics of the reparative osteogenesis in the presence of osteoplastic materials of different classes. In group 1, the filling of the defect with newly formed bone tissue is not uniform; in group 2, the filling of the defect with newly formed bone tissue is uniform; in group 3 the filling of the defect with non-formed bone tissue is uneven due to the pronounced hyperostosis; in the control group, the filling of the defect with newly formed bone tissue is not happening.Conclusion. Stimulation, the dynamics of reparative osteogenesis and the three-dimensional organization of bone regenerate depend on the osteoplastic material class, which requires further study of the dynamics and three-dimensional organization of bone regenerate to select the optimal bone-replacing agent.

scholarly journals Dual-Drug-Loaded Silk Fibroin/PLGA Scaffolds for Potential Bone Regeneration Applications

2019 ◽  
Vol 2019 ◽  
pp. 1-16 ◽  
Author(s):  
Jihang Yao ◽  
Yilong Wang ◽  
Wendi Ma ◽  
Wenying Dong ◽  
Mei Zhang ◽  
...  
Keyword(s):  
Drug Release ◽  
Sustained Release ◽  
Bone Regeneration ◽  
Silk Fibroin ◽  
Bone Morphogenetic Protein 2 ◽  
Burst Release ◽  
Nanofiber Membrane ◽  
Differentiation Potential ◽  
Dual Drug

Developing scaffold materials with excellent biocompatibility, mechanical properties, and controlled drug release properties is vital to tissue engineering. In this study, we fabricated silk fibroin (SF)/poly(lactide-co-glycolide) (PLGA) nanofiber scaffolds containing recombinant human bone morphogenetic protein 2 (rhBMP2) and dexamethasone (DXM) via coaxial electrospinning, which were used in in vitro bone formation with rat bone marrow mesenchymal stem cells (rBMSCs). An in vitro drug release study was adopted to evaluate the sustained release potential of the core-shell structured nanofibers. Furthermore, we detected the potential of the SF/PLGA nanofiber membrane in vitro. In vitro studies showed that rhBMP2 still remained active on the nanofiber membrane. In addition, the dual-drug-loaded nanofiber membrane showed an early burst release of DXM and late sustained release of rhBMP2. rhBMP2 and DXM exhibited strong osteogenic differentiation potential when they acted on rBMSCs. Therefore, the SF/PLGA nanofiber membrane loaded with rhBMP2 and DXM has great potential for the enhancement of bone regeneration.

Fabrication and in-vitro biocompatibility of freeze-dried CTS-nHA and CTS-nBG scaffolds for bone regeneration applications

2020 ◽  
Vol 149 ◽  
pp. 1-10 ◽  
Author(s):  
Pawan Kumar ◽  
Meenu Saini ◽  
Brijnandan S. Dehiya ◽  
Ahmad Umar ◽  
Anil Sindhu ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
In Vitro Biocompatibility ◽  
Freeze Dried

scholarly journals The Effects of Atmospheric Pressure Argon Plasma Treated Bovine Bone Substitute on Bone Regeneration

Coatings ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 790
Author(s):  
Jong-Ju Ahn ◽  
Ji-Hyun Yoo ◽  
Eun-Bin Bae ◽  
Gyoo-Cheon Kim ◽  
Jae Joon Hwang ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Atmospheric Pressure ◽  
Bone Substitute ◽  
Argon Plasma ◽  
Atmospheric Pressure Plasma ◽  
Regeneration Ability ◽  
Bovine Bone ◽  
Significant Difference

This study was undertaken to compare new bone formation between non-expired and expired bovine-derived xenogeneic bone substitute (expired, out-of-use period) and to evaluate the efficacy of argon (Ar)-based atmospheric pressure plasma (APP) treatment on expired bone substitute in rat calvarial defect. The groups were divided into (1) Non/Expired group (Using regular xenografts), (2) Expired group (Using expired xenografts), and (3) Ar/Expired group (Using Ar-based APP treated expired xenografts). Surface observation and cell experiments were performed in vitro. Twelve rats were used for in vivo experiment and the bony defects were created on the middle of the cranium. The bone substitute of each group was implanted into the defective site. After 4 weeks, all the rats were sacrificed, and the volumetric, histologic, and histometric analyses were performed. In the results of osteogenic differentiation and mineralization, Non/Expired and Ar/Expired groups were significantly higher than Expired group (p < 0.05). However, there was no significant difference between groups in the animal study (p > 0.05). Within the limitations of this study, the surface treatment of Ar-based APP has a potential effect on the surface modification of bone grafts. However, there was no significant difference in bone regeneration ability between groups in vivo; thus, studies on APP to enhance bone regeneration should be carried out in the future.

scholarly journals The Influence of Eggshell on Bone Regeneration in Preclinical In Vivo Studies

Biology ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 476
Author(s):  
Horia Opris ◽  
Cristian Dinu ◽  
Mihaela Baciut ◽  
Grigore Baciut ◽  
Ileana Mitre ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Animal Studies ◽  
Meta Analysis ◽  
Bone Matrix ◽  
In Vivo Studies ◽  
Defect Model ◽  
Exclusion Criteria ◽  
Freeze Dried ◽  
Grafting Materials

The aim of this study is to systemically review the available evidence on the in vivo behavior of eggshell as a guided bone regeneration substitute material. Five databases (PubMed, Cochrane, Web of Science, Scopus, EMBASE) were searched up to October 2020. In vivo animal studies with a bone defect model using eggshell as a grafting material were included. Risk of bias was assessed using SYRCLE tool and the quality assessment using the ARRIVE guidelines. Overall, a total of 581 studies were included in the study, 187 after duplicate removal. Using the inclusion and exclusion criteria 167 records were further excluded. The full text of the remaining 20 articles was assessed for eligibility and included in the qualitative and quantitative assessment synthesis. There were different methods of obtaining eggshell grafting materials. Eggshell is a biocompatible grafting material, with osteoconduction proprieties. It forms new bone similar to Bio-Oss and demineralized freeze-dried bone matrix. It can be combined with other materials to enhance its proprieties. Due to the high variability of the procedures, animals, production and assessment methods, no meta-analysis could be performed. Eggshell might be considered a promising biomaterial to be used in bone grafting procedures, though further research is needed.

Porous Silk Fibroin Membrane as a Potential Scaffold for Bone Regeneration

2008 ◽  
Vol 396-398 ◽  
pp. 187-190 ◽  
Author(s):  
Raquel Farias Weska ◽  
Grínia M. Nogueira ◽  
Wellington C. Vieira ◽  
Marisa Masumi Beppu
Keyword(s):  
Bone Regeneration ◽  
Silk Fibroin ◽  
Biomedical Application ◽  
Early Stage ◽  
Bone Matrix ◽  
Octacalcium Phosphate ◽  
Ion Concentration ◽  
Phosphate Deposits

The requirements for scaffolds for bone tissue engineering include appropriate chemistry, morphology and structure to promote cell adhesion and synthesis of new bone matrix. Silk fibroin (SF) represents an important biomaterial for biomedical application, due to its suitable mechanical properties, biodegradability, biocompatibility, and versatility in processing. Our group has developed a new method to obtain a porous SF membrane, and the study of its potential for use as a scaffold for bone regeneration was the aim of this study. Porous membranes were obtained from SF solution, through the compression of a material generated by phase separation. For in vitro calcification experiments, porous SF membrane samples were immersed in SBF at pH 7.4 placed in polyethylene flasks. The experiments were carried out for seven days, at 36.5±0.5 °C. After 48 and 96h, the solutions were changed for fresh SBF with the ion concentration 1.5-fold higher than that of the standard one, to accelerate the calcification process. The characterization of morphology and composition of samples was performed by using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS), respectively. The SEM micrographs indicated that the porous SF membranes presented calcium phosphate deposits after undergoing in vitro calcification. These results were confirmed by EDS spectra, which showed a stoichiometric molar Ca/P ratio ranging from 1.27 to 1.52. This fact may suggest that calcification deposits consisted of mixtures of HAP (Ca/P ratio = 1.67) and transient HAP precursor phases, such as octacalcium phosphate (Ca/P = 1.33) and dicalcium phosphate dehydrate (Ca/P = 1), indicating early stage mineralization. The porous silk fibroin membrane analysed in the current study is a promising material to be used as scaffolds for bone regeneration.

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