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 Effect of Azithromycin on Mineralized Nodule Formation in MC3T3-E1 Cells

2021 ◽  
Vol 43 (3) ◽  
pp. 1451-1459
Author(s):  
Kengo Kato ◽  
Manami Ozaki ◽  
Kumiko Nakai ◽  
Maki Nagasaki ◽  
Junya Nakajima ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Type I Collagen ◽  
Inflammatory Diseases ◽  
Bone Matrix ◽  
Nodule Formation ◽  
Type I ◽  
Alizarin Red ◽  
Odontogenic Infections ◽  
Low Concentrations

Azithromycin displays immunomodulatory and anti-inflammatory effects in addition to broad-spectrum antimicrobial activity and is used to treat inflammatory diseases, including respiratory and odontogenic infections. Few studies have reported the effect of azithromycin therapy on bone remodeling processes. The aim of this study was to examine the effects of azithromycin on the osteogenic function of osteoblasts using osteoblast-like MC3T3-E1 cells. Cells were cultured in the presence of 0, 0.1, 1, and 10 µg/mL azithromycin, and cell proliferation and alkaline phosphatase (ALPase) activity were determined. In vitro mineralized nodule formation was detected with alizarin red staining. The expression of collagenous and non-collagenous bone matrix protein was determined using real-time PCR or enzyme-linked immunosorbent assays. In cells cultured with 10 µg/mL azithromycin, the ALPase activity and mineralized nodule formation decreased, while the type I collagen, bone sialoprotein, osteocalcin, and osteopontin mRNA expression as well as osteopontin and phosphorylated osteopontin levels increased. These results suggest that a high azithromycin concentration (10 µg/mL) suppresses mineralized nodule formation by decreasing ALPase activity and increasing osteopontin production, whereas low concentrations (≤l.0 µg/mL) have no effect on osteogenic function in osteoblastic MC3T3-E1 cells.

scholarly journals Decreased C-Src Expression Enhances Osteoblast Differentiation and Bone Formation

2000 ◽  
Vol 151 (2) ◽  
pp. 311-320 ◽  
Author(s):  
Marilena Marzia ◽  
Natalie A. Sims ◽  
Susanne Voit ◽  
Silvia Migliaccio ◽  
Anna Taranta ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Matrix Protein ◽  
Bone Matrix ◽  
Osteogenic Cells ◽  
Alp Activity ◽  
Pthrp Receptor ◽  
Bone Matrix Protein

c-src deletion in mice leads to osteopetrosis as a result of reduced bone resorption due to an alteration of the osteoclast. We report that deletion/reduction of Src expression enhances osteoblast differentiation and bone formation, contributing to the increase in bone mass. Bone histomorphometry showed that bone formation was increased in Src null compared with wild-type mice. In vitro, alkaline phosphatase (ALP) activity and nodule mineralization were increased in primary calvarial cells and in SV40-immortalized osteoblasts from Src−/− relative to Src+/+ mice. Src-antisense oligodeoxynucleotides (AS-src) reduced Src levels by ∼60% and caused a similar increase in ALP activity and nodule mineralization in primary osteoblasts in vitro. Reduction in cell proliferation was observed in primary and immortalized Src−/− osteoblasts and in normal osteoblasts incubated with the AS-src. Semiquantitative reverse transcriptase-PCR revealed upregulation of ALP, Osf2/Cbfa1 transcription factor, PTH/PTHrP receptor, osteocalcin, and pro-alpha 2(I) collagen in Src-deficient osteoblasts. The expression of the bone matrix protein osteopontin remained unchanged. Based on these results, we conclude that the reduction of Src expression not only inhibits bone resorption, but also stimulates osteoblast differentiation and bone formation, suggesting that the osteogenic cells may contribute to the development of the osteopetrotic phenotype in Src-deficient mice.

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.

Alterations of Bone Matrix Protein mRNA Expression in Rat Aorta In Vitro

1995 ◽  
Vol 15 (9) ◽  
pp. 1474-1480 ◽  
Author(s):  
Hiroyuki Hao ◽  
Seiichi Hirota ◽  
Yoshitane Tsukamoto ◽  
Masami Imakita ◽  
Hatsue Ishibashi-Ueda ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Matrix Protein ◽  
Bone Matrix ◽  
Rat Aorta ◽  
Bone Matrix Protein

Preparation and In Vitro Characterization of Polycaprolactone and Demineralized Bone Matrix Scaffolds

2012 ◽  
Vol 1417 ◽  
Author(s):  
Titilayo Moloye ◽  
Christopher Batich
Keyword(s):  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Apatite Formation ◽  
Alizarin Red ◽  
Salt Leaching ◽  
Synthetic Materials ◽  
Calcium And Phosphorus ◽  
Demineralized Bone

ABSTRACTCylindrical porous polycaprolactone (PCL) scaffolds containing 25, 35, and 50 wt% demineralized bone matrix (DBM) were fabricated using a salt-leaching method for application in bone engineering. In the present work, PCL-DBM scaffolds were monitored for calcium and phosphorus deposition in both deionized (DI) water and simulated body fluid (SBF) for time periods of 5, 10, 15, and 20 days at 37°C under constant rotation. An in vitro assessment of the bioactivity of synthetic materials using SBF under physiological conditions can be used as a barometer of scaffold behavior in vivo. DBM, an osteoinductive material, was used to gauge if there was a correlation between the concentration of DBM within a scaffold and the apatite formation on its surface. Biochemical assays, alizarin red S staining, and scanning electron microscopy (SEM) with elemental analysis of calcium and phosphorus were consistent in that they confirmed that PCL scaffolds containing 35 wt% DBM in SBF at 14 days post-immersion showed signs of early apatite formation.

scholarly journals DMP1 mutations in autosomal recessive hypophosphatemia implicate a bone matrix protein in the regulation of phosphate homeostasis

2006 ◽  
Vol 38 (11) ◽  
pp. 1248-1250 ◽  
Author(s):  
Bettina Lorenz-Depiereux ◽  
Murat Bastepe ◽  
Anna Benet-Pagès ◽  
Mustapha Amyere ◽  
Janine Wagenstaller ◽  
...  
Keyword(s):  
Autosomal Recessive ◽  
Matrix Protein ◽  
Bone Matrix ◽  
Phosphate Homeostasis ◽  
Bone Matrix Protein

scholarly journals Method for Detecting the Expression of Bone Matrix Protein by in situ Hybridization Using Decalcified Mineralized Tissue.

1993 ◽  
Vol 26 (4) ◽  
pp. 303-309 ◽  
Author(s):  
SHINTARO NOMURA ◽  
KIMIAKI HIRAKAWA ◽  
JUNSUKE NAGOSHI ◽  
SEIICHI HIROTA ◽  
HYUNG-MIN KIM ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Matrix Protein ◽  
Bone Matrix ◽  
Mineralized Tissue ◽  
Bone Matrix Protein

Induction of Bone Matrix Protein Expression by Native Bone Matrix Proteins in C2C12 Culture

2009 ◽  
Vol 22 (2) ◽  
pp. 164-169 ◽  
Author(s):  
Zhen-Ming HU ◽  
Sean A.F. PEEL ◽  
Stephen K.C. HO ◽  
George K.B. SANDOR ◽  
Cameron M.L. CLOKIE
Keyword(s):  
Protein Expression ◽  
Matrix Protein ◽  
Bone Matrix ◽  
Matrix Proteins ◽  
Native Bone ◽  
Bone Matrix Proteins ◽  
Bone Matrix Protein
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