In vitro re-mineralization of demineralized bone matrix in human serum

2006 ◽  
Vol 44 (1) ◽  
Author(s):  
Metin Manouchehr Eskandari ◽  
Hatice Gulcin Eskandari ◽  
Melih Aktas ◽  
Ugur Atik

AbstractThe aim of this study was to develop an in vitro re-mineralization model in human serum. For this purpose a commercially produced demineralized human bone matrix (DBM) was incubated in samples of human serum pools obtained from two physiologically different groups. The first group consisted of young males and the second of older females. After incubation periods of 4 and 7days at 37°C, changes in the levels of calcium and inorganic phosphate content of the serum and DBM samples were measured. The results of the study showed that the change in mineral content of serum and DBM samples in both study groups was statistically significant. The decrease in serum calcium content and increase in DBM inorganic phosphate content were significant in the young group for longer incubation times. In the older group, both serum calcium and inorganic phosphate decreased and DBM mineral content increased for the same incubation time. When the two physiological groups were compared, statistically significant differences were identified for changes in mineral levels in both serum and the DBM samples. These data indicate that the mineral content of human serum decreases and that of DBM increases when these two materials are incubated together. These changes provide evidence for the re-mineralization of DBM. The model described here could also detect a difference in re-mineralization capability between two different groups of human sera.

2005 ◽  
Vol 288-289 ◽  
pp. 63-66 ◽  
Author(s):  
Lei Cui ◽  
Dong Li ◽  
Xiang Dong Liu ◽  
Fanfan Chen ◽  
Wei Liu ◽  
...  

Objective The purpose of this study is to explore the growth, differentiation and osteogeneration of bone marrow stromal cells (BMSCs) on partially demineralized bone matrix (pDBM) and to generate bone tissue by tissue engineering approach in vivo. Methods Demineralized bone was processed from femur head of Shanghai white swine. Calcium content, porosity and pore size was measured respectively. In vitro osteogenic differentiated human BMSCs of passage 3 were seeded in pDBM. Adhesive rate of cells to pDBM was calculated 24hours after seeding. Distribution, growth and proliferation of BMSCs on pDBM were observed with fluorescent DiI labeling. Matrix disposition was analyzed with SEM observation. Cell-material complex was implanted subcutaneously in nude mice. The implants were harvested at 8, 12 weeks post surgery and samples were observed by H&E staining. Results BMSCs adhered well on the material and the distribution of cells was uniform. The adhesive rate is 99.1%±1%. New bone formation was observed in implant of 8, 12 weeks respectively. The newly formed bone was generated on the surface of the residual material and a layer of cells with typical characteristic of osteoblast was observed to adhere on the surface of the new bone. Conclusion With good biocompatibility to hBMSCs, pDBM could serve as ideal scaffold for bone tissue engineering both in vitro and in vivo.


2012 ◽  
Vol 1417 ◽  
Author(s):  
Titilayo Moloye ◽  
Christopher Batich

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.


2021 ◽  
Author(s):  
Yutong Wu ◽  
Hongbo Ai ◽  
Yuchi Zou ◽  
Jianzhong Xu

Abstract Small extracellular vesicles (sEVs) are considered to play critical roles in intercellular communications during normal and pathological processes since they are enriched with miRNAs and other signal molecules. In bone remodeling, osteoclasts generate large amounts of sEVs. However, there is very little research about whether and how osteoclast-derived sEVs (OC-sEVs) affect surrounding cells. In our study, microarray analysis identified miR-106a-5p highly enriched in OC-sEV. Further experiments confirmed that OC-sEVs inhibited Fam134a through miR-106a-5p and significantly promoted bone mesenchymal stem cell (BMSC) osteogenic mineralization in vitro. Next, we prepared sEV-modified demineralized bone matrix (DBM) as a repair scaffold, and used a calvarial defect mouse model to evaluate the pro-osteogenic activities of the scaffold. In vivo result indicated DBM modified with miR-106a-5p-sEVs showed an enhanced capacity of bone regeneration. This important finding further emphasizes that sEV-mediated miR-106a-5p transfer play critical roles in osteogenesis and indicate a novel communication mode between osteoclasts and BMSCs.


Materials ◽  
2019 ◽  
Vol 12 (9) ◽  
pp. 1360 ◽  
Author(s):  
Monica Mattioli-Belmonte ◽  
Francesca Montemurro ◽  
Caterina Licini ◽  
Iolanda Iezzi ◽  
Manuela Dicarlo ◽  
...  

Decellularized bone matrix is receiving much attention as biological scaffolds and implantable biomaterials for bone tissue regeneration. Here, we evaluated the efficacy of a cell-free demineralized bone matrix on mesenchymal stem cells (MSCs) survival and differentiation in vitro. The seeding of human umbilical cord-derived MSCs (hUC-SCs) on decellularized bone matrices up to 14 days was exploited, assessing their capability of scaffold colonization and evaluating gene expression of bone markers. Light and Scanning Electron Microscopies were used. The obtained cell-free decalcified structures showed elastic moduli attributable to both topology and biochemical composition. Morphological observation evidenced an almost complete colonization of the scaffolds after 14 days of culture. Moreover, in hUC-SCs cultured on decalcified scaffolds, without the addition of any osteoinductive media, there was an upregulation of Collagen Type I (COL1) and osteonectin (ON) gene expression, especially on day 14. Modifications in the expression of genes engaged in stemness were also detected. In conclusion, the proposed decellularized bone matrix can induce the in vitro hUC-SCs differentiation and has the potential to be tested for in in vivo tissue regeneration.


2011 ◽  
Vol 13 (4) ◽  
pp. 653-661 ◽  
Author(s):  
William S. Pietrzak ◽  
Michael Dow ◽  
Jerry Gomez ◽  
Meghan Soulvie ◽  
George Tsiagalis

2012 ◽  
Vol 113 (4) ◽  
pp. 619-626 ◽  
Author(s):  
Zhan Kong ◽  
Jianjun Li ◽  
Qun Zhao ◽  
Zhendong Zhou ◽  
Xiangnan Yuan ◽  
...  

Neovascularization is required for bone formation and successful fracture healing. In the process of neovascularization, endothelial progenitor cells (EPCs) play an important role and finish vascular repair through reendothelialization to promote successful fracture healing. In this study, we found that dynamic compression can promote the proliferation and capillary-like tube formation of EPCs in the demineralized bone matrix (DBM) scaffold seed. EPCs isolated from the bone marrow of rats have been cultured in DBM scaffolds before dynamic compression and then seeded in the DBM scaffolds under dynamic conditions. The cells/scaffold constructs were subjected to cyclic compression with 5% strain and at 1 Hz for 4 h/day for 7 consecutive days. By using MTT and real-time PCR, we found that dynamic compression can significantly induce the proliferation of EPCs in three-dimensional culture with an even distribution of cells onto DBM scaffolds. Both in vitro and in vivo, the tube formation assays in the scaffolds showed that the loaded EPCs formed significant tube-like structures. These findings suggest that dynamic compression promoted the vasculogenic activities of EPCs seeded in the scaffolds, which would benefit large bone defect tissue engineering.


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