scholarly journals The Possible Roles of Biological Bone Constructed with Peripheral Blood Derived EPCs and BMSCs in Osteogenesis and Angiogenesis

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Li Wu ◽  
Xian Zhao ◽  
Bo He ◽  
Jie Jiang ◽  
Xiao-Jie Xie ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Coculture System ◽  
Polymerase Chain

This study aimed to determine the possible potential of partially deproteinized biologic bone (PDPBB) seeded with bone marrow stromal cells (BMSCs) and endothelial progenitor cells (EPCs) in osteogenesis and angiogenesis. BMSCs and EPCs were isolated, identified, and coculturedin vitro, followed by seeding on the PDPBB. Expression of osteogenesis and vascularization markers was quantified by immunofluorescence (IF) staining, immunohistochemistry (IHC), and quantitive real-time polymerase chain reaction (qRT-PCR). Scanning electron microscope (SEM) was also employed to further evaluate the morphologic alterations of cocultured cells in the biologic bone. Results demonstrated that the coculture system combined with BMSCs and EPCs had significant advantages of (i) upregulating the mRNA expression of VEGF, Osteonectin, Osteopontin, and Collagen Type I and (ii) increasing ALP and OC staining compared to the BMSCs or EPCs only group. Moreover, IHC staining for CD105, CD34, and ZO-1 increased significantly in the implanted PDPBB seeded with coculture system, compared to that of BMSCs or EPCs only, respectively. Summarily, the present data provided evidence that PDPBB seeded with cocultured system possessed favorable cytocompatibility, provided suitable circumstances for different cell growth, and had the potential to provide reconstruction for cases with bone defection by promoting osteogenesis and angiogenesis.

scholarly journals Pro-fibrotic phenotype of bone marrow stromal cells in Modic type 1 changes

2021 ◽  
Vol 41 ◽  
pp. 648-667
Author(s):  
I Heggli ◽  
◽  
S Epprecht ◽  
A Juengel ◽  
R Schuepbach ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Collagen Type ◽  
Collagen Type I ◽  
Marrow Stromal Cells ◽  
Type I ◽  
Cell Contractility ◽  
Modic Type

Modic type 1 changes (MC1) are painful vertebral bone marrow lesions frequently found in patients suffering from chronic low-back pain. Marrow fibrosis is a hallmark of MC1. Bone marrow stromal cells (BMSCs) are key players in other fibrotic bone marrow pathologies, yet their role in MC1 is unknown. The present study aimed to characterise MC1 BMSCs and hypothesised a pro-fibrotic role of BMSCs in MC1. BMSCs were isolated from patients undergoing lumbar spinal fusion from MC1 and adjacent control vertebrae. Frequency of colony-forming unit fibroblast (CFU-F), expression of stem cell surface markers, differentiation capacity, transcriptome, matrix adhesion, cell contractility as well as expression of pro-collagen type I alpha 1, α-smooth muscle actin, integrins and focal adhesion kinase (FAK) were compared. More CFU-F and increased expression of C-X-C-motif-chemokine 12 were found in MC1 BMSCs, possibly indicating overrepresentation of a perisinusoidal BMSC population. RNA sequencing analysis showed enrichment in extracellular matrix proteins and fibrosis-related signalling genes. Increases in pro-collagen type I alpha 1 expression, cell adhesion, cell contractility and phosphorylation of FAK provided further evidence for their pro-fibrotic phenotype. Moreover, a leptin receptor high expressing (LEPRhigh) BMSC population was identified that differentiated under transforming growth factor beta 1 stimulation into myofibroblasts in MC1 but not in control BMSCs. In conclusion, pro-fibrotic changes in MC1 BMSCs and a LEPRhigh MC1 BMSC subpopulation susceptible to myofibroblast differentiation were found. Fibrosis is a hallmark of MC1 and a potential therapeutic target. A causal link between the pro-fibrotic phenotype and clinical characteristics needs to be demonstrated.

Repair of large cranial defects byhBMP-2expressing bone marrow stromal cells: Comparison between alginate and collagen type I systems

2010 ◽  
Vol 9999A ◽  
pp. NA-NA ◽  
Author(s):  
Sophia Chia Ning Chang ◽  
Hui-Ying Chung ◽  
Ching-Lung Tai ◽  
Philips Kuo Ting Chen ◽  
Tsung-Min Lin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Collagen Type ◽  
Collagen Type I ◽  
Marrow Stromal Cells ◽  
Type I

The Effect Orthosilicic Acid on Collagen Type I, Alkaline Phosphatase and Osteocalcin mRNA Expression in Human Bone-Derived Osteoblasts In Vitro

2006 ◽  
Vol 309-311 ◽  
pp. 121-124 ◽  
Author(s):  
Meera Q. Arumugam ◽  
D.C. Ireland ◽  
Roger A. Brooks ◽  
Neil Rushton ◽  
William Bonfield
Keyword(s):  
Alkaline Phosphatase ◽  
Mrna Expression ◽  
Messenger Rna ◽  
Collagen Type ◽  
Collagen Type I ◽  
Orthosilicic Acid ◽  
Type I ◽  
Mrna Levels ◽  
Polymerase Chain

The object of this study was to investigate the effect of the concentration of orthosilicic acid (0, 0.5, 1, 5 and 10µM) on gene expression in human osteoblast cells isolated from trabecular bone. This was measured using reverse transcriptase-polymerase chain reaction (RT-PCR) to quantify messenger RNA (mRNA) levels for collagen type I, alkaline phosphatase and osteocalcin. Results showed that while collagen type I mRNA expression was increased by the addition of up to 10µM orthosilicic acid, ALP message was suppressed over time and osteocalcin levels were decreased.

scholarly journals Comparative study of the effect of bFGF and plasma rich in growth factors on cryopreserved multipotent mesenchymal stromal cells from bone marrow and tendon of rats

2017 ◽  
Vol 5 (2) ◽  
pp. 170-175 ◽  
Author(s):  
N. Volkovа ◽  
M. Yukhta ◽  
A. Goltsev
Keyword(s):  
Bone Marrow ◽  
Growth Factors ◽  
Stromal Cells ◽  
Collagen Type ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Collagen Type I ◽  
Type I ◽  
And Migration ◽  
Proliferation And Migration

The purpose of study was to investigate in vitro effects of growth factors, known as cell proliferation stimulants, to determine the most suitable agent for enhancing the proliferation and migration activity of cryopreserved multipotent mesenchymal stromal cells (MMSCs) derived from bone marrow and tendon tissue.Materials and methods. MMSCs were obtained from bone marrow and tendon tissues of rats. Cryopreservation was carried out under the protection of 10 % DMSO with the addition of 20 % fetal bovine serum at a cooling rate of 1°C/min to -80°C and subsequent freeze in liquid nitrogen. During the cultivation of the cryopreserved MMSCs, basis fibroblast growth factor (bFGF) and plasma rich in growth factors were used. The ability to proliferation (MTT assay), migration (in vitro scratch assay), and the synthesis of collagen type I (immunocytochemical study of collagen type I expression) were evaluated.Results. The use of plasma rich in growth factors contributes to increasing the ability of cryopreserved MMSCs from bone marrow to proliferate and migrate, associated with decreasing in the relative number of cells that express collagen type I. Cultures of cryopreserved MMSCs from the tendon tissue exhibit greater sensitivity to the bFGF compared to the plasma rich in growth factors that have a manifestation in the increasing of cell proliferation and migration ability.Conclusions. bFGF and plasma rich in growth factors can be used as stimulants for stromal cell cultures.

scholarly journals Relative rates of biosynthesis of collagen type I, type V and type VI in calf cornea

1991 ◽  
Vol 274 (2) ◽  
pp. 615-617 ◽  
Author(s):  
P Kern ◽  
M Menasche ◽  
L Robert
Keyword(s):  
Type I Collagen ◽  
Collagen Type ◽  
Corneal Stroma ◽  
Collagen Type I ◽  
Type I ◽  
Type Vi Collagen ◽  
Sds Page ◽  
Proline Incorporation ◽  
Type V

The biosynthesis of type I, type V and type VI collagens was studied by incubation of calf corneas in vitro with [3H]proline as a marker. Pepsin-solubilized collagen types were isolated by salt fractionation and quantified by SDS/PAGE. Expressed as proportions of the total hydroxyproline solubilized, corneal stroma comprised 75% type I, 8% type V and 17% type VI collagen. The rates of [3H]proline incorporation, linear up to 24 h for each collagen type, were highest for type VI collagen and lowest for type I collagen. From pulse-chase experiments, the calculated apparent half-lives for types I, V and VI collagens were 36 h, 10 h and 6 h respectively.

scholarly journals Plant Recombinant Human Collagen Type I Hydrogels for Corneal Regeneration

2021 ◽  
Author(s):  
Michel Haagdorens ◽  
Elle Edin ◽  
Per Fagerholm ◽  
Marc Groleau ◽  
Zvi Shtein ◽  
...  
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Collagen Fibrils ◽  
Type I ◽  
Safe Alternative ◽  
Human Donor ◽  
Human Collagen ◽  
Donor Corneas

Abstract Purpose To determine feasibility of plant-derived recombinant human collagen type I (RHCI) for use in corneal regenerative implants Methods RHCI was crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form hydrogels. Application of shear force to liquid crystalline RHCI aligned the collagen fibrils. Both aligned and random hydrogels were evaluated for mechanical and optical properties, as well as in vitro biocompatibility. Further evaluation was performed in vivo by subcutaneous implantation in rats and corneal implantation in Göttingen minipigs. Results Spontaneous crosslinking of randomly aligned RHCI (rRHCI) formed robust, transparent hydrogels that were sufficient for implantation. Aligning the RHCI (aRHCI) resulted in thicker collagen fibrils forming an opaque hydrogel with insufficient transverse mechanical strength for surgical manipulation. rRHCI showed minimal inflammation when implanted subcutaneously in rats. The corneal implants in minipigs showed that rRHCI hydrogels promoted regeneration of corneal epithelium, stroma, and nerves; some myofibroblasts were seen in the regenerated neo-corneas. Conclusion Plant-derived RHCI was used to fabricate a hydrogel that is transparent, mechanically stable, and biocompatible when grafted as corneal implants in minipigs. Plant-derived collagen is determined to be a safe alternative to allografts, animal collagens, or yeast-derived recombinant human collagen for tissue engineering applications. The main advantage is that unlike donor corneas or yeast-produced collagen, the RHCI supply is potentially unlimited due to the high yields of this production method. Lay Summary A severe shortage of human-donor corneas for transplantation has led scientists to develop synthetic alternatives. Here, recombinant human collagen type I made of tobacco plants through genetic engineering was tested for use in making corneal implants. We made strong, transparent hydrogels that were tested by implanting subcutaneously in rats and in the corneas of minipigs. We showed that the plant collagen was biocompatible and was able to stably regenerate the corneas of minipigs comparable to yeast-produced recombinant collagen that we previously tested in clinical trials. The advantage of the plant collagen is that the supply is potentially limitless.

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