scholarly journals Comparative Evaluation of Fibrin Gel and Platelet-Rich Fibrin on Stem Cells of the Apical Papilla Fate Using a Dentin-Based Model

2022 ◽  
Author(s):  
Pegah Sarraf ◽  
Razieh Sadat Moayeri ◽  
Noushin Shokouhinejad ◽  
Mehrfam Khoshkhounejad ◽  
Roya Karimi ◽  
...  
Keyword(s):  
Mtt Assay ◽  
Matrix Protein ◽  
Scratch Test ◽  
Collagen Type I ◽  
Type I ◽  
Fibrin Gel ◽  
Special Equipment ◽  
Great Ability ◽  
Dentin Matrix Protein ◽  
And Migration

Abstract Background: PRF as one of the favorable scaffolds in Regenerative Endodontic Treatment (RET), has several limitations such as the need for blood sampling and special equipment. High available commercial scaffolds such as fibrin are able to meet all the necessary requirements of dentin tissue engineering. The present study was designed to evaluate the effect of PRF and fibrin gel, with and without the presence of EDTA-treated radicular dentin segments on SCAP viability, proliferation, migration, and differentiation.Methods: Radicular dentin were prepared from extracted teeth and treated by EDTA 17% .The samples were divided into 6 groups: Dentin/PRF/Cell, Dentin/Fibrin/Cell, Dentin/Cell, PRF/Cell, Fibrin/Cell and Cell (Control). SCAP viability was assessed using MTT assay. Gene expression levels of odontogenic markers [Dentin sialophosphoprotein (DSPP), Dentin matrix protein 1(DMP1), Collagen type I Alpha 1(COL 1A1) and Alkaline phosphatase (ALP) were assessed using qrt-PCR. Cell migration were also evaluated by means of scratch test. Results: The results of MTT assay at showed that the viability of SCAP significantly increased after 7 days for both groups containing fibrin (P <0.05). The viability of SCAP seeded on Dentin/PRF and PRF significantly decreased after 7 days (P <0.001). The odontogenic markers were significantly expressed for both scaffolds in the presence of dentin segment (p<0.05). Significant decrease in scratch area was seen in Fibrin/Dentin group (p < 0.001)Conclusions:Fibrin beside EDTA-treated dentin showed great ability in survival, proliferation, differentiation, and migration of SCAP rather than PRF.

Asporin competes with decorin for collagen binding, binds calcium and promotes osteoblast collagen mineralization

2009 ◽  
Vol 423 (1) ◽  
pp. 53-59 ◽  
Author(s):  
Sebastian Kalamajski ◽  
Anders Aspberg ◽  
Karin Lindblom ◽  
Dick Heinegård ◽  
Åke Oldberg
Keyword(s):  
Matrix Protein ◽  
Collagen Type I ◽  
Full Length ◽  
Extracellular Matrix Protein ◽  
Type I ◽  
Similar System ◽  
Collagen Binding ◽  
Osteoblastic Activity ◽  
Collagen Mineralization

The interactions of the ECM (extracellular matrix) protein asporin with ECM components have previously not been investigated. Here, we show that asporin binds collagen type I. This binding is inhibited by recombinant asporin fragment LRR (leucine-rich repeat) 10–12 and by full-length decorin, but not by biglycan. We demonstrate that the polyaspartate domain binds calcium and regulates hydroxyapatite formation in vitro. In the presence of asporin, the number of collagen nodules, and mRNA of osteoblastic markers Osterix and Runx2, were increased. Moreover, decorin or the collagen-binding asporin fragment LRR 10–12 inhibited the pro-osteoblastic activity of full-length asporin. Our results suggest that asporin and decorin compete for binding to collagen and that the polyaspartate in asporin directly regulates collagen mineralization. Therefore asporin has a role in osteoblast-driven collagen biomineralization activity. We also show that asporin can be expressed in Escherichia coli (Rosetta-gami™) with correctly positioned cysteine bridges, and a similar system can possibly be used for the expression of other SLRPs (small LRR proteoglycans/proteins).

Investigation of the effects of prostaglandin E2 on equine superficial digital flexor tendon fibroblasts in vitro

2010 ◽  
Vol 23 (06) ◽  
pp. 417-423 ◽  
Author(s):  
J. M. Cissell ◽  
S. C. Milton ◽  
L. A. Dahlgren
Keyword(s):  
Gene Expression ◽  
Matrix Protein ◽  
Collagen Type ◽  
Flexor Tendon ◽  
Cell Metabolism ◽  
Collagen Type I ◽  
Type I ◽  
Matrix Metalloproteinase 1 ◽  
Superficial Digital Flexor Tendon

Summary Objectives: To evaluate the effects of pros-taglandin E2 (PGE2) treatment on the metabolism of equine tendon fibroblasts in vitro to aid in investigating the response of tendon fibroblasts to injury and novel therapeutics. Methods: Superficial digital flexor tendon fibroblasts isolated via collagenase digestion from six young adult horses were grown in monolayer in four concentrations of PGE2 (0, 10, 50, 100 ng/ml) for 48 hours. Cells and medium were harvested for gene expression (collagen types I and III, cartilage oligomeric matrix protein [COMP], decorin, and matrix metalloproteinase-1, –3, and –13), biochemical analysis (glycosaminoglycan, DNA, and collagen content), and cytological staining. Results: Gene expression for collagen type I was significantly increased at 100 ng/ml PGE2 compared to 10 and 50 ng/ml. There were not any significant differences detected for gene expression of collagen type III, COMP or dec-orin or for biochemical content and cell morphology. Clinical significance: Under the conditions investigated, exogenous treatment of equine tendon fibroblasts with PGE2 failed to alter cell metabolism in a manner useful as a model of tendon injury. A model that applies cyclic strain to a three dimensional construct seeded with tendon fibroblasts may prove to be a more useful model and merits further investigation for this purpose. The ability to assess cellular responses in an environment where the cells are supported within the extracellular matrix may prove beneficial.

scholarly journals Effect of a Collagen-Based Compound on Morpho-Functional Properties of Cultured Human Tenocytes

Cells ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 246 ◽  
Author(s):  
Filippo Randelli ◽  
Alessandra Menon ◽  
Alessio Giai Via ◽  
Manuel Mazzoleni ◽  
Fabio Sciancalepore ◽  
...  
Keyword(s):  
Matrix Metalloproteinases ◽  
Collagen Type ◽  
Clinical Investigation ◽  
Pain Syndrome ◽  
Collagen Type I ◽  
Type I ◽  
Mrna Levels ◽  
Collagen Turnover ◽  
And Migration ◽  
Proliferation And Migration

Background: Greater Trochanter Pain Syndrome (GTPS) is the main reason for recalcitrant lateral hip pain. Gluteus medius and minimus tendinopathy plays a key role in this setting. An injectable medical compound containing collagen type I (MD-Tissue, Guna) has been produced with the aim to counteract the physiological and pathological degeneration of tendons. In this study we aimed at characterizing the effect of this medical compound on cultured human gluteal tenocytes, focusing on the collagen turnover pathways, in order to understand how this medical compound could influence tendon biology and healing. Methods: Tenocytes were obtained from gluteal tendon fragments collected in eight patients without any gluteal tendon pathology undergoing total hip replacement through an anterior approach. Cell proliferation and migration were investigated by growth curves and wound healing assay, respectively. The expression of genes and proteins involved in collagen turnover were analysed by real-time PCR, Slot blot and SDS-zymography. Results: Our data show that tenocytes cultured on MD-Tissue, compared to controls, have increased proliferation rate and migration potential. MD-Tissue induced collagen type I (COL-I) secretion and mRNA levels of tissue inhibitor of matrix metalloproteinases (MMP)-1 (TIMP-1). Meanwhile, lysyl hydroxylase 2b and matrix metalloproteinases (MMP)-1 and -2, involved, respectively, in collagen maturation and degradation, were not affected. Conclusions: Considered as a whole, our results suggest that MD-Tissue could induce in tenocytes an anabolic phenotype by stimulating tenocyte proliferation and migration and COL-I synthesis, maturation, and secretion, thus favouring tendon repair. In particular, based on its effect on gluteal tenocytes, MD-Tissue could be effective in the discouraging treatment of GTPS. From now a rigorous clinical investigation is desirable to understand the real clinical potentials of this compound.

scholarly journals In situ localization and chromosomal mapping of the AG1 (Dmp1) gene.

1994 ◽  
Vol 42 (12) ◽  
pp. 1527-1531 ◽  
Author(s):  
A George ◽  
J Gui ◽  
N A Jenkins ◽  
D J Gilbert ◽  
N G Copeland ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Type I Collagen ◽  
Chromosomal Mapping ◽  
Human Tooth ◽  
Type I ◽  
Dentin Matrix Protein ◽  
Biomineralization Process ◽  
Tooth Mineralization ◽  
The Matrix

Dentinogenesis is being used as a model for understanding the biomineralization process. The odontoblasts synthesize a structural matrix comprised of Type I collagen fibrils which define the basic architecture of the tissue. The odontoblasts also synthesize and deliver a number of dentin-specific acidic macromolecules into the extracellular compartment. These acidic macromolecules may be involved in regulating the ordered deposition of hydroxyapatite crystals within the matrix. AG1 is the first tooth-specific acidic macromolecule to have been cloned and sequenced. To identify which cells of the rat incisor pulp/odontoblast complex were responsible for synthesis of AG1, in situ hybridization was used. Digoxigenin labeled sense and anti-sense AG1 riboprobes were prepared. The AG1 mRNA was found to be expressed in the mature secretory odontoblasts. Neither pulp cells nor pre-odontoblasts showed any staining with the anti-sense probes. Chromosomal localization studies placed the AG1 gene on mouse chromosome 5q21, in tight linkage with Fgf5. AG1 has been renamed Dmp1 (dentin matrix protein 1) in accordance with present chromosomal nomenclature. Mouse 5q21 corresponds to the 4q21 locus in humans. This is the locus for the human tooth mineralization disorder dentinogenesis imperfecta Type II (DI-II). These data suggest that the Dmp1 gene is involved in mineralization and is a candidate gene for DI-II.

β1 integrin-extracellular matrix protein interaction modulates the migratory response to chemokine stimulation

2001 ◽  
Vol 79 (4) ◽  
pp. 399-407 ◽  
Author(s):  
Priti S Shenoy ◽  
Shashi Uniyal ◽  
Kohei Miura ◽  
Christopher McColl ◽  
Tamas Oravecz ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Collagen Type ◽  
Cell Movement ◽  
Collagen Type I ◽  
Matrix Proteins ◽  
Extracellular Matrix Protein ◽  
Type I ◽  
Migratory Response ◽  
Cc Chemokine Receptor 5

It is well established that chemokines have a major role in the stimulation of cell movement on extracellular matrix (ECM) substrates. However, it is also clear that ECM substrates may influence the ability of cells to undergo migration. Using the migration chamber method, we assessed the migratory response of human embryonic kidney-293 (HEK) transfectant cells expressing the CC chemokine receptor 5 (CCR5) (HEK-CCR5) to stimulation by chemokines (macrophage inflamatory protein (MIP)-1α, MIP-1β, and regulated on activation normal-T cell expressed and secreted (RANTES)) on ECM substrates (collagen type I and fibronectin). Using filters coated with collagen (20 µg/mL), results showed that the chemokines differed in their ability to elicit cell movement according to the order MIP-1β > RANTES [Formula: see text] MIP-1α. In contrast, using filters coated with fibronectin (20 µg/mL), all three chemokines were similar in their ability to stimulate migration of HEK-CCR5 cells. In addition, the migratory response with respect to the concentrations of ECM substrates appeared biphasic; thus, chemokine-stimulated cell movement was inhibited at high ECM concentrations (100 µg/mL). To determine the involvement of β1 integrins, results showed that the migratory response to chemokine stimulation on collagen was largely inhibited by monoclonal antibody (mAb) to α2β1; however, complete inhibition required a combination of mAbs to α1β1 and α2β1. In comparison, migration on fibronectin was inhibited by mAb to α3β1 and α5β1. Our results suggest that the migratory response to CCR5 stimulation may vary quantitatively with both the CCR5 ligand (MIP-1α, MIP-1β, and RANTES), as well as the nature and concentration of the ECM substrate involved.Key words: chemokines, integrins, cell movement, extracellular matrix proteins, CCR5.

Type I collagen regulated dentin matrix protein-1 (Dmp-1) and osteocalcin (OCN) gene expression of rat dental pulp cells

2003 ◽  
Vol 88 (6) ◽  
pp. 1112-1119 ◽  
Author(s):  
Morimichi Mizuno ◽  
Tetsuro Miyamoto ◽  
Keinoshin Wada ◽  
Sanae Watatani ◽  
Gui Xia Zhang
Keyword(s):  
Gene Expression ◽  
Dental Pulp ◽  
Matrix Protein ◽  
Type I Collagen ◽  
Type I ◽  
Dental Pulp Cells ◽  
Dentin Matrix Protein 1 ◽  
Dentin Matrix Protein ◽  
Pulp Cells ◽  
Dentin Matrix

scholarly journals X-radiation enhances the collagen type I strap formation and migration potentials of colon cancer cells

Oncotarget ◽  
2016 ◽  
Vol 7 (44) ◽  
pp. 71390-71399 ◽  
Author(s):  
Stephanie Blockhuys ◽  
Na Liu ◽  
Nisha Rani Agarwal ◽  
Annika Enejder ◽  
Vesa Loitto ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cancer Cells ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Colon Cancer Cells ◽  
And Migration

scholarly journals Immunocytochemical detection of dentin matrix proteins in primary teeth from patients with dentinogenesis imperfecta associated with osteogenesis imperfecta

2014 ◽  
Author(s):  
G. Orsini ◽  
A. Majorana ◽  
A. Mazzoni ◽  
A. Putignano ◽  
M. Falconi ◽  
...  
Keyword(s):  
Osteogenesis Imperfecta ◽  
Matrix Protein ◽  
Primary Teeth ◽  
Type I Collagen ◽  
Immunohistochemical Analysis ◽  
Matrix Proteins ◽  
Type I ◽  
Dentinogenesis Imperfecta ◽  
Dentin Matrix Protein ◽  
Dentin Matrix

Dentinogenesis imperfecta determines structural alterations of the collagen structure still not completely elucidated. Immunohistochemical analysis was used to assay Type I and VI collagen, various non-collagenous proteins distribution in human primary teeth from healthy patients or from patients affected by type I dentinogenesis imperfecta (DGI-I) associated with osteogenesis imperfecta (OI). In sound primary teeth, an organized well-known ordered pattern of the type I collagen fibrils was found, whereas atypical and disorganized fibrillar structures were observed in dentin of DGI-I affected patients. Expression of type I collagen was observed in both normal and affected primary teeth, although normal dentin stained more uniformly than DGI-I affected dentin. Reactivity of type VI collagen was significantly lower in normal teeth than in dentin from DGI-I affected patients (P<0.05). Expressions of dentin matrix protein (DMP)-1 and osteopontin (OPN) were observed in both normal dentin and dentin from DGI-I affected patients, without significant differences, being DMP1 generally more abundantly expressed. Immunolabeling for chondroitin sulfate (CS) and biglycan (BGN) was weaker in dentin from DGI-I-affected patients compared to normal dentin, this decrease being significant only for CS. This study shows ultrastructural alterations in dentin obtained from patients affected by DGI-I, supported by immunocytochemical assays of different collagenous and non-collagenous proteins.

Histology and immunohistochemistry study of ovine tendon grafted with cBMSCs and BMMNCs after collagenase-induced tendinitis

2008 ◽  
Vol 21 (04) ◽  
pp. 329-336 ◽  
Author(s):  
L. Lacitignola ◽  
E. Francioso ◽  
G. Rossi ◽  
A. Crovace
Keyword(s):  
Bone Marrow ◽  
Matrix Protein ◽  
Mononuclear Cells ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Sham Control ◽  
Type Iii ◽  
Histology And Immunohistochemistry

Summary Objectives: The aim of this study was to compare the regeneration abilities of cultured bone marrow mesenchymal cells (cBMSC) and bone marrow mononuclear cells (BMMNC) with fibrin glue, saline solution and sham control in collagenase–induced tendinitis of the Achilles tendon in sheep. Methods: Six sheep were recruited randomly to each group: cBMSC, BMMNC, fibrin, saline and sham control. Each group received the relative treatment two weeks after inducing lesions (T0). After eight weeks (T8) of treatment, the tendons were harvested and evaluated for histomorphology, Collagen type I, III, Cartilage Oligomeric Matrix Protein (COMP) and CD34 positive cells expression. Results: Histology and immunohistochemistry showed similar capabilities of cBMSC and BMMNC to restore the architecture of fibres and Extra Cellular Matrix (ECM), with a high expression of collagen type I and COMP and a very low expression of collagen type III in treated tendons. The complete architectural disruption of fibres, dramatic reduction of collagen Type I and COMP expression and increase collagen type III expression were commonly observed in tendons treated with fibrin or saline only. The presence of CD34 positive cells was appreciable in the BMMNC group while few cBMSC showed this cluster of differentiation, not expressed in tendons treated with fibrin or saline. Clinical significance: The data in this study show the efficacy of cBMSC and BMMNC in regenerating tendon tissue after collagenase–induced tendinitis.

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