Cell-matrix interaction in bone: type I collagen modulates signal transduction in osteoblast-like cells

1995 ◽  
Vol 268 (5) ◽  
pp. C1090-C1103 ◽  
Author(s):  
J. Green ◽  
S. Schotland ◽  
D. J. Stauber ◽  
C. R. Kleeman ◽  
T. L. Clemens
Keyword(s):  
Signal Transduction ◽  
Signaling Pathways ◽  
Type I Collagen ◽  
Primary Cultures ◽  
Neonatal Rat ◽  
Type I ◽  
Osteosarcoma Cell ◽  
Umr 106 ◽  
In Cells

Cell interaction with extracellular matrix (ECM) modulates cell growth and differentiation. By using in vitro culture systems, we tested the effect of type I collagen (Coll-I) on signal transduction mechanisms in the osteosarcoma cell line UMR-106 and in primary cultures from neonatal rat calvariae. Cells were cultured for 72 h on Coll-I gel matrix and compared with control cells plated on plastic surfaces. Agonist-dependent and voltage-dependent rises in cytosolic Ca2+ concentration ([Ca2+]i; measured by fura 2 fluorometry) were significantly blunted in cells cultured on Coll-I compared with cells grown on plastic. In UMR-106 cells, the collagen matrix effect was mimicked by 24-h incubation with soluble Coll-I or short peptides containing the arginine-glycine-aspartate motif. Accumulation of cellular adenosine 3',5'-cyclic monophosphate (cAMP) stimulated by parathyroid hormone, cholera toxin, and forskolin was augmented (50-150%) in cells plated on Coll-I vs. control. The collagen effect on both [Ca2+]i- and adenylate cyclase-signaling pathways in UMR-106 cells was abrogated in the presence of protein kinase C (PKC) depletion or inhibition. Also, Coll-I induced a twofold increase in membrane-bound PKC without changing cytosolic PKC activity. Thus, by altering PKC activity, Coll-I modulates the [Ca2+]i- and cAMP-signaling pathways in osteoblasts. This, in turn, may influence bone remodeling processes.

Morphological and Functional Characteristics of Three-Dimensional Engineered Bone-Ligament-Bone Constructs Following Implantation

2009 ◽  
Vol 131 (10) ◽  
Author(s):  
Jinjin Ma ◽  
Kristen Goble ◽  
Michael Smietana ◽  
Tatiana Kostrominova ◽  
Lisa Larkin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Type I Collagen ◽  
Three Dimensional ◽  
Functionally Graded ◽  
Neonatal Rat ◽  
Ligament Injury ◽  
Type I ◽  
Tangent Moduli

The incidence of ligament injury has recently been estimated at 400,000/year. The preferred treatment is reconstruction using an allograft, but outcomes are limited by donor availability, biomechanical incompatibility, and immune rejection. The creation of an engineered ligament in vitro solely from patient bone marrow stromal cells (has the potential to greatly enhance outcomes in knee reconstructions. Our laboratory has developed a scaffoldless method to engineer three-dimensional (3D) ligament and bone constructs from rat bone marrow stem cells in vitro. Coculture of these two engineered constructs results in a 3D bone-ligament-bone (BLB) construct with viable entheses, which was successfully used for medial collateral ligament (MCL) replacement in a rat model. 1 month and 2 month implantations were applied to the engineered BLBs. Implantation of 3D BLBs in a MCL replacement application demonstrated that our in vitro engineered tissues grew and remodeled quickly in vivo to an advanced phenotype and partially restored function of the knee. The explanted 3D BLB ligament region stained positively for type I collagen and elastin and was well vascularized after 1 and 2 months in vivo. Tangent moduli of the ligament portion of the 3D BLB 1 month explants increased by a factor of 2.4 over in vitro controls, to a value equivalent to those observed in 14-day-old neonatal rat MCLs. The 3D BLB 1 month explants also exhibited a functionally graded response that closely matched native MCL inhomogeneity, indicating the constructs functionally adapted in vivo.

scholarly journals Macromolecular Organization and In Vitro Growth Characteristics of Scaffold-free Neocartilage Grafts

2007 ◽  
Vol 55 (8) ◽  
pp. 853-866 ◽  
Author(s):  
Anthony J. Hayes ◽  
Amanda Hall ◽  
Liesbeth Brown ◽  
Ross Tubo ◽  
Bruce Caterson
Keyword(s):  
Articular Cartilage ◽  
Type I Collagen ◽  
Primary Cultures ◽  
Surface Zone ◽  
Type I ◽  
Focal Lesions ◽  
Electron Microscopic ◽  
Chondrocyte Phenotype ◽  
Macromolecular Organization

Recent advances in tissue engineering offer considerable promise for the repair of focal lesions in articular cartilage. Here we describe (1) the macromolecular organization of tissue-engineered neocartilage grafts at light and electron microscopic levels, (2) their in vitro development, and (3) the effect of chondrocyte dedifferentiation, induced by monolayer expansion, on their resultant structure. We show that grafts produced from primary cultures of chondrocytes are hyaline in appearance with identifiable zonal strata as evidenced by cell morphology, matrix organization, and immunohistochemical composition. Like native articular cartilage, their surface zone contains type I collagen, surface zone proteoglycan, biglycan and decorin with type II collagen, aggrecan, chondroitin sulfate, chondroitin-4-sulfate, and keratan sulfate, becoming more prominent with depth. Assessment of cell viability by Live/Dead staining and cell-cycle analysis with BrDU suggest that the in vitro tissue has a high cellular turnover and develops through both appositional and interstitial growth mechanisms. Meanwhile, cell-tracker studies with CMFDA (5-chloromethyl-fluorescein diacetate) demonstrate that cell sorting in vitro is not involved in their zonal organization. Finally, passage expansion of chondrocytes in monolayer culture causes progressive reductions in graft thickness, loss of zonal architecture, and a more fibrocartilaginous tissue histology, consistent with a dedifferentiating chondrocyte phenotype. (J Histochem Cytochem 55: 853–866, 2007)

Bioactive Glass Templates for the Synthesis of Bone-Like Tissue In Vitro

1993 ◽  
Vol 331 ◽  
Author(s):  
A. Ei-Ghannam ◽  
P. Ducheyne ◽  
I. M. Shapiro
Keyword(s):  
Bioactive Glass ◽  
Porous Glass ◽  
Type I Collagen ◽  
Neonatal Rat ◽  
Bone Lesions ◽  
Type I ◽  
Sds Page ◽  
Biochemical Analyses ◽  
Ca:P Ratio

AbstractPorous glass templates of pore size 75–200 14m and 20–30% porosity were synthesized. The glass disks were conditioned in a modified tris buffer (SBF) for 48 hrs and then treated with tissue culture medium (TCM) for 1 hr at 37°C. Other porous glass templates were treated with either SBF or TCM. The conditioned glass disks were seeded with 106 neonatal rat calvaria osteoblasts and maintained in culture for 2, 5 or 7 days. It was found that the bioactive glass was rapidly invaded by cells which colonized the porous template. Morphological and biochemical analyses suggested that a bone-like tissue was formed inside templates conditioned with both SBF and TCM. By 7 days, the cells exhibited high alkaline phosphatase activity and synthesized osteocalcin. SDS-PAGE indicated the presence of type I collagen. SEM-EDAX analysis showed the formation of a bone-like tissue throughout the entire sample, while inspection of the Ca:P ratio indicated that the cells synthesized a mineral phase. Indeed, the FTIR spectra of the mineral confirmed that it was a biological hydroxyapatite. The rapid formation of the bone-like material within the template in vitro suggests that this template may be valuable clinically for the repair of bone lesions.

Independent expression of type I collagen and fibronectin by normal fibroblast-like cells

1983 ◽  
Vol 63 (1) ◽  
pp. 233-244
Author(s):  
N.S. Connor ◽  
J.E. Aubin ◽  
J. Sodek
Keyword(s):  
Type I Collagen ◽  
Primary Cultures ◽  
Gingival Tissue ◽  
Normal Fibroblast ◽  
Type I ◽  
Gingival Fibroblasts ◽  
Connective Tissues ◽  
Double Label ◽  
Attachment Mechanism

A double-label immunofluorescence technique was used to determine whether all normal fibroblast-like cells cultured from porcine connective tissues simultaneously express both type I collagen and fibronectin (FN). Cells surrounding explants in primary cultures from gingival and periodontal ligament (PL) tissue were heterogeneous in the expression of collagen and FN. Mass population cells at first and second subculture after primary explanting of gingival tissue were also heterogeneous in the expression of collagen and FN. When clonal populations were analyzed, four distinct phenotypes were observed: Coll+/FN+;Coll+/FN-;Coll-/FN+;Coll-/FN-. Mass populations of PL cells were more homogeneous than gingival fibroblasts and when clones were analyzed only two phenotypes were observed. These data suggest that normal fibroblast-like cells in vitro are capable of expressing collagen and FN independently and that FN-negative cells may utilize a FN-independent attachment mechanism in culture.

Collagen fibrillogenesis in vitro: interaction of types I and V collagen

1986 ◽  
Vol 44 ◽  
pp. 210-211
Author(s):  
Arthur J. Wasserman ◽  
Kathy C. Kloos ◽  
David E. Birk
Keyword(s):  
Type I Collagen ◽  
Corneal Stroma ◽  
Minor Component ◽  
Homogeneous Distribution ◽  
Type I ◽  
Type V Collagen ◽  
Fibril Morphology ◽  
Type V ◽  
In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

Hyperactivation of RAP1 and JAK/STAT Signaling Pathways Contributes to Fibrosis during the Formation of Nasal Capsular Contraction

2021 ◽  
pp. 1-12
Author(s):  
Meng Wu ◽  
Ming Li ◽  
Hong-Ju Xie ◽  
Hong-Wei Liu
Keyword(s):  
Signaling Pathways ◽  
Type I Collagen ◽  
Ex Vivo ◽  
Capsular Contracture ◽  
Silicone Implant ◽  
Type I ◽  
Loss Of Function ◽  
Sequencing Data ◽  
Functional Changes ◽  
Stat Signaling

Silicone implant-based augmentation rhinoplasty or mammoplasty induces capsular contracture, which has been acknowledged as a process that develops an abnormal fibrotic capsule associated with the immune response to allogeneic materials. However, the signaling pathways leading to the nasal fibrosis remain poorly investigated. We aimed to explore the molecular mechanism underlying the pathogenesis of nasal capsular contracture, with a specific research interest in the signaling pathways involved in fibrotic development at the advanced stage of contracture. By examining our recently obtained RNA sequencing data and global gene expression profiling between grade II and grade IV nasal capsular tissues, we found that both the RAP1 and JAK/STAT signaling pathways were hyperactive in the contracted capsules. This was verified on quantitative real-time PCR which demonstrated upregulation of most of the representative component signatures in these pathways. Loss-of-function assays through siRNA-mediated Rap1 silencing and/or small molecule-directed inhibition of JAK/STAT pathway in ex vivo primary nasal fibroblasts caused a series of dramatic behavioral and functional changes, including decreased cell viability, increased apoptosis, reduced secretion of proinflammatory cytokines, and synthesis of type I collagen, compared to control cells, and indicating the essential role of the RAP1 and JAK/STAT signaling pathways in nasal capsular fibrosis. Our results sheds light on targeting downstream signaling pathways for the prevention and therapy of silicone implant-induced nasal capsular contracture.

scholarly journals Restoration of Osteogenesis by CRISPR/Cas9 Genome Editing of the Mutated COL1A1 Gene in Osteogenesis Imperfecta

2021 ◽  
Vol 10 (14) ◽  
pp. 3141
Author(s):  
Hyerin Jung ◽  
Yeri Alice Rim ◽  
Narae Park ◽  
Yoojun Nam ◽  
Ji Hyeon Ju
Keyword(s):  
Osteogenesis Imperfecta ◽  
Type I Collagen ◽  
Disease Modeling ◽  
Bone Fragility ◽  
Osteogenic Potential ◽  
Type I ◽  
Gene Correction ◽  
Col1a1 Gene ◽  
Collagen Formation

Osteogenesis imperfecta (OI) is a genetic disease characterized by bone fragility and repeated fractures. The bone fragility associated with OI is caused by a defect in collagen formation due to mutation of COL1A1 or COL1A2. Current strategies for treating OI are not curative. In this study, we generated induced pluripotent stem cells (iPSCs) from OI patient-derived blood cells harboring a mutation in the COL1A1 gene. Osteoblast (OB) differentiated from OI-iPSCs showed abnormally decreased levels of type I collagen and osteogenic differentiation ability. Gene correction of the COL1A1 gene using CRISPR/Cas9 recovered the decreased type I collagen expression in OBs differentiated from OI-iPSCs. The osteogenic potential of OI-iPSCs was also recovered by the gene correction. This study suggests a new possibility of treatment and in vitro disease modeling using patient-derived iPSCs and gene editing with CRISPR/Cas9.

scholarly journals Effects of chitosan-collagen dressing on wound healing in vitro and in vivo assays

2021 ◽  
Vol 19 ◽  
pp. 228080002198969
Author(s):  
Min-Xia Zhang ◽  
Wan-Yi Zhao ◽  
Qing-Qing Fang ◽  
Xiao-Feng Wang ◽  
Chun-Ye Chen ◽  
...  
Keyword(s):  
Wound Healing ◽  
Wound Dressing ◽  
Type I Collagen ◽  
Inhibition Zone ◽  
Negative Control ◽  
Type I ◽  
Nih3t3 Cells ◽  
Post Operation

The present study was designed to fabricate a new chitosan-collagen sponge (CCS) for potential wound dressing applications. CCS was fabricated by a 3.0% chitosan mixture with a 1.0% type I collagen (7:3(w/w)) through freeze-drying. Then the dressing was prepared to evaluate its properties through a series of tests. The new-made dressing demonstrated its safety toward NIH3T3 cells. Furthermore, the CCS showed the significant surround inhibition zone than empty controls inoculated by E. coli and S. aureus. Moreover, the moisture rates of CCS were increased more rapidly than the collagen and blank sponge groups. The results revealed that the CCS had the characteristics of nontoxicity, biocompatibility, good antibacterial activity, and water retention. We used a full-thickness excisional wound healing model to evaluate the in vivo efficacy of the new dressing. The results showed remarkable healing at 14th day post-operation compared with injuries treated with collagen only as a negative control in addition to chitosan only. Our results suggest that the chitosan-collagen wound dressing were identified as a new promising candidate for further wound application.

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