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)

Generation of Differentiating Human Epidermal Cells on Basement Membrane Matrigel: Potential for use as Autografts in Recurrent Ulcerative Lesions

1987 ◽  
Vol 110 ◽  
Author(s):  
Dorothy G. Walker-Jones ◽  
C. A. Reindorf ◽  
E. Massac ◽  
A. D. Adekile ◽  
M. G. Hall ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Sickle Cell ◽  
Type I Collagen ◽  
Epidermal Cells ◽  
Type I ◽  
Electron Microscopic ◽  
Human Epidermal Cells ◽  
Ulcerative Lesions ◽  
Rapid Generation

AbstractThe successful use of in vitro-generated skin autografts in the treatment of burns in humans (Green and O'Connor, 1984) has advanced the potential for application in the treatment of recurrent ulcerative lesions that occur in some patients with sickle cell disease and other disorders. Conventional grafts frequently break down in these patients, requiring repeated grafting that may eventually result in the lack of suitable donor sites. Preliminary studies show that Type I collagen (Collistat) promotes healing of sickle cell-lesions but with extensive scarring and loss of pigmentation (Adekile et al., 1987). Therefore, the primary aim of this study was the development of a culture procedure for the rapid generation of differentiating human epidermal cells on a biomatrix for use in conjunction with the Collistat, and later, the establishment of a frozen culture bank. Dispasefollowed by trypsin-dissociated inner epidermal cells from dermatome sections of skin of adult patients undergoing elective plastic surgery were cultured on basement membrane-derived Matrigel (Collaborative Research, Inc.) coated onto plastic coverslips. Growth was established in RPMI1640 or IMEM supplemented with insulin, transferrin, hydrocortisone and bovine pituitary extract or Keratinocyte Growth Medium (KGM)a nd from 0.5–2.0% fetal bovine serum. The epidermal cells exhibited active migration and the formation of bi- and tri-layered aggregates by day 3. Large multilayered aggregates were produced in KGM with 1.16 mM CaCl2 added. Electron microscopic observations revealed increased numbers of desmosomal junctions, interdigitations and apical-basal polarity in these aggregates. A distinguishing feature ofthese and other cells grown on Matrigel is the absence of the degree of spreading that is characteristic of cells grown on plastic. It is noteworthy that the epidermal cells did not invade through the Matrigel.

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.

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.

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.

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.

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