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.

scholarly journals Effect of acute resistance exercise and sex on human patellar tendon structural and regulatory mRNA expression

2009 ◽  
Vol 106 (2) ◽  
pp. 468-475 ◽  
Author(s):  
Bridget E. Sullivan ◽  
Chad C. Carroll ◽  
Bozena Jemiolo ◽  
Scott W. Trappe ◽  
S. Peter Magnusson ◽  
...  
Keyword(s):  
Resistance Exercise ◽  
Mrna Expression ◽  
Patellar Tendon ◽  
Type I Collagen ◽  
Collagen Type ◽  
Collagen Type I ◽  
Tissue Inhibitors Of Metalloproteinases ◽  
Type I ◽  
Collagen Type Iii ◽  
Type Iii

Tendon is mainly composed of collagen and an aqueous matrix of proteoglycans that are regulated by enzymes called matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Although it is known that resistance exercise (RE) and sex influence tendon metabolism and mechanical properties, it is uncertain what structural and regulatory components contribute to these responses. We measured the mRNA expression of tendon's main fibrillar collagens (type I and type III) and the main proteoglycans (decorin, biglycan, fibromodulin, and versican) and the regulatory enzymes MMP-2, MMP-9, MMP-3, and TIMP-1 at rest and after RE. Patellar tendon biopsy samples were taken from six individuals (3 men and 3 women) before and 4 h after a bout of RE and from a another six individuals (3 men and 3 women) before and 24 h after RE. Resting mRNA expression was used for sex comparisons (6 men and 6 women). Collagen type I, collagen type III, and MMP-2 were downregulated ( P < 0.05) 4 h after RE but were unchanged ( P > 0.05) 24 h after RE. All other genes remained unchanged ( P > 0.05) after RE. Women had higher resting mRNA expression ( P < 0.05) of collagen type III and a trend ( P = 0.08) toward lower resting expression of MMP-3 than men. All other genes were not influenced ( P > 0.05) by sex. Acute RE appears to stimulate a change in collagen type I, collagen type III, and MMP-2 gene regulation in the human patellar tendon. Sex influences the structural and regulatory mRNA expression of tendon.

scholarly journals Connective tissue, glial and neuronal expressions in testis of the African giant rat (Cricetomys gambianus)

2017 ◽  
Vol 34 (03) ◽  
pp. 186-193
Author(s):  
T. Falade ◽  
M. Olude ◽  
O. Mustapha ◽  
E. Mbajiorgu ◽  
A. Ihunwo ◽  
...  
Keyword(s):  
Connective Tissue ◽  
Type I Collagen ◽  
Collagen Type ◽  
Neuronal Cells ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Neuronal Markers ◽  
Type Iii ◽  
African Giant Rat

Abstract Introduction: This study was carried out to investigate the expression of connective tissue (Collagens I and III), glia and neuronal markers in the testis of the African giant rat using histology and immunohistochemistry techniques. Materials and Methods: Eight (8) apparently healthy wild male African giant rats were used for this experiment, divided into 2 groups (juvenile and adult) of 4 animals each. The testes were harvested following intracardial perfusion of the rats and histology was performed using Haematoxylin-Eosin stain and Mallory-Heideinhain rapid one- step staining for connective tissue. Immunohistochemical identification was achieved using the following antibodies: anti-collagen type I, anti-collagen type III, anti-glial fibrillary acidic protein and anti-p75 nerve growth factor for the expression of collagen type I, collagen type III, astrocyte-like cell and neuronal cells respectively. Photomicrography was achieved using Axioskop® microscope and quantitative data were analyzed using student t-test. Results: The cyto-architecture of the testis was typical in the African giant rat. The connective tissue expressed in the juvenile and adult group, signaling of glial-like cells were seen in the perivascular region across the experimental groups. Immuno-localization of neuronal cells were seen in the interstitial spaces across all the groups, but with more expressions in the juvenile. Conclusion: This work has provided a clear description of the expression of connective tissue, neuronal and glial cells in the testis of the African giant rat and their possible relationships across juvenile and adult groups.

Enhancement by Heparin of Affinity between Cold-Insoluble Globulin and Native Collagen Type III

1979 ◽  
Author(s):  
H. Hörmann ◽  
F. Jilek
Keyword(s):  
Collagen Type ◽  
Weight Ratio ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Type Iii ◽  
Soluble Collagen ◽  
Native Collagen ◽  
Collagen Molecules ◽  
Cold Insoluble Globulin

Affinity between collagen and cold-insoluble globulin was measured by complexing soluble 125-J labelled collagen preparations with the globulin. Precipitates containing considerable activity were formed at 4°C and 22°C by denatured soluble collagen, type I and type III, but only little by native soluble collagen. The precipitation of native collagen, type III, by cold-insoluble globulin was enhanced by heparin. Under optimal conditions at a weight ratio or heparin and cold-insoluble globulin of about 1:1 up to 60% of the collagen applied was insolubilized. Native collagen, type I, was complexed far less effectively even in presence of heparin. Electronmicroscopic and precipitation experiments using 125-J labelled cold-insoluble globulin indicated that heparin might induce a partial conversion of cold-insoluble globulin to a fibrillar derivative which exhibited improved binding properties for the rod-like native collagen molecules. – Supported by Deutsche Forschungsgemeinschaft, Project Ho 740/1.

scholarly journals Immunohistochemical distribution of collagens types IV, V, and VI and of pro-collagens types I and III in human alveolar bone and dentine.

1986 ◽  
Vol 34 (11) ◽  
pp. 1417-1429 ◽  
Author(s):  
J Becker ◽  
D Schuppan ◽  
H Benzian ◽  
T Bals ◽  
E G Hahn ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Collagen Type ◽  
Alveolar Bone ◽  
Organic Matrix ◽  
Collagen Type I ◽  
Basement Membranes ◽  
Type I ◽  
Collagen Type Iii ◽  
Type Iii ◽  
Hard Tissues

The aim of the present study was to characterize the composition of the organic matrix in alveolar jaw bone and dentine using antibodies against pro-collagens Types I and III and collagens Types IV, V, and VI. After demineralization of oral hard tissues in 0.2 N HCl, antigenicity was well preserved and the distribution of the pro-collagens and collagens could be demonstrated. Staining for pro-collagen Type I was prominent around osteoblasts and in pre-dentine, indicating active de novo synthesis of Type I pro-collagen. Pro-collagen Type I was ubiquitous but was less abundant in bone and dentine, whereas pro-collagen Type III was seen only in areas of bone remodeling, in peritubular spaces, and in pre-dentine. Type IV collagen was limited to the basement membranes of vessels in osteons and bone marrow. Type V collagen was detected neither in pre-dentine nor in bone. In contrast, Type VI collagen was found in dentine and bone, showing a faint but homogeneous staining which, similarly to pro-collagen Type III, was pronounced around osteoblasts and in pre-dentine, areas of active bone and dentine formation. This study showed that the organic matrix of dentine and bone contains Type VI as well as Type I collagen. Pro-collagen Type III (and to a lesser extent collagen Type VI) is transiently produced during new formation and remodeling of oral hard tissues, and disappears once the matrix calcifies. Type I pro-collagen qualifies as a general marker protein for increased osteoblastic activity. We conclude that immunostaining for the different collagen/pro-collagen types can be used to assess normal or abnormal stages of bone/dentine formation.

Topological Differences in the Expression of Collagen Type I and Collagen Type III mRNAs in the Rat Gingiva

1994 ◽  
Vol 65 (8) ◽  
pp. 776-780 ◽  
Author(s):  
Meir Redlich ◽  
Itzhak Peleg ◽  
Helena Cooperman ◽  
Shmuel Shoshan
Keyword(s):  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Type Iii

Long-Term in Vitro Cytokine-Free and Serum-Free Culture of Human Cord Blood Mononuclear Cells in a Three-Dimensional Scaffold.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 503-503
Author(s):  
Teresa Mortera-Blanco ◽  
Athanasios Mantalaris ◽  
Alexander Bismarck ◽  
Nicki Panoskaltsis
Keyword(s):  
Bone Marrow ◽  
Cord Blood ◽  
Clinical Application ◽  
Mononuclear Cells ◽  
Collagen Type ◽  
Three Dimensional ◽  
Collagen Type I ◽  
Type I ◽  
Serum Free

Abstract Abstract 503 The ability to expand cord blood (CB) cells ex vivo overcomes an important limitation to its wider clinical application in cellular therapies. The current practice of hematopoietic cell culture is based on two-dimensional (2D) tissue culture flasks or well plates which require either co-culture with allogeneic or xenogeneic stromal cells and the exogenous provision of several cytokines. This 2D culture environment is artificial and lacks the 3D cellular niches that characterise the in vivo hematopoietic inductive microenvironment. Specifically, the cultured cells are exposed to abnormally high cytokine concentrations, which may result in differentiation and loss of pluripotency. We have previously developed a 3D bone marrow biomimicry through the use of synthetic scaffolds made of poly (D,L-lactide-co-glycolide) (PLGA) and polyurethane (PU) coated with collagen type I. Our previous work has shown that these scaffolds, which were seeded with cord blood (CB) mononuclear cells (MNCs) at a cell density of 3-6×106cells per scaffold (5×5×5mm3), could successfully support long-term culture in the absence of exogenous growth factors for over 4 weeks. Specifically, the 3D biomimicry facilitated a 53-fold total MNC expansion, with an increase in the BFU-E and CFU-GM progenitor cell population. However, these cultures, although cytokine-free, contained 20-30% (v/v) fetal calf serum which can have both conducive and inhibitory effects on hematopoietic cell cultures due to the unknown composition and concentration of humoral factors contained within. Inclusion of serum in expansion-type cultures can limit the clinical application of the derived product. The serum-free and cytokine-free culture and expansion of hematopoietic cells has not been achieved until now. Herein, we report that for at least 4 weeks the polyurethane (PU) scaffolds coated with collagen type I were able to maintain and expand human CB MNCs. Furthermore the progenitor population, as determined by the colony forming unit assay, was also maintained and preferentially directed towards the granulocytic lineage, even though the CFU-GEMMs declined. Immunophenotypic analysis of the extracted cells confirmed the presence of erythroid precursors (CD71+CD45-) as well as early maturing myeloid cells. In contrast, the 2D cytokine- and serum-free cultures collapsed within 3-4 days. We hypothesized that the 3D biomimicry was able to facilitate serum- and cytokine-free conditions because it can recapitulate the three-dimensional architecture of the human bone marrow. This hypothesis was supported by scanning electron microscopy of the central sections of the scaffolds that showed the migration of cells within the pores and establishment of “niche-like” structures. In conclusion, this novel 3D culture system is capable of long-term, cytokine- and serum-free expansion of haematopoietic cells from cord blood, enabling the study of haematopoiesis as well as facilitating the expansion of cells for future clinical applications. Disclosures: No relevant conflicts of interest to declare.

Imbalance in the Synthesis of Collagen Type I and Collagen Type III in Smooth Muscle Cells Derived from Human Varicose Veins

2001 ◽  
Vol 38 (6) ◽  
pp. 560-568 ◽  
Author(s):  
Patricia Sansilvestri-Morel ◽  
Alain Rupin ◽  
Cécile Badier-Commander ◽  
Patrick Kern ◽  
Jean-Noël Fabiani ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Collagen Type ◽  
Varicose Veins ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Type Iii
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