scholarly journals Encapsulation of Rat Bone Marrow Derived Mesenchymal Stem Cells (rBMMSCs) in Collagen Type I Containing Platelet-Rich Plasma for Osteoarthritis Treatment in Rat Model

2021 ◽  
Author(s):  
somayeh Ebrahimi-barough ◽  
Md Shahidul Islam ◽  
Mamun Al Mahtab ◽  
Sadegh Shirian ◽  
Hamid Reza Aghayan ◽  
...  
Keyword(s):  
Knee Joint ◽  
Rat Model ◽  
Collagen Type ◽  
Platelet Rich Plasma ◽  
Cartilage Damage ◽  
Collagen Type I ◽  
Joint Disease ◽  
Type I ◽  
Regenerative Effect

Abstract Osteoarthritis (OA) is the most common form of degenerative joint disease, affecting more than 25% of the adult though prevalent in the elderly population. Most of the current therapeutic modalities aim at symptomatic treatment and lingering the disease progression. In recent years, regenerative medicine such as stem cell transplantation and tissue engineering has been suggested as a potential curative intervention for OA. The objective of current study was to assess the safety and efficacy of an injectable tissue-engineered construct composed of BMMSCs, PRP, and Collagen type I in rat model of OA.To produce collagen type I, PRP and BMMSCs, male Wistar rats were ethically euthanized. After expansion and characterization of rat BMMSCs (rBMMSCs), tissue-engineered construct was formed by combination of appropriate amount of collagen type I, PRP and rBMMSCs. In vitro studies were conducted to evaluate the effect of PRP on chondrogenic differentiation capacity of encapsulated cells. Then tissue-engineered construct was injected in knee joint of rat model of OA (24 rat in 4 groups:OA, OA+MSC, ‎OA+Collagen+MSC+PRP, OA+MSC+Collagen).After 6 weeks, the animals were euthanized and knee joint histopathology examinations were performed to evaluate the effect of each treatment on OA.Tissue-engineered construct was successfully manufactured and in vitro assays demonstrated that relevant chondrogenic genes and proteins expression were higher in PRP group than the others. Histopathological findings of the knee joint samples showed favorable regenerative effect of rBMMSCs+PRP+Collagengroup comparing to others.In this study, we introduced an injectable tissue-engineered product composed of rBMMSCs+PRP+Collagenwith potential regenerative effect on cartilage damage caused by OA.

Rat Bone Marrow Derived Mesenchymal Stem Cells (rBMMSCs) Encapsulated in Collagen Type I Containing Platelet-Rich Plasma for Osteoarthritis Treatment in Rat model

2021 ◽  
Author(s):  
Md Shahidul Islam ◽  
Somayeh Ebrahimi-Barough ◽  
Mamun Al Mahtab ◽  
Sadegh Shirian ◽  
Hamid Reza Aghayan ◽  
...  
Keyword(s):  
Knee Joint ◽  
Rat Model ◽  
Collagen Type ◽  
Cartilage Damage ◽  
Collagen Type I ◽  
Joint Disease ◽  
Type I ◽  
Regenerative Effect ◽  
Collagen Group

Abstract Osteoarthritis (OA) is the most common form of degenerative joint disease, affecting more than 25% of the adult though prevalent in the elderly population. Most of the current therapeutic modalities aim at symptomatic treatment and lingering the disease progression. In recent years, regenerative medicine such as stem cell transplantation and tissue engineering has been suggested as a potential curative intervention for OA. The objective of current study was to assess the safety and efficacy of an injectable tissue-engineered construct composed of BMMSCs, PRP, and Collagen type I in rat model of OA. To produce collagen type I, PRP and BMMSCs, male Wistar rats were ethically euthanized. After expansion and characterization of rat BMMSCs (rBMMSCs), tissue-engineered construct was formed by combination of appropriate amount of collagen type I, PRP and rBMMSCs. In vitro studies were conducted to evaluate the effect of PRP on chondrogenic differentiation capacity of encapsulated cells. Then tissue-engineered construct was injected in knee joint of rat model of OA (24 rat in 4 groups: OA, OA+MSC, ‎OA+Collagen+MSC+PRP, OA+MSC+Collagen). After 6 weeks, the animals were euthanized and knee joint histopathology examinations were performed to evaluate the effect of each treatment on OA. Tissue-engineered construct was successfully manufactured and in vitro assays demonstrated that relevant chondrogenic genes and proteins expression were higher in PRP group than the others. Histopathological findings of the knee joint samples showed favorable regenerative effect of rBMMSCs+PRP+Collagen group comparing to others. In this study, we introduced an injectable tissue-engineered product composed of rBMMSCs+PRP+Collagen with potential regenerative effect on cartilage damage caused by OA.

scholarly journals Redifferentiation of Articular Chondrocytes by Hyperacute Serum and Platelet Rich Plasma in Collagen Type I Hydrogels

2019 ◽  
Vol 20 (2) ◽  
pp. 316 ◽  
Author(s):  
Vivek Jeyakumar ◽  
Eugenia Niculescu-Morzsa ◽  
Christoph Bauer ◽  
Zsombor Lacza ◽  
Stefan Nehrer
Keyword(s):  
Collagen Type ◽  
Articular Chondrocytes ◽  
Platelet Rich Plasma ◽  
Collagen Type I ◽  
Cartilage Defects ◽  
Type I ◽  
New Approach ◽  
Sgag Content ◽  
Col2a1 Expression

Matrix-assisted autologous chondrocyte transplantation (MACT) for focal articular cartilage defects often fails to produce adequate cartilage-specific extracellular matrix in vitro and upon transplantation results in fibrocartilage due to dedifferentiation during cell expansion. This study aimed to redifferentiate the chondrocytes through supplementation of blood-products, such as hyperacute serum (HAS) and platelet-rich plasma (PRP) in vitro. Dedifferentiated monolayer chondrocytes embedded onto collagen type I hydrogels were redifferentiated through supplementation of 10% HAS or 10% PRP for 14 days in vitro under normoxia (20% O2) and hypoxia (4% O2). Cell proliferation was increased by supplementing HAS for 14 days (p < 0.05) or by interchanging from HAS to PRP during Days 7–14 (p < 0.05). Sulfated glycosaminoglycan (sGAG) content was deposited under both HAS, and PRP for 14 days and an interchange during Days 7–14 depleted the sGAG content to a certain extent. PRP enhanced the gene expression of anabolic markers COL2A1 and SOX9 (p < 0.05), whereas HAS enhanced COL1A1 production. An interchange led to reduction of COL1A1 and COL2A1 expression marked by increased MMP13 expression (p < 0.05). Chondrocytes secreted less IL-6 and more PDGF-BB under PRP for 14 days (p < 0.0.5). Hypoxia enhanced TGF-β1 and BMP-2 release in both HAS and PRP. Our study demonstrates a new approach for chondrocyte redifferentiation.

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.

scholarly journals WNT-5A regulates TGF-β-related activities in liver fibrosis

2017 ◽  
Vol 312 (3) ◽  
pp. G219-G227 ◽  
Author(s):  
Leonie Beljaars ◽  
Sara Daliri ◽  
Christa Dijkhuizen ◽  
Klaas Poelstra ◽  
Reinoud Gosens
Keyword(s):  
Liver Fibrosis ◽  
Functional Role ◽  
Collagen Type ◽  
Collagen Type I ◽  
Matrix Proteins ◽  
Type I ◽  
Human Livers

WNT-5A is a secreted growth factor that belongs to the noncanonical members of the Wingless-related MMTV-integration family. Previous studies pointed to a connection between WNT-5A and the fibrogenic factor TGF-β warranting further studies into the functional role of WNT-5A in liver fibrosis. Therefore, we studied WNT-5A expressions in mouse and human fibrotic livers and examined the relation between WNT-5A and various fibrosis-associated growth factors, cytokines, and extracellular matrix proteins. WNT-5A gene and protein expressions were significantly increased in fibrotic mouse and human livers compared with healthy livers. Regression or therapeutic intervention in mice resulted in decreased hepatic WNT-5A levels paralleled by lower collagen levels. Immunohistochemical analysis showed WNT-5A staining in fibrotic septa colocalizing with desmin staining indicating WNT-5A expression in myofibroblasts. In vitro studies confirmed WNT-5A expression in this cell type and showed that TGF-β significantly enhanced WNT-5A expression in contrast to PDGF-BB and proinflammatory cytokines IL-1β and TNF-α. Additionally, TGF-β induces the expression of the WNT receptors FZD2 and FZD8. After silencing of WNT-5A, reduced levels of collagen type I, vimentin, and fibronectin in TGF-β-stimulated myofibroblasts were measured compared with nonsilencing siRNA-treated controls. Interestingly, the antifibrotic cytokine IFNγ suppressed WNT-5A in vitro and in vivo. IFNγ-treated fibrotic mice showed significantly less WNT-5A expression compared with untreated fibrotic mice. In conclusion, WNT-5A paralleled collagen I levels in fibrotic mouse and human livers. WNT-5A expression in myofibroblasts is induced by the profibrotic factor TGF-β and plays an important role in TGF-β-induced regulation of fibrotic matrix proteins, whereas its expression can be reversed upon treatment, both in vitro and in vivo. NEW & NOTEWORTHY This study describes the localization and functional role of WNT-5A in human and mouse fibrotic livers. Hepatic WNT-5A expression parallels collagen type I expression. In vivo and in vitro, the myofibroblasts were identified as the key hepatic cells producing WNT-5A. WNT-5A is under control of TGF-β and its activities are primarily profibrotic.

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.

Mysterious path of Borrelia spielmanii: spreading without morphological alteration of collagen type I and IV

2019 ◽  
Vol 14 (17) ◽  
pp. 1469-1475
Author(s):  
Pavle Banović ◽  
Ivan Čapo ◽  
Dejan Ogorelica ◽  
Nenad Vranješ ◽  
Verica Simin ◽  
...  
Keyword(s):  
Collagen Type ◽  
Erythema Migrans ◽  
Collagen Type I ◽  
Punch Biopsy ◽  
Morphological Alteration ◽  
Type I ◽  
Contributing Factors ◽  
Metalloproteinase 9 ◽  
Serology Testing

The majority of suggested mechanisms of Borrelia spreading inside erythema migrans (EM) are developed from in vitro studies and animal models. This report is the first to describe pathomorphological substrate of EM caused by Borrelia spielmanii in humans, addressing the hypothesis of enhanced Borrelia penetration through extracellular matrix. In the process of ruling out of atypical Masters’ disease, we conducted a punch biopsy of suspected EM and a two-tier serology testing for Lyme borreliosis, where we registered antibodies against B. spielmanii. Skin biopsy showed CD4+ and CD8+ lymphocyte involvement and high activity of matrix metalloproteinase 9. No alterations were detected in distribution and morphology of collagen type I and IV. Therefore, it is suggested that other mechanisms should be considered as major contributing factors to local spreading of B. spielmanii.

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