Tissue Integration and Degradation of a Porous Collagen-Based Scaffold Used for Soft Tissue Augmentation

Collagen-based scaffolds hold great potential for tissue engineering, since they closely mimic the extracellular matrix. We investigated tissue integration of an engineered porous collagen-elastin scaffold developed for soft tissue augmentation. After implantation in maxillary submucosal pouches in 6 canines, cell invasion (vimentin), extracellular matrix deposition (collagen type I) and scaffold degradation (cathepsin k, tartrate-resistant acid phosphatase (TRAP), CD86) were (immuno)-histochemically evaluated. Invasion of vimentin+ cells (scattered and blood vessels) and collagen type I deposition within the pores started at 7 days. At 15 and 30 days, vimentin+ cells were still numerous and collagen type I increasingly filled the pores. Scaffold degradation was characterized by collagen loss mainly occurring around 15 days, a time point when medium-sized multinucleated cells peaked at the scaffold margin with simultaneous labeling for cathepsin k, TRAP, and CD86. Elastin was more resistant to degradation and persisted up to 90 days in form of packages well-integrated in the newly formed soft connective tissue. In conclusion, this collagen-based scaffold maintained long-enough volume stability to allow an influx of blood vessels and vimentin+ fibroblasts producing collagen type I, that filled the scaffold pores before major biomaterial degradation and collapse occurred. Cathepsin k, TRAP and CD86 appear to be involved in scaffold degradation.

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14 Characterization of adipose tissue extracellular matrix component mRNA expression during porcine adipogenesis using real-time PCR

Journal of Animal Science ◽

10.1093/jas/skz397.080 ◽

2020 ◽

Vol 98 (Supplement_2) ◽

pp. 35-35

Author(s):

Maegan A Reeves ◽

Courtney E Charlton ◽

Terry D Brandebourg

Keyword(s):

Extracellular Matrix ◽

Adipose Tissue ◽

Mrna Expression ◽

Real Time ◽

Real Time Pcr ◽

Collagen Type ◽

Primary Cultures ◽

Collagen Type I ◽

Type I ◽

Matrix Metallopeptidase

Abstract Given adipose tissue is histologically classified as connective tissue, we hypothesized expression of extracellular matrix (ECM) components are significantly altered during adipogenesis. However, little is known about the regulation of the ECM during adipose tissue development in the pig. Therefore, the objective of this study was to characterize expression of ECM components during porcine adipogenesis. Primary cultures of adipose tissue stromal-vascular cells were harvested from 3-day-old neonatal pigs (n=6) and preadipocytes induced to differentiate in vitro for 8 days in the presence of insulin, hydrocortisone, and rosiglitazone. Total RNA was extracted from these cultures on days 0 and 8 post-induction. Real-time PCR was then utilized to determine changes in mRNA expression for collagen type I alpha 1 chain (COL1A), collagen type I alpha 2 chain (COL2A), collagen type I alpha 3 chain (COL3A), collagen type I alpha 4 chain (COL4A), collagen type I alpha 6 chain (COL6A), biglycan, fibronectin, laminin, nitogen-1 (NID1), matrix metallopeptidase 2 (MMP2), matrix metallopeptidase 9 (MMP9), metallopeptidase inhibitor 3 (TIMP3). The mRNA abundances of COL1A, COL3A and MMP2 were significantly downregulated 2.86-fold (P < 0.05), 16.7-fold (P < 0.01) and 3.1-fold (P < 0.05) respectively in day 8 (differentiated) compared to day 0 (undifferentiated) cultures. Meanwhile, mRNA abundances were significantly upregulated during adipogenesis for the COL2A (2.82-fold; P < 0.05), COL4A (2.01-fold; P < 0.05), COL6A (2.8-fold; P < 0.05), biglycan (49.9- fold; P < 0.001), fibronectin (452-fold; P < 0.001), laminin (6.1-fold; P < 0.05), NID1(47.4-fold; P < 0.01), MMP9 (76.8- fold; P < 0.01), and TIMP3(3.04-fold; P < 0.05) genes. These data support the hypothesis that significant changes in ECM components occur during porcine adipogenesis. Modulating adipose tissue ECM remodeling might be a novel strategy to manipulate adiposity in the pig.

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Precision-Cut Kidney Slices as a Tool to Understand the Dynamics of Extracellular Matrix Remodeling in Renal Fibrosis

Biomarker Insights ◽

10.4137/bmi.s38439 ◽

2016 ◽

Vol 11 ◽

pp. BMI.S38439 ◽

Cited By ~ 7

Author(s):

Federica Genovese ◽

Zsolt S. Kàrpàti ◽

Signe H. Nielsen ◽

Morten A. Karsdal

Keyword(s):

Extracellular Matrix ◽

Collagen Type ◽

Interstitial Fibrosis ◽

Ex Vivo ◽

Collagen Type I ◽

Extracellular Matrix Remodeling ◽

Type I ◽

Matrix Remodeling ◽

Renal Interstitial Fibrosis ◽

Ex Vivo Model

The aim of this study was to set up an ex vivo model for renal interstitial fibrosis in order to investigate the extracellular matrix (ECM) turnover profile in the fibrotic kidney. We induced kidney fibrosis in fourteen 12-week-old male Sprague Dawley rats by unilateral ureteral obstruction (UUO) surgery of the right ureter. The left kidney (contralateral) was used as internal control. Six rats were sham operated and used as the control group. Rats were terminated two weeks after the surgery; the kidneys were excised and precision-cut kidney slices (PCKSs) were cultured for five days in serum-free medium. Markers of collagen type I formation (P1NP), collagen type I and III degradation (C1M and C3M), and α-smooth muscle actin (αSMA) were measured in the PCKS supernatants by enzyme-linked immunosorbent assay. P1NP, C1M, C3M, and α-SMA were increased up to 2- to 13-fold in supernatants of tissue slices from the UUO-ligated kidneys compared with the contralateral kidneys ( P < 0.001) and with the kidneys of sham-operated animals ( P < 0.0001). The markers could also reflect the level of fibrosis in different animals. The UUO PCKS ex vivo model provides a valuable translational tool for investigating the extracellular matrix remodeling associated with renal interstitial fibrosis.

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Collagen Type I/III Matrix (Mucograft) for the plastic coverage of soft tissue defects after tumor resection in the oral cavity and oropharynx. Field report from 21 patient cases

10.1055/s-0041-1728580 ◽

2021 ◽

Author(s):

S Balster ◽

M Gröger ◽

E Endemann ◽

S Ghanaati ◽

T Stöver

Keyword(s):

Soft Tissue ◽

Oral Cavity ◽

Collagen Type ◽

Tumor Resection ◽

Collagen Type I ◽

Type I ◽

Soft Tissue Defects ◽

Field Report ◽

Tissue Defects

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Tissue Response to a Porous Collagen Matrix Used for Soft Tissue Augmentation

Materials ◽

10.3390/ma12223721 ◽

2019 ◽

Vol 12 (22) ◽

pp. 3721 ◽

Cited By ~ 3

Author(s):

Jordi Caballé-Serrano ◽

Sophia Zhang ◽

Luca Ferrantino ◽

Massimo Simion ◽

Vivianne Chappuis ◽

...

Keyword(s):

Soft Tissue ◽

Blood Vessels ◽

Collagen Matrix ◽

Giant Cells ◽

Tissue Response ◽

Soft Tissue Augmentation ◽

Proliferating Cells ◽

Tissue Integration ◽

Inflammatory Phase ◽

Tissue Augmentation

A short inflammatory phase and fast ingrowth of blood vessels and mesenchymal cells are essential for tissue integration of a biomaterial. Macrophages play a key role in this process. We investigated invasion of macrophages, blood vessels, and proliferating cells into a highly porous and volume-stable collagen matrix (VCMX) used for soft tissue augmentation around teeth and dental implants. The biomaterial was implanted in submucosal pouches in the canine maxilla, and the tissue response was analyzed at six different time points. Immunohistochemistry was done for proliferating cells (PCNA), macrophages (MAC387), multinucleated giant cells (CD86), and blood vessels (TGM2). Blood rapidly filled the VCMX pores. During the first week, MAC387+ cells populated the VCMX pores, blood vessels and PCNA+ cells invaded the VCMX, and CD86+ scattered cells were observed. At 15 days, MAC387+ cells were scanty, blood vessels had completely invaded the VCMX, the number of proliferating cells peaked, and fibroblasts appeared. At 30 days, MAC387+ were absent, the numbers of proliferating and CD86+ cells had declined, while blood vessel and fibroblast numbers were high. At 90 days, residual VCMX was well-integrated in soft connective tissue. In conclusion, the VCMX elicited a short inflammatory phase followed by rapid tissue integration.

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Dopamine D1 receptor stimulates cathepsin K-dependent degradation and resorption of collagen I in lung fibroblasts

Journal of Cell Science ◽

10.1242/jcs.248278 ◽

2020 ◽

Vol 133 (23) ◽

pp. jcs248278 ◽

Cited By ~ 1

Author(s):

Ana M. Diaz-Espinosa ◽

Patrick A. Link ◽

Delphine Sicard ◽

Ignasi Jorba ◽

Daniel J. Tschumperlin ◽

...

Keyword(s):

Collagen Type ◽

Cathepsin K ◽

Collagen Type I ◽

D1 Receptor ◽

Collagen I ◽

Lung Fibroblasts ◽

Type I ◽

Cultured Fibroblasts ◽

Normal Wound Healing

ABSTRACTMatrix resorption is essential to the clearance of the extracellular matrix (ECM) after normal wound healing. A disruption in these processes constitutes a main component of fibrotic diseases, characterized by excess deposition and diminished clearance of fibrillar ECM proteins, such as collagen type I. The mechanisms and stimuli regulating ECM resorption in the lung remain poorly understood. Recently, agonism of dopamine receptor D1 (DRD1), which is predominantly expressed on fibroblasts in the lung, has been shown to accelerate tissue repair and clearance of ECM following bleomycin injury in mice. Therefore, we investigated whether DRD1 receptor signaling promotes the degradation of collagen type I by lung fibroblasts. For cultured fibroblasts, we found that DRD1 agonism enhances extracellular cleavage, internalization and lysosomal degradation of collagen I mediated by cathepsin K, which results in reduced stiffness of cell-derived matrices, as measured by atomic force microscopy. In vivo agonism of DRD1 similarly enhanced fibrillar collagen degradation by fibroblasts, as assessed by tissue labeling with a collagen-hybridizing peptide. Together, these results implicate DRD1 agonism in fibroblast-mediated collagen clearance, suggesting an important role for this mechanism in fibrosis resolution.This article has an associated First Person interview with the first author of the paper.

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Changes in extracellular matrix composition regulate cyclooxygenase-2 expression in human mesangial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00176.2010 ◽

2011 ◽

Vol 300 (4) ◽

pp. C907-C918 ◽

Cited By ~ 6

Author(s):

Matilde Alique ◽

Laura Calleros ◽

Alicia Luengo ◽

Mercedes Griera ◽

Miguel Ángel Iñiguez ◽

...

Keyword(s):

Extracellular Matrix ◽

Collagen Type ◽

Mesangial Cells ◽

Collagen Type I ◽

Collagen I ◽

Type I ◽

Camp Response Element ◽

Response Element ◽

Human Mesangial Cells ◽

Cox 2

Glomerular diseases are characterized by a sustained synthesis and accumulation of abnormal extracellular matrix proteins, such as collagen type I. The extracellular matrix transmits information to cells through interactions with membrane components, which directly activate many intracellular signaling events. Moreover, accumulating evidence suggests that eicosanoids derived from cyclooxygenase (COX)-2 participate in a number of pathological processes in immune-mediated renal diseases, and it is known that protein kinase B (AKT) may act through different transcription factors in the regulation of the COX-2 promoter. The present results show that progressive accumulation of collagen I in the extracellular medium induces a significant increase of COX-2 expression in human mesangial cells, resulting in an enhancement in PGE2 production. COX-2 overexpression is due to increased COX-2 mRNA levels. The study of the mechanism implicated in COX-2 upregulation by collagen I showed focal adhesion kinase (FAK) activation. Furthermore, we observed that the activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway by collagen I and collagen I-induced COX-2 overexpression was abolished by PI3K and AKT inhibitors. Additionally, we showed that the cAMP response element (CRE) transcription factor is implicated. Finally, we studied COX-2 expression in an animal model, NG-nitro-l-arginine methyl ester hypertensive rats. In renal tissue and vascular walls, COX-2 and collagen type I content were upregulated. In summary, our results provide evidence that collagen type I increases COX-2 expression via the FAK/PI3K/AKT/cAMP response element binding protein signaling pathway.

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Modelling fibrillogenesis of collagen-mimetic molecules

10.1101/2020.06.08.140061 ◽

2020 ◽

Author(s):

A. E. Hafner ◽

N. G. Gyori ◽

C. A. Bench ◽

L. K. Davis ◽

A. Šarić

Keyword(s):

Extracellular Matrix ◽

Electrostatic Interactions ◽

Collagen Type ◽

Collagen Type I ◽

Coarse Grained ◽

Cell Phenotype ◽

Type I ◽

Collagen Scaffolds ◽

Crucial Component ◽

Self Assembled

One of the most robust examples of self-assembly in living organisms is the formation of collagen architectures. Collagen type I molecules are a crucial component of the extracellular-matrix where they self-assemble into fibrils of well defined striped patterns. This striped fibrilar pattern is preserved across the animal kingdom and is important for the determination of cell phenotype, cell adhesion, and tissue regulation and signalling. The understanding of the physical processes that determine such a robust morphology of self-assembled collagen fibrils is currently almost completely missing. Here we develop a minimal coarse-grained computational model to identify the physical principles of the assembly of collagen-mimetic molecules. We find that screened electrostatic interactions can drive the formation of collagen-like filaments of well-defined striped morphologies. The fibril pattern is determined solely by the distribution of charges on the molecule and is robust to the changes in protein concentration, monomer rigidity, and environmental conditions. We show that the fibril pattern cannot be easily predicted from the interactions between two monomers, but is an emergent result of multi-body interactions. Our results can help address collagen remodelling in diseases and ageing, and guide the design of collagen scaffolds for biotechnological applications.Statement of SignificanceCollagen type I protein is the most abundant protein in mammals. It is a crucial component of the extracellular-matrix where it robustly self-assembles into fibrils of specific striped architectures that are crucial for the correct collagen function. The molecular features that determine such robust fibril architectures are currently not well understood. Here we develop a minimal coarse-grained model to connect the design of collagen-like molecules to the architecture of the resulting self-assembled fibrils. We find that the pattern of charged residues on the surface of molecules can drive the formation of collagen-like fibrils and fully control their architectures. Our findings can help understand changes in collagen architectures observed in diseases and guide the design of synthetic collagen scaffolds.

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Morphometric analysis of the human endoneurial extracellular matrix components during aging

Archives of Biological Sciences ◽

10.2298/abs201214006k ◽

2021 ◽

Vol 73 (1) ◽

pp. 103-110

Author(s):

Braca Kundalic ◽

Sladjana Ugrenovic ◽

Ivan Jovanovic ◽

Vladimir Petrovic ◽

Aleksandar Petrovic ◽

...

Keyword(s):

Extracellular Matrix ◽

Collagen Type ◽

Linear Regression Analysis ◽

Collagen Type I ◽

Nerve Fibers ◽

Collagen Type Iv ◽

Type I ◽

Type Iv ◽

Ecm Proteins ◽

Extracellular Matrix Components

The aim of this study was to analyze the expression of extracellular matrix (ECM) proteins in human endoneurium during aging. We harvested 15 cadaveric sural nerves, distributed in 3 age groups (I: 25-44, II: 45-64, III: 65-86 years old). Histological sections were stained immunohistochemically for the presence of collagen type I, type IV and laminin, and the ImageJ processing program was used in morphometrical analysis to determine the percentages of these endoneurial proteins. In two younger groups, the endoneurial matrix of the sural nerve was composed from about equal proportions of these proteins, which may be considered a favorable microenvironment for the regeneration of nerve fibers. Linear regression analysis showed a significant increase in endoneurial collagen type IV with age, while collagen type I and laminin significantly decreased during the aging process. In cases older than 65 years, remodeling of the endoneurial matrix was observed to be significantly higher for the presence of collagen type IV, and lower for the expression of collagen type I and laminin. This age-related imbalance of ECM proteins could represent a disadvantageous microenvironment for nerve fiber regeneration in older adults. Our findings contribute to the development of therapeutic approaches for peripheral nerve regeneration.

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