Regeneration of Collagen Fibrils at the Papillary Dermis by Reconstructing Basement Membrane at the Dermal-Epidermal Junction

Abstract The epidermal basement membrane deteriorates with aging. We previously reported that basement membrane reconstruction not only serves to maintain epidermal stem/progenitor cells in the epidermis, but also increases collagen fibers in the papillary dermis. Here, we investigated the mechanism of the latter action. Collagen fibrils in the papillary dermis were increased in organotypic human skin culture treated with matrix metalloproteinase and heparinase inhibitors. The expression levels of COL5A1 and COL1A1 genes (encoding collagen type V α 1 chain and collagen type I α 1 chain, respectively) were increased in fibroblasts cultured with conditioned medium from a skin equivalent model cultured with the inhibitors and in keratinocytes cultured on laminin-511 fragment-coated plates. We then examined cytokine expression, and found that the inhibitors increased the expression of PDGF-BB (platelet-derived growth factor consisting of two B subunits) in epidermis. Expression of COL5A1 and COL1A1 genes was increased in cultured fibroblasts stimulated with PDGF-BB. Further, the bifunctional inhibitor hydroxyethyl imidazolidinone (HEI) increased skin elasticity and the thickness of the papillary dermis in the skin equivalent. Taken together, our data suggests that reconstructing the basement membrane promotes secretion of PDGF-BB by epidermal keratinocytes, leading to increased collagen expression at the papillary dermis.

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Plant Recombinant Human Collagen Type I Hydrogels for Corneal Regeneration

Regenerative Engineering and Translational Medicine ◽

10.1007/s40883-021-00220-3 ◽

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.

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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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Immunoelectron microscopic localization of collagens type I, V, VI and of procollagen type III in human periodontal ligament and cementum.

Journal of Histochemistry & Cytochemistry ◽

10.1177/39.1.1983870 ◽

1991 ◽

Vol 39 (1) ◽

pp. 103-110 ◽

Cited By ~ 51

Author(s):

J Becker ◽

D Schuppan ◽

J P Rabanus ◽

R Rauch ◽

U Niechoy ◽

...

Keyword(s):

Periodontal Ligament ◽

Collagen Type ◽

Ultrastructural Localization ◽

Collagen Type I ◽

Collagen Fibrils ◽

Type I ◽

Type Iii ◽

Procollagen Type ◽

Collagen Type Vi ◽

Type V

We examined the ultrastructural localization of collagens Type I, V, VI and of procollagen Type III in decalcified and prefixed specimens of the periodontal ligament and cementum, by immunoelectron microscopy using ultra-thin cryostat sections. Immunostaining for collagen Type I was pronounced on the major cross-striated fibrils entering cementum and in cementum proper, whereas staining for procollagen Type III was almost exclusively observed on the major fibrils in the periodontal ligament situated more remote from cementum. Reactivity for collagen Type V was limited to aggregated, unbanded filamentous material of about 12 nm diameter that was found mainly in larger spaces between bundles of cross-striated collagen fibrils and occasionally on single microfibrils that apparently originated from the ends of the major collagen fibrils, which may support the concept of this collagen as a component of core fibrils. Collagen Type VI was present as microfilaments appearing to interconnect single cross-striated fibrils. In the densely packed fibril bundles of the periodontal ligament, no collagen type VI was detected. Neither Type V or Type VI collagen was observed in cementum.

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Eisenia bicyclis Extract Repairs UVB-Induced Skin Photoaging In Vitro and In Vivo: Photoprotective Effects

Marine Drugs ◽

10.3390/md19120693 ◽

2021 ◽

Vol 19 (12) ◽

pp. 693

Author(s):

Se-In Choi ◽

Hee-Soo Han ◽

Jae-Min Kim ◽

Geonha Park ◽

Young-Pyo Jang ◽

...

Keyword(s):

Collagen Type ◽

Collagen Type I ◽

Skin Aging ◽

Dermal Fibroblasts ◽

Tissue Inhibitors Of Metalloproteinases ◽

Epidermal Keratinocytes ◽

Heme Oxygenase 1 ◽

Type I ◽

Skin Damage ◽

Eisenia Bicyclis

Chronic exposure to ultraviolet B (UVB) is a major cause of skin aging. The aim of the present study was to determine the photoprotective effect of a 30% ethanol extract of Eisenia bicyclis (Kjellman) Setchell (EEB) against UVB-induced skin aging. By treating human dermal fibroblasts (Hs68) with EEB after UVB irradiation, we found that EEB had a cytoprotective effect. EEB treatment significantly decreased UVB-induced matrix metalloproteinase-1 (MMP-1) production by suppressing the activation of mitogen-activated protein kinase (MAPK)/activator protein 1 (AP-1) signaling and enhancing the protein expression of tissue inhibitors of metalloproteinases (TIMPs). EEB was also found to recover the UVB-induced degradation of pro-collagen by upregulating Smad signaling. Moreover, EEB increased the mRNA expression of filaggrin, involucrin, and loricrin in UVB-irradiated human epidermal keratinocytes (HaCaT). EEB decreased UVB-induced reactive oxygen species (ROS) generation by upregulating glutathione peroxidase 1 (GPx1) and heme oxygenase-1 (HO-1) expression via nuclear factor erythroid-2-related factor 2 (Nrf2) activation in Hs68 cells. In a UVB-induced HR-1 hairless mouse model, the oral administration of EEB mitigated photoaging lesions including wrinkle formation, skin thickness, and skin dryness by downregulating MMP-1 production and upregulating the expression of pro-collagen type I alpha 1 chain (pro-COL1A1). Collectively, our findings revealed that EEB prevents UVB-induced skin damage by regulating MMP-1 and pro-collagen type I production through MAPK/AP-1 and Smad pathways.

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Platelet Collagen Activation by Basement Membrane Collagens

10.1055/s-0039-1684661 ◽

1979 ◽

Author(s):

L. Balleisen ◽

J. Rauterberg

Keyword(s):

Basement Membrane ◽

Collagen Type ◽

Normal Subjects ◽

Collagen Type I ◽

Short Chain ◽

Collagen Type Iv ◽

Type I ◽

Type Iv ◽

Aggregation Activity ◽

Type V

The collagen component of the vessel intima consists of collagen type I, III and basement membrane collagens. For type III a particularly potent platelet aggregation activity was found earlier.Three different basement membrane collagens were isolated from call placenta. Occurence of type IV and V(A/B collagen) in the intimal and medial layer of vessel walls has been shown by chemical and immunhistological means(1). The third basement membrane collagen is probably isentical with the 55000 molecular weight component described by CHUNG et al. (2).Platelets from normal subjects were tested for aggregation and spreading on Zaponlack with type IV, V, the separated A and B chains of type V and the short chain collagen. Complete aggregation was obtained only with native type V collagen. Spreading of platelets was induced only by collagen type IV, the A chain of type V and the short chain collagen.The data indicate that all basement membrane collagens activate platelets but in different respects.1. Rauterberg J. et al.: Coll. Int. Cent. Nat. Rech. Sc. 287:220(1978)2. Chung E. et al.: Bioehem. Biophys. Res. Comm. 71: 1167(1976)

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