A New Method to Measure Type I and III Collagen Synthesis in Human Skin In Vivo: Demonstration of Decreased Collagen Synthesis After Topical Glucocorticoid Treatment

We have developed a novel cell stretching device (called Cell Gym) capable of applying physiologically relevant low magnitude strains to tenocytes on a collagen type I coated membrane. We validated our device thoroughly on two levels: (1) substrate strains, (2) cell level strains. Our cell level strain results showed that the applied stretches were transferred to cells accurately (∼90%). Our gene expression data showed that mechanically stimulated tenocytes (4%) expressed a lower level of COL I gene. COX2 gene was increased but did not reach statistical significance. Our device was then tested to see if it could reproduce results from an in vivo study that measured time-dependent changes in collagen synthesis. Our results showed that collagen synthesis peaked at 24 hrs after exercise and then decreased, which matched the results from the in vivo study. Our study demonstrated that it is important to incorporate physiologically relevant low strain magnitudes in in vitro cell mechanical studies and the need to validate the device thoroughly to operate the device at small strains. This device will be used in designing novel tendon tissue engineering scaffolds in the future.

Download Full-text

Sequential extracts of human bone show differing collagen synthetic rates

Biochemical Society Transactions ◽

10.1042/bst0300061 ◽

2002 ◽

Vol 30 (2) ◽

pp. 61-65 ◽

Cited By ~ 39

Author(s):

J. Babraj ◽

D.J. Cuthbertson ◽

P. Rickhuss ◽

W. Meier-Augenstein ◽

K. Smith ◽

...

Keyword(s):

Acetic Acid ◽

Collagen Synthesis ◽

Type I Collagen ◽

Human Bone ◽

Bone Collagen ◽

Type I ◽

Human Beings ◽

Collagen Turnover ◽

Bone Collagen Synthesis

Type I collagen is the major bone protein. Little is known quantitatively about human bone collagen synthesis in vivo, despite its importance for the understanding of bone formation and turnover. Our aim was to develop a method that could be used for the physiological and pathophysiological investigation of human bone collagen synthesis. We have carried out preliminary studies in patients undergoing hip replacement and in pigs to validate the use of the flooding dose method using 13C- or 15N-labelled proline and we have now refined our techniques to allow them to be used in a normal clinical or physiological setting. The results show that the application of a flooding dose causes bone free-proline labelling to equilibrate with that of blood in pigs and human beings, so that only 150 mg of bone will provide enough sample to prepare and measure the labelling of three fractions of bone collagen (dissolved in NaCl, acetic acid and pepsin/acetic acid) which have the same relative labelling (1.0:0.43:0.1) as measured by GC-combustion-isotope ratio MS. The rates of incorporation were substantially faster than in skeletal muscle samples taken at the same time. The results suggest that different fractions of human bone collagen turnover at markedly higher rates than had been previously considered. This approach should allow us to discover how growth and development, food, activity and drugs affect bone collagen turnover and to measure the effects on it of ageing and bone disease.

Download Full-text

Systemic glucocorticoids decrease the synthesis of type I and type III collagen in human skin in vivo, whereas isotretinoin treatment has little effect

British Journal of Dermatology ◽

10.1111/j.1365-2133.1994.tb04978.x ◽

1994 ◽

Vol 131 (5) ◽

pp. 660-663 ◽

Cited By ~ 49

Author(s):

P. AUTIO ◽

A. OIKARINEN ◽

J. MELKKO ◽

J. RISTELI ◽

L. RISTELI

Keyword(s):

Human Skin ◽

Type Iii Collagen ◽

Type I ◽

Type Iii ◽

Systemic Glucocorticoids

Download Full-text

Age-Associated Differences in Infection of Human Skin in the SCID Mouse Model of Varicella-Zoster Virus Pathogenesis

Journal of Virology ◽

10.1128/jvi.00002-18 ◽

2018 ◽

Vol 92 (11) ◽

Cited By ~ 3

Author(s):

Leigh Zerboni ◽

Phillip Sung ◽

Gordon Lee ◽

Ann Arvin

Keyword(s):

Herpes Zoster ◽

Varicella Zoster Virus ◽

Human Skin ◽

Inflammatory Responses ◽

Skin Lesions ◽

Type I ◽

Older Individuals ◽

Varicella Zoster ◽

Adult Skin

ABSTRACTVaricella-zoster virus (VZV) is the skin-tropic human alphaherpesvirus responsible for both varicella-zoster and herpes zoster. Varicella-zoster and herpes zoster skin lesions have similar morphologies, but herpes zoster occurs disproportionally in older individuals and is often associated with a more extensive local rash and severe zoster-related neuralgia. We hypothesized that skin aging could also influence the outcome of the anterograde axonal transport of VZV to skin. We utilized human skin xenografts maintained in immunodeficient (SCID) mice to study VZV-induced skin pathologyin vivoin fetal and adult skin xenografts. Here we found that VZV replication is enhanced in skin from older compared to younger adults, correlating with clinical observations. In addition to measures of VZV infection, we examined the expression of type I interferon (IFN) pathway components in adult skin and investigated elements of the cutaneous proliferative and inflammatory response to VZV infectionin vivo. Our results demonstrated that VZV infection of adult skin triggers intrinsic IFN-mediated responses such as we have described in VZV-infected fetal skin xenografts, including MxA as well as promyelocytic leukemia protein (PML), in skin cells surrounding lesions. Further, we observed that VZV elicited altered cell signaling and proliferative and inflammatory responses that are involved in wound healing, driven by follicular stem cells. These cellular changes are consistent with VZV-induced activation of STAT3 and suggest that VZV exploits the wound healing process to ensure efficient delivery of the virus to keratinocytes. Adult skin xenografts offer an approach to further investigate VZV-induced skin pathologiesin vivo.IMPORTANCEVaricella-zoster virus (VZV) is the agent responsible for both varicella-zoster and herpes zoster. Herpes zoster occurs disproportionally in older individuals and is often associated with a more extensive local rash and severe zoster-related neuralgia. To examine the effect of skin aging on VZV skin lesions, we utilized fetal and adult human skin xenografts maintained in immunodeficient (SCID) mice. We measured VZV-induced skin pathology, examined the expression of type I interferon (IFN) pathway components in adult skin, and investigated elements of the cutaneous proliferative and inflammatory response to VZV infectionin vivo. Our results demonstrate that characteristics of aging skin are preserved in xenografts; that VZV replication is enhanced in skin from older compared to younger adults, correlating with clinical observations; and that VZV infection elicits altered cell signaling and inflammatory responses. Adult skin xenografts offer an approach to further investigate VZV-induced skin pathologiesin vivo.

Download Full-text

Effect of Hydrocortisone, Methylprednisolone Aceponate and Momethasone Furoate on Collagen Synthesis in Human Skin in vivo

Skin Pharmacology and Physiology ◽

10.1159/000211513 ◽

1997 ◽

Vol 10 (5-6) ◽

pp. 261-264 ◽

Cited By ~ 29

Author(s):

K.-M. Haapasaari ◽

J. Risteli ◽

J. Karvonen ◽

A. Oikarinen

Keyword(s):

Human Skin ◽

Collagen Synthesis ◽

Methylprednisolone Aceponate

Download Full-text

Model for human skin reconstructed in vitro composed of associated dermis and epidermis

Sao Paulo Medical Journal ◽

10.1590/s1516-31802006000200005 ◽

2006 ◽

Vol 124 (2) ◽

pp. 71-76 ◽

Cited By ~ 27

Author(s):

Luís Ricardo Martinhão Souto ◽

Jussara Rehder ◽

José Vassallo ◽

Maria Letícia Cintra ◽

Maria Helena Stangler Kraemer ◽

...

Keyword(s):

Human Skin ◽

Collagen Type ◽

Human Fibroblasts ◽

Skin Grafting ◽

Human Keratinocytes ◽

Collagen Type I ◽

Type I ◽

Bovine Collagen

CONTEXT AND OBJECTIVE: The technique of obtaining human skin with dermis and epidermis reconstructed from cells isolated from patients can enable autologous skin grafting on patients with few donor sites. It also enables in vitro trials on chemicals and drugs. The objective of this work was to demonstrate a method for obtaining human skin composed of associated dermis and epidermis, reconstructed in vitro. DESIGN AND SETTING: Experimental laboratory study, in the Skin Cell Culture Laboratory of Faculdade de Ciências Médicas, Universidade Estadual de Campinas. METHODS: Cells from human fibroblast cultures are injected into bovine collagen type I matrix and kept immersed in specific culturing medium for fibroblasts. This enables human dermis reconstruction in vitro. On this, by culturing human keratinocytes and melanocytes, differentiated epidermis is formed, leading to the creation of human skin composed of associated dermis and epidermis, reconstructed in vitro. RESULTS: We showed that human skin composed of associated dermis and epidermis can be successfully reconstructed in vitro. It is histologically formed in the same way as human skin in vivo. Collagen tissue can be identified in the dermis, with cells and extracellular matrix organized in parallel to multilayer epidermis. CONCLUSIONS: It is possible to obtain completely differentiated human skin composed of associated dermis and epidermis, reconstructed in vitro, from injection of human fibroblasts into bovine collagen type I matrix and culturing of human keratinocytes and melanocytes on this matrix.

Download Full-text