A spatiotemporal analysis using second harmonic generation reveals enhancement of type I collagen polymerization by dermatan sulfate glycosaminoglycans

Keshav Kumar Jha; Purba Sarkar; Rabindra Biswas; Ramray Bhat; Varun Raghunathan

doi:10.1117/12.2526896

22 July 2019 A spatiotemporal analysis using second harmonic generation reveals enhancement of type I collagen polymerization by dermatan sulfate glycosaminoglycans

Keshav Kumar Jha, Purba Sarkar, Rabindra Biswas, Ramray Bhat, Varun Raghunathan

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Proceedings Volume 11076, Advances in Microscopic Imaging II; 110761G (2019) https://doi.org/10.1117/12.2526896
Event: European Conferences on Biomedical Optics, 2019, Munich, Germany

Abstract

Type 1 Collagen is the most abundant member of the family of collagens, which are the dominant proteins constituting the extracellular matrix (ECM) of multicellular organisms. Within tissues, Type 1 collagen exhibits a fibrillar geometry that serves as a mechanical scaffold for cells. The latter remodel the collagen through the secretion of proteoglycans (proteins with long chains of sulfated glycosaminoglycans (GAGs)), both within physiological and pathological contexts. The dermatan sulfate proteoglycans (DS-PGs) are abundantly present within the developing organs and are known to be dysregulated in diseases such as cancer. How DS alters the fibrillar architecture of collagen is however, not well known. Herein, we have used second harmonic generation (SHG) microscopy to dissect the effects of DS GAGs on Type1 Collagen polymerization. We observe that the presence of DS during polymerization enhances the width and number of the fibers, the surface occupancy (which we define as the ability of the collagen matrix to fill a given volume) and the mean SHG signals. We then image polymerizing collagen matrices at temporal intervals: at very early time points (<6 h), the SHG signals in both control and DS-treated polymerizing Type-1 collagen are low and do not show any difference. However, there is a sudden increase in SHG signals 6 h onwards, with a sharper and significantly increased enhancement in the presence of DS. Our results suggest the presence of DS kinetically alters the collagen polymerization leading to significant changes in its eventual architecture.

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Keshav Kumar Jha, Purba Sarkar, Rabindra Biswas, Ramray Bhat, and Varun Raghunathan "A spatiotemporal analysis using second harmonic generation reveals enhancement of type I collagen polymerization by dermatan sulfate glycosaminoglycans", Proc. SPIE 11076, Advances in Microscopic Imaging II, 110761G (22 July 2019); https://doi.org/10.1117/12.2526896

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KEYWORDS

Collagen

Polymerization

Second-harmonic generation

Harmonic generation

Image enhancement

Microscopy

Image filtering

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