High intensity narrow spectrum (HINS) light has been shown to have bactericidal effects on a range of medically important bacteria[1]. HINS technology could potentially be useful as a method for disinfecting medical implants, tissue engineered constructs and wounds. The fibroblast populated collagen lattice (FPCL) was used as an in vitro model to investigate the effect of HINS light on the wound contraction phase of wound healing

Anderson, J.G.

Grant, M.H.

MacGregor, S.J.

Maclean, M.

McDonald, R.

English

University of Strathclyde Institutional Repository

Strathprints Institutional RepositoryMcDonald, R. and MacGregor, S.J. and Maclean, M. and Anderson, J.G. and Grant, M.H. (2009)Effect of HINS light on the contraction of fibroblast populated collagen lattices. European Cells andMaterials, 18 (Suppl.). p. 79. ISSN 1473-2262Strathprints is designed to allow users to access the research output of the University of Strathclyde.Copyright c© and Moral Rights for the papers on this site are retained by the individual authorsand/or other copyright owners. You may not engage in further distribution of the material for anyprofitmaking activities or any commercial gain. You may freely distribute both the url (http://strathprints.strath.ac.uk/) and the content of this paper for research or study, educational, ornot-for-profit purposes without prior permission or charge.Any correspondence concerning this service should be sent to Strathprints administrator:mailto:strathprints@strath.ac.ukhttp://strathprints.strath.ac.uk/  McDonald, R. and MacGregor, S.J. and Maclean, M. and Anderson, J.G. and Grant, M.H. (2009) Effect of HINS light on the contraction of fibroblast populated collagen lattices. European Cells and Materials, 18 (Suppl. 2). p. 79. ISSN 1473-2262    http://strathprints.strath.ac.uk/18716/       Strathprints is designed to allow users to access the research output of the University of Strathclyde. Copyright © and Moral Rights for the papers on this site are retained by the individual authors and/or other copyright owners. You may not engage in further distribution of the material for any profitmaking activities or any commercial gain. You may freely distribute both the url (http://strathprints.strath.ac.uk) and the content of this paper for research or study, educational, or not-for-profit purposes without prior permission or charge. You may freely distribute the url (http://strathprints.strath.ac.uk) of the Strathprints website.  Any correspondence concerning this service should be sent to The Strathprints Administrator: eprints@cis.strath.ac.uk  European Cells and Materials Vol. 18. Suppl. 2, 2009 (page 79)                                      ISSN 1473-2262                                                                                      http://www.ecmjournal.org  Effect of HINS light on the contraction of fibroblast populated collagen lattices R McDonald1, S J MacGregor2 , M Maclean2 , JG Anderson2 & M H Grant1 1Bioengineering Unit, University of Strathclyde, Glasgow, G4 0NW, UK.  2The Robertson Trust Laboratory for Electronic Sterilisation Technologies, University of Strathclyde, Glasgow, G1 1XW, UK  INTRODUCTION: High intensity narrow spectrum (HINS) light has been shown to have bactericidal effects on a range of medically important bacteria[1].  HINS technology could potentially be useful as a method for disinfecting medical implants, tissue engineered constructs and wounds.  The fibroblast populated collagen lattice (FPCL) was used as an in vitro model to investigate the effect of HINS light on the wound contraction phase of wound healing. METHODS:  Collagen lattices (0.3% (w/v) type I rat tail collagen) were seeded with 3T3 mouse fibroblasts cells at a density of 2.5x104 cells/cm2 and allowed to contract freely.  FPCLs were treated with HINS light at 0.1, 1, and 10mW/cm2 for 1 h, equivalent to a dose of 0.36, 3.6 or 36 J/cm2 respectively.  The contraction of FPCLs was measured prior to, and for up to 7 days following treatment. At 24, 48 and 120 h post treatment, cells were counted using the MTT assay, after using collagenase to release the cells from the lattices.  At these same time points, FPCLs were stained with propidium iodide (PI) and acridine orange (AO) to assess cell viability by fluorescence microscopy. RESULTS:  Figure 1 shows that no significant difference was observed between contraction rates of untreated FPCLs and those treated at 0.1mW/cm2 and 1mW/cm2.  FPCLs treated at 10mW/cm2 stopped contracting immediately and did not recover significantly within 7 days. -100102030405060700 20 40 60 80 100 120 140 160hours after treatment% contraction* * * * Fig. 1: Effect of HINS light on the contraction of FPCLs.  Treatments were carried out, at 0.1, 1 and 10mW/cm2 (squares, triangles and crosses respectively) for 1 hour. Control is indicated by circles.  The percentage contraction, of the FPCL area, was calculated from the point of treatment. *indicates significant difference from control at each time point (P>0.05, ANOVA followed by Dunnett’s test)  The MTT assay results show that for up to 120 h post treatment, there was no significant difference in cell number when treating FPCLs at 0.1mW/cm2 and 1 mW/cm2 (see Figure 2).  Treatment at 10mW/cm2 for 1 h caused an approximate 80% decrease in cell number after 24 h.  By 120 h post treatment, cells in FPCLs treated at 10mW/cm2 did not show significant recovery.   02040608010012014016024 48 120time (hours following treatment)% control** *†Fig. 2: MTT assay results for FPCLs treated at 0.1, 1 and 10mW/cm2 (grey, shaded and black respectively) at 24, 48 and 120 hours post treatment. *P<0.5, comparing between treatments at each time point, †P<0.5, comparing each individual intensity over the time points (ANOVA followed by Fisher’s test for both comparisons) Microscopic assessment of cell viability using PI and AO staining confirmed these results. DISCUSSION & CONCLUSIONS:  The results show that HINS light treatment at values of 0.1 and 1mW/cm2 have no detrimental effect on FPCL contraction.  This technology has considerable potential to augment efforts to disinfect medical devices, tissue engineered constructs and implants. REFERENCES: 1 M. Maclean, S.J. MacGregor, J.G. Anderson, et al (2008) FEMS Microbiol Lett Aug;285(2):227-32  ACKNOWLEDGEMENTS:  RM is supported by the ESPRC Doctoral Training Centre (DTC) in Medical Devices.   

FEMS Microbiol Lett Aug;285(2):227-32 ACKNOWLEDGEMENTS: RM is supported by the ESPRC Doctoral Training Centre (DTC) in Medical Devices.

Effect of HINS light on the contraction of fibroblast populated collagen lattices

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Effect of HINS light on the contraction of fibroblast populated collagen lattices

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