Vellore Institute of Technology (VIT) is a private university located in&nbsp;Tamil Nadu, India. Founded in 1984, as Vellore Engineering College, the institution offers 20 undergraduate, 34 postgraduate, four integrated and four research programs. It has campuses in Vellore, Amravati, Bhopal and Chennai.

VIT is one of the top ranked private universities in India according to NIRF, THE and QS Rankings.&nbsp;Govt. of India has recognized&nbsp;VIT, Vellore as an&nbsp;Institution of Eminence. This has allowed VIT to take independent quality initiatives and move up in world ranking.

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Honey is recognized traditionally for its medicinal properties and also appreciated as a topical healing agent for infected and noninfected wounds. This study evaluates impact of honey-based occlusive dressing on nonhealing (nonresponding to conventional antibiotics) traumatic lower limb wounds (n=34) through clinicopathological and immunohistochemical (e.g., expression of p63, E-cadherin, and Collagen I and III) evaluations to enrich the scientific validation. Clinical findings noted the nonadherence of honey dressing with remarkable chemical debridement and healing progression within 11-15 days of postintervention. Histopathologically, in comparison to preintervention biopsies, the postintervention tissues of wound peripheries demonstrated gradual normalization of epithelial and connective tissue features with significant changes in p 63 + epithelial cell population, reappearance of membranous E-cadherin (P&lt;.0001), and optimum deposition of collagen I and III (P&lt;.0001). Thus, the present study for the first time reports the impact of honey on vital protein expressions in epithelial and connective tissues during repair of nonhealing lower limb wounds. Copyright © 2011 Ananya Barui et al.

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Centre for Biomaterials, Cellular and Molecular Theranostics

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Evidence-based Complementary and Alternative Medicine

Immunohistochemical evaluation of p63, E-cadherin, Collagen i and III expression in lower limb wound healing under honey

Direct noninvasive visualization of wound bed with depth information is important to understand the tissue repair. We correlate skin swept-source-optical coherence tomography (OCT) with histopathological and immunohistochemical evaluation on traumatic lower limb wounds under honey dressing to compare and assess the tissue repair features acquired noninvasively and invasively. Analysis of optical biopsy identifies an uppermost brighter band for stratum corneum with region specific thickness (p 0.0001) and gray-level intensity (p 0.0001) variation. Below the stratum corneum, variation in optical intensities is remarkable in different regions of the wound bed. Correlation between OCT and microscopic observations are explored especially in respect to progressive growth and maturation of the epithelial and subepithelial components. Characteristic transition of uniform hypolucid band in OCT image for depigmented zone to wavy highly lucid band in the pigmented zone could be directly correlated with the microscopic findings. The transformation of prematured epithelium of depigmented area, with low expression of E-cadherin, to matured epithelium with higher E-cadherin expression in pigmented zone, implicated plausible change in their optical properties as depicted in OCT. This correlated evaluation of multimodal images demonstrates applicability of swept-source-OCT in wound research and importance of integrated approach in validation of new technology. © 2011 Society of Photo-Optical Instrumentation Engineers (SPIE).

Journal of Biomedical Optics

Swept-source optical coherence tomography of lower limb wound healing with histopathological correlation

Imaging bio-molecules especially in tissues, cells and even in in-vitro live conditions require expertise not only in staining and tagging of the particular molecules of interest but also their imaging so as to make any meaningful analysis and quantification of the obtained images. This necessitates a microscopic facility which can grab fluorescent images with minimum bleaching of the dyes and gives clear and crisp images without the blur caused by out of focus light. In this context ApoTome provides an alternative with metal halide light source and structural illumination with the help of grids, along with integrated image processing modality to generate images with digital interface. In the current study ApoTome was used to grab images of p63 and E-cadherin on histological sections of Oral Submucous Firosis an oral pre-cancer. The obtained images could aptly localize the expression of p63 and E-cadherin and distinguish the early and advanced stages of the precancer and did not show any effect of photo-bleaching. This demonstrated the applicability of ApoTome and its potential impact on diagnostics. © 2011 IEEE.

Proceedings of the 2011 IEEE/NIH Life Science Systems and Applications Workshop, LiSSA 2011

Low cost semi-confocal molecular imaging with cold light source

Natural product like honey is well known for its compositional diversity, medicinal properties including anti-inflammatory role, wound repair efficacy and nutritional competence. This multi-dimensional character of honey attracts regenerative medicine researchers. However, flow behavior of honey restricts its applications as a sustainable biomaterial matrix. In this juncture this study develops honey-biomaterial formulations and fabricates fibrous scaffold through wet spinning technique towards regenerative medicine applications. The cell culture study demonstrates the bio-compatibility of the fabricated constructs for fibroblasts (3T3) and keratinocyte (HaCaT) through MTT assay and by scanning electron microscopy regarding adhesion and penetration of cells on the scaffold. Better performance has been exhibited by the honey-biomaterial based scaffolds in comparison to alginate fibrous construct. This study also shows better expression of prime molecules like collagen I and III in 3T3 and p63 and E-cadherin in HaCaT on honey based matrix than that of alginate matrix or control group. Thus present work develops a honey-based fibrous matrix with biocompatible interface for plausible tissue engineering applications. © 2011 IEEE.

Honey based fibrous scaffold for tissue engineering application

Background: The assessment of malignant potential of oral submucous fibrosis grades vis-à-vis their progression towards malignancy is associated with expression of possible multiple molecular markers. Aims: To analyse p63, E-cadherin and CD105 expression in this premalignant pathosis with a view to unravel and understand the expression of these molecules as markers. Methods: The oral mucosal biopsies (normal, oral submucous fibrosis with and without dysplasia) were studied with routine H&E, and by immunohistochemistry for p63, E-cadherin and CD105 expression. p63 was assessed as percentage of positive nuclei. E-cadherin expression was estimated through (i) distance between basement membrane and E-cadherin expression initiation point, (ii) ratio between epithelial thickness and epithelial thickness displaying E-cadherin, and (iii) E-cadherin intensity variation along the expression path. CD105 expression was assessed qualitatively. Results: The p63+ cells were highest in severely dysplastic tissues followed by other dysplastic grades, normal oral mucosa and non-dysplastic conditions. However, the p63+ cells displayed the feature of progressive maturation only in normal mucosa. There was a loss of membranous E-cadherin in basal layers of all diseased conditions; it was highest in severe dysplasia. There was significant variation (p<0.0001) in E-cadherin intensity within and between the tissues (normal and diseased). CD105 expression increased abruptly in dysplasia. Conclusions: The malignant potential of this pre-cancerous condition is likely to be correlated with an increase in p63 and CD105 expression and a concomitant loss of membranous E-cadherin. This may lead to marker identification through greater validation.

Journal of Clinical Pathology

Assessment of malignant potential of oral submucous fibrosis through evaluation of p63, E-cadherin and CD105 expression

Non-invasive visualization of wound bed with depth information is important to understand the tissue repair under therapeutic interventions. In this context, present study performs skin swept source-optical coherence tomography (i.e. optical biopsy/OCT) and correlates findings with histo-pathological observations on different types of traumatic non-healing wounds under honey dressing. Correlation between OCT and microscopic observations were explored especially in respect to progressive growth and maturation of the epithelial and sub-epithelial components. Characteristic transition of uniform hypo-lucid band for de-pigmented zone to wavy highly lucid band for the pigmented zone in the OCT images could be directly correlated with the microscopic findings on transformation of flattened pre-mature epithelium of de-pigmented area to thicker and matured epithelium with rete-pegs in pigmented zone. This correlated evaluation of multimodal images demonstrates the applicability of SS-OCT in wound research and importance of integrated approach in validation of newer technology. © 2010 IEEE.

International Conference on Systems in Medicine and Biology, ICSMB 2010 - Proceedings

Correlating optical biopsy with histopathology of wounds under topical intervention with honey

Investigation of electrical properties along with molecular expressions of the keratinocyte and fibroblast under different physiological conditions can provide insight into their functioning which can contribute significantly towards skin tissue engineering application. For electrical characterization, cell based biosensors are usually designed with photolithographically patterned electrodes which are of the dimensions of cells. This increases cost and complexity of the analysis. In this paper, we report electrical characterization of keratinocyte and fibroblast cells, cultured in DMEM-F12 and DMEM media with a low cost and sensitive macroporous silicon platform using simple electrode geometries for the first time. Impedance spectroscopy results show that, there is a distinct difference between the electrical properties like effective capacitance and resistance of HaCaT (keratinocyte) and 3T3 (fibroblast) cells in the frequency range from 100Hz to 1MHz. The keratinocyte cells are observed to possess lower impedance than the fibroblast cells. The differences in the electrical properties have been correlated with the immunocytochemically detected prime molecular expressions of these cells. This multimodal analysis will help in the estimation of the degree of normalcy of these cells through a less complex and low cost electrical route. © 2010 IEEE.

Simple cytosensor based electrical characterization of keratinocytes and fibroblasts with prime molecular expressions towards skin tissue engineering applications

Late-diagnosis of malignant potentiality of oral precancers like Oral Sub-mucous Fibrosis (OSF) contributes towards high mortality of oral cancer patients. Current study intends to differentiate Normal Oral Mucosa (NOM) from OSF stages in terms of expression of a prime candidate gene (p63), correlated with epithelial proliferation and maturation, for better assessment of malignant potentiality of the pre-cancer. Fifty three oral biopsies (i.e. 8 NOM, 45 OSF patients with and without dysplasia) were histopathologically evaluated by hematoxylin and eosin, whereas, p63+ nuclei were identified immunohistochemically. The differences in cellular arrangement, especially in epithelial basal layer of diseased epithelium were observed. In respect to p63 expression the percentage of nuclei positive to this molecule and their size were studied. The p63 expression was higher in dysplastic epithelium and distribution of p63+ cells and their nuclear shape features demonstrated remarkable differences between NOM and OSF stages. © 2010 IEEE.