Embryonic Environment Research Articles

Nrf2a modulates the embryonic antioxidant response to perfluorooctanesulfonic acid (PFOS) in the zebrafish, Danio rerio

The glutathione redox system undergoes precise and dynamic changes during embryonic development, protecting against and mitigating oxidative insults. The antioxidant response is coordinately largely by the transcription factor Nuclear factor erythroid-2 (Nrf2), an endogenous sensor for cellular oxidative stress. We have previously demonstrated that impaired Nrf family signaling disrupts the glutathione redox system in the zebrafish embryo, and that impaired Nrf2 function increases embryonic sensitivity to environmental toxicants. Here, we investigated the persistent environmental toxicant and reported pro-oxidant perfluorooctanesulfonic acid (PFOS), and its impact on the embryonic glutathione-mediated redox environment. We further examined whether impaired Nrf2a function exacerbates PFOS-induced oxidative stress and embryotoxicity in the zebrafish, and the potential for Nrf2-PPAR crosstalk in the embryonic adaptive response. Wild-type and nrf2afh318−/− mutant embryos were exposed daily to 0 (0.01% v/v DMSO), 16, 32, or 64 μM PFOS beginning at 3 h post fertilization (hpf). Embryonic glutathione and cysteine redox environments were examined at 72 hpf. Gross embryonic toxicity, antioxidant gene expression, and apoptosis were examined at 96 hpf. Mortality, pericardial edema, and yolk sac utilization were increased in wild-type embryos exposed to PFOS. Embryonic glutathione and cysteine redox couples and gene expression of Nrf2 pathway targets were modulated by both exposure and genotype. Apoptosis was increased in PFOS-exposed wild-type embryos, though not in nrf2a mutants. In silico examination of putative transcription factor binding site suggested potential crosstalk between Nrf2 and PPAR signaling, since expression of PPARs and gene targets was modulated by both PFOS exposure and Nrf2a genotype. Overall, this work demonstrates that nrf2a modulates the embryonic response to PFOS, and that PPAR signaling may play a role in the embryonic adaptive response to PFOS.

Wnt-signaling enhances neural crest migration of melanoma cells and induces an invasive phenotype

BackgroundDuring embryonic development Wnt family members and bone morphogenetic proteins (BMPs) cooperatively induce epithelial-mesenchymal transition (EMT) in the neural crest. Wnt and BMPs are reactivated during malignant transformation in melanoma. We previously demonstrated that the BMP-antagonist noggin blocked the EMT phenotype of melanoma cells in the neural crest and malignant invasion of melanoma cells in the chick embryo; vice-versa, malignant invasion was induced in human melanocytes in vivo by pre-treatment with BMP-2.ResultsAlthough there are conflicting results in the literature about the role of β-catenin for invasion of melanoma cells, we found Wnt/β-catenin signaling to be analogously important for the EMT-like phenotype of human metastatic melanoma cells in the neural crest and during invasion: β-catenin was frequently expressed at the invasive front of human primary melanomas and Wnt3a expression was inversely correlated with survival of melanoma patients. Accordingly, cytoplasmic β-catenin levels were increased during invasion of melanoma cells in the rhombencephalon of the chick embryo. Fibroblast derived Wnt3a reduced melanoma cell adhesion and enhanced migration, while the β-catenin inhibitor PKF115–584 increased adhesion and reduced migration in vitro and in the chick embryonic neural crest environment in vivo. Similarly, knockdown of β-catenin impaired intradermal melanoma cell invasion and PKF115–584 efficiently reduced liver metastasis in a chick chorioallantoic membrane model. Our observations were accompanied by specific alterations in gene expression which are linked to overall survival of melanoma patients.ConclusionWe present a novel role for Wnt-signaling in neural crest like melanoma cell invasion and metastasis, stressing the crucial role of embryonic EMT-inducing neural crest signaling for the spreading of malignant melanoma.

Simulation of Embryonic Environments Produced by the Effect of Estradiol on Induced Pluripotent Stem Cells

Estradiol (E2) is one of the sex steroid hormones, whose various well-known functions include control of the female reproductive system, causing breast cancers, and inhibiting arterial sclerosis. In this study to investigate how E2 affected cell differentiation of induced pluripotent stem cells (iPSCs), we used different methods to culture hiPSCs, the undifferentiated state, longterm culture without passages (natural differentiation), embryoid bodies (EBs), and accelerated differentiation by activin, focusing on marker gene expression, including Brachyury, hepatocyte nuclear factor (HNF) 3β, estrogen receptor α, and other markers. When iPSCs began to differentiate by responding to E2, Brachyury expression decreased regardless of the amount of E2, cell line, or protocol. Other markers were up or down-regulated, but did not show any tendency except in the early stage of EBs. The definite timing that E2 had the strongest effect on the aspect of differentiation was the EB stage analogous to gastrulation, in which E2 repressed Brachyury expression and induced HNF3β at its restricted amount. These results suggest that the first target of E2 is the gastrulation stage.

Severity of Premenstrual Symptoms Predicted by Second to Fourth Digit Ratio.

Women of reproductive age often experience a variety of unpleasant symptoms prior to the onset of menstruation. While genetics may influence the variability of these symptoms and their severity among women, the exact causes remain unknown. We hypothesized that symptom variability originates from differences in the embryonic environment and thus development caused by variation in exposure to sex hormones. We measured the second to fourth digit ratios (2D:4D) in 402 young women and investigated the potential relationships of this ratio premenstrual symptoms using a generalized linear model. We found that two models (one with two predictors such as both hands’ digit ratios and the other with the difference between the two digit ratios, Dr-l) were significantly different from the constant model as assessed by chi-square test. The right digit ratio and Dr-l were negatively related to the symptom scores, and the left digit ratio was related to the scores. When premenstrual symptoms were classified into eight categories, five categories, including pain, concentration, autonomic reaction, negative affect, and control were associated with the digit ratios and Dr-l. Behavioral changes and water retention were not predicted by them. Arousal was predicted by Dr-l. The right 2D:4D is thought to be determined by the balance of testosterone and estrogen levels during early embryogenesis and is not affected by postpartum levels of sex hormones, while the left 2D:4D might be affected by the other prenatal environmental factors. We conclude that the embryonic environment, including the relative concentration of sex hormones an embryo is exposed to, is associated with the severity of premenstrual symptoms once menarche is reached.

How does calcium interact with the cytoskeleton to regulate growth cone motility during axon pathfinding?

The precision with which neurons form connections is crucial for the normal development and function of the nervous system. The development of neuronal circuitry in the nervous system is accomplished by axon pathfinding: a process where growth cones guide axons through the embryonic environment to connect with their appropriate synaptic partners to form functional circuits. Despite intense efforts over many years to understand how this process is regulated, the complete repertoire of molecular mechanisms that govern the growth cone cytoskeleton and hence motility, remain unresolved. A central tenet in the axon guidance field is that calcium signals regulate growth cone behaviours such as extension, turning and pausing by regulating rearrangements of the growth cone cytoskeleton. Here, we provide evidence that not only the amplitude of a calcium signal is critical for growth cone motility but also the source of calcium mobilisation. We provide an example of this idea by demonstrating that manipulation of calcium signalling via L-type voltage gated calcium channels can perturb sensory neuron motility towards a source of netrin-1. Understanding how calcium signals can be transduced to initiate cytoskeletal changes represents a significant gap in our current knowledge of the mechanisms that govern axon guidance, and consequently the formation of functional neural circuits in the developing nervous system.

Abstract 36: Cardiac Progenitor Cell Lineage Tracing During Embryonic Cardiomyogenesis

Introduction: Stem cell therapy represents great promises to myocardium regeneration. Multipotent c-Kit pos cardiac progenitor cells (CPCs) are able to differentiate into endothelial cells, smooth muscle cells, and cardiomyocytes. However, fundamental knowledge of CPC biology remains incomplete. Studies in rodent myocardial infarction model revealed that CPCs have poor long-term survival and engraftment after adoptive transfer, perhaps due to the severely damaged host environment. Therefore, it is critical to understand how CPCs interface with the recipient environment following transfer in order to enhance their true regenerative potentials. Hypothesis: Adoptively transferred stem cells are thought to survive and engraft best in an environment closely resembling their original habitat. Thus, we hypothesized that the embryonic environment provides the optimal spatiotemporal conditions to promote CPCs engraftment and commitment to cardiac fate. Methods: CPCs isolated from adult mouse hearts were expanded, fluorescence-tagged, and injected into blastocysts at E3.75 and in utero at E15.5. Embryos were analyzed following cardiogenesis by immunofluorescence for presence of CPC-derived tissues. Additionally, CPCs were injected intramyocardially at various stages from P0 to P7, to follow long-term adoptive transfer and assess CPCs lineage commitment. Results and Conclusions: At 48 hours post injection, donor CPCs were found anchoring in blastocoel and trophoblasts at E5.5, and were detected within the host myocardium at E17.5 predominantly at perivascular regions (n=4). Interestingly, CPCs also integrated into aminochorionic sac, indicating a novel non-cardiogenic fate of CPCs (n=5). CPCs injected at P3 stably engrafted into left ventricular myocardium by 14 days post injection (n=4), sharing gap junction proteins (ZO-1, Connexin-43) with neighboring cells. In conclusion, this study provides vivid evidence for the first time of CPC engraftment and survival in vivo under homeostasis during cardiogenesis. Future studies will assess the permissive environmental conditions, which may optimize their use in therapeutic applications, and the cardiogenic potential of CPCs in order to provide fundamental insights on CPCs biology.

Intraspecific Variation in and Environment-Dependent Resource Allocation to Embryonic Development Time in Common Terns.

Embryonic development time is thought to impact life histories through trade-offs against life-history traits later in life, yet the inference is based on interspecific comparative analyses only. It is largely unclear whether intraspecific variation in embryonic development time that is not caused by environmental differences occurs, which would be required to detect life-history trade-offs. Here we performed a classical common-garden experiment by incubating fresh eggs of free-living common terns (Sterna hirundo) in a controlled incubation environment at two different temperatures. Hatching success was high but was slightly lower at the lower temperature. While correcting for effects of year, incubation temperature, and laying order, we found significant variation in the incubation time embryos required until hatching and in their heart rate. Embryonic heart rate was significantly positively correlated within clutches, and a similar tendency was found for incubation time, suggesting that intrinsic differences in embryonic development rate between offspring of different parents exist. Incubation time and embryonic heart rate were strongly correlated: embryos with faster heart rates required shorter incubation time. However, after correction for heart rate, embryos still required more time for development at the lower incubation temperature. This suggests that processes other than development require a greater share of resources in a suboptimal environment and that relative resource allocation to development is, therefore, environment dependent. We conclude that there is opportunity to detect intraspecific life-history trade-offs with embryonic development time and that the resolution of trade-offs may differ between embryonic environments.

Biotagging of Specific Cell Populations in Zebrafish Reveals Gene Regulatory Logic Encoded in the Nuclear Transcriptome

SummaryInterrogation of gene regulatory circuits in complex organisms requires precise tools for the selection of individual cell types and robust methods for biochemical profiling of target proteins. We have developed a versatile, tissue-specific binary in vivo biotinylation system in zebrafish termed biotagging that uses genetically encoded components to biotinylate target proteins, enabling in-depth genome-wide analyses of their molecular interactions. Using tissue-specific drivers and cell-compartment-specific effector lines, we demonstrate the specificity of the biotagging toolkit at the biochemical, cellular, and transcriptional levels. We use biotagging to characterize the in vivo transcriptional landscape of migratory neural crest and myocardial cells in different cellular compartments (ribosomes and nucleus). These analyses reveal a comprehensive network of coding and non-coding RNAs and cis-regulatory modules, demonstrating that tissue-specific identity is embedded in the nuclear transcriptomes. By eliminating background inherent to complex embryonic environments, biotagging allows analyses of molecular interactions at high resolution.

Embryonic environment and transgenerational effects in quail

BackgroundEnvironmental exposures, for instance to chemicals, are known to impact plant and animal phenotypes on the long term, sometimes across several generations. Such transgenerational phenotypes were shown to be promoted by epigenetic alterations such as DNA methylation, an epigenetic mark involved in the regulation of gene expression. However, it is yet unknown whether transgenerational epigenetic inheritance of altered phenotypes exists in birds. The purpose of this study was to develop an avian model to investigate whether changes to the embryonic environment had a transgenerational effect that could alter the phenotypes of third-generation offspring. Given its impact on the mammalian epigenome and the reproductive system in birds, genistein was used as an environment stressor.ResultsWe compared several third-generation phenotypes of two quail “epilines”, which were obtained from genistein-injected eggs (Epi+) or from untreated eggs (Epi−) from the same founders. A “mirrored” crossing strategy was used to minimize between-line genetic variability by maintaining similar ancestor contributions across generations in each line. Three generations after genistein treatment, a significant difference in the sexual maturity of the females, which, after three generations, could not be attributed to direct maternal effects, was observed between the lines, with Epi+ females starting to lay eggs later. Adult body weight was significantly affected by genistein treatment applied in a previous generation, and a significant interaction between line and sex was observed for body weight at 3 weeks. Behavioral traits, such as evaluating the birds’ reaction to social isolation, were also significantly affected by genistein treatment. Yet, global methylation analyses revealed no significant difference between the epilines.ConclusionsThese findings demonstrate that embryonic environment affects the phenotype of offspring three generations later in quail. While one cannot rule out the existence of some initial genetic variability between the lines, the mirrored animal design should have minimized its effects, and thus, the observed differences in animals of the third generation may be attributed, at least partly, to transgenerational epigenetic phenomena.

Reprogramming Malignant Cancer Cells toward a Benign Phenotype following Exposure to Human Embryonic Stem Cell Microenvironment

The embryonic microenvironment is well known to be non-permissive for tumor development because early developmental signals naturally suppress the expression of proto-oncogenes. In an analogous manner, mimicking an early embryonic environment during embryonic stem cell culture has been shown to suppress oncogenic phenotypes of cancer cells. Exosomes derived from human embryonic stem cells harbor substances that mirror the content of the cells of origin and have been reported to reprogram hematopoietic stem/progenitor cells via horizontal transfer of mRNA and proteins. However, the possibility that these embryonic stem cells-derived exosomes might be the main effectors of the anti-tumor effect mediated by the embryonic stem cells has not been explored yet. The present study aims to investigate whether exosomes derived from human embryonic stem cells can reprogram malignant cancer cells to a benign stage and reduce their tumorigenicity. We show that the embryonic stem cell-conditioned medium contains factors that inhibit cancer cell growth and tumorigenicity in vitro and in vivo. Moreover, we demonstrate that exosomes derived from human embryonic stem cells display anti-proliferation and pro-apoptotic effects, and decrease tumor size in a xenograft model. These exosomes are also able to transfer their cargo into target cancer cells, inducing a dose-dependent increase in SOX2, OCT4 and Nanog proteins, leading to a dose-dependent decrease of cancer cell growth and tumorigenicity. This study shows for the first time that human embryonic stem cell-derived exosomes play an important role in the tumor suppressive activity displayed by human embryonic stem cells.

The influence of prenatal environmental conditions on the weight of newborn domestic pigs

The weight of newborn piglets in domestic pigs depends on the maternal environment. Using an original algorithm based on the comparison of the coefficients of correlation between the half-nests (the same average genotype and the same conditions of the embryonic environment) and the twin sockets (the same genotype, different conditions of the embryonic environment), it was possible to estimate the relative contribution of the various components to the weight of the newborn in the domestic pigs. Our investigation found that the relative contribution of environmental conditions during prenatal development into newborn weight variation is equal to 0.4717, the contribution of genotypic component is 0.4032; the contribution of interaction between environmental conditions and ambivalence of the “genotype–phenotype” relationships is 0.0846, indeterminacy of phenotypic realization of the littermate’s genotypes is 0.0405. Maternal resource plays the main role amongst factors forming the environmental conditions of prenatal development. The gene pool will remain unchanged in any number of generations, despite a change in the population parameters of the trait. In principle, it can be assumed that when a population reaches a certain level, when the mean value of the trait and the gene pool are optimal (the selection pressure vectors are mutually balanced), selection becomes less effective in transforming the average phenotype than changing environmental conditions. Obviously, therefore, livestock maintenance and feeding are paid as much attention as breeding, and breeds of farmed animals are zoned.

Establishment of the Vertebrate Germ Layers.

The process of germ layer formation is a universal feature of animal development. The germ layers separate the cells that produce the internal organs and tissues from those that produce the nervous system and outer tissues. Their discovery in the early nineteenth century transformed embryology from a purely descriptive field into a rigorous scientific discipline, in which hypotheses could be tested by observation and experimentation. By systematically addressing the questions of how the germ layers are formed and how they generate overall body plan, scientists have made fundamental contributions to the fields of evolution, cell signaling, morphogenesis, and stem cell biology. At each step, this work was advanced by the development of innovative methods of observing cell behavior in vivo and in culture. Here, we take an historical approach to describe our current understanding of vertebrate germ layer formation as it relates to the long-standing questions of developmental biology. By comparing how germ layers form in distantly related vertebrate species, we find that highly conserved molecular pathways can be adapted to perform the same function in dramatically different embryonic environments.

Habitat- and season-specific temperatures affect phenotypic development of hatchling lizards

Embryonic environments influence phenotypic development, but relatively few experiments have explored the effects of natural environmental variation. We incubated eggs of the lizard Anolis sagrei under conditions that mimicked natural spatial and temporal thermal variation to determine their effects on offspring morphology and performance. Incubation temperatures mimicked two microhabitats (open, shade) at two different times of the incubation season (April, July). Egg survival, incubation duration and offspring size were influenced by interactions between habitat- and season-specific nest temperatures, and locomotor performance was influenced primarily by temporal factors. These findings highlight the importance of spatial and temporal environmental variation in generating variation in fitness-related phenotypes.

P2006 Transgenerational effects of modifications of the embryonic environment in quail

P2006 Transgenerational effects of modifications of the embryonic environment in quail

Tumorigenic potential is restored during differentiation in fusion-reprogrammed cancer cells.

Detailed understanding of the mechanistic steps underlying tumor initiation and malignant progression is critical for insights of potentially novel therapeutic modalities. Cellular reprogramming is an approach of particular interest because it can provide a means to reset the differentiation state of the cancer cells and to revert these cells to a state of non-malignancy. Here, we investigated the relationship between cellular differentiation and malignant progression by the fusion of four independent mouse cancer cell lines from different tissues, each with differing developmental potentials, to pluripotent mouse embryonic stem (ES) cells. Fusion was accompanied by loss of differentiated properties of the four parental cancer cell lines and concomitant emergence of pluripotency, demonstrating the feasibility to reprogram the malignant and differentiative properties of cancer cells. However, the original malignant and differentiative phenotypes re-emerge upon withdrawal of the fused cells from the embryonic environment in which they were maintained. cDNA array analysis of the malignant hepatoma progression implicated a role for Foxa1, and silencing Foxa1 prevented the re-emergence of malignant and differentiation-associated gene expression. Our findings support the hypothesis that tumor progression results from deregulation of stem cells, and our approach provides a strategy to analyze possible mechanisms in the cancer initiation.

Low Incubation Temperature Induces DNA Hypomethylation in Lizard Brains.

Developmental stress can have organizational effects on suites of physiological, morphological, and behavioral characteristics. In lizards, incubation temperature is perhaps the most significant environmental variable affecting embryonic development. Wall lizards (Podarcis muralis) recently introduced by humans from Italy to England experience stressfully cool incubation conditions, which we here show reduce growth and increase the incidence of scale malformations. Using a methylation-sensitive AFLP protocol optimized for vertebrates, we demonstrate that this low incubation temperature also causes hypomethylation of DNA in brain tissue. A consistent pattern across methylation-susceptible AFLP loci suggests that hypomethylation is a general response and not limited to certain CpG sites. The functional consequences of hypomethylation are unknown, but it could contribute to genome stability and regulation of gene expression. Further studies of the effects of incubation temperature on DNA methylation in ectotherm vertebrates may reveal mechanisms that explain why the embryonic thermal environment often has physiological and behavioral consequences for offspring.

Mono-2-ethylhexyl phthalate disrupts neurulation and modifies the embryonic redox environment and gene expression.

Mono-2-ethylhexl phthalate (MEHP) is the primary metabolite of di-2-ethylhexyl phthalate (DEHP), a ubiquitous contaminant in plastics. This study sought to determine how structural defects caused by MEHP in mouse whole embryo culture were related to temporal and spatial patterns of redox state and gene expression. MEHP reduced morphology scores along with increased incidence of neural tube defects. Glutathione (GSH) and cysteine (Cys) concentrations fluctuated spatially and temporally in embryo (EMB) and visceral yolk sac (VYS) across the 24h culture. Redox potentials (Eh) for GSSG/GSH were increased by MEHP in EMB (12h) but not in VYS. CySS/CyS Eh in EMB and VYS were significantly increased at 3h and 24h, respectively. Gene expression at 6h showed that MEHP induced selective alterations in EMB and VYS for oxidative phosphorylation and energy metabolism pathways. Overall, MEHP affects neurulation, alters Eh, and spatially alters the expression of metabolic genes in the early organogenesis-stage mouse conceptus.

The influence of thermal signals during embryonic development on intrasexual and sexually dimorphic gene expression and circulating steroid hormones in American alligator hatchlings (Alligator mississippiensis)

Incubation temperatures experienced by developing embryos exert powerful influences over gonadal sex determination and differentiation in many species. However, the molecular mechanisms controlling these impacts remain largely unknown. We utilize the American alligator to investigate the sensitivity of the reproductive system to thermal signals experienced during development and ask specifically whether individuals of the same sex, yet derived from different incubation temperatures display persistent variation in the expression patterns of sex biased transcripts and plasma sex hormones. Our analysis focuses on assessments of circulating sex steroids and transcript abundance in brain and gonad, two tissues that display sexually dimorphic gene expression and directly contribute to diverse sexually dimorphic phenotypes. Whereas our results identify sexually dimorphic patterns for several target gonadal genes in postnatal alligators, sex linked variation in circulating 17β-estradiol, testosterone, and expression of two brain transcripts (aromatase and gonadotropin releasing hormone) was not observed. Regarding intrasexual variation, we found that AMH transcript abundance in hatchling testes is positively correlated with temperatures experienced during sexual differentiation. We also describe highly variable patterns of gene expression and circulating hormones within each sex that are not explained by the intensity of embryonic incubation temperatures. The magnitude of sexually dimorphic gene expression, however, is directly associated with temperature for SOX9 and AMH, two transcripts with upstream roles in Sertoli cell differentiation. Collectively, our findings regarding temperature linked variation provide new insights regarding the connections between embryonic environment and persistent impacts on sexual differentiation in a reptile species that displays temperature dependent sex determination.

An empirical examination of endogenous ownership in Chinese private enterprises

ABSTRACTThe relationship between ownership structure and firm performance in small and medium enterprises (SMEs) in developing and transition countries has been largely ignored. Drawing on endogenous ownership theory, this paper seeks to understand the relationship between ownership structure and firm performance in SMEs in China, which represents an embryonic market environment. Using survey data collected from 189 private firms in Shantou City, China, we find that ownership variables do not have a statistically significant relationship with firm performance. We examine the determinants of changing ownership shares and find that firm size and business instability are related to changes in ownership shares. Our findings provide empirical support for the central tenets of endogenous ownership theory. As such, the results can be interpreted as meaning that even in a developing market environment, such as China, market forces are strong enough to dominate ownership structure change or that the market mechanism is functioning well in some areas of emerging markets.

Regenerative potential of characterised honey in wound healing

Event Abstract Back to Event Regenerative potential of characterised honey in wound healing Satarupa Sarkar1, Amrita Chaudhary1, Swarnendu Bag1, Bhuvaneshwaran Subramanian1, Amit K. Das2, Provas Banerjee3 and Jyotirmoy Chatterjee1 1 Indian Institute of Technology, Kharagpur, School of Medical Science & Technology, India 2 Indian Institute of Technology, Kharagpur, Department of Biotechnology, India 3 Banerjee's Biomedical Research Foundation, India Introduction: Outright harmonization amongst the events such as infiltration of inflammatory cells, granulation tissues constitution along with angiogenesis, proliferation and migration of fibroblasts and keratinocytes maintains dynamicity of ideal and rapid wound repair. Treatment with honey has regained popularity in recent times as because a number of small clinical trials have been completed which corroborate clinical improvement with regenerative potentiality on topical application of honey on wound. Honey is thought to be providing regenerative ambience in wound bed leading to the development of an embryonic environment with its own system intelligence. In this context present study performs exhaustive estimation of honey’s regenerative potential on topical application in wound model with a special emphasis on elucidation based on histopathological study. Materials and Methods: 10 male swiss albino mice (Mus musculus) were housed individually in a pathogen free environment and the study was performed under the Guidelines of the Institutional Animal Care and Use Committee. Two full thickness wounds were created on each animal after anaesthetizing with xylazine and ketamine hydrochloride and subsequent shaving of the dorsal part. One wound was regarded as control (povidone-iodine treated) and another was treated with physico-chemically characterized honey. The wounds were examined clinically at specific time points along with the collection of biopsy samples followed by Hematoxylin-Eosin (H and E), Periodic Acid–Schiff (PAS) and Van Gieson’s (VG) staining. Results and Discussion: As shown in figure 1, the total wound area (pixel value) for honey treated wound as calculated by ImageJ software was found to be significantly reduced when compared to control. Higher level of significance ( p= 5.6*E-05) is noted in case of honey treated wound as an indicator of wound contraction. The long lasting effect of honey on the release of TNF-α and IL-1β and IL-6 production may explain its therapeutic interface in the harmonized process of wound healing. On clinical evaluation, the honey treated wounds were found to show accelertaed healing comparitively with respect to control. The healing progression of skin biopsies were observed by histopathological techniques On 10th and 20th days after treatment, the microscopic observations under H and E staining explored remarkably improved histological features with progressive maturation of epithelial and connective tissues. The VG staining illustrated upgraded collagen density and formation of normal skin orientation. Conclusion: Topical application of honey escalates wound healing with exquisite dynamicity via apposite wound contraction and re-epithilialization in wound model. Results of this study may serve as a crucial step for exploration of honey in varied regenerative applications. This work is carried out under the project "Noninvasive Characterization Of Healing And Nonhealing Wounds And Development Of Honey-Biomaterial And Stem Cell Based Wound Therapy" (NIC), sponsored by MHRD, Department of higher Education, New Delhi, Government of India. IIT Kharagpur has been acknowledged for providing infrastructural facility