Mechanisms of repigmentation induced by photobiomodulation therapy in vitiligo

Photobiomodulation (PBM) therapy is based on the exposure of biological tissues to low‐level laser light (coherent light) or light‐emitting diodes (LEDs; noncoherent light), leading to the modulation of cellular functions, such as proliferation and migration, which result in tissue regeneration. PBM therapy has important clinical applications in regenerative medicine. Vitiligo is an acquired depigmentary disorder resulting from disappearance of functional melanocytes in the involved skin. Vitiligo repigmentation depends on available melanocytes derived from (a) melanocyte stem cells located in the bulge area of hair follicles and (b) the epidermis at the lesional borders, which contains a pool of functional melanocytes. Since follicular melanoblasts (MBs) are derived from the melanocyte stem cells residing at the bulge area of hair follicle, the process of vitiligo repigmentation presents a research model for studying the regenerative effect of PBM therapy. Previous reports have shown favourable response for treatment of vitiligo with a low‐energy helium‐neon (He‐Ne) laser. This review focuses on the molecular events that took place during the repigmentation process of vitiligo triggered by He‐Ne laser (632.8 nm, red light). Monochromatic radiation in the visible and infrared A (IRA) range sustains matrix metalloproteinase (MMP), improves mitochondrial function, and increases adenosine triphosphate (ATP) synthesis and O₂ consumption, which lead to cellular regenerative pathways. Cytochrome c oxidase in the mitochondria was reported to be the photoacceptor upon which He‐Ne laser exerts its effects. Mitochondrial retrograde signalling is responsible for the cellular events by red light. This review shows that He‐Ne laser initiated mitochondrial retrograde signalling via a Ca²⁺‐dependent cascade. The impact on cytochrome c oxidase within the mitochondria, an event that results in activation of CREB (cyclic‐AMP response element binding protein)‐related cascade, is responsible for the He‐Ne laser promoting functional development at different stages of MBs and boosting functional melanocytes. He‐Ne laser irradiation induced (a) melanocyte stem cell differentiation; (b) immature outer root sheath MB migration; (c) differentiated outer root sheath MB melanogenesis and migration; and (d) perilesional melanocyte migration and proliferation. These photobiomodulation effects result in perifollocular and marginal repigmentation in vitiligo.     

The pig as a model system for investigating the recruitment and contribution of myofibroblasts in skin healing

In the skin-healing field, porcine models are regarded as a useful analogue for human skin due to their numerous anatomical and physiological similarities. Despite the widespread use of porcine models in skin healing studies, the initial origin, recruitment and transition of fibroblasts to matrix-secreting contractile myofibroblasts are not well defined for this model. In this review, we discuss the merit of the pig as an animal for studying myofibroblast origin, as well as the challenges associated with assessing their contributions to skin healing. Although a variety of wound types (incisional, partial thickness, full thickness, burns) have been investigated in pigs in attempts to mimic diverse injuries in humans, direct comparison of human healing profiles with regards to myofibroblasts shows evident differences. Following injury in porcine models, which often employ juvenile animals, myofibroblasts are described in the developing granulation tissue at 4 days, peaking at Days 7–14, and persisting at 60 days post-wounding, although variations are evident depending on the specific pig breed. In human wounds, the presence of myofibroblasts is variable and does not correlate with the age of the wound or clinical contraction. Our comparison of porcine myofibroblast-mediated healing processes with those in humans suggests that further validation of the pig model is essential. Moreover, we identify several limitations evident in experimental design that need to be better controlled, and standardisation of methodologies would be beneficial for the comparison and interpretation of results. In particular, we discuss anatomical location of the wounds, their size and depth, as well as the healing microenvironment (wet vs. moist vs. dry) in pigs and how this could influence myofibroblast recruitment. In summary, although a widespread model used in the skin healing field, further research is required to validate pigs as a useful analogue for human healing with regards to myofibroblasts.     

The role of CFU-GEMM in human hemopoiesis

Summary The assay for CFU-GEMM has provided a measurement for pluripotent hemopoietic precursors in normal and abnormal hemopoiesis. While these cells are able to express the functional repertoire that includes not only myelopoiesis but also lymphopoiesis attempts to determine their self-renewal have shown little or no self-renewal capability. It is currently not known whether this observation reflects culture conditions favouring differentiation processes and surpressing self-renewal, or whether the observation made in culture truly reflects the potential of cells in vivo. Recent advances in molecular biology have lead to the identification of the genomic sequences of at least one of the hemopoietic growth factors thus confirming their importance as regulators.     

Effect of laminin environments and tumor factors on the biology of myeloid dendritic cells

Dendritic cells (DCs) are immune cells that surveil the organism for infections or malignancies and activate specific T lymphocytes initiating specific immune responses. Contrariwise, DCs have been show to participate in the development of diseases, among them some types of cancer by inducing angiogenesis or immunosuppression. The ultimate fate of DC functions regarding their role in disease or health is prompted by signals from the microenvironment. We have previously shown that the interaction of DCs with various extracellular matrix components modifies the immune properties and angiogenic potential of these cells.The objective of the current studies was to investigate the angiogenic and immune profile of murine myeloid DCs upon interaction with laminin environments, with a particular emphasis on ovarian cancer.Our results show that murine ovarian tumors produce several types of laminins, as determined by PCR analysis, and also that tumor-associated DCs, both from ascites or solid tumors express adhesion molecules capable of interacting with these molecules as determined by flow cytometry and PCR analysis. Further, we established that DCs cultured on laminin upregulate both AKT and MEK signaling pathways, and that long-term culture on laminin surfaces decreases the immunological capacities of these cells when compared to the same cells cultured on synthetic substrates. In addition, we observed that tumor conditioned media was able to modify the metabolic status of these cells, and also reprogram the development of DCs from bone marrow precursors towards the generation of myeloid-derived suppressor cells.Overall, these studies demonstrate that the interaction between soluble factors and extracellular matrix components of the ovarian cancer microenvironment shape the biology of DCs and thus help them become co-conspirators of tumor growth.     

Rat Osteoclast Precursors In Vivo Express a Vitronectin Receptor and a Chloride-Bicarbonate Exchanger

In vivo osteoclast precursors, which are mononuclear, were previously found to express TRAP (tartrate-resistant acid phosphatase) and CTR (calcitonin receptor), like multi-nucleated osteoclasts. In vitro, they were found to express, in addition, VNR (vitronectin receptor) and CBE (chloride-bicarbonate exchanger). In order to ascertain that osteoclast precursors in vivo express VNR and CBE like their in vitro counterparts, we used immunohistochemistry to localize these molecules in developing long bones of neonatal rats. Frozen sections of metatarsals and phalanges of 1-2 day-old rats were stained for TRAP and mineralization using histochemistry or were reacted with polyclonal antibodies specific for either the β₃ chain of the VNR or synthetic sequences of the CBE. Both mature, multinucleated osteoclasts within the forming marrow cavity of metatarsals (as shown previously) and mononuclear osteoclast precursors located outside the bony collar of the phalangeal calcified rudiment (as shown here for the first time) expressed both TRAP, VNR and CBE. These findings suggest that mononuclear osteoclast precursors express many of the phenotypical markers of multinucleated osteoclasts prior to their fusion and multinucleation which may allow them to resorb bone, as suggested by in vitro observations of pit formation by preosteoclasts cultured on resorbable substances.     

Three-dimensional hydrogel scaffolds facilitate in vitro self-renewal of human skin-derived precursors

Skin-derived precursors (SKPs) are multipotent cells with dermal stem cell properties. These easily available cells possess the capacity to reconstitute the skin in vivo, as well as a broader differentiation potential in vitro, which endows them with great prospects in regenerative medicine. However, the present authors’ group and others previously found that adult human SKPs (hSKPs) expanded deficiently in vitro, which largely counteracted their research and practical values. Taking the physiological micro-environment of hSKPs into consideration, the authors sought to establish a hydrogel scaffold-based three-dimensional (3-D) culture system for hSKPs in the present study. After comparing their morphology, growth characteristics, signature gene expression and differentiation potential in different hydrogels, the present authors found that a chemically defined hyaluronic acid and denatured collagen-based hydrogel system that mimicked the natural niche of hSKPs in the dermis could alleviate hSKP senescence, support hSKP proliferation as spheres, while largely retaining their properties and potential. This study suggested that recapitulating the in vivo stem cell niche by providing them with 3-D extracellular matrix environments could help them achieve better self-renewal in vitro. In addition, the animal-origin-free and biocompatible 3-D hydrogel system will certainly benefit fundamental research and clinical applications of hSKPs in the near future.     

Pathology of flat bladder lesions with emphasis on putative precursors

Flat bladder lesions comprise a spectrum of morphologic changes ranging from reactive atypia to carcinoma in situ (CIS). Differentiating these lesions is important because of differences in patient management and clinical outcome. The precise nature of precursor lesions of bladder cancer remains incompletely understood. Urothelial CIS is the most definitely characterized precursor lesion of high grade bladder cancer. Atypia of unknown significance (AUS) is somewhat controversial. For practical purposes, AUS and reactive urothelial changes should be considered a single entity, since neither lesion has established preneoplastic potential. Simple hyperplasia and papillary hyperplasia are recently identified putative preneoplastic lesions. More recent molecular data also support the precursor nature of intestinal metaplasia and keratinizing squamous metaplasia. In this review, we also discuss the utility of molecular ancillary studies in establishing premalignant lesions, diagnosis, and differential diagnosis of flat bladder lesions.