Targeting Wnt/ß-catenin signaling and its interplay with TGF-ß and Notch signaling pathways for the treatment of chronic wounds.

Wound healing is a tightly regulated process that ensures tissue repair and normal function following injury. It is modulated by activation of pathways such as the transforming growth factor-beta (TGF-ß), Notch, and Wnt/ß-catenin signaling pathways. Dysregulation of this process causes poor wound healing, which leads to tissue fibrosis and ulcerative wounds. The Wnt/ß-catenin pathway is involved in all phases of wound healing, primarily in the proliferative phase for formation of granulation tissue. This review focuses on the role of the Wnt/ß-catenin signaling pathway in wound healing, and its transcriptional regulation of target genes. The crosstalk between Wnt/ß-catenin, Notch, and the TGF-ß signaling pathways, as well as the deregulation of Wnt/ß-catenin signaling in chronic wounds are also considered, with a special focus on diabetic ulcers. Lastly, we discuss current and prospective therapies for chronic wounds, with a primary focus on strategies that target the Wnt/ß-catenin signaling pathway such as photobiomodulation for healing diabetic ulcers.

The effect of photomodulation on fibroblast growth factor and the Ras/MAPK signalling pathway: a review.

OBJECTIVE: Current therapies and technologies used to treat hard-to-heal diabetic wounds are limited to a 50% healing rate. The rise in the percentage of lower limb non-traumatic amputations in patients with diabetes has caused an increased demand for alternative, effective and safe treatment modalities. Photobiomodulation therapy (PBMT) utilises light to induce physiological changes and provide therapeutic benefits and has been shown to increase the healing of hard-to-heal wounds through the release of growth factors. The aim of this narrative review is to investigate the effect of photobiomodulation (PBM) on fibroblast growth factor (FGF) and the role of the Ras/MAPK signalling pathway in diabetic wound healing. METHOD: Relevant journal articles were obtained through PubMed and Google Scholar. RESULTS: Experimental and clinical findings from the review show that PBM can stimulate the release of growth factors, including FGF, an essential cytokine in wound healing, and one which is present at lower concentrations in diabetic wounds. There is also activation of the Ras/MAPK signalling pathway. CONCLUSION: One mechanism through which healing may be stimulated by PBM is via the FGF-Ras/MAPK signalling pathway, although strong evidence under hyperglycaemic conditions is lacking.

Oxidative Stress in Ageing and Chronic Degenerative Pathologies: Molecular Mechanisms Involved in Counteracting Oxidative Stress and Chronic Inflammation.

Ageing and chronic degenerative pathologies demonstrate the shared characteristics of high bioavailability of reactive oxygen species (ROS) and oxidative stress, chronic/persistent inflammation, glycation, and mitochondrial abnormalities. Excessive ROS production results in nucleic acid and protein destruction, thereby altering the cellular structure and functional outcome. To stabilise increased ROS production and modulate oxidative stress, the human body produces antioxidants, "free radical scavengers", that inhibit or delay cell damage. Reinforcing the antioxidant defence system and/or counteracting the deleterious repercussions of immoderate reactive oxygen and nitrogen species (RONS) is critical and may curb the progression of ageing and chronic degenerative syndromes. Various therapeutic methods for ROS and oxidative stress reduction have been developed. However, scientific investigations are required to assess their efficacy. In this review, we summarise the interconnected mechanism of oxidative stress and chronic inflammation that contributes to ageing and chronic degenerative pathologies, including neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease (PD), cardiovascular diseases CVD, diabetes mellitus (DM), and chronic kidney disease (CKD). We also highlight potential counteractive measures to combat ageing and chronic degenerative diseases.

Rubus Capped Zinc Oxide Nanoparticles Induce Apoptosis in MCF-7 Breast Cancer Cells.

Rubus fairholmianus (RF) has widely been used to treat various ailments, including pain, diabetes, and cancer. Zinc oxide nanoparticles (ZnO NPs) have drawn attention in modern healthcare applications. Hence, we designed this study to synthesize zinc oxide (ZnO) nanoparticles using R. fairholmianus root extract to investigate its synergistic cytotoxic effect on MCF-7 cells and explore the possible cell death mechanism. ZnO NPs were synthesized via green synthesis using R. fairholmianus root extract, and the effect on MCF-7 cells was determined by looking at cellular morphology, proliferation, cytotoxicity, apoptosis, and reactive oxygen species (ROS). The results showed that cellular proliferation was reduced following treatment with R. fairholmianus capped zinc oxide nanoparticles (RFZnO NPs), while cytotoxicity and ROS were increased. There was also an increase in apoptosis as indicated by the significant increase in cytoplasmic cytochrome c and caspase 3/7 (markers of apoptosis), as well as increased levels of pro-apoptotic proteins (p53, Bax) and decreased levels of anti-apoptotic protein (Bcl-2). In conclusion, these results showed that RFZnO NPs induce apoptosis in breast cancer cells via a mitochondria-mediated caspase-dependent apoptotic pathway and suggest the use of acetone root extract of R. fairholmianus for the treatment of cancer-related ailments.

Effect of photobiomodulation on cellular migration and survival in diabetic and hypoxic diabetic wounded fibroblast cells.

A disrupted wound repair process often leads to the development of chronic wounds, and pose a major physical, social and economic inconvenience on patients and the public health sector. Chronic wounds are a common complication seen in diabetes mellitus (DM), and often the severity necessitates amputation of the lower limbs. Recently, there has been increasing evidence that photobiomodulation (PBM) initiates wound healing, including increased protein transcription for cell proliferation, viability, migration and tissue reepithelialisation. Here, the hypothesis that PBM at a wavelength of 660 nm and energy density of 5 J/cm2 regulates wound repair in diabetic wounded and hypoxic diabetic wounded fibroblasts by enhancing cell migration and survival was investigated. PBM increased migration and survival in diabetic wounded and hypoxic diabetic wounded fibroblasts. Our findings suggest that PBM enhances migration and survival in diabetic wounded and hypoxic diabetic wounded fibroblasts, indicating that this therapeutic method may be beneficial against chronic wounds in diabetic patients.

Cellular Signalling and Photobiomodulation in Chronic Wound Repair.

Photobiomodulation (PBM) imparts therapeutically significant benefits in the healing of chronic wounds. Chronic wounds develop when the stages of wound healing fail to progress in a timely and orderly frame, and without an established functional and structural outcome. Therapeutic benefits associated with PBM include augmenting tissue regeneration and repair, mitigating inflammation, relieving pain, and reducing oxidative stress. PBM stimulates the mitochondria, resulting in an increase in adenosine triphosphate (ATP) production and the downstream release of growth factors. The binding of growth factors to cell surface receptors induces signalling pathways that transmit signals to the nucleus for the transcription of genes for increased cellular proliferation, viability, and migration in numerous cell types, including stem cells and fibroblasts. Over the past few years, significant advances have been made in understanding how PBM regulates numerous signalling pathways implicated in chronic wound repair. This review highlights the significant role of PBM in the activation of several cell signalling pathways involved in wound healing.

Synthesis of Zinc Oxide Nanoparticles Using Rubus fairholmianus Root Extract and Their Activity against Pathogenic Bacteria.

Recently, the biosynthesis of zinc oxide nanoparticles (ZnO NPs) from crude extracts and phytochemicals has attracted much attention. Green synthesis of NPs is cost-effective, eco-friendly, and is a promising alternative for chemical synthesis. This study involves ZnO NPs synthesis using Rubus fairholmianus root extract (RE) as an efficient reducing agent. The UV spectrum of RE-ZnO NPs exhibited a peak at 357 nm due to intrinsic bandgap absorption and an XRD pattern that matches the ZnO crystal structure (JCPDS card no: 36-1451). The average particle size calculated from the Debye-Scherrer equation is 11.34 nm. SEM analysis showed that the RE-ZnO NPs spherical in shape with clusters (1-100 nm). The antibacterial activity of the NPs was tested against Staphylococcus aureus using agar well diffusion, minimum inhibitory concentration, and bacterial growth assay. The R. fairholmianus phytochemicals facilitate the synthesis of stable ZnO NPs and showed antibacterial activity.

Photobiomodulation at 660 nm Stimulates Fibroblast Differentiation.

BACKGROUND AND OBJECTIVES: Among many of the different complications that diabetic patients suffer, foot ulcers are the most challenging, and in many cases result in non-traumatic lower limb amputation and permanent disability. To alleviate this burden, new interventions such as photobiomodulation (PBM) have been utilized. However, the cellular pathways affected by PBM have not yet been fully recognized. The differentiation of fibroblasts into myofibroblasts forms a vital part of wound healing and is often impaired under diabetic conditions. Therefore, this study sought to investigate the effects of PBM at 660 nm on the transforming growth factor-ß1 (TGF-ß1)/Smad pathway and the differentiation of fibroblasts into myofibroblasts. STUDY DESIGN/MATERIALS AND METHODS: WS1 fibroblasts were treated with PBM using a wavelength of 660 nm at a fluence of 5 J/cm2 in normal, normal wounded, diabetic, and diabetic wounded models. Post-irradiation cellular responses were observed at 24, 48, and 72 hours to ascertain morphological changes and cell viability, and the expression of fibroblast differentiation markers (Thy-1 or CD90, extra domain A fibronectin or EDA-FN and α-smooth muscle actin or α-SMA), TGF-ß1, phosphorylated (p)TGF-ß receptor 1 (R1), and p-Smad2/3. RESULTS: There was a significant increase in cell viability in all irradiated cell models, and no real significant changes in TGF-ß1, pTGF-ß1R1, and p-Smad2/3. As incubation time post-irradiation increased, Thy-1 (CD90) decreased, while EDA-FN and α-SMA increased in wounded models. CONCLUSIONS: PBM at 660 nm with 5 J/cm2 was successful in stimulating the differentiation of fibroblasts into myofibroblasts in diabetic wounded cells, which was independent of the TGF-ß1/Smad pathway. Fibroblast transition into myofibroblasts is vital to wound healing, failure of which results in impaired healing; PBM is able to foster such a transition. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

The "in's and outs" of laser hair removal: a mini review.

Conventional treatments for excessive hair are tedious and time consuming. Laser hair removal has become the leading therapy option for long-term results. It works on the principle of selective photothermolysis, whereby photons destroy the hair follicle while sparing the surrounding tissue. As demand increases, there has been an increase in the regulation of these treatments. Laser hair removal is not risk-free and side effects are associated with the treatment. Adequate training is vital to minimise adverse side effects. Certain countries have regulatory boards that laser operators need to register with, as well as the completion of mandatory hours. Others require laser operators to register with a Care Quality Commission (CQC) to ensure that minimum training and safety standards are met. Currently, in South Africa, the lack of regulatory boards and mandatory hours poses a risk to the public as anyone with little or no qualification is allowed to perform laser hair removal treatments, placing patients at risk. This review looks at some of the devices used and basic mechanisms of action of laser hair removal, its associated risks, side effects and current regulation.

Shedding light on a new treatment for diabetic wound healing: a review on phototherapy.

Impaired wound healing is a common complication associated with diabetes with complex pathophysiological underlying mechanisms and often necessitates amputation. With the advancement in laser technology, irradiation of these wounds with low-intensity laser irradiation (LILI) or phototherapy, has shown a vast improvement in wound healing. At the correct laser parameters, LILI has shown to increase migration, viability, and proliferation of diabetic cells in vitro; there is a stimulatory effect on the mitochondria with a resulting increase in adenosine triphosphate (ATP). In addition, LILI also has an anti-inflammatory and protective effect on these cells. In light of the ever present threat of diabetic foot ulcers, infection, and amputation, new improved therapies and the fortification of wound healing research deserves better prioritization. In this review we look at the complications associated with diabetic wound healing and the effect of laser irradiation both in vitro and in vivo in diabetic wound healing.

Effect of photobiomodulation at 830 nm on gene expression correlated with JAK/STAT signalling in wounded and diabetic wounded fibroblasts in vitro.

Treatment of chronic diabetic wounds is an ongoing socio-economic challenge. Dysregulated signalling pathways characterise cells from chronic diabetic wounds. Photobiomodulation (PBM) stimulates healing by eliciting photochemical effects that affect gene regulation. JAK/STAT signalling is a primary signal transduction pathway involved in wound healing. This in vitro study aimed to determine if PBM at 830 nm and a fluence of 5 J/cm2 regulates genes related to JAK/STAT signalling in wounded and diabetic wounded fibroblast cells. A continuous wave diode laser (12.53 mW/cm2 ) was used to irradiate cells. Forty-eight hours post-PBM, RT-qPCR was used to analyse 84 genes related to JAK/STAT signalling. Five genes were upregulated and four downregulated in wounded cell models, while six genes were downregulated in diabetic wounded models. The results show drastic gene expression differences between wounded and diabetic wounded cell models in response to PBM using 830 nm.

Phototoxic effect of photodynamic therapy on lung cancer cells grown as a monolayer and three dimensional multicellular spheroids.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) is a minimally invasive therapeutic modality for the treatment of neoplastic and non-neoplastic diseases. The photochemical interaction of light, photosensitizer (PS) and molecular oxygen produces singlet oxygen which induces cell death. The effectiveness of zinc sulfophthalocyanine (ZnPcSmix ) has been shown on A549 monolayers and not on MCTS. The objective of this study was to investigate whether the same pattern of cell death would be induced in lung cancer cells (A549) grown as monolayer cells compared to three dimensional multicellular tumor spheroids (MCTS) using the same laser parameters. MATERIALS AND METHODS: Commercially purchased A549 cells used in this study were cultured as monolayers and as MCTS. ZnPcSmix at different concentrations [0, 5, 10, 20, and 40 µM] was used and activated at a wavelength of 680 nm with 5 J/cm2. Lysosomal and mitochondrial damage after PDT and reactive oxygen species (ROS) production were determined by fluorescent microscopy. Changes in cellular responses were evaluated using cell morphology, viability, proliferation, cytotoxicity, and cell death analysis. RESULTS: Cells exposed to neither laser light nor PS, showed no changes in cell morphology, proliferation, cytotoxicity, and ROS production. However, cells treated with light activated ZnPcSmix resulted in a significant production of ROS and a dose dependant decrease in viability and proliferation as well as an increase in cell membrane damage in both monolayer and MCTSs. CONCLUSION: ZnPcSmix PS used in this study induced damage to vital organelles and was effective in inducing apoptosis in lung cancer cells grown as a monolayer and MTCS through ROS production.

Photobiomodulation at 660 nm Stimulates In Vitro Diabetic Wound Healing via the Ras/MAPK Pathway.

Diabetic foot ulcers (DFUs) are open chronic wounds that affect diabetic patients due to hyperglycaemia. DFUs are known for their poor response to treatment and frequently require amputation, which may result in premature death. The present study evaluated the effect of photobiomodulation (PBM) at 660 nm on wound healing via activation of Ras/MAPK signalling in diabetic wounded cells in vitro. This study used four human skin fibroblast cell (WS1) models, namely normal (N), wounded (W), diabetic (D), and diabetic wounded (DW). Cells were irradiated at 660 nm with 5 J/cm2. Non-irradiated cells (0 J/cm2) served as controls. Cells were incubated for 24 and 48 h post-irradiation, and the effect of PBM on cellular morphology and migration rate, viability, and proliferation was assessed. Basic fibroblast growth factor (bFGF), its phosphorylated (activated) receptor FGFR, and phosphorylated target proteins (Ras, MEK1/2 and MAPK) were determined by enzyme-linked immunosorbent assay (ELISA) and Western blotting; nuclear translocation of p-MAPK was determined by immunofluorescence. PBM resulted in an increase in bFGF and a subsequent increase in FGFR activation. There was also an increase in downstream proteins, p-Ras, p-MEK1/2 and p-MAPK. PBM at 660 nm led to increased viability, proliferation, and migration as a result of increased bFGF and subsequent activation of the Ras/MAPK signalling pathway. Therefore, this study can conclude that PBM at 660 nm stimulates in vitro diabetic wound healing via the bFGF-activated Ras/MAPK pathway.

Effect of photobiomodulation therapy on inflammatory cytokines in healing dynamics of diabetic wounds: a systematic review of preclinical studies.

CONTEXT: Delayed wound healing in diabetes mellitus (DM) is due to the overlapping phases of the healing process. The prolonged inflammation and altered levels of inflammatory cytokines lead to deformed cell proliferation. Photobiomodulation alleviates the expression of inflammatory cytokines and promotes tissue repair, thereby restoring the wound healing process. OBJECTIVE: To find out the effect of photobiomodulation therapy (PBMT) in the healing dynamics of diabetic wounds with particular emphasis on interleukin-6, interleukin-1ß, and tumour necrosis factor-α. METHODS: Scientific databases searched using keywords of the population: DM, intervention: PBMT, and outcomes: inflammatory cytokines. RESULTS: We have included five preclinical studies in the present systematic review for qualitative analysis. These studies evaluated the effect of PBMT at different wavelengths, dosage, and time on wound healing in DM. CONCLUSIONS: The systematic review concludes that PBMT regulates inflammatory cytokines levels, enhances cell proliferation, and migration, thereby improving the wound healing properties.

Low-intensity laser irradiation at 660 nm stimulates cytochrome c oxidase in stressed fibroblast cells.

BACKGROUND AND OBJECTIVE: Low-intensity laser irradiation (LILI) has been used to modulate a variety of biological processes, including diabetic wound healing. The mechanism of action is thought to exist primarily with the mitochondria. This study aimed to determine the effect of irradiation on normal, diabetic, and ischemic mitochondrial electron transport chain (ETC) complexes. MATERIALS AND METHODS: Normal, diabetic and ischemic human skin fibroblast mitochondria were irradiated in vitro at a wavelength of 660 nm and a fluence of either 5 or 15 J/cm(2). Non-irradiated mitochondria served as controls. Enzyme activities of mitochondrial complexes I, II, III, and IV were determined immediately post-irradiation. Normal, diabetic, and ischemic cells were irradiated and adenosine triphosphate (ATP) and active mitochondria were determined by luminescence and fluorescent microscopy, respectively. RESULTS: Irradiated diabetic mitochondria at a fluence of 15 J/cm(2) showed a significant decrease in complex III activity (P < 0.05). Normal (P < 0.01) and diabetic (P < 0.05) mitochondria irradiated at either 5 or 15 J/cm(2) showed a significant increase in complex IV activity. ATP results showed a significant increase in irradiated normal cells (5 J/cm(2); P < 0.05) and diabetic cells (15 J/cm(2); P < 0.01). There was a higher accumulation of active mitochondria in irradiated cells than non-irradiated cells. CONCLUSION: Irradiation at 660 nm has the ability to influence mitochondrial enzyme activity, in particular cytochrome c oxidase. This leads to increased mitochondrial activity and ATP synthesis.

In Vitro Wound Healing Potential of Photobiomodulation Is Possibly Mediated by Its Stimulatory Effect on AKT Expression in Adipose-Derived Stem Cells.

Increasing evidence suggests that adipose-derived stem cells (ADSCs) serve as a therapeutic approach for wound healing. The aim of this study was to determine the effect of photobiomodulation (PBM) on antioxidant enzymes in ADSCs. Four ADSC cell models, namely, normal, wounded, diabetic, and diabetic wounded, were irradiated with 660 nm (fluence of 5 J/cm2 and power density of 11.2 mW/cm2) or 830 nm (fluence of 5 J/cm2 and power density of 10.3 mW/cm2). Nonirradiated cells served as controls. Cell morphology and wound migration were determined using light microscopy. Cell viability was determined by the trypan blue exclusion assay. The enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of antioxidants (superoxide dismutase (SOD), catalase (CAT), and heme oxygenase (HMOX1)). AKT activation and FOXO1 levels were determined by immunofluorescence and western blotting. The gaps (wound) in PBM-treated wounded and diabetic wounded cell models closed faster than the controls. PBM treatment significantly increased antioxidant levels in all cell models. This reflects that oxidative stress is reduced on the counterpart of increased antioxidant levels. This might be due to the activation of the AKT signaling pathway as evidenced by the increased AKT signals via western blotting and immunofluorescence. This data suggests that PBM promotes wound healing by increasing antioxidant levels by activating AKT signaling.

Levels of Cyclooxygenase 2, Interleukin-6, and Tumour Necrosis Factor-α in Fibroblast Cell Culture Models after Photobiomodulation at 660 nm.

Chemicals and signaling molecules released by injured cells at the beginning of wound healing prompt inflammation. In diabetes, prolonged inflammation is one of the probable causes for delayed wound healing. Increased levels of cyclooxygenase-2 (cox-2), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF-α) are associated with the inflammatory response and in diabetes, and increased levels of these contribute to chronic wounds that do not heal. Rising levels of cox-2, IL-6, and TNF-α have also been associated with increased oxidative stress. Photobiomodulation (PBM) may impact wound healing processes by affecting the signaling pathways and molecules pertinent to tissue repair. In the present study, the effect of PBM (wavelength: 660 nm; energy density: 5 J/cm2) on levels of cox-2, IL-6, and TNF-α was determined in fibroblast cell culture models. Four WS1 models (normal, normal wounded, diabetic, and diabetic wounded) were irradiated at 660 nm, and the culture media was collected at 0, 24, and 48 h postirradiation. Cells that were not irradiated (0 J/cm2) served as the controls. The following parameters were determined postirradiation: cell morphology using light microscopy, cell viability using the Trypan Blue exclusion assay, and levels of the inflammatory markers cox-2, IL-6, and TNF-α were measured using ELISA. Cell migration increased in the wounded groups over the 48 h interval after PBM; viability improved postirradiation in the diabetic wounded groups at 0 and 24 h (P ≤ 0.05 and P ≤ 0.01, respectively); levels of cox-2 decreased in normal and diabetic wounded groups at 0 h (P ≤ 0.001) and increased in the diabetic and diabetic wounded groups at 48 h postirradiation (P ≤ 0.05 and P ≤ 0.01, respectively), while levels of IL-6 decreased in the normal (P ≤ 0.01), diabetic (P ≤ 0.05), and diabetic wounded (P ≤ 0.001) groups at 24 h and in the diabetic and diabetic wounded groups at 48 h (P ≤ 0.05) postirradiation. TNF-α was decreased in the normal wounded groups (P ≤ 0.05) at 48 h. Through its effect on decreased IL-6 levels in diabetic cell models, PBM at 660 nm may be successful at decreasing oxidative stress; however, the present study also found an increase in cox-2 levels at 48 h postirradiation.

Irradiation at 830 nm stimulates nitric oxide production and inhibits pro-inflammatory cytokines in diabetic wounded fibroblast cells.

BACKGROUND AND OBJECTIVE: Wound healing in diabetic patients remains a chief problem in the clinical setting and there is a strong need for the development of new, safe, reliable therapies. This study aimed to establish the effect of irradiating diabetic wounded fibroblast cells (WS1) in vitro on pro-inflammatory cytokines and the production of nitric oxide (NO). MATERIALS AND METHODS: Normal, wounded and diabetic wounded WS1 cells were exposed to an 830 nm laser with 5 J/cm(2) and incubated for a pre-determined amount of time. Changes in cellular viability, proliferation and apoptosis were evaluated by the Trypan blue assay, VisionBlue fluorescence assay and caspase 3/7 activity respectively. Changes in cytokines (interleukin--IL-6, IL-1 beta and tumour necrosis factor-alpha, TNF-alpha) were determined by ELISA. NO was determined spectrophotometrically and reactive oxygen species (ROS) was evaluated by immunofluorescent staining. RESULTS: Diabetic wounded WS1 cells showed no significant change in viability, a significant increase in proliferation at 24 and 48 hours (P<0.001 and P<0.01 respectively) and a decrease in apoptosis 24 hours post-irradiation (P<0.01). TNF-alpha levels were significantly decreased at both 1 and 24 hours (P<0.05), while IL-1 beta was only decreased at 24 hours (P<0.05). There was no significant change in IL-6. There was an increase in ROS and NO (P<0.01) 15 minutes post-irradiation. CONCLUSION: Results show that irradiation of diabetic wounded fibroblast cells at 830 nm with 5 J/cm(2) has a positive effect on wound healing in vitro. There was a decrease in pro-inflammatory cytokines (IL-1 beta and TNF-alpha) and irradiation stimulated the release of ROS and NO due to what appears to be direct photochemical processes.

The qualification and training of laser/intense pulse light hair removal operators within South Africa.

BACKGROUND: Lasers and intense pulse light (IPL) sources are powerful devices that can cause skin burns, pigmentary changes, and scarring if used incorrectly. Adequate training is essential, and regulations are required to limit complications. AIMS: The purpose was to investigate the qualifications and training obtained by laser hair removal operators in South Africa. METHODS: Questionnaires were distributed and information gathered from owners/managers of laser clinics, suppliers of laser devices in South Africa, individuals in the workplace performing laser hair removal procedures, and accredited tertiary institutions. RESULTS: A majority of clinic owners/managers (94.45%) felt that more emphasis should be placed on laser hair removal training at a tertiary level, and 66.67% outsource additional training provided by the manufacturer of laser devices. Based on the survey to manufacturers, 50% did not require any formal qualification as a minimum requirement, while 33.33% indicated laser hair removal training is incorporated at a NQF level 4 (National/Senior Certificate). The majority of individuals (68.89%) received training from tertiary institutions; however, they did not receive any practical training, and 60.87% felt the amount of training was insufficient. According to the survey sent to tertiary institutions in South Africa, only 27.78% offer laser hair removal training, and of these, 20% offer no practical training and 50% indicated that no practical examination is provided. CONCLUSIONS: There is a clear lack of training in laser hair removal in South Africa. The industry should have standard requirements in terms of minimum practical and theoretical hours with regards to the therapy.

DNA damage after phototherapy in wounded fibroblast cells irradiated with 16 J/cm(2).

BACKGROUND AND OBJECTIVE: Phototherapy or biomodulation is a remarkable therapy that has become more popular and widely used in the treatment of a variety of medical conditions, such as slow to heal wounds, pain, soft tissue injuries and skin trauma. It has been shown to induce DNA damage; however this damage appears to be repairable. This study aimed to determine the effects of phototherapy induced DNA damage and activation of the DNA repair gene methylpurine DNA glycosylase (MPG). MATERIALS AND METHODS: DNA integrity was assessed using the comet assay, with and without formamidopyrimidine glycosylase (Fpg). For the comet assay, wounded human skin fibroblast cells (WS1) were irradiated twice, once at 30 min and again at 72 h with 5 or 16 J/cm(2) using a diode laser at 636 nm and cellular responses were assessed 1 or 24h post-irradiation. Real time reverse transcriptase polymerase chain reaction (RT-PCR) assessed MPG expression and three reference genes namely; beta Actin (ACTB), Glyceraldehyde three phosphate dehydrogenase (GAPDH) and Ubiquitin c (UBC). Wounded cells were irradiated once (30 min) with 16 J/cm(2), and MPG expression was assessed at 0, 3 and 8h post-laser irradiation. RESULTS: At both 1 and 24h, wounded cells irradiated with 5 J/cm(2) showed insignificant DNA damage compared to control cells, while irradiation with 16 J/cm(2) showed significant damage. However, 24h post-irradiation these cells showed a significant decrease in damage compared to cells left to incubate for 1h. This observation was attributed to activation of DNA repair mechanisms. Real time RT-PCR showed that ACTB was not influenced by cell culture conditions or laser irradiation, and MPG expression was not detected. CONCLUSION: In conclusion, irradiation with 5 J/cm(2) did not produce additional DNA damage, while damage to cells irradiated with 16 J/cm(2) was repairable by mechanisms other than MPG. This study also showed that ACTB can be used as a reference gene in laser experiments, using parameters set out in this study.