BASIC PHOTOBIOLOGY IN REGENERATIVE MEDICINE. Lect. Dr.Nisamanee Charoenchon

Transcription

1 BASIC PHOTOBIOLOGY IN REGENERATIVE MEDICINE Lect. Dr.Nisamanee Charoenchon Edited:

2 Introduction deals with the local and systemic effects of incident radiation on living organisms. This lecture was focused on cutaneous photobiology and effects of ultraviolet (UV) radiation (UVR), both from its natural source (ie, the sun) and artificial sources (ie, those used in phototherapy).

3 Cosmic Rays Cosmic rays are energetic, subatomic particles that arrive from outside the Earth's atmosphere. The lowest energy cosmic rays are produced by ordinary stars like the Sun. For example, during a solar flare many particles are ejected from the Sun.

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5 Photobiology is the study of the effects of ultraviolet, visible and infrared radiation on living organisms. It includes topics such as photosynthesis, vision, bioluminescence, circadian rhythms and photodynamic therapy. Ref: (retrieved )

6 Regenerative medicine is a branch of translational research in tissue engineering and molecular biology which deals with the "process of replacing, engineering or regenerating human cells, tissues or organs to restore or establish normal function" Tissue engineering evolved from the field of biomaterials development and refers to the practice of combining scaffolds, cells, and biologically active molecules into functional tissues.

7 Photomedicine is an interdisciplinary branch of medicine that involves the study and application of light with respect to health and disease. various fields of medicine including dermatology, surgery, interventional radiology, optical diagnostics, cardiology, circadian rhythm sleep disorders and oncology.

8 Chromophores Molecules that absorb light and have reaction within molecule. If molecule doesn t absorb the particular wavelength, the reaction won t happen. Visible light that hits the chromophore can thus be absorbed by exciting an electron from its ground state into an excited state.

9 Chromophores in skin are coloured molecules the term refers to molecules able to absorb light and whose accumulation is readily perceived by the eyes. The two most common cutaneous chromophores are a brown chromophore: melanin, produced by melanocytes and stored in neighbouring keratinocytes, and a red chromophore: the haemoglobin contained in the blood

10 When an electron is in a higher orbit, an excited state, it wants to return to the ground state - a process called 'decay'. To do so, it needs to lose energy, and this often happens by giving off a photon - in other words emitting light.

11 Potential pathways of UVR-induced protein degradation Following exposure to UVR radiation, ECM remodelling in human skin may occur as a result of: i) cell mediated mechanisms via the synthesis of ECM proteases such as MMPs ii) ii) acellular pathways. Whilst cellular mechanisms undoubtedly play a role in downstream ECM remodelling. Physiologically attainable doses of UVR are capable of differentially degrading key ECM components in a cell-free environment. It remains to be determined whether this protein degradation occurs as a consequence of direct photon absorption by amino acid residues or the indirect action of UVR-induced ROS. Chemical consequences of cutaneous photoageing, 2012

12 Chromospheres in skin Elma D.Baron, 2014 Antony R Young, 1997 Different wavelength(s) target different chromophores, which results in a variety of cutaneous effects. Chromophores can be cellular/molecular components, such as amino acids, nucleotides, lipids, and 7-dehydrocholesterol (a vitamin D precursor). Epidermal chromophores These include 1. nucleic acids 2. urocanic acid 3. aromatic amino acids (proteins) 4. melanins and their precursors 5. vitamin D precursor

13 1.nucleic acids Significant mutagenic waveband is in UVC ( nm) UVA and UVB clearly cause DNA damage UVB (300 nm) and UVA (350 nm) If DNA is not repaired, it will result in cell death. And mutation induce carcinogenic event

14 2.urocanic acid Neil K. Gibbs, 2008 Urocanic acid (UCA), produced in the upper layers of mammalian skin, is a major absorber of ultraviolet radiation (UVR). 1 It is synthesised as the trans-isomer (trans-uca) from histidine in a deamination reaction Trans-UCA absorbs erythemogenic UVB ( nm) radiation and was initially proposed to play a role in natural photoprotection Originally thought to be a 'natural sunscreen', studies conducted a quarter of a century ago proposed that UCA may be a chromophore for the immunosuppression that follows exposure to UVR (cis- form). The photoisomerisation of naturally occurring trans (E)-urocanic acid to cis (Z)-urocanic acid.

15 3. Proteins/ amino acids Absorbent length (UVB) Cellular DNA was visualised with diamidino-2-phenylindole (DAPI) which stains cell nuclei with blue fluorescence. In both photoprotected and photoexposed skin the cellular population is concentrated in the epidermis whilst in the dermis, fibroblasts are sparsely distributed. Photoprotected (a)and photoaged (b) skin biopsies collected from the buttock and forearm respectively, of a 75 year old individual were immunofluorescently stained for the key elastic fibre component fibrillin-1 using a primary monoclonal antibody (clone 11C1.3) As a consequence, marked ECM remodelling in photoexposed skin (in this case loss of the red fibrillin-1 fluorescence) may be spatially separated from cells and hence from potential cell-derived mediators of tissue homeostasis. Chemical consequences of cutaneous photoageing, 2012

16 Sensitive amino acids (Cys and His, Phe, Trp and Tyr residues)

17 Differential amino acid composition of major dermal ECM components. In contrast, the major structural components: dermal fibrillar collagen and elastin are almost devoid of UVR. The fibrillins in particular, are enriched in Cys residues. Furthermore, most of these in latter proteins are associated with the disulphide bonded microfibrils which are: i) degraded in the papillary dermis of mildly photoaged skin and ii) abundantly distributed in the elastotic material which characterises the deeper dermis of severely photoaged ski Chemical consequences of cutaneous photoageing, 2012

18 4. Melanin Natural sunscreen, stops at least some of the UVR energy penetrating deeper into the skin. Absorb nm. UV stimulates melanogenesis

19 5. Vitamin D precursor Vitamin D from the diet or skin synthesis is biologically inactive; enzymatic conversion (hydroxylation) in the liver and kidney is required for activation. absorbs UV B radiation (280 to 315 nm)

20 Consequences in photomedicine The transfer of light energy through photon absorption can lead to many different aspect Diseases Caused by Light Therapy by Light Phototherapy is the use of nonionizing radiation to treat cutaneous disease. Various types of artificial light sources are used for photo testing and phototherapy For more than a century, phototherapy has played a pivotal role in the treatment of dermatologic diseases.

21 Photobiology in therapeutic purposes is broadly defined involving non-ionizing radiation. Non-ionizing radiation is the term given to radiation in the part of the electromagnetic spectrum where there does not have enough energy to remove an electron from an atom and turn it into an ion.

22 Non-ionizing electromagnetic radiation Non-ionizing radiations (NIR) World Health Organization, 2003 encompass the long wavelength (> 100 nm), low photon energy (<12.4 ev)

23 Laser: definition Laser, a device that stimulates atoms or molecules to emit light at particular wavelengths and amplifies that light, typically producing a very narrow beam of radiation. The emission generally covers an extremely limited range of visible, infrared, or ultraviolet wavelengths.

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25 Lasic- commonly referred to as laser eye surgery or laser vision correction psorasis medical/home use portable handheld NB UVB lamp

26 Laser and light therapy in melasma Melasma is a dysregulation of the homeostatic mechanisms that control skin pigmentation and excess pigment is produced. Melasma is a common and welldescribed dermatological condition that primarily affects female patients. It involves hyperpigmentation that is chronic, relapsing, and characterized by symmetric, brownish-grey macules and patches on the face and sometimes the neck, chest, and forearm. Melasma has also been referred to as chloasma or the mask of pregnancy because the condition is often associated with women who are pregnancy. A review of laser and light therapy in melasma, 2017

27 The three most common types of melasma are centrofacial, (forehead, cheeks, nose and upper lips) malar, (cheeks and nose) mandibular, (jawline) which describe the patterns of facial involvement.

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29 (A) Malar type melasma in patient 1. (B) The melanocytes are markedly increased in number and show pendulous change (NKIbeteb, 200), (A) Malar type melasma in patient 2. (B) The lesional skin shows increased melanocytes in the epidermis and dermal dendritic pigmented cells Two Cases of Melasma with Unusual Histopathologic Findings, 2006

30 Laser- and light-based treatments One key point of patient counseling prior to laser- and light-based treatment is that these therapies have the potential to speed up the removal of melasma-related hyperpigmentation but they are not cures for melasma. Optimal treatment management of difficult cases should include a combination therapy whereby a topical regimen inhibits melanin production and/or melanosome transfer and a procedure accelerates melanin removal. A review of laser and light therapy in melasma, 2017

31 The five broad categories of laser and light therapy intense pulsed light (IPL), Q-switched lasers, picosecond lasers, Non-ablative fractionated resurfacing lasers, and ablative fractionated resurfacing lasers.

32 Photoepilation and Skin Photorejuvenation: An Update, 2017 The IPL wavelength window is about 590 to 1,200 nm. The 694-nm ruby laser is no longer used in photoepilation because of strong absorption by melanin and high risk of adverse effects in tanned and darker skin types and hypopigmented areas observed even in skin type II Diagram illustrating the overall relationship between wavelengths in the visible and near-infrared spectrum and electromagnetic energy absorption by melanin.

33 1. Intense pulsed light IPL therapy uses a flash lamp light source that emits non-coherent light with wavelengths between 515 nm and 1200 nm. Filter sets allow for the targeting of selective chromophores (melanin vs. hemoglobin) and has been used to treat various pigmentary disorders. target both epidermal and dermal melasma The size of the IPL head is larger than most laser spot sizes, which allows for the rapid treatment of large areas.

34 efficacy and safety of a new intense pulsed light (IPL) device in the treatment of melasma in Chinese patients. (89 women) Mexameter results demonstrated a significant decrease in the degree of pigmentation and erythema beneath the melasma lesions. Patients with the epidermal-type melasma responded better to treatment than the mixed type. Efficacy and Safety of Intense Pulsed Light in Treatment of Melasma in Chinese, 2008 A representative photograph of a melasma patient before and after IPL treatments: (Top row) Left side of face (A) Pretreatment (C) at 3-month follow-up visit (bottom row) right side of face (B) after 4 sessions

35 After treatment completion, the group of 18 patients that was treated with IPL plus hydroquinone demonstrated Intense Pulsed Light for the Treatment of Refractory Melasma in Asian Persons, 2014 with 11.6% for the patients in the control, hydroquinone-only a reduction of 39.8% in relative melanin index hydroquinone It is used to lighten the skin where there are changes in color. Side effects include: Mild skin irritation and sensitization (e.g., burning, erythema, stinging). ท ำให ระคำยเค อง,ท ำให ค น,ท ำให แสบ Exogenous ochronosis (EO) is a cutaneous disorder characterized by blueblack pigmentation resulting as a complication of long-term application of skin-lightening creams containing hydroquinone Exogenous ochronosis hydroquinone induced: a report of four cases, 2010

36 Hyaluronan (HA) Recently, age-de- pendent change in the metabolism of epidermal hyaluronan (HA) is suggested to be responsible for these epidermal dysfunctions by activating CD44 signaling. Expression of CD44 and hyaluronan synthases (HAS) was also shown to be reduced in aged skin

37 Intense pulsed light (IPL) treatment increases hyaluronan (HA) production and hyaluronan synthases (HAS) 3 expression in young and old aged mouse epidermis and cultured human keratinocytes. Sections of young and aged mouse skin treated with IPL were stained for HA binding protein (HABP) 4 and examined by confocal microscope. Dashed line is the dermal-epidermal junction. Magnified image shows example of accumulated HA in the intercellular (HAo) and intracellular space (HAi) ( 400). Intense Pulsed Light Increases Hyaluronan and CD44 in Epidermal Keratinocytes and Improves Age- Related Epidermal Structure Defects in Mice, 2017

38 2. Q-switched lasers Q-switched lasers produce high intensity laser beams with very short pulse durations. The speed of a Q-switched laser pulse is approximately one million times that of an IPL pulse. Q-switched lasers that target melanin are available in multiple wavelengths including ruby (694 nm), alexandrite (755 nm), and neodymium-doped yttrium aluminum garnet (Nd:YAG; 532 nm or 1064 nm). Because these lasers are standard therapies for the removal of birthmarks, กระแดด solar lentigines, and tattoos, they were expected to be effective for the treatment of patients with melasma as well.

39 The regeneration of thermal wound on mice skin (Mus Musculus) after Q-Switch Nd: YAG laser irradiation for cancer therapy candidate, 2017 The average score of reepithelization The average score of angiogenesis The average distribution score of collagen In summary, Q-switched laser therapy at the fluences that is used to treat benign pigmented lesions has not been effective. A review of laser and light therapy in melasma, 2017

40 case being removal, separation, or taking away 5. ablative fractionated resurfacing lasers Ablative fractionated resurfacing lasers (AFL) such as CO2 lasers and erbium:yag lasers have been reported for the treatment of patients with melasma (Morais et al., 2013). Fractional resurfacing can be broadly categorized into ablative fractional laser (AFL) and nonablative fractional laser (NAFL) The CO 2 laser emits a 10,600-nm wavelength, which is strongly absorbed by water in the skin cells. The penetration depth is dependent on the water content and independent of either melanin or hemoglobin. A fractionated approach decreases the amount of epidermal injury and therefore results in fewer side effects and less risk of dyspigmentation.

41 It is hypothesized that CO 2 lasers cause immediate contraction of the ablated areas by denaturing existing old collagen. 3 This stimulates new collagen, and collagen content continues to increase well after the procedure. Current Laser Resurfacing Technologies: A Review that Delves Beneath the Surface, 2012 Although ablative lasers result in far more down time and a more difficult recovery process, they remain the lasers that produce the most dramatic outcomes. For more severe facial wrinkles, dyspigmentation, and textural skin challenges, the ablative laser is often the treatment of choice. Facelift scar. (A) Before and (B) after fractional resurfacing. Two passes of Active FX at 90 mj, density 4.

42 It has also been suggested that the microscopic injury zones that are caused by fractionated ablation allow for the transport of necrotic epidermal debris including melanin through the DEJ.

43 4. non-ablative fractionated resurfacing lasers recovery is more rapid and theoretically the resulting inflammation is lower, which lessens the risk for scarring or dyspigmentation NAFL devices target water-containing tissues but create columns of coagulative damage within the dermis The stratum corneum is intact throughout the treatment and a visible wound does not occur.

44 The use of fractional ablative lasers to deliver bioactive agent(s) to a patient via channels of predetermined depth, into cutaneous tissue, has wide clinical implications. This technology platform uses fractional lasers to predictably disrupt the barrier properties of the skin, creating deep channels that allow for the delivery of cellular materials and agents through the disrupted barrier.

45 bfgf is a glycoprotein, which is widely used in treating wounds and ulcers Although allergic contact dermatitis (ACD) and irritated contact dermatitis (ICD) are a major concern after laser resurfacing, this particular compound has been shown to improve wound healing without adverse event of dermatitis. Additional studies are needed to validate this observation. This is the first study to show that Vitamin C, E, and Ferulic acid correlates the wound healing clinical effect seen in patients with a specific molecular mechanism of action. Split face photo of patients at day 5 after fractional ablative laser and daily application of CEF. Laser-assisted delivery of vitamin C, vitamin E, and ferulic acid formula serum decreases fractional laser postoperative recovery by increased beta fibroblast growth factor expression, 2015

46 5. Picosecond lasers Shorter laser pulse durations result in pigment fragmentation that is more a result of photoacoustic than photothermal effects.

47 Picosecond lasers are currently available with laser outputs of 532 nm, 755 nm, and 1064 nm. More recently, fractionated picosecond handpieces have been developed for the purpose of resurfacing and rejuvenation. The average treatment pain score was 4.2 on a 1 10 scale. Picosecond laser with specialized optic for facial rejuvenation using a compressed treatment interval, 2016 The 1,064 and 532 nm picosecond-domain laser incorporating a holographic beam-splitting handpiece was found to be safe and effective for the treatment of facial acne scars. Treatment of acne scarring with a novel fractionated, dualwavelength, picosecond-domain laser incorporating a novel holographic beam-splitter, 2017 before 532 nm 3 months following the fourth and final treatment 1,064 nm

48 Mechanisms of Action of Fractionated 532nm and 1064nm Picosecond Laser for Skin Rejuvenation Habbema s proposed technique for skin rejuvenation has since been demonstrated by others via studies showing improvement in the appearance of facial acne scars and facial wrinkles

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50 Psorisis is a long-lasting autoimmune disease characterized by patches of abnormal skin. These skin patches are typically red, itchy, and scaly. Psoriasis varies in severity from small, localized patches to complete body coverage.

51 Learn about different types of light therapy. Sunlight Ultraviolet light B (UVB) Psoralen + UVA (PUVA) Laser Treatments Tanning beds

52 1. UVB generally used= narrow band UVB phototherapy 311 and 313 nanometers an effective treatment slows the growth of affected skin cells. broad band and narrow band release a smaller range

53 narrow-band lessening UVB clears psoriasis faster and produces longer remissions than broad-band UVB. fewer treatments per week than broad-band UVB. he ability of NB- UVB or BB-UVB to deplete T cells from psoriatic lesions was studied with CD3 antibodies and quantitative image analysis using immunohistoche mistry 312-nanometer Ultraviolet B Light (Narrow-Band UVB) Induces Apoptosis of T Cells within Psoriatic Lesions, 1999

54 2. Psoralen + UVA (PUVA) UVA is relatively ineffective unless used with a light-sensitizing medication psoralen, which is administered topically or orally. This process, called PUVA, slows down excessive skin cell growth and can clear psoriasis symptoms Stable plaque psoriasis, guttate psoriasis, and psoriasis of the palms and soles are most responsive to PUVA treatment. เป นหยด

55 3.Laser Treatments for treating chronic, localized psoriasis plaques emits a highintensity beam of ultraviolet light B (UVB). 4. Tanning beds Tanning beds in commercial salons emit mostly UVA light, not UVB. The beneficial effect for psoriasis is attributed primarily to UVB light. Indoor tanning raises the risk of melanoma The ultraviolet radiation from these devices can damage the skin, cause premature aging and increase the risk of skin cancer.

56 How effective are the different types of light therapy? Light therapy is often an effective treatment for psoriasis: It is estimated that the symptoms improve noticeably or go away completely for a while in 50 to 90 out of 100 people. It is currently thought that PUVA using psoralen tablets is more effective than narrow band UVB phototherapy. But side effects are also more common and the associated risk of skin cancer may be greater. So it is a good idea to talk to your doctor and carefully weigh the pros and cons of PUVA therapy using psoralen tablets.

57 Commercial LED mask

58 Non-thermal LED light is called photobiomodulation (PBM) or low-level light therapy (LLLT) In contrast to traumatic ablative (e.g., laser resurfacing) and nonablative (e.g., intense pulsed light [IPL]) skin rejuvenation modalities that induce secondary tissue repair by causing controlled damage to either the epidermis or the dermis, is atraumatic, and bypasses the initial destructive step by directly stimulating regenerative processes in the skin.

59 Acne Acne vulgaris is a common skin condition affecting 70 percent of adolescents 2 and frequently continues into adulthood P.acne (Propionibacterium acnes)

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61 P. acnes produce mainly coproporphyrin III, which has an absorption spectrum peak at 415nm. 9 In-vitro irradiation of P. acnes colonies with blue visible light leads to photo-excitation of endogenous bacterial porphyrin, singlet oxygen production, and bacterial destruction. 10

62 This study was an evaluation of the performance of self-applied, blue light, light-emitting diode therapy in the treatment of mild-to-moderate inflammatory acne on the face Clinical Efficacy of Self-applied Blue Light Therapy for Mild-to-Moderate Facial Acne, 2009 This 28-year old male patient had the first outbreak of acne at the age of 23. Baseline (a) and after the eight-week treatment period (b). The total number of lesions had reduced by 94 percent. The number of open comedones was reduced by 100 percent, the number of closed comedones was reduced by 100 percent, and the number of papules was reduced by 84 percent. There were no pustules or nodules present at the start and end of treatment.

63 Results Subjects evaluated self administration of the blue light treatment according to the device s labeling as being safe and effective. The study showed that daily self treatment using the device for mildto-moderate inflammatory acne reduced the number of acne lesions significantly. This female patient had her first outbreak of acne at the age of 17.The total number of lesions had reduced by 62 percent. Baseline (a) and after the eightweek treatment period (b)

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66 Red light technology (RLT) 136 volunteers. Collagen ultrasonography examples significant improvement in the intervention groups compared with the control. shows two series of collagen ultrasonography scans, demonstrating the collagen density increase from t0 to t30 for one subject each in the RLT group A Controlled Trial to Determine the Efficacy of Red and Near-Infrared Light Treatment in Patient Satisfaction, Reduction of Fine Lines, Wrinkles, Skin Roughness, and Intradermal Collagen Density Increase, 2014

67 Conclusions with either or nm polychromatic light (normalized to 9 J/cm 2 in the range of nm) For non-thermal photorejuvenation, laser and LED light sources have been demonstrated to be safe and effective. However, lasers and LEDs may offer some disadvantages because of dot-shaped (punctiform) emission characteristics and their narrow spectral bandwidths. Because the action spectra for tissue regeneration and repair consist of more than one wavelength if it is favorable to apply a polychromatic spectrum covering a broader spectral region for skin rejuvenation and repair.