Just Accepted by Journal of Cosmetic and Laser Therapy
Biophotonic effect of diode laser irradiance on tensile strength of diabetic rats Pik Suan Lau, Noriah Bidin, Ganesan Krishnan, Zaleha Nassir, Harzi Bahktiar doi: 10.3109/14764172.2014.968587
J Cosmet Laser Ther Downloaded from informahealthcare.com by Ryerson University on 10/10/14 For personal use only.
Abstract Low energy laser irradiance at certain wavelengths able to stimulate the tissue bio-reaction and enhanced the healing process. Collagen deposition is one of the important aspects in healing process because it can increase the strength of the skin. This study was designed to examine the biophotonic effect of irradiance on collagen production of diabetic wound in rat model. The tensile strength of skin was employed as parameter to describe the wound. Diabetic rat model were induced by streptozotocin (STZ) via intravenous injection. Skin-breaking strength was measured by using an Instron tensile test machine. The experimental animals were treated with 808nm diode laser at two different powers that are 0.1 W/cm2 and 0.5 W/cm2 and 30 s, 60 s and 120 s for each session. The tensile strength was optimized after treated with high power diode laser. The photostimulation effect was revealed in accelerating healing process and enhanced the tensile strength of wound. Laser photostimulation on tensile strength in diabetic wound suggests that such therapy facilitates collagen production in manner that diabetic wound healing.
© 2014 Informa UK, Ltd. This provisional PDF corresponds to the article as it appeared upon acceptance. Fully formatted PDF and full text (HTML) versions will be made available soon. DISCLAIMER: The ideas and opinions expressed in the journal’s Just Accepted articles do not necessarily reflect those of Informa Healthcare (the Publisher), the Editors or the journal. The Publisher does not assume any responsibility for any injury and/or damage to persons or property arising from or related to any use of the material contained in these articles. The reader is advised to check the appropriate medical literature and the product information currently provided by the manufacturer of each drug to be administered to verify the dosages, the method and duration of administration, and contraindications. It is the responsibility of the treating physician or other health care professional, relying on his or her independent experience and knowledge of the patient, to determine drug dosages and the best treatment for the patient. Just Accepted articles have undergone full scientific review but none of the additional editorial preparation, such as copyediting, typesetting, and proofreading, as have articles published in the traditional manner. There may, therefore, be errors in Just Accepted articles that will be corrected in the final print and final online version of the article. Any use of the Just Accepted articles is subject to the express understanding that the papers have not yet gone through the full quality control process prior to publication.
Biophotonic effect of diode laser irradiance on tensile strength of diabetic rats
Pik Suan Lau1, Noriah Bidin1,*, Ganesan Krishnan1, Zaleha Nassir2, Harzi Bahktiar1 1
Advance Photonic Science Institute,Faculty of science,Universiti Teknologi Malaysia, 81310,UTM Johor Bahru,Johor
PT ED
Darul Ta’azim, 2Klinik Perubatan, UTM Johor, Universiti Teknologi Malaysia,81310 UTM Johor Bahru, Johor,Malaysia
Short title: Laser therapy on diabetic rats Abstract
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Low energy laser irradiance at certain wavelengths able to stimulate the tissue bio-reaction and enhanced the healing process. Collagen deposition is one of the important aspects in healing process because it can increase the strength of the skin. This study was designed to examine the biophotonic effect of irradiance on collagen production of diabetic wound in rat model. The tensile strength of skin was employed as parameter to describe the wound. Diabetic rat model were induced by streptozotocin (STZ) via intravenous injection. Skin-breaking strength was measured by using an Instron tensile test machine. The experimental animals were treated with 808nm diode laser at two different powers that are 0.1 W/cm2 and 0.5 W/cm2 and 30 s, 60 s and 120 s for each session. The tensile strength was optimized after treated with high power diode laser. The photostimulation effect was revealed in accelerating healing process and enhanced the tensile strength of wound. Laser photostimulation on tensile strength in diabetic wound suggests that such therapy facilitates collagen production in manner that diabetic wound healing. Key word: diode laser, biophotonic, diabetic wound, collagen, healing, tensile strength
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*Corresponding Author: Noriah Bidin, Advance Photonic Science Institute,Faculty of science,Universiti Teknologi Malaysia, 81310,UTM Johor Bahru,Johor Darul Ta’azim. E-mail: [email protected]
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Biophysics is an knowledge that applied the fundamental of physics to study biological system, which lead to the new field that study of life1 . The one of the studies is biophotonic, it denotes a combination interaction of biology and photonic2. The energy of photon involve the intermolecular interaction, conformational changes and molecular transport that will activates the biological process3-4. Therefore, biophotonic technique with lights has been used for variety of medical therapies such as wound healing. Photo-irradiation at low energy levels absorbed by photo agent in living cell able to generates significant bio-effects either in biostimulation or bioinhibition5-9. Low level laser therapy (LLLT) with output power 5-500mW has been found to promote the healing process in term of reduce the inflammation and enhanced the proliferation phase10-16. The effective of treatment was depending on laser parameter, the complexity of choosing was reported by several authors 17-20. Hawkins et al.21 have report that the visible red and infrared wavelength revealed to have highly absorbent and unique therapeutic effects in living tissues. Gungormus et al. 22 evaluated the inflammation and re-epithelialization using near infrared laser to exposure cells in vivo stated that the treatment schedule of irradiation are more important than the total energy dose in biostimulatory. The most commonly stated mechanism for enhancement of laser therapy on wound healing is that intracellular energy transfer motivated adenosine diphosphate (ADP) transform to adenosine triphosphate (ATP) in mitochondria vice versa 23-25. ADP-ATP cycling provides the chemical energy within cells for induce metabolism and collagen production by fibroblasts26-27. Fibroblast is synthesizes the extracellular matrix and collagen, it is the most common type of cell found in connective tissue. Fibroblast is a main feature to create collagen in proliferation phase during the healing process. Then, in phase of wound repair regulation of excess collagen deposition enable to enhanced the quality of wound28. The total collagen and insoluble collagen as well as the collagen ratio indirectly represent the tensile strength of skin29. Tensile strength is important in healing closed incision, it will restoration the strength of skin. Many of the chronic complications of diabetes involve defects in connective tissue such as poor healing and collagen production30-31. These have results to demonstrate the tensile strength of diabetic lower than normal control group32. In this study was undertaken to investigate the interaction between the LLLT and tensile strength in diabetic model. 2.0 Material and method Twenty-one laboratory male rats, weight in the range of 250-300g were employed as experimental animals. The procedures are following the policy for care and use of experimental animals by Animal Ethics Committee, in Universiti Kebangsaan Malaysia (approval project code: UTM/APSI/2013/NORIAH/20-MARCH/502-MARCH-2013-JAN-2015). All of rats were allowed to have free access to sterile food and water. The rats were then divided randomly into one control group and two treated groups, the treat groups were further sub-divided into three groups. Each rat was inducing diabetes through the administration of streptozotocin (STZ) by single intravenous injection of 45mg/kg body weight 33. The fur on the dorsal surface of rat was shaved off using an electric mouse clipper and anesthetized with Ketamine (10mg/kg body weight). The shaved surface was first cleaned with an alcohol pad for skin disinfection. This experiment was investigated the optimum laser dose and time exposure for single treatment to exhibit the best tissue regeneration potential. 808nm diode laser in continuous wave mode was irradiance to skin. The spot size of the beam at focus point was a 0.47mm2. In order to cover wide exposed area as well as to preserve from direct thermal damage, the target tissue is placed 5 mm away from the focal point. The rats were exposed by scanning a single laser beam on the rat skin. The rats were divided into three groups, one serve as control group and two treat groups with power of 0.1 W/cm2 and 0.5 W/cm2 respectively. The different treatment schedule was verified within the 30 s, 60 s and 120 s for each session. For allowing the collagen biostimulation effect, 1 hour post irradiance all the diabetic rats were sacrifice. The samples skin was cut with dimension of around 2 × 20 mm2. Skin-breaking strength was measured by using an Instron tensile test machine. The holder was pulls the sample vertically and moving at a constant speed of 5 mm/min until the broken skin occurs. Maximal load for pulling sample skin is recorded in kilo-newton (kN), the value obtained will divide by cross section area (m2). It is considered to be maximal wound breaking strength and also called Young’s modulus tensile strength (E) in unit MPa. 3.0 Result and discussion
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1.0 Introduction
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The findings from current study indicated that the overall group of power 0.5 W/cm2 enhanced the skin tensile strength in several treatment schedules compare with control group and the group of power 0.1 W/cm2. To understand the collagen capacity of each laser dose and treatment schedule, a comparison was display in Figure 1. The tensile strength E of skin for group of rat which exposed after 30 s interaction time with power 0.5 W/cm2 was increased to be as E=12.50 MPa, but reduced to E=12.125 MPa after longer treatment time of 120s. A single scanning of diode laser with 0.5 W/cm2 exposure within interaction time of 60 second showed a significantly enhancement in tensile strength of E=15.25 MPa. For comparison purpose the tensile strength of control skin was measured to be as E=10.81 MPa. Apparently, no significant change was noticed in the tensile strength rat skin after exposed with diode laser with power of 0.1 W/cm2. In fact after treatment time of 30s and 60s, the tensile strength was realized to be reduced as E=6.88 MPa and E=9.56 MPa respectively. However after exposure with longer period of interaction time of 120s a significant enhancement was revealed to be as 13.63 MPa. The reduction in tensile strength is also means that the decreasing the collagen and fibroblast in the derma layer of skin. In this case the lower power of diode laser deactivate the collagen production instead of stimulated as desire. It is not clear the reason for such reduction. May be further experiment require to be carried to understand the cause of weakening the tensile strength consequently decreasing the collagen in the skin. Based on analysis results, the laser power at 0.5 W/cm2 has been stimulation the collagen to enhance the tensile strength of skin. The ability to achieve successful stimulation collagen is of paramount importance for wound healing process. The increment is almost 41% better than the untreated skin. Thus the diode laser 808 nm has offering a new way of treatment, beneficial for render wound strong sooner, which able to reduce the risk of inflammation and increase the healing process that is benefit for surgery and trauma. Low level laser therapy is one of technique in biophotonic in medical field. It involves the application of low power monochromatic and coherent light to wound and lesion. The results are conflicting in different studies and may depend on the method of application and other parameter of the laser 34-36. However, our group and other have shown that LLLT are capable of motivate fibroblast and collagen to accelerating healing and enhance the tensile strength. The use of low power of near infrared light for enhance tensile strength has been reported by Stadler et al.37, 830nm diode laser was employed to irradiance to diabetic and nondiabetic groups of mice with 85mW and 5J/cm2 fluence. The overall wound strength was enhanced in both groups. Whereas several investigators have mention the effect of stimulation was occurs in low doses of laser radiation and suppressive in high doses of LLLT. In our study, power of 0.5 W/cm2 was illustrate biostimulation at short exposure time and bioinhibit at longer exposure time, which was similar to research by Skopin 38. Still, low levels of laser within 0.1 W/cm2 have much better effects on stimulating and repairing in bio-molecular than higher levels of light in longer exposure time. Photobiostimualtion is not only dose dependence, it also wavelength dependence. Do Nascimento et al.39 have indicated that inversely proportional relationship between wavelength and intensity. The combining higher intensity with short wavelength or lower intensity wither higher wavelength were more effective. This vivo study is still in preliminary stage that suggests photostimulation is capable to increase collagen for enhanced tensile strength of skin. The results demonstrated that collagen synthesis is increasing by photo-radiation at 808 nm with power 0.5 W/cm2 at 60s. It is a plenty room for improvement. For future works the relationship between laser parameters and skin properties will be performed to establish the optimum activation of collagen. 4.0 Conclusion Diode laser 808 nm capable to enhance the tensile strength of the rat skin. The optimum power and treatment interaction time is achieved with 0.5 W/cm2 and 60s respectively. The lower power laser diode at 0.1 W/cm2 is also manage to activate a stronger tensile strength but required a longer period of exposure. The enhancement of the tensile strength after diode laser at 808 nm is due to stimulation of collagen and fibroblast in the dermal layer of skin, which responsible to strengthen the elasticity properties of the skin.
Acknowledgements The authors would like to thank MyBrain15 for the scholarship and Malaysian Ministry of Higher Education and Universiti Teknologi Malaysia for their financial funding through FRGS grant 40. Reference
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M., Couppé, C., Chow, R. T., Turner, J., and Ljunggren, E. A.A systematic review of low level laser therapy with location-specific doses for pain from chronic joint disorders, Australian Journal of Physiotherapy 49,(2003), 107-122. Correa, F., Martins, R. A. B. L., Correa, J. C., Iversen, V. V., Joenson, J., and Bjordal, J. M.Low-level laser therapy (GaAs λ= 904 nm) reduces inflammatory cell migration in mice with lipopolysaccharide-induced peritonitis, Photomedicine and laser surgery 25,(2007), 245-249. Stein, A., Benayahu, D., Maltz, L., and Oron, U.Low-level laser irradiation promotes proliferation and differentiation of human osteoblasts in vitro, Photomedicine and Laser Therapy 23,(2005), 161-166. Walsh, L.The current status of low level laser therapy in dentistry, Part 1. Soft tissue applications, Australian dental journal 42,(1997), 247-254. Al-Watban, F. A., Zhang, X. Y., and Andres, B. L.Low-level laser therapy enhances wound healing in diabetic rats: a comparison of different lasers, Photomedicine and laser surgery 25,(2007), 72-77. Herascu, N., Velciu, B., Calin, M., Savastru, D., and Talianu, C.Low-level laser therapy (LLLT) efficacy in postoperative wounds, Photomedicine and Laser Therapy 23,(2005), 70-73. Hamblin, M. R., and Demidova, T. N. Mechanisms of low level light therapy, In Biomedical Optics 2006, pp 614001-614001-614012, International Society for Optics and Photonics.2006. Huang, Y.-Y., Chen, A. C.-H., Carroll, J. D., and Hamblin, M. R.Biphasic dose response in low level lightherapy, Dose-Response 7,(2009), 358-383. Hamblin, M. R., Demidova-Rice, T. N., Weersink, R., White, R., Lilge, L., Eduardo, F. P., Mehnert, D. U., Monezi, T. A., Zezell, D. M., and Schubert, M. M.Mechanisms for Low-Light Therapy III (Proceedings Volume),(2008). Krishnan, G., and Bidin, N.Determination of a stimulated emission cross section gradient on the basis of the performance of a diode-end-pumped Nd: YVO4 laser, Laser Physics Letters 10,(2013), 055007. Hawkins, D., Houreld, N., and Abrahamse, H.Low level laser therapy (LLLT) as an effective therapeutic modality for delayed wound healing, Annals of the New York Academy of Sciences 1056,(2005), 486-493. Güngörmüş, M., and Akyol, U. K.Effect of biostimulation on wound healing in diabetic rats, Photomedicine and laser surgery 27,(2009), 607-610. Tohgi, H., Takahashi, H., Watanabe, K., Kuki, H., and Shirasawa, Y.Development of large platelet aggregates from small aggregates as determined by laser-light scattering: effects of aggregant concentration and antiplatelet medication, Thrombosis and haemostasis 75,(1996), 838-843. Oron, U., Ilic, S., De Taboada, L., and Streeter, J.Ga-As (808 nm) laser irradiation enhances ATP production in human neuronal cells in culture, Photomedicine and laser surgery 25,(2007), 180-182. Passarella, S., Ostuni, A., Atlante, A., and Quagliariello, E.Increase in the ADP/ATP exchange in rat liver mitochondria irradiated in vitro by helium-neon laser, Biochemical and biophysical research communications 156,(1988), 978-986. Weksler, B. B., Marcus, A. J., and Jaffe, E. A.Synthesis of prostaglandin I2 (prostacyclin) by cultured human and bovine endothelial cells, Proceedings of the National Academy of Sciences 74,(1977), 3922-3926.
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Murugappa, S., and Kunapuli, S. P.The role of ADP receptors in platelet function, Frontiers in bioscience: a journal and virtual library 11,(2006), 1977. Witte, M. B., Thornton, F. J., Kiyama, T., Efron, D. T., Schulz, G. S., Moldawer, L. L., and Barbul, A.Metalloproteinase inhibitors and wound healing: a novel enhancer of wound strength, Surgery 124,(1998), 464470. Vogel, H.Correlation between tensile strength and collagen content in rat skin. Effect of age and cortisol treatment, Connective tissue research 2,(1974), 177-182. Fahey III, T. J., Sadaty, A., Jones II, W. G., Barber, A., Smoller, B., and Shires, G. T.Diabetes impairs the late inflammatory response to wound healing, Journal of Surgical Research 50,(1991), 308-313. Spanheimer, R. G., Umpierrez, G. E., and Stumpf, V.Decreased collagen production in diabetic rats, Diabetes 37,(1988), 371-376. Riou, J.-P. A., Cohen, J. R., and Johnson Jr, H.Factors influencing wound dehiscence, The American journal of surgery 163,(1992), 324-330. Kang, K. P., Lee, J. E., Lee, A. S., Jung, Y. J., Lee, S., Park, S. K., Kim, W., Pokrywczynska, M., Jundzill, A., and Krzyzanowska, S.Diabetes-Experimental, Nephrology Dialysis Transplantation 27,(2012), ii159-ii166. Santos, N. R., dos Santos, J. N., dos Reis Jr, J. A., Oliveira, P. C., de Sousa, A. P. C., de Carvalho, C. M., Soares, L. G., Marques, A. M., and Pinheiro, A. L. B.Influence of the Use of Laser Phototherapy (λ660 or 790 nm) on the Survival of Cutaneous Flaps on Diabetic Rats, Photomedicine and laser surgery 28,(2010), 483-488. Kaviani, A., Djavid, G. E., Ataie-Fashtami, L., Fateh, M., Ghodsi, M., Salami, M., Zand, N., Kashef, N., and Larijani, B.A randomized clinical trial on the effect of low-level laser therapy on chronic diabetic foot wound healing: a preliminary report, Photomedicine and laser surgery 29,(2011), 109-114. Kawalec, J. S., Hetherington, V. J., Pfennigwerth, T. C., Dockery, D. S., and Dolce, M.Effect of a diode laser on wound healing by using diabetic and nondiabetic mice, The Journal of foot and ankle surgery 43,(2004), 214-220. Stadler, I., Lanzafame, R. J., Evans, R., Narayan, V., Dailey, B., Buehner, N., and Naim, J. O.830‐nm irradiation increases the wound tensile strength in a diabetic murine model*, Lasers in surgery and medicine 28,(2001), 220226. Skopin, M. D., and Molitor, S. C.Effects of near‐infrared laser exposure in a cellular model of wound healing, Photodermatology, Photoimmunology & Photomedicine 25,(2009), 75-80. Do Nascimento, P. M., Pinheiro, A. L. B., Ângelo Castilho Salgado, M., and Pedreira Ramalho, L. M.A preliminary report on the effect of laser therapy on the healing of cutaneous surgical wounds as a consequence of an inversely proportional relationship between wavelength and intensity: histological study in rats, Photomedicine and Laser Therapy 22,(2004), 513-518.
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Figure Legends
Figure 1. The comparison between treated and untreated skin
Biophotonic effect of diode laser irradiance on tensile strength of diabetic rats Pik Suan Lau, Noriah Bidin, Ganesan Krishnan, Zaleha Nassir, Harzi Bahktiar doi: 10.3109/14764172.2014.968587
J Cosmet Laser Ther Downloaded from informahealthcare.com by Ryerson University on 10/10/14 For personal use only.
Abstract Low energy laser irradiance at certain wavelengths able to stimulate the tissue bio-reaction and enhanced the healing process. Collagen deposition is one of the important aspects in healing process because it can increase the strength of the skin. This study was designed to examine the biophotonic effect of irradiance on collagen production of diabetic wound in rat model. The tensile strength of skin was employed as parameter to describe the wound. Diabetic rat model were induced by streptozotocin (STZ) via intravenous injection. Skin-breaking strength was measured by using an Instron tensile test machine. The experimental animals were treated with 808nm diode laser at two different powers that are 0.1 W/cm2 and 0.5 W/cm2 and 30 s, 60 s and 120 s for each session. The tensile strength was optimized after treated with high power diode laser. The photostimulation effect was revealed in accelerating healing process and enhanced the tensile strength of wound. Laser photostimulation on tensile strength in diabetic wound suggests that such therapy facilitates collagen production in manner that diabetic wound healing.
© 2014 Informa UK, Ltd. This provisional PDF corresponds to the article as it appeared upon acceptance. Fully formatted PDF and full text (HTML) versions will be made available soon. DISCLAIMER: The ideas and opinions expressed in the journal’s Just Accepted articles do not necessarily reflect those of Informa Healthcare (the Publisher), the Editors or the journal. The Publisher does not assume any responsibility for any injury and/or damage to persons or property arising from or related to any use of the material contained in these articles. The reader is advised to check the appropriate medical literature and the product information currently provided by the manufacturer of each drug to be administered to verify the dosages, the method and duration of administration, and contraindications. It is the responsibility of the treating physician or other health care professional, relying on his or her independent experience and knowledge of the patient, to determine drug dosages and the best treatment for the patient. Just Accepted articles have undergone full scientific review but none of the additional editorial preparation, such as copyediting, typesetting, and proofreading, as have articles published in the traditional manner. There may, therefore, be errors in Just Accepted articles that will be corrected in the final print and final online version of the article. Any use of the Just Accepted articles is subject to the express understanding that the papers have not yet gone through the full quality control process prior to publication.
Biophotonic effect of diode laser irradiance on tensile strength of diabetic rats
Pik Suan Lau1, Noriah Bidin1,*, Ganesan Krishnan1, Zaleha Nassir2, Harzi Bahktiar1 1
Advance Photonic Science Institute,Faculty of science,Universiti Teknologi Malaysia, 81310,UTM Johor Bahru,Johor
PT ED
Darul Ta’azim, 2Klinik Perubatan, UTM Johor, Universiti Teknologi Malaysia,81310 UTM Johor Bahru, Johor,Malaysia
Short title: Laser therapy on diabetic rats Abstract
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Low energy laser irradiance at certain wavelengths able to stimulate the tissue bio-reaction and enhanced the healing process. Collagen deposition is one of the important aspects in healing process because it can increase the strength of the skin. This study was designed to examine the biophotonic effect of irradiance on collagen production of diabetic wound in rat model. The tensile strength of skin was employed as parameter to describe the wound. Diabetic rat model were induced by streptozotocin (STZ) via intravenous injection. Skin-breaking strength was measured by using an Instron tensile test machine. The experimental animals were treated with 808nm diode laser at two different powers that are 0.1 W/cm2 and 0.5 W/cm2 and 30 s, 60 s and 120 s for each session. The tensile strength was optimized after treated with high power diode laser. The photostimulation effect was revealed in accelerating healing process and enhanced the tensile strength of wound. Laser photostimulation on tensile strength in diabetic wound suggests that such therapy facilitates collagen production in manner that diabetic wound healing. Key word: diode laser, biophotonic, diabetic wound, collagen, healing, tensile strength
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*Corresponding Author: Noriah Bidin, Advance Photonic Science Institute,Faculty of science,Universiti Teknologi Malaysia, 81310,UTM Johor Bahru,Johor Darul Ta’azim. E-mail: [email protected]
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Biophysics is an knowledge that applied the fundamental of physics to study biological system, which lead to the new field that study of life1 . The one of the studies is biophotonic, it denotes a combination interaction of biology and photonic2. The energy of photon involve the intermolecular interaction, conformational changes and molecular transport that will activates the biological process3-4. Therefore, biophotonic technique with lights has been used for variety of medical therapies such as wound healing. Photo-irradiation at low energy levels absorbed by photo agent in living cell able to generates significant bio-effects either in biostimulation or bioinhibition5-9. Low level laser therapy (LLLT) with output power 5-500mW has been found to promote the healing process in term of reduce the inflammation and enhanced the proliferation phase10-16. The effective of treatment was depending on laser parameter, the complexity of choosing was reported by several authors 17-20. Hawkins et al.21 have report that the visible red and infrared wavelength revealed to have highly absorbent and unique therapeutic effects in living tissues. Gungormus et al. 22 evaluated the inflammation and re-epithelialization using near infrared laser to exposure cells in vivo stated that the treatment schedule of irradiation are more important than the total energy dose in biostimulatory. The most commonly stated mechanism for enhancement of laser therapy on wound healing is that intracellular energy transfer motivated adenosine diphosphate (ADP) transform to adenosine triphosphate (ATP) in mitochondria vice versa 23-25. ADP-ATP cycling provides the chemical energy within cells for induce metabolism and collagen production by fibroblasts26-27. Fibroblast is synthesizes the extracellular matrix and collagen, it is the most common type of cell found in connective tissue. Fibroblast is a main feature to create collagen in proliferation phase during the healing process. Then, in phase of wound repair regulation of excess collagen deposition enable to enhanced the quality of wound28. The total collagen and insoluble collagen as well as the collagen ratio indirectly represent the tensile strength of skin29. Tensile strength is important in healing closed incision, it will restoration the strength of skin. Many of the chronic complications of diabetes involve defects in connective tissue such as poor healing and collagen production30-31. These have results to demonstrate the tensile strength of diabetic lower than normal control group32. In this study was undertaken to investigate the interaction between the LLLT and tensile strength in diabetic model. 2.0 Material and method Twenty-one laboratory male rats, weight in the range of 250-300g were employed as experimental animals. The procedures are following the policy for care and use of experimental animals by Animal Ethics Committee, in Universiti Kebangsaan Malaysia (approval project code: UTM/APSI/2013/NORIAH/20-MARCH/502-MARCH-2013-JAN-2015). All of rats were allowed to have free access to sterile food and water. The rats were then divided randomly into one control group and two treated groups, the treat groups were further sub-divided into three groups. Each rat was inducing diabetes through the administration of streptozotocin (STZ) by single intravenous injection of 45mg/kg body weight 33. The fur on the dorsal surface of rat was shaved off using an electric mouse clipper and anesthetized with Ketamine (10mg/kg body weight). The shaved surface was first cleaned with an alcohol pad for skin disinfection. This experiment was investigated the optimum laser dose and time exposure for single treatment to exhibit the best tissue regeneration potential. 808nm diode laser in continuous wave mode was irradiance to skin. The spot size of the beam at focus point was a 0.47mm2. In order to cover wide exposed area as well as to preserve from direct thermal damage, the target tissue is placed 5 mm away from the focal point. The rats were exposed by scanning a single laser beam on the rat skin. The rats were divided into three groups, one serve as control group and two treat groups with power of 0.1 W/cm2 and 0.5 W/cm2 respectively. The different treatment schedule was verified within the 30 s, 60 s and 120 s for each session. For allowing the collagen biostimulation effect, 1 hour post irradiance all the diabetic rats were sacrifice. The samples skin was cut with dimension of around 2 × 20 mm2. Skin-breaking strength was measured by using an Instron tensile test machine. The holder was pulls the sample vertically and moving at a constant speed of 5 mm/min until the broken skin occurs. Maximal load for pulling sample skin is recorded in kilo-newton (kN), the value obtained will divide by cross section area (m2). It is considered to be maximal wound breaking strength and also called Young’s modulus tensile strength (E) in unit MPa. 3.0 Result and discussion
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1.0 Introduction
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The findings from current study indicated that the overall group of power 0.5 W/cm2 enhanced the skin tensile strength in several treatment schedules compare with control group and the group of power 0.1 W/cm2. To understand the collagen capacity of each laser dose and treatment schedule, a comparison was display in Figure 1. The tensile strength E of skin for group of rat which exposed after 30 s interaction time with power 0.5 W/cm2 was increased to be as E=12.50 MPa, but reduced to E=12.125 MPa after longer treatment time of 120s. A single scanning of diode laser with 0.5 W/cm2 exposure within interaction time of 60 second showed a significantly enhancement in tensile strength of E=15.25 MPa. For comparison purpose the tensile strength of control skin was measured to be as E=10.81 MPa. Apparently, no significant change was noticed in the tensile strength rat skin after exposed with diode laser with power of 0.1 W/cm2. In fact after treatment time of 30s and 60s, the tensile strength was realized to be reduced as E=6.88 MPa and E=9.56 MPa respectively. However after exposure with longer period of interaction time of 120s a significant enhancement was revealed to be as 13.63 MPa. The reduction in tensile strength is also means that the decreasing the collagen and fibroblast in the derma layer of skin. In this case the lower power of diode laser deactivate the collagen production instead of stimulated as desire. It is not clear the reason for such reduction. May be further experiment require to be carried to understand the cause of weakening the tensile strength consequently decreasing the collagen in the skin. Based on analysis results, the laser power at 0.5 W/cm2 has been stimulation the collagen to enhance the tensile strength of skin. The ability to achieve successful stimulation collagen is of paramount importance for wound healing process. The increment is almost 41% better than the untreated skin. Thus the diode laser 808 nm has offering a new way of treatment, beneficial for render wound strong sooner, which able to reduce the risk of inflammation and increase the healing process that is benefit for surgery and trauma. Low level laser therapy is one of technique in biophotonic in medical field. It involves the application of low power monochromatic and coherent light to wound and lesion. The results are conflicting in different studies and may depend on the method of application and other parameter of the laser 34-36. However, our group and other have shown that LLLT are capable of motivate fibroblast and collagen to accelerating healing and enhance the tensile strength. The use of low power of near infrared light for enhance tensile strength has been reported by Stadler et al.37, 830nm diode laser was employed to irradiance to diabetic and nondiabetic groups of mice with 85mW and 5J/cm2 fluence. The overall wound strength was enhanced in both groups. Whereas several investigators have mention the effect of stimulation was occurs in low doses of laser radiation and suppressive in high doses of LLLT. In our study, power of 0.5 W/cm2 was illustrate biostimulation at short exposure time and bioinhibit at longer exposure time, which was similar to research by Skopin 38. Still, low levels of laser within 0.1 W/cm2 have much better effects on stimulating and repairing in bio-molecular than higher levels of light in longer exposure time. Photobiostimualtion is not only dose dependence, it also wavelength dependence. Do Nascimento et al.39 have indicated that inversely proportional relationship between wavelength and intensity. The combining higher intensity with short wavelength or lower intensity wither higher wavelength were more effective. This vivo study is still in preliminary stage that suggests photostimulation is capable to increase collagen for enhanced tensile strength of skin. The results demonstrated that collagen synthesis is increasing by photo-radiation at 808 nm with power 0.5 W/cm2 at 60s. It is a plenty room for improvement. For future works the relationship between laser parameters and skin properties will be performed to establish the optimum activation of collagen. 4.0 Conclusion Diode laser 808 nm capable to enhance the tensile strength of the rat skin. The optimum power and treatment interaction time is achieved with 0.5 W/cm2 and 60s respectively. The lower power laser diode at 0.1 W/cm2 is also manage to activate a stronger tensile strength but required a longer period of exposure. The enhancement of the tensile strength after diode laser at 808 nm is due to stimulation of collagen and fibroblast in the dermal layer of skin, which responsible to strengthen the elasticity properties of the skin.
Acknowledgements The authors would like to thank MyBrain15 for the scholarship and Malaysian Ministry of Higher Education and Universiti Teknologi Malaysia for their financial funding through FRGS grant 40. Reference
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Figure Legends
Figure 1. The comparison between treated and untreated skin
