Effects of Cryolipolysis on the Conversion of White Adipose Tissue: Pilot Study.

INTRODUCTION: Cryolipolysis (CLL) is a widely employed noninvasive procedure for body fat reduction. It operates by inducing cooling, leading to the crystallization of cytoplasmic lipids, loss of cellular integrity, and apoptosis/necrosis of adipocytes, accompanied by local inflammation. Ongoing discussions revolve around CLL's potential to transform white adipocytes into brown adipocytes, potentially yielding more significant effects compared to alternative procedures. OBJECTIVE: Thus, this randomized, blinded clinical study aimed to investigate the effects of CLL on adipose tissue and elucidate the mechanisms involved in its application and capacity for adipocyte conversion. METHODOLOGY: Tissue samples from six patients were assessed at intervals of 45, 60, and 90 days following the application of the CLL protocol during abdominoplasty surgeries. RESULTS: The samples underwent immunohistochemical analyses targeting various markers, revealing higher expression of PPAR-gamma, PPAR-alpha, and UCP-1 markers in CLL-treated samples. CONCLUSION: Therefore, the present study suggests that CLL has the ability to intervene in adipocyte conversion.

Plasma Jet versus Electrocarbonization in the Treatment of Wrinkles of the Upper Palpebral Region.

Background: The plasma jet is a non-surgical and minimally invasive procedure that acts by heating the superficial region of the skin, providing rejuvenation of the region. Objective: We sought to compare the clinical and histological effects of direct plasma jet versus electrocarbonization without plasma in the treatment of wrinkles in the upper palpebral region. Methods: This is a clinical trial in which 20 volunteers participated and divided into two groups: electrocarbonization (EG) and plasma jet (JPG), which were clinically evaluated before and after treatment through evaluation protocols, photographs, and questionnaires. Three treatment sessions were performed in the upper eyelid region with an interval of 30 days. After the treatment, a surgical procedure of upper blepharoplasty was performed on two volunteers to remove a skin fragment and later histological analysis. Results: The JPG showed better clinical responses in rejuvenation. The EG promoted an increase in the number of fibroblasts, the number of blood vessels and the amount of inflammation. The JPG showed an increase in the number of fibroblasts and blood vessels. It was observed that the JPG generated activation of T lymphocytes (CD3), macrophages (CD68), and plasmocytes (CD138); in addition to reducing the number of positive cells for CD57 (NK cells). The satisfaction analysis shows that 100 percent in the JPG considered themselves satisfied with the treatment. Conclusion: We verified that in comparison with electrocarbonization, the use of a direct plasma jet promoted tissue improvement at the histological level, in addition to fewer adverse reactions.

Innovation of the protocol for the application of cryolipolysis: Effects and mechanisms of action.

BACKGROUND: In recent years, aesthetic procedures aiming at body remodeling and have grown exponentially. Cryolipolysis (CLL) has stood out as a noninvasive resource that acts directly on the subcutaneous adipose tissue promoting a significant reduction of adipose tissue through of cooling that could lead to the crystallization cytoplasmic lipids, loss of cellular integrity, apoptosis/necrosis of adipocytes, and local inflammation, producing selective loss of adipose tissue. Thus, the objective of the present study was to evaluate the effects of a specific technique of CLL application on the inflammatory reactions of the target tissue in different post-application times. METHODS: This is a randomized, blind clinical study that evaluated the tissue sample of six patients after 45, 60, and 90 days of an innovative protocol for the application of CLL, with samples collected through abdominoplasty surgeries. The samples were evaluated by immunohistochemical analyses of several markers. RESULTS: A significantly greater increase in fibroblasts was observed at 45 days and greater phagocytic action at 60 days. Regarding the apoptosis process, the expression of caspase 3 and cleaved caspase 3 markers varied at different times, with cleaved caspase 3 being higher at 45 and 90 days after CLL application. CONCLUSION: The protocol of the CLL presented in this study was able to induce inflammatory responses in addition to confirming the selective apoptotic action at the different times studied.

Effect of photobiomodulation on the behaviour of mesenchymal stem cells in three-dimensional cultures.

Photobiomodulation (PBM) has been proposed as a strategy to improve the regenerative capacity of human adipose-derived stem cells (hASCs). Yet, this effect has been proved in 2D culture conditions. To analyze the effect of different doses of laser irradiation (660 nm) with different levels of energy (1 J, 2 J and 6 J) on hASCs cultured at 2D and 3D conditions. We used gellan gum spongy-like hydrogels as a biomaterial to 3D culture hASCs. Different doses (1-7 daily irradiations) and energy levels (1-6 J) of PBM were applied, and the metabolic activity, viability, proliferation, and release of ROS and IL-8 was evaluated up to 7 days. In 3D, cell proliferation increased at high energy (6 J) and after a single dose of irradiation, while in 2D, metabolic activity and proliferation was enhanced only after 3 doses and independently of the energy. More than 1 dose was needed to promote ROS secretion both in 2D and 3D culture conditions. Interestingly, a decrease of IL-8 secretion was detected only in 3D after 3-7 daily irradiations. Overall, hASCs response to PBM was not only dependent on the energy level and the number of applied stimuli, but also on the in vitro culture conditions.

Bone substitutes and photobiomodulation in bone regeneration: A systematic review in animal experimental studies.

In general, bone fractures are able of healing by itself. However, in critical situations such as large bone defects, poor blood supply or even infections, the biological capacity of repair can be impaired, resulting in a delay of the consolidation process or even in non-union fractures. Thus, technologies able of improving the process of bone regeneration are of high demand. In this context, ceramic biomaterials-based bone substitutes and photobiomodulation (PBM) have been emerging as promising alternatives. Thus, the present study performed a systematic review targeting to analyze studies in the literature which investigated the effects of the association of ceramic based bone substitutes and PBM in the process of bone healing using animal models of bone defects. The search was conducted from March and April of 2019 in PubMed, Web of Science and Scopus databases. After the eligibility analyses, 16 studies were included in this review. The results showed that the most common material used was hydroxyapatite (HA) followed by Biosilicate associated with infrared PBM. Furthermore, 75% of the studies demonstrated positive effects to stimulate bone regeneration from association of ceramic biomaterials and PBM. All studies used low-level laser therapy (LLLT) device and the most studies used LLLT infrared. The evidence synthesis was moderate for all experimental studies for the variable histological analysis demonstrating the efficacy of techniques on the process of bone repair stimulation. In conclusion, this review demonstrates that the association of ceramic biomaterials and PBM presented positive effects for bone repair in experimental models of bone defects.

Collagen from Marine Sources and Skin Wound Healing in Animal Experimental Studies: a Systematic Review.

Collagen (Col) from marine organisms has been emerging as an important alternative for commercial Col and it has been considered highly attractive by the industry. Despite the positive effects of Col from marine origin, there is still limited understanding of the effects of this natural biomaterial in the process of wound healing in animal studies. In this context, the purpose of this study was to perform a systematic review of the literature to examine the effects of Col from different marine species in the process of skin tissue healing using experimental models of skin wound. The search was carried out according to the orientations of Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA), and the descriptors of the Medical Subject Headings (MeSH) were defined: "marine collagen," "spongin," "spongin," "skin," and "wound." A total of 42 articles were retrieved from the databases PubMed and Scopus. After the eligibility analyses, this review covers the different marine sources of Col reported in 10 different papers from the beginning of 2011 through the middle of 2019. The results were based mainly on histological analysis and it demonstrated that Col-based treatment resulted in a higher deposition of granulation tissue, stimulation of re-epitalization and neoangiogenesis and increased amount of Col of the wound, culminating in a more mature morphological aspect. In conclusion, this review demonstrates that marine Col from different species presented positive effects on the process of wound skin healing in experimental models used, demonstrating the huge potential of this biomaterial for tissue engineering proposals.

Evaluation of the In Vivo Biological Effects of Marine Collagen and Hydroxyapatite Composite in a Tibial Bone Defect Model in Rats.

One of the most promising strategies to improve the biological performance of bone grafts is the combination of different biomaterials. In this context, the aim of this study was to evaluate the effects of the incorporation of marine spongin (SPG) into Hydroxyapatite (HA) for bone tissue engineering proposals. The hypothesis of the current study is that SPG into HA would improve the biocompatibility of material and would have a positive stimulus into bone formation. Thus, HA and HA/SPG materials were produced and scanning electron microscopy (SEM) analysis was performed to characterize the samples. Also, in order to evaluate the in vivo tissue response, samples were implanted into a tibial bone defect in rats. Histopathological, immunohistochemistry, and biomechanical analyses were performed after 2 and 6 weeks of implantation to investigate the effects of the material on bone repair. The histological analysis demonstrated that composite presented an accelerated material degradation and enhanced newly bone formation. Additionally, histomorphometry analysis showed higher values of %BV/TV and N.Ob/T.Ar for HA/SPG. Runx-2 immunolabeling was higher for the composite group and no difference was found for VEGF. Moreover, the biomechanical analysis demonstrated similar values for all groups. These results indicated the potential of SPG to be used as an additive to HA to improve the biological performance for bone regeneration applications. However, further long-term studies should be carried out to provide additional information regarding the material degradation and bone regeneration.

Biphasic Dose/Response of Photobiomodulation Therapy on Culture of Human Fibroblasts.

Objective: The objective of this study was to evaluate the effects of application of different fluences and energies of laser in the 24-, 48-, and 72-h periods in fibroblasts originating from human skin (HFF-1). Methods: The cell used as a template for cell proliferation was HFF-1. For the photobiomodulation (PBM) application, a 660 nm laser with a power of 40 mW and energies of 0.84, 1.40, 5.88, and 6.72 J was used. Five experimental groups were studied: one control group (CG) with simulated PBM and four groups that received PBM in different doses. The changes observed after laser irradiation were evaluated by cell viability (trypan blue) and proliferation [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)] tests. Intergroup comparisons were performed using two-way analysis of variance and the Tukey post hoc test (software GraphPad Prism 7.0). Results: In the trypan blue test, the total number of cells was significantly different between the irradiated groups and the CG at all times studied. The total number of cells increased in laser group (LG)1 (0.84 J) and LG2 (1.40 J) and decreased in LG4 (6.72 J). The mitochondrial activity increased significantly in LG1 and LG2 at 48 and 72 h and decreased in LG3 (5.88 J) and LG4 (6.72 J) compared with CG. Conclusions: The results indicate that the lower doses (0.45 and 0.75 J/cm2) of PBM induce the highest mitochondrial activity and cellular viability.

Effect of photobiomodulation associated with cell therapy in the process of cutaneous regeneration in third degree burns in rats.

Due to the complexity involved in the healing process of full thickness burns, the literature looks for alternatives to optimize tissue reconstruction. The objective of this study was to explore the action of photobiomodulation therapy associated with MSCs in the healing process of third degree burns. A total of 96 male Wistar rats were used, distributed in four groups with 24 animals each: Control Group, Laser Group, Cell Therapy Group, and Laser Group and Cell Therapy. The burn was performed with aluminum plate (150 °C). We performed analysis of wound contraction, histology, immunohistochemistry, birefringence analysis, and immunoenzymatic assay to evaluate tissue quality. Our results demonstrate that the association of the techniques is able to accelerate the repair process, modulating the inflammatory process, presenting a cutaneous tissue with better quality. Thus, we conclude that the use of photobiomodulation therapy associated with cell therapy is a promising treatment in the repair of total thickness burns.

Photobiomodulation therapy and transcutaneous electrical nerve stimulation on chronic neck pain patients: Study protocol clinical trial (SPIRIT Compliant).

INTRODUCTION: Chronic neck pain is a common musculoskeletal disorder that is associated with functional disability and decreased of quality of life. Electrophysical agents are commonly used to relieve pain, however the effects of combined use of these agents are little studied. The objective is to investigate the efficacy of photobiomodulation and electrical stimulation to relieve pain, both in isolation and combined. MATERIALS AND METHODS: This a 4-arm randomized placebo-controlled trial with patient and evaluator blinded. This study will be performed in Department of Physical Therapy at Federal University of São Carlos, São Carlos/SP, Brazil. One hundred and forty-four patients with chronic neck pain will be randomized into 4 groups: active photobiomodulation therapy with active electrical stimulation, active photobiomodulation therapy, active electrical stimulation, or placebo treatment. They will receive 10 sessions of treatment. PRIMARY OUTCOME: pain intensity (measured by pain numerical rating scale) posttreatment. SECONDARY OUTCOMES: pain during movement, neck disability, range of motion, pressure pain threshold, temporal summation, conditioned pain modulation, depressive symptoms, pain catastrophizing, quality of life, analgesic intake, and global perceived effect at posttreatment (10 sessions). Pain intensity and global perceived effect will also be measured after 6 weeks randomization. DISCUSSION: The findings of this study might clarify the importance of using the photobiomodulation therapy and transcutaneous electrical nerve stimulation for patients with chronic neck pain. TRIAL REGISTRATION: NCT04020861. https://clinicaltrials.gov/ct2/show/NCT04020861?term=NCT04020861&draw=2&rank=1.

Photobiomodulation effect on the proliferation of adipose tissue mesenchymal stem cells.

The use of mesenchymal stem cells (MSCs) in tissue engineering has been extensively investigated. The greater the proliferation of this cellular group, the greater the regenerative and healing capacity of the tissue to which they belong. In this context, photobiomodulation (PBM) is an efficient technique in proliferation of distinct cell types. However, its parameters and mode of action are still unclear and require further investigation. This study aimed to evaluate the PBM action with different energies in MSCs of adipose tissue (hASCs). We used hASCs, seeded in 24-well plates, with 3 × 104 cells per well, in culture media. We used a total of four experimental groups, one with hASCs and simulated PBM and three other groups, which received PBM irradiation at 24, 48, and 72 h, with a 660-nm laser and power of 40 mW and energy of 0.56, 1.96, and 5.04 J. We performed analyses of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor) and trypan blue to evaluate cell proliferation and viability, 1 h after PBM irradiation. Software Graph PadPrism 7.0 was used. Intergroup comparisons were performed with ANOVA two-way and we used the Tukey post hoc test. Mitochondrial activity evaluated by MTT revealed the statistical difference in the first 24 h for group with more high energy when compared to control group; and in the 72 h for two irradiated groups when compared to the control group. The trypan blue test showed significant differences at the end of the experiment for two irradiated groups LG1 (4.52 × 104 ± 0.2) and LG2 (4.85 × 104 ± 0.8), when compared to the control group (1.87 × 104 ± 0.7). Both tests failed to be statistically different at the end of the experiment for groups LG1 and LG2 and observed a reduction in cellular mitochondrial growth and activity for group LG3. We conclude that PBM with energy close to 0.56 and 1.96 J promote proliferation of hASCs, and higher energy, such as 5.04 J, can be harmful.

Photobiomodulation on critical bone defects of rat calvaria: a systematic review.

Bone defects following trauma represent a high impact on the quality of life of millions of people around the world. The aim of this study was to review photobiomodulation (PBM) action in the treatment of bone critical defects in rat calvaria, related to evaluation of the current protocols applied. One hundred and forty-seven articles related to the subject were found by searching the main databases (Pubmed, Lilacs, Web of Science, and Scopus) considering the period of publication until the year 2017, and only 14 corresponded the inclusion criteria established for this systematic review. The main parameters of the PBM were expressed in Table 1. In addition, it was possible to observe the use of two different wavelengths (red and infrared), which are considered therapeutic. Most of the evaluated articles presented positive results that describe a greater amount of neoformed bone, an increase in collagen synthesis, and a contribution to microvascular reestablishment. However, two studies report no effect on the repair process when the PBM was used. In addition, we observed considerable variations between the values of power, fluence, and total energy, which make it difficult to compare the results presented between the selected studies. It was possible to conclude that the infrared laser was more effective in positively stimulating the bone repair process of critical defects. Furthermore, a discrepancy was found in the parameter values used, which made it difficult to choose the best protocol for the treatment of this type of lesion.

Evaluation of the low-level laser therapy application parameters for skin burn treatment in experimental model: a systematic review.

Burn is defined as a traumatic injury of thermal origin, which affects the organic tissue. Low-level laser therapy (LLLT) has gained great prominence as a treatment in this type of injury; however, the application parameters are still controversial in the literature. The aims of this study were to review the literature studies that use LLLT as a treatment in burns conducted in an experimental model, discuss the main parameters used, and highlight the benefits found in order to choose an appropriate therapeutic window to be applied in this type of injury. The selection of the studies related to the theme was carried out in the main databases (PubMed, Cochrane Library, LILACS, Web of Science, and Scopus in the period from 2001 to 2017). Subsequently, the articles were then chosen that fell within the inclusion criteria previously established. In the end, 22 were evaluated, and the main parameters were presented. The analyzed studies presented both LLLT use in continuous and pulsed mode. Differences between the parameters used (power, fluence, and total energy) were observed. In addition, the protocols are distinct as to the type of injury and the number of treatment sessions. Among the results obtained by the authors are the improvements in the local microcirculation and cellular proliferation; however, a study reported no effects with LLLT as a treatment. LLLT is effective in accelerating the healing process. However, there is immense difficulty in establishing the most adequate protocol, due to the great discrepancy found in the applied dosimetry values.

Effect of photobiomodulation therapy (808 nm) in the control of neuropathic pain in mice.

Neuropathic pain can be defined as the pain initiated or caused by a primary lesion or dysfunction of the central or peripheral nervous system. Photobiomodulation therapy (PBM) stands out among the physical therapy resources used for analgesia. However, application parameters, especially the energy density, remain controversial in the literature. Therefore, this study aimed to investigate the PBM effect, in different energy densities to control neuropathic pain in mice. Fifty (50) mice were induced to neuropathy by chronic constriction surgery of the sciatic nerve (CCI), treated with PBM (808 nm), and divided into five groups: GP (PBM simulation), GS (sham), GL10, GL20, GL40 (energy density of 10, 20, and 40 J/cm2, respectively). The evaluations were carried out using the hot plate test and Randall and Selitto test, before and after the CCI surgery, every 15 days during the 90 days experiment. ß-Endorphin blood dosage was also tested. For both the hot plate and Randall and Selitto tests, the GL20 and GL40 groups presented reduction of the nociceptive threshold from the 30th day of treatment, the GL10 group only after day 75, and the GP group did not show any improvement throughout the experiment. The ß-endorphin dosage was higher for all groups when compared to the GP group. However, only the GL20 group and GL40 presented a significant increase. This study demonstrates that PBM in higher energy density (20, 40 J/cm2) is more effective in the control of neuropathic pain.

Use of low level laser therapy to control neuropathic pain: A systematic review.

Neuropathic pain can be defined as pain initiated or caused by a primary lesion or dysfunction in the central or peripheral nervous system. The low level laser therapy (LLLT) has gained great prominence as a treatment in this type of pain; however, the application parameters are still controversial in the literature. This study aimed to review the literature on the use of LLLT in neuropathic pain with the goal of establishing a "therapeutic window" for the effective use of this treatment. We analyzed 14 articles, 10 in experimental animals and 4 in humans. The results are presented in three tables, the first being for comparison of the studies' application parameters, the second showing the average and median parameters experimental studies and third showing the clinical studies embodiment. The experimental studies revealed better results for LLLT and infrared laser powers above 70mW. Clinical studies are inconclusive as to the application parameters, due to the discrepancy; however all demonstrate the effectiveness of LLLT. According to the data presented, it was concluded that LLLT has positive effects on the control of analgesia for neuropathic pain, but further studies with high scientific rigor are needed in order to define treatment protocols that optimize the action LLLT in neuropathic pain.