Effect of laser phototherapy on wound healing following cerebral ischemia by cryogenic injury

Laser phototherapy emerges as an alternative or auxiliary therapy for acute ischemic stroke, traumatic brain injury, degenerative brain disease, spinal cord injury, and peripheral nerve regeneration, but its effects are still controversial. We have previously found that laser phototherapy immunomodulates the response to focal brain damage. Following direct cortical cryogenic injury the effects of laser phototherapy on inflammation and repair was assessed after cryogenic injury (CI) to the central nervous system (CNS) of rats. The laser phototherapy was carried out with a 780 nm AlGaAs diode laser. The irradiation parameters were: power of 40 mW, beam area of 0.04 cm(2), energy density of 3 J/cm(2) (3s) in two points (0.12 J per point). Two irradiations were performed at 3 h-intervals, in contact mode. Rats (20 non-irradiated - controls and 20 irradiated) were used. The wound healing in the CNS was followed in 6 h, 1, 7 and 14 days after the last irradiation. The size of the lesions, the neuron cell viability percentages and the amount of positive GFAP labeling were statistically compared by ANOVA complemented by Tukey's test (p<0.05). The distribution of lymphocytes, leukocytes and macrophages were also analyzed. CI created focal lesions in the cortex represented by necrosis, edema, hemorrhage and inflammatory infiltrate. The most striking findings were: lased lesions showed smaller tissue loss than control lesions in 6 h. During the first 24 h the amount of viable neurons was significantly higher in the lased group. There was a remarkable increase in the amount of GFAP in the control group by 14 days. Moreover, the lesions of irradiated animals had fewer leukocytes and lymphocytes in the first 24 h than controls. Considering the experimental conditions of this study it was concluded that laser phototherapy exerts its effect in wound healing following CI by controlling the brain damage, preventing neuron death and severe astrogliosis that could indicate the possibility of a better clinical outcome.     

Effect of low intensity helium-neon (HeNe) laser irradiation on experimental paracoccidioidomycotic wound healing dynamics

The effect of HeNe laser on the extracellular matrix deposition, chemokine expression and angiogenesis in experimental paracoccidioidomycotic lesions was investigated. At days 7, 8 and 9 postinfection the wound of each animal was treated with a 632.8 nm HeNe laser at a dose of 3 J cm(-2). At day 10 postinfection, the wounds were examined by using histologic and immunohistochemical methods. Results revealed that laser-treated lesions were lesser extensive than untreated ones, and composed mainly by macrophages and lymphocytes. High IL-1beta expression was shown in the untreated group whereas in laser-treated animals the expression was scarce. On the other hand, the expression of CXCL-10 was found to be reduced in untreated animals and quite intensive and well distributed in the laser-treated ones. Also, untreated lesions presented vascular endothelial growth factor (VEGF) in a small area near the center of the lesion and high immunoreactivity for hypoxia-inducible factor-1 (HIF-1), whereas laser-treated lesions expressed VEGF surrounding blood vessels and little immunoreactivity for HIF-1. Laser-treated lesions presented much more reticular fibers and collagen deposition when compared with the untreated lesion. Our results show that laser was efficient in minimizing the local effects observed in paracoccidioidomycosis and can be an efficient tool in the treatment of this infection, accelerating the healing process.     

Effect of Andrographis paniculata leaf extract on wound healing in rats

This work was carried out to study the effect of topical application of Andrographis paniculata on the rate of wound enclosure and its histological features. A wound was created in four groups of rat in posterior neck region. Blank placebo was applied topically to the wounds of Group 1. Groups 2 and 3 were dressed with placebo containing 5% and 10% extracts of A. paniculata, respectively. Intrasite gel was applied topically to the wounds of Group 4. Macroscopical examination revealed that the rate of wound healing was significantly accelerated in the wound dressed with A. paniculata extract compared to the blank placebo. The wounds dressed with 10% extract or Intrasite gel healed earlier compared to the wounds dressed with placebo containing 5% A. paniculata extract. Histologically, wounds dressed with A. paniculata extracts showed markedly less scar width and contained large amounts of fibroblast proliferation. More collagen and less angiogenesis with absence of inflammatory cells were seen for wounds dressed with 10% A. paniculata compared to the blank placebo. Conclusion, A. paniculata extracts significantly enhanced rate of wound healing in rats.     

Influence of laser photobiomodulation upon connective tissue remodeling during wound healing

The modulation of collagen fibers during experimental skin wound healing was studied in 112 Wistar rats submitted to laser photobiomodulation treatment. A standardized 8mm-diameter wound was made on the dorsal skin of all animals. In half of them, 0.2ml of a silica suspension was injected along the border of the wound in order to enhance collagen deposition and facilitate observation. The others received saline as vehicle. The treatment was carried out by means of laser rays from an aluminum-gallium arsenide diode semiconductor with 9mW applied every other day (total dose=4J/cm(2)) on the borders of the wound. Tissue sections obtained from four experimental groups representing sham-irradiated animals, laser, silica and the association of both, were studied after 3, 7, 10, 15, 20, 30 and 60 days from the laser application. The wounded skin area was surgically removed and submitted to histological, immunohistochemical, ultrastructural, and immunofluorescent studies. Besides the degree and arrangement of collagen fibers and of their isotypes, the degree of edema, the presence of several cell types especially pericytes and myofibroblasts, were described and measured. The observation of Sirius-red stained slides under polarized microscopy revealed to be of great help during the morphological analysis of the collagen tissue dynamic changes. It was demonstrated that laser application was responsible for edema regression and a diminution in the number of inflammatory cells (p<0.05). An evident increase in the number of actin-positive cells was observed in the laser-treated wounds. Collagen deposition was less than expected in silica-treated wounds, and laser treatment contributed to its better differentiation and modulation in all irradiated groups. Thus, laser photobiomodulation was able to induce several modifications during the cutaneous healing process, especially in favoring newly-formed collagen fibers to be better organized and compactedly disposed.     

Evaluation of mitochondrial respiratory chain activity in wound healing by low-level laser therapy

Laser therapy is used in many biomedical sciences to promote tissue regeneration. Many studies involving low-level laser therapy have shown that the healing process is enhanced by such therapy. In this work, we evaluated mitochondrial respiratory chain complexes II and IV and succinate dehydrogenase activities in wounds after irradiation with low-level laser. The animals were divided into two groups: group 1, the animals had no local nor systemic treatment and were considered as control wounds; group 2, the wounds were treated immediately after they were made and every day after with a low-level laser (AsGa, wavelength of 904 nm) for 10 days. The results showed that low-level laser therapy improved wound healing. Besides, our results showed that low-level laser therapy significantly increased the activities of complexes II and IV but did not affect succinate dehydrogenase activity. These findings are in accordance to other works, where cytochrome c oxidase (complex IV) seems to be activated by low-level laser therapy. Besides, we showed, for the first time, that complex II activity was also activated. More studies are being carried out in order to evaluate other mitochondrial enzymes activities after different doses and irradiation time of low-level laser.     

Janus hydrogel with dual antibacterial and angiogenesis functions for enhanced diabetic wound healing

Anti-infection and neovascularization at the wound site are two vital factors that accelerate diabetic wound healing. However, for a wound healing dressing, the two functions need to work at different sites(inner and outer), giving big challenges for dressing design. In this study, we fabricated a novel sodium alginate/chitosan(SA/CS) Janus hydrogel dressing by the assembly of SA hydrogel loaded with silver nanoparticles(Ag NPs) and CS hydrogel impregnated with L-arginine loaded sodium alginate ...     

Electrospun cellulose acetate wound dressings loaded with Pramipexole for diabetic wound healing: an in vitro and in vivo study

In the current study, a Pramipexole-loaded wound dressing was produced via electrospinning of cellulose acetate solution. Pramipexole was added to cellulose acetate solution at 3, 5, and 10% w/w concentrations and then electrospun. The produced wound dressings were studied regarding their physicochemical and biological properties. Results of cell viability assay and cytoprotection studies showed that cellulose acetate wound dressings containing 3% w/w Pramipexole had significantly higher cell viability compared with other concentrations. The wound healing potential of dressings incorporated with 3% drug was studied in a rat model of diabetic wound. Study showed that the cellulose acetate/3% Pramipexole scaffolds had significantly higher percentage of wound closure, epithelial thickness, and collagen deposition compared with drug-free dressings and control group. Gene expression study showed that the drug-loaded wound dressings could reduce oxidative stress and alleviate inflammation at significantly higher extent compared with other groups.

     

Effect of electrospun non-woven mats of dibutyryl chitin/poly(lactic acid) blends on wound healing in hairless mice

The aim of this study was to examine the proliferative ability of dibutyryl chitin (DBC) on scratch wounds in HaCaT keratinocytes and to evaluate the effect of nanoporous non-woven mat (DBCNFM) on skin wound healing in hairless mice using the advantages of DBCNFM, such as high porosity and high surface area to volume. The cell spreading activity of DBC was verified through a cell spreading assay in scratched human HaCaT keratinocytes. Scratch wound experiments showed that DBC notably accelerates the spreading rate of HaCaT keratinocytes in a dose dependent manner. The molecular aspects of the healing process were also investigated by hematoxylin & eosin staining of the healed skin, displaying the degrees of reepithelialization and immunostaining on extracellular matrix synthesis and remodeling of the skin. Topical application of DBCNFM significantly reduced skin wound rank scores and increased the skin remodeling of the wounded hairless mice in a dose dependent way. Furthermore, DBCNFM notably increased the expression of the type 1 collagen and filaggrin. These results demonstrate that DBC efficiently accelerates the proliferation of HaCaT keratinocytes and DBCNFM notably increases extracellular matrix synthesis on remodeling of the skin, and these materials are a good candidate for further evaluation as an effective wound healing agent.     

Low-power laser irradiation improves histomorphometrical parameters and bone matrix organization during tibia wound healing in rats

The influence of daily energy doses of 0.03, 0.3 and 0.9 J of He-Ne laser irradiation on the repair of surgically produced tibia damage was investigated in Wistar rats. Laser treatment was initiated 24 h after the trauma and continued daily for 7 or 14 days in two groups of nine rats (n=3 per laser dose and period). Two control groups (n=9 each) with injured tibiae were used. The course of healing was monitored using morphometrical analysis of the trabecular area. The organization of collagen fibers in the bone matrix and the histology of the tissue were evaluated using Picrosirius-polarization method and Masson's trichrome. After 7 days, there was a significant increase in the area of neoformed trabeculae in tibiae irradiated with 0.3 and 0.9 J compared to the controls. At a daily dose of 0.9 J (15 min of irradiation per day) the 7-day group showed a significant increase in trabecular bone growth compared to the 14-day group. However, the laser irradiation at the daily dose of 0.3 J produced no significant decrease in the trabecular area of the 14-day group compared to the 7-day group, but there was significant increase in the trabecular area of the 15-day controls compared to the 8-day controls. Irradiation increased the number of hypertrophic osteoclasts compared to non-irradiated injured tibiae (controls) on days 8 and 15. The Picrosirius-polarization method revealed bands of parallel collagen fibers (parallel-fibered bone) at the repair site of 14-day-irradiated tibiae, regardless of the dose. This organization improved when compared to 7-day-irradiated tibiae and control tibiae. These results show that low-level laser therapy stimulated the growth of the trabecular area and the concomitant invasion of osteoclasts during the first week, and hastened the organization of matrix collagen (parallel alignment of the fibers) in a second phase not seen in control, non-irradiated tibiae at the same period. The active osteoclasts that invaded the regenerating site were probably responsible for the decrease in trabecular area by the fourteenth day of irradiation.     

Topical propolis improves wound healing in patients with diabetic foot ulcer: a randomized controlled trial

In this randomized controlled trial, diabetic patients with foot ulcers (Wagner grades 1 and 2) were randomly assigned to conventional therapies for diabetic foot ulcer plus topical propolis ointment (5%; twice daily) or conventional therapies alone. The process of ulcer healing was observed during 4 weeks and compared between the two groups regarding the size, erythema, exudates, white blood cell (WBC) count and erythrocyte sedimentation rate (ESR). The process of ulcer size reduction during the four-week period of study was significantly different between the groups. However, this difference was not significant between the third and fourth weeks. There was no significant difference between two groups regarding erythema and exudate reduction as well as WBC count and ESR. Administration of topical propolis ointment in addition to the conventional treatments of diabetic foot ulcer could reduce the size of ulcers with Wagner grades 1 and 2.     

Effect of low-level helium-neon laser therapy on the healing of third-degree burns in rats

This paper presents the results of a study on the effects of low-level helium-neon laser therapy (LL He-Ne LT) on the healing of burns. Seventy-eight adult male rats, having been subjected to third-degree burns, were randomly divided into four groups: two laser treated groups (n=20, each), one control group (n=19) and one nitrofurazone treated group (n=19). In the two laser treated groups, the burns were treated on a daily basis with LL He-Ne LT with an energy density of 1.2 and 2.4 J/cm(2), respectively. The response to treatment was assessed histologically at 7, 16 and 30 days after burning, and microbiologically at day 15. Analysis of variance showed that the mean of blood vessel sections in the 1.2J/cm(2) laser group was significantly higher than those in the other groups and the mean of the depth of new epidermis in the 2.4 J/cm(2) laser group on day 16 was significantly lower than in the nitrofurazone treated group (P=0.025, P=0.047, respectively). When Staphylococcus aureus and Pseudomonas aeruginosa grew in more than 50% of samples obtained from control group, there were no S. aureus and P. aeruginosa in the samples of 2.4 J/cm(2) laser group. It is concluded that LL He-Ne LT induced the destruction of S. aureus and P. aeruginosa in third-degree burns of rats, yet at the same time our histological findings showed that LL He-Ne LT caused a significant increase in the mean of blood vessel sections on day 7 after third degree burns and a decrease in the mean of the depth of new epidermis on day 16 after the same burns in rats.     

Effect of a topical formulation containing Calophyllum brasiliense Camb. extract on cutaneous wound healing in rats

This study evaluated the wound healing effects of topical application of an emulsion containing the HPLC-standardised extract from Calophyllum brasiliense Cambess (Clusiaceae) leaves in rats. The macroscopic analysis demonstrated that the wounds treated with the C. brasiliense emulsion healed earlier than the wounds treated with emulsion base and Dersani®. The percentage of wound healing in the group treated with the C. brasiliense emulsion was significantly higher than in the other groups at 7 and 14 days. On day 14, the animals treated with the C. brasiliense emulsion exhibited a 90.67% reduction of the wound areas. The histological evaluation revealed that on day 21, the group treated with the C. brasiliense emulsion exhibited a significant increase in fibroblasts compared with the other groups. Thus, the C. brasiliense emulsion had healing properties in the topical treatment of wounds and accelerated the healing process.     

The effect of 880 nm low level laser energy on human fibroblast cell numbers: a possible role in hypertrophic wound healing

Low level lasers (LLLs) have been shown to induce therapeutic effects in wound healing. However, there have been few LLL studies on burn wounds which may become unsightly, hypertrophic and impair function. Inhibitory effects on the healing of fibrotic wounds, prone to hypertrophy may be expected to reasonably reduce the problems accompanying hypertrophic scarring. The effects of LLL wavelengths and treatment parameters on wound healing cells in vitro often demonstrate meaningful results and without concurrent ethical difficulties of clinical trials. This experiment investigated the effect of an 880 nm, 16 mW GaAlAs diode at 2.4 and 4 J/cm(2) on cell numbers of two human fibroblast cell lines, derived from hypertrophic scar (HF) and normal dermal explants (NF), respectively. After irradiation by 880 nm LLL, cell numbers were measured utilising methylene blue bioassay and read by the spectrophotometer in the same microculture plates. HF and NF exhibited decreased cell numbers as compared to sham-irradiated controls. HF cell number, after 2.4 J/cm(2), was significantly lower on day 5 (P<0.05). The NF cell numbers were significantly lower on day 4 and/or day 5 (P<0.05). The results have implications on hypertrophic wound healing and further studies are required.     

Antioxidant, antimicrobial and wound healing activities of Boesenbergia rotunda

The ethanolic extract of Boesenbergia rotunda (L.) Mansf was studied for its wound-healing potential. Since wound healing is interrelated with microbial infection and reactive oxygen species (ROS), this study was conducted to evaluate the antimicrobial and antioxidant activity of B. rotunda. The antimicrobial activity of B. rotunda was studied against six bacterial and two yeast strains using disc diffusion, minimum inhibitory concentration (MIC), and minimum microbicidal concentration (MMC). The B. rotunda extract displayed potential antimicrobial and antifungal activities by inhibiting the Gram-positive bacteria Staphylococcus aureus (ATCC 25923), S. epidermidis, and Bacillus subtilis (ATCC 6633), and the yeasts Candida albicans (ATCC 10231), and Saccharomyces cerevisiae. MIC and MMC values varied from 0.04 to 25 mg/mL and from 0.16 to 25 mg/mL, respectively. The antioxidant activity of B. rotunda was evaluated by measuring the Ferric Reducing/Antioxidant Power (FRAP) and DPPH free radical scavenging activity. The FRAP and DPPH values were 22.2 microM/microg and 76.3 mg/mL, respectively. In the wound-healing studies, the topical application of the B. rotunda extract indicated a significantly increased percentage of wound contraction on day 12 compared with the control group. Histological studies showed the complete epidermis and found collagen fibers and hair follicles in the dermis. The results of the present study support the continued and expanded utilization of B. rotunda in Thai folk medicine.     

Effect of low-level laser therapy on the healing of second-degree burns in rats: a histological and microbiological study

This paper presents the results of a study on the effects of two different doses of low-level laser therapy on healing of deep second-degree burns. Sixty rats were randomly allocated to one of four groups. A deep second-degree burn was inflicted in each rat. In the control group burns remained untreated; in two laser treated groups the burns were irradiated daily with low-level helium-neon laser with energy densities of 1.2 and 2.4 J/cm2, respectively. In the fourth group the burns were treated topically with 0.2% nitrofurazone cream every day. The response to treatments was assessed histologically at 7, 16 and 30 days after burning, and microbiologically at Day 15. The number of macrophages at day 16, and the depth of new epidermis at day 30, was significantly less in the laser treated groups in comparison with control and nitrofurazone treated groups (P=0.000). Staphylococcus epidermidis was found in the 70% of rat wounds in the laser treated groups in comparison with 100% of rats in the control group. S. aureus was found in the 40% rat wounds in the nitrofurazone treated group, but there was not found in the wounds of laser treated, and control groups. It is concluded that low-level laser therapy of deep second-degree burn caused significant decrease in the number of macrophage and depth of new epidermis. In addition, it decreased incidence of S. epidermidis and S. aureus.     

Antimicrobial and wound healing activities of electrospun nanofibers based on functionalized carbohydrates and proteins

The anti-adhesion, anti-growth and the anti-penetration of bacteria, specifically multidrug-resistant bacteria, should be taken into consideration when designing promising wound dressings for infected wounds such as diabetic foot ulcers. Wound dressings composed of natural polymeric nanofibers such as functionalized cellulose, chitosan, alginate, hyaluronic acid, dextrin and cyclodextrin with appropriate antimicrobial and skin reconstruction properties are suitable alternatives that can accelerate wound healing and remove microbial infections. For instance, to improve the release profile of antibacterial agents such as metal nanoparticles and antibiotics, water-soluble polymers like polyethylene oxide and polyvinylpyrrolidone may be incorporated into polymeric nanofiber scaffolds. This review, therefore, addresses the current status and future challenges of antibacterial activities of nanofiber scaffolds composed of some of the natural occurring polymers.

     

Synergy of light-controlled Pd nanozymes with NO therapy for biofilm elimination and diabetic wound treatment acceleration

The presence of bacteria, existing as highly organized biofilm communities, in chronic non-healing wounds has been identified as a significant impediment for wound healing. Nanozymes, with unique antimicrobial mechanisms, as a new alternative for antibiotics, have the potential to synergize with nitric oxide (NO) with enhanced antibacterial and antibiofilm ability. However, the always-on state of nanozymes and the reactivity of NO limit their clinical applications. In this context, an intelligent and multifunctional Pd-MOF@PAzo@SNP nanoplatform was fabricated using UiO-66 as a palladium (Pd) nanozyme-loading vehicle, then a surface modification with photosensitive polyazobenzene (PAzo), and the adsorption of the NO donor sodium nitroprusside (SNP) via a host-guest interaction between β-cyclodextrin-modified hyaluronic acid (β-CD-HA) and azobenzene. The activity of Pd-nanozyme was easily controlled via ultraviolet (UV) light, and its photosensitivity was regulated by changing the side-chain unit length of PAzo. Furthermore, NO was released in response to the UV irradiation and played a synergistic role with the peroxidase activity of Pd nanozyme, exhibiting excellent antibacterial and antibiofilm activity in the presence of 0.01 mM hydrogen peroxide (H₂O₂). In vivo, Pd-MOF@PAzo@SNP accelerated the healing of a biofilm-infected diabetic wound by dispersing the biofilm, reducing bacterial burden, and promoting angiogenesis and collagen deposition. Overall, the nanoplatform provides a reliable and highly efficient strategy to develop an intelligent nanozyme synergy with NO therapy in chronic wound management.     

Application of titanium nanoparticles containing natural compounds in cutaneous wound healing

The aim of the study was the rapid green synthesis of titanium nanoparticles using the aqueous extract of Falcaria vulgaris leaves (TiNPs@FV) and exploring their antioxidant, cytotoxicity, antifungal, antibacterial, and cutaneous wound healing activities under in vitro and in vivo condition. These nanoparticles were characterized by UV-Vis, Fourier transform-infrared(FT-IR), X-ray diffraction XRD), field emission-scanning electron microscopy FE-SEM), and transmission electron microscopy TEM) analyses. The synthesized TiNPs@FV had great cell viability on human umbilical vein endothelial cells and indicted this method was nontoxic. DPPH (2,2-diphenyl-1-picrylhydrazyl) test revealed similar antioxidant potentials for F. vulgaris, TiNPs@FV, and butylated hydroxytoluene. All data of antibacterial, antifungal, and cutaneous wound healing tests were analyzed by SPSS 22 software. In the antimicrobial part of this study, TiNPs@FV indicated higher antifungal and antibacterial effects than all standard antibiotics (p ≤ 0.01). Minimal inhibitory concentration (MIC) and minimal fungicidal concentration of TiNPs@FV against all fungi were at 2–4 mg/mL and 2-8 mg/mL ranges, respectively. But, MIC and minimal bactericidal concentration of TiNPs@FV against all bacteria were at 2-8 mg/mL and 2-16 mg/mL ranges, respectively. In the part of cutaneous wound healing, use of TiNPs@FV ointment significantly (p ≤ 0.01) raised the wound contracture, vessel, hydroxyl proline, hexuronic acid, hexosamine, fibrocyte, and fibrocytes/fibroblast rate and significantly (p ≤ 0.01) decreased the wound area, total cells, neutrophil, and lymphocyte compared to other groups in rats. The results of FT-IR, UV-Vis, XRD, TEM, and FE-SEM confirm that the aqueous extract of F. vulgaris leaves can be used to yield titanium nanoparticles with a notable amount of remedial effects.