Photobiomodulation therapy was more effective than photobiomodulation plus arginine on accelerating wound healing in an animal model of delayed healing wound.

The combined and individual influences of photobiomodulation therapy (PBMT) and arginine on wound strength, stereological parameters, and gene expressions of some related growth factors in ischemic and delayed healing wounds in rats were analyzed. We divided 108 rats into six groups: control, lower energy density (LOW)-PBMT, 2% arginine ointment (Arg 2%), LOW-PBMT + Arg 2%, high energy density (HIGH)-PBMT, and HIGH-PBMT + Arg 2%. First, we generated an ischemic and delayed healing wound model in each rat. We examined wound strength, stereological parameters, and gene expressions of basic fibroblast growth factor (bFGF), vascular endothelial growth factor A (VEGF-A), and stromal cell-derived factor 1 (SDF-1) by quantitative real-time polymerase chain reaction (qRT-PCR). PBMT alone and PBMT + Arg 2% considerably increased wound strength compared to the control and Arg 2% groups during the inflammatory and proliferative steps of wound healing (p < 0.05). In these steps, PBMT alone significantly induced an anti-inflammatory effect and increased fibroblast counts; Arg 2% alone induced an inflammatory response (p < 0.05). Concurrently, PBMT and PBMT + Arg 2% significantly increased keratinocyte counts and volume of the new dermis (p < 0.05). At the remodeling step, the Arg 2% groups had significantly better wound strength than the other groups (p < 0.05). In this step, PBMT and PBMT + Arg 2% significantly decreased inflammation, and increased fibroblast counts, vascular length, and the volume of new epidermis and dermis compared to the control and Arg 2% groups (p < 0.05). In all cases of gene analysis, there were statistically better results in the PBMT and PBMT + Arg 2% groups compared with the Arg 2% and control groups (p < 0.05). The anti-inflammatory and repairing effects of PBMT on an ischemic and delayed healing wound model in rats were shown by significant improvements in wound strength, stereological parameters, and gene expressions of bFGF, VEGF-A, and SDF-1α.

Combined Treatment of Photobiomodulation and Arginine on Chronic Wound Healing in an Animal Model.

Introduction: Herein, the individual and combined effects of photobiomodulation (PBM) and arginine (ARG) on the wound healing course of an experimental model of a slow healing wound (ulcer) in rats were assessed. Methods: A total of 108 male rats were divided into 6 groups: control; lower energy density (low)-PBM; arginine ointment (ARG); low-PBM+ARG; high energy density (high)-PBM; and high-PBM+ARG. In each rat, one ischemic wound in the center of a bipedicle flap and one non-ischemic wound out of the flap were created. Both wounds were treated in the experimental groups. Microbial growth, wound area, and wound strength were assessed on days 0, 5, 10, 15, and 20 after wound infliction. Results: All non-ischemic wounds closed before day 15. High-PBM+ARG and ARG significantly increased wound closure rates compared to the control group (LSD test, P = 0.000, and P = 0.001, respectively) on day 10. All slow healing wounds were open on day 15 but closed completely before day 20. Low-PBM+ARG and high-PBM significantly increased wound strength (stress high load, SHL) on day 10 compared to the control group (LSD test, P = 0.001, and P = 0.000, respectively). ARG, high-PBM, and low-PBM+ARG significantly increased wound closure rates on day 15 relative to the control group (LSD test, P = 0.000, P = 0.000, and P = 0.001, respectively). Conclusion: High-PBM and low-PBM+ARG have biostimulatory and antibacterial effects on slow-healing wounds, which were shown by significant increases in wound closure rates, wound strength, and inhibition of Staphylococcus aureus growth.

The effect of photobiomodulation therapy on antioxidants and oxidative stress profiles of adipose derived mesenchymal stem cells in diabetic rats.

We studied the effects of photobiomodulation therapy (PBMT) on adipose-derived mesenchymal stem cells (ADSCs) which were extracted from streptozotocin (STZ) induced diabetic rats. Adipose tissue was extracted from the hypodermis of diabetic rats, and diabetic ADSCs were extracted, characterized, and cultured. There were two in vitro groups: control-diabetic ADSCs, and PBMT-diabeticADSCs. We used 630 nm and 810 nm laser at 1.2 J/cm2 with 3 applications 48 h apart. We measured cell viability, apoptosis, population doubling time (PDT), and reactive oxygen species (ROS) by flow cytometry. Gene expression of antioxidants, including cytosolic copper-zinc superoxide dismutase (SOD1), catalase (CAT), total antioxidant capacity (TAC), and oxidative stress biomarkers (NADPH oxidase 1 and 4) by quantitative real time (qRT) - PCR. In this study, data were analyzed using t-test. Viability of PBMT-diabetic- ADSC group was higher than control- diabetic-ADSC (p = 0.000). PDT and apoptosis of PBMT- diabetic-ADSC group were lower than control-diabetic -ADSC (p = 0.001, p = 0.02). SOD1 expression and TAC of PBMT- diabetic-ADSC group were higher than control -diabetic -ADSC (p = 0.018, p = 0.005). CAT of PBMT -diabetic-ADSC group was higher than control-diabetic -ADSC. ROS, NOX1, and NOX4 of PBMT- diabetic -ADSC group were lower than control-diabetic-ADSC (p = 0.002, p = 0.021, p = 0.017). PBMT may improve diabetic- ADSC function in vitro by increasing levels of cell viability, and gene expression of antioxidant agents (SOD1, CAT, and TAC), and significantly decreasing of levels of PDT, apoptosis, ROS, and gene expression of oxidative stress biomarkers (NOX1 and NOX4).

Combined effects of photobiomodulation and curcumin on mast cells and wound strength in wound healing of streptozotocin-induced diabetes in rats.

We investigated the probable involvement of mast cell degranulation and their numbers in the remodeling step of wound healing in a diabetic ischemic skin wound model treated with photobiomodulation plus curcumin. A total of 108 adult male Wistar rats were randomized into one healthy control and five diabetic groups. Type I diabetes was inflicted in 90 of the 108 rats. After 1 month, an excisional wound was generated in each of the 108 rats. There were one healthy group (group 1) and five diabetic groups as follows: group 2 was the untreated diabetic control group and group 3 rats were treated with sesame oil. Rats in group 4 were treated with photobiomodulation (890 nm, 890 ± 10 nm, 80 Hz, 0.2 J/cm2) and those in group 5 received curcumin dissolved in sesame oil. Group 6 rats were treated with photobiomodulation and curcumin. We conducted stereological and tensiometric tests on days 4, 7, and 15 after treatment. The results indicated that photobiomodulation significantly improved wound strength in the diabetic rats and significantly decreased the total numbers of mast cells. The diabetic control group had significantly reduced tensiometric properties of the healing wounds and a significant increase in the total numbers of mast cells. Photobiomodulation significantly improved the healing process in diabetic animals and significantly decreased the total number of mast cells. The increased numbers of mast cells in the diabetic control group negatively affected tensiometric properties of the ischemic skin wound.

Transplantation of photobiomodulation-preconditioned diabetic stem cells accelerates ischemic wound healing in diabetic rats.

BACKGROUND: Diabetic foot ulcer is the most costly and complex challenge for patients with diabetes. We hereby assessed the effectiveness of different preconditioned adipose-derived mesenchymal stem cells (AD-MSCs) and photobiomodulation protocols on treating an infected ischemic wound in type 1 diabetic rats. METHODS: There were five groups of rats: (1) control, (2) control AD-MSCs [diabetic AD-MSCs were transplanted (grafted) into the wound bed], (3) AD-MSC + photobiomodulation in vivo (diabetic AD-MSCs were grafted into the wound, followed by in vivo PBM treatment), (4) AD-MSCs + photobiomodulation in vitro, and (5) AD-MSCs + photobiomodulation in vitro + in vivo. RESULTS: Diabetic AD-MSCs preconditioned with photobiomodulation had significantly risen cell function compared to diabetic AD-MSC. Groups 3 and 5 had significantly decreased microbial flora correlated to groups 1 and 2 (all, p = 0.000). Groups 2, 3, 4, and 5 had significantly improved wound closure rate (0.4, 0.4, 0.4, and 0.8, respectively) compared to group 1 (0.2). Groups 2-5 had significantly increased wound strength compared to group 1 (all p = 0.000). In most cases, group 5 had significantly better results than groups 2, 3, and 4. CONCLUSIONS: Preconditioning diabetic AD-MSCs with photobiomodulation in vitro plus photobiomodulation in vivo significantly hastened healing in the diabetic rat model of an ischemic infected delayed healing wound.

Preconditioning adipose-derived stem cells with photobiomodulation significantly increased bone healing in a critical size femoral defect in rats.

We assessed the combined impacts of human demineralized bone matrix (hDBM) scaffold, adipose-derived stem cells (hADS), and photobiomodulation (PBM) on bone repair of a critical size femoral defect (CSFD) in 72 rats. The rats were divided into six groups: control (group 1); ADS (group 2 - ADS transplanted into hDBM); PBM (group 3 - PBM-treated CSFDs); ADS + PBM in vivo (group 4 - ADS transplanted into hDBM and the CSFDs were treated with PBM in vivo); ADS + PBM in vitro (group 5 - ADS were treated with PBM in vitro, then seeded into hDBM); and ADS + PBM in vitro+in vivo (group 6 - PBM-treated ADS were seeded into hDBM, and the CSFDs were treated with PBM in vivo. At the anabolic phase (2 weeks after surgery), bone strength parameters of the groups 5, 6, and 4 were statistically greater than the control, ADS, and PBM in vivo groups (all, p = 0.000). Computed tomography (CT) scans during the catabolic phase (6 weeks after surgery) of bone healing revealed that the Hounsfield unit (HU) of CSFD in the groups 2 (p = 0.000) and 5 (p = 0.019) groups were statistically greater than the control group. The groups 5, 4, and 6 had significantly increased bone strength parameters compared with the PBM in vivo, control, and ADS groups (all, p = 0.000). The group 5 was statistically better than the groups 4, and 6 (both, p = 0.000). In vitro preconditioned of hADS with PBM significantly increased bone repair in a rat model of CSFD in vivo.

Combined therapy of photobiomodulation and adipose-derived stem cells synergistically improve healing in an ischemic, infected and delayed healing wound model in rats with type 1 diabetes mellitus.

OBJECTIVE: We assessed the therapeutic effects of photobiomodulation (PBM) and adipose-derived stem cell (ADS) treatments individually and together on the maturation step of repairing of a delayed healing wound model in rats with type 1 diabetes mellitus (DM1). RESEARCH DESIGN AND METHODS: We randomly assigned 24 rats with DM1 to four groups (n=6 per group). Group 1 was the control (placebo) group. In group 2, allograft human ADSs were transplanted. Group 3 was subjected to PBM (wavelength: 890 nm, peak power output: 80 W, pulse frequency: 80 Hz, pulsed duration: 180 ns, duration of exposure for each point: 200 s, power density: 0.001 W/cm2, energy density: 0.2 J/cm2) immediately after surgery, which continued for 6 days per week for 16 days. Group 4 received both the human ADS and PBM. In addition, we inflicted an ischemic, delayed healing, and infected wound simulation in all of the rats. The wounds were infected with methicillin-resistant Staphylococcus aureus (MRSA). RESULTS: All three treatment regimens significantly decreased the amount of microbial flora, significantly increased wound strength and significantly modulated inflammatory response and significantly increased angiogenesis on day 16. Microbiological analysis showed that PBM+ADS was significantly better than PBM and ADS alone. In terms of wound closure rate and angiogenesis, PBM+ADS was significantly better than the PBM, ADS and control groups. CONCLUSIONS: Combination therapy of PBM+ADS is more effective that either PBM or ADS in stimulating skin injury repair, and modulating inflammatory response in an MRSA-infected wound model of rats with DM1.

Photobiomodulation plus Adipose-derived Stem Cells Improve Healing of Ischemic Infected Wounds in Type 2 Diabetic Rats.

In this study, we sought to investigate the impact of photobiomodulation and adipose-derived stem cells (ADS), alone and in combination, on the maturation step of wound healing in an ischemic infected delayed healing wound model in rats with type 2 diabetes mellitus (DM2). We randomly divided 24 adult male rats into 4 groups (n = 6 per group). DM2 plus an ischemic delayed healing wound were induced in all rats. The wounds were infected with methicillin-resistant Staphylococcus aureus. Group 1 was the control (placebo) group. Group 2 received only photobiomodulation (890 nm, 80 Hz, 0.324 J/cm2, and 0.001 W/cm2). Group 3 received only the allograft ADS. Group 4 received allograft ADS followed by photobiomodulation. On days 0, 4, 8, 12, and 16, we performed microbiological examination (colony forming units, [CFU]), wound area measurement, wound closure rate, wound strength, and histological and stereological examinations. The results indicated that at day 16, there was significantly decreased CFU (Analysis of variance, p = 0.001) in the photobiomodulation + ADS (0.0 ± 0.0), ADS (1350 ± 212), and photobiomodulation (0.0 ± 0.0) groups compared with the control group (27250 ± 1284). There was significantly decreased wound area (Analysis of variance, p = 0.000) in the photobiomodulation + ADS (7.4 ± 1.4 mm2), ADS (11 ± 2.2 mm2), and photobiomodulation (11.4 ± 1.4 mm2) groups compared with the control group (25.2 ± 1.7). There was a significantly increased tensiometeric property (stress maximal load, Analysis of variance, p = 0.000) in the photobiomodulation + ADS (0.99 ± 0.06 N/cm2), ADS (0.51 ± 0.12 N/cm2), and photobiomodulation (0.35 ± 0.15 N/cm2) groups compared with the control group (0.18 ± 0.04). There was a significantly modulated inflammatory response in (Analysis of variance, p = 0.049) in the photobiomodulation + ADS (337 ± 96), ADS (1175 ± 640), and photobiomodulation (69 ± 54) treatments compared to control group (7321 ± 4099). Photobiomodulation + ADS gave significantly better improvements in CFU, wound area, and wound strength compared to photobiomodulation or ADS alone. Photobiomodulation, ADS, and their combination significantly hastened healing in ischemic methicillin-resistant Staphylococcus aureus infected delayed healing wounds in rats with DM2. Combined application of photobiomodulation plus ADS demonstrated an additive effect.

Combined effects of metformin and photobiomodulation improve the proliferation phase of wound healing in type 2 diabetic rats.

We determined the impact of Photobiomodulation (PBM) and metformin administration alone and combined on the inflammation and proliferation steps of wound healing of incisions in type two diabetes mellitus (T2DM) rats. 40 rats were divided into 4 groups (n = 10 each group). A non-genetic model of T2DM was induced in all rats, and an incision was made on each rat. There were 4 groups as follows: Group 1 was control group. Group 2 received PBM alone (890 nm, 80 Hz, 0.324 J/cm2, daily). Group 3 received metformin alone (50 mg/kg, i.p., daily) and the fourth group received combination of PBM + metformin. At inflammation (day 4) and proliferation (day 7) steps, tensiometerical, stereological, and immunohistochemical examinations were performed. PBM and PBM + metformin treatments significantly increased wound strength at inflammation and proliferation steps of wound healing respectively. PBM, metformin, and PBM + metformin groups significantly decreased inflammatory cells at inflammation and proliferation steps of wound healing. PBM, metformin, and PBM + metformin groups significantly improved granulation tissue formation by increasing fibroblasts, and new blood vessel formation at inflammation and proliferation steps of wound healing. Metformin significantly increased M2 macrophages than other treatment groups at inflammation and proliferation steps of wound healing. Simultaneously, PBM significantly decreased M2 macrophages than control group. We concluded PBM and PBM + metformin treatments significantly hastened repair at the inflammation and proliferation steps of repairing skin injury in a non-genetic model of T2 DM. PBM + metformin showed a synergistic impact. There were not a positive relation between M2 macrophage number and wound strength in the studied groups. The details of the molecular mechanisms of PBM, and PBM + metformin treatments of repairing wounds in animals, and treatment of DFUs of patients with T2 DM should be elucidated by further research.

Effects of pentoxifylline and alendronate on fracture healing in ovariectomy-induced osteoporosis in rats.

Osteoporosis is determined by decreased bone strength that increases the threat of fractures. The aim of this study was to evaluate the effects of pentoxifylline (PTX) and alendronate (ALN), on the stereological parameters, and gene expression in callus of fracture in an experimental rat model of ovariectomy-induced osteoporosis (OVX). The OVX was induced in 90 female rats. Fourteen weeks later, a complete fracture on the right femur was made. Rats were divided into five groups: 1) control: no treatment; 2) sham: received daily distilled water; 3) daily 3.00 mg kg-1 ALN subcutaneously (SC); 4) daily 200 mg kg-1 PTX (SC) and 5) daily PTX (SC) + ALN (same doses). The osteoclast count was significantly lower in all treatment groups, at 21 and 56 days post-surgery, compared to the control and sham groups. The PTX significantly increased total callus volume at 21 and 56 days post-surgery, compared to the other groups. The PTX+ALN treatment significantly increased both cortical bone volume on day 21, and osteocyte and osteoblast numbers on day 56, compared to the control and sham groups. It can be concluded that PTX and ALN have antiresorptive effects, in OVX rats. Also, PTX has increased the extracellular matrix on both 21 and 56 days after surgery, compared to the other groups. PTX+ALN elevated cortical bone volume on day 21, and osteocyte and osteoblast numbers compared to the control and sham groups on day 56.

An improvement in acute wound healing in rats by the synergistic effect of photobiomodulation and arginine.

In this probe, at first we examined the best route and dosage of arginine administration on wound healing in an excisional wound model in rats. Next, we intend to assess the impact of photobiomodulation (PBM) and arginine, individually and together, on the wound healing. In the pilot study, an excisional wound was made in each of 24 rats. There were 4 groups. Group 1 was the control group. In groups 2 and 3, wounds were topically treated with arginine ointments (ARG.) 2% and 5%, respectively. In group 4, arginine was injected (ARG. INJ.,i.p.). In the main phase, in 24 new rats, an excisional wound was made. There were 4 groups: group 5 served as the control. Wounds in group 6 were topically treated with ARG 2%. Wounds in group 7 were subjected to PBM. Wounds in group 8 were treated with PBM+ARG. 2%. On day 15, wound area measurement, wound strength, and stereological examination were performed. In the pilot study, we found that the ARG 2% ointment significantly decreased wound area than ARG. 5%, ARG. INJ. and control groups, and significantly increased wound strength compared to the control and ARG.5% groups. In the main phase, a significant decrease of wound area in all treatment regimens was induced. PBM + ARG. 2% and PBM treatment regimens significantly improved wound strength and almost all stereological parameters, compared to the control and ARG. 2% groups. PBM + ARG. 2% induced anti-inflammatory and angiogenic activities, and hastened the wound healing process in an excisional wound model in rats.