Efficacy and Safety of Intra-articular Injections of Hyaluronic Acid Combined With Polydeoxyribonucleotide in the Treatment of Knee Osteoarthritis

OBJECTIVE: To assess the clinical efficacy and safety of intra-articular injection of hyaluronic acid (HA) combined with polydeoxyribonucleotide (PDRN) in patients with knee osteoarthritis in comparison with that of HA alone. METHODS: The current single-center, prospective, randomized, double-blind, controlled study was conducted in 36 patients with knee osteoarthritis at our medical institution. All the eligible patients (n=30) were equally assigned to two treatment arms (trial group ‘HA+PDRN’ and control group ‘HA’). For efficacy assessment, the patients were evaluated for the visual analogue scale (VAS) scores, the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the Knee Society Scores (KSS), all of which served as efficacy outcome measures. We monitored time-dependent changes in efficacy outcome measures at baseline and 1, 3 and 6 months. Subsequently, we compared differences in changes in efficacy outcome measures at 6 months from baseline between the two groups. Moreover, we assessed the safety based on the treatment-emergent adverse events (TEAEs), adverse drug reactions (ADRs) and any other complications serving as safety outcome measures. RESULTS: There were significant differences in changes in the VAS scores, the WOMAC scores in all domains, except ‘Stiffness’, the total WOMAC scores, and the KSS scores in all the domains at 6 months from baseline between the two groups (p<0.05). In our series, there were no TEAEs, ADRs, and any other complications. CONCLUSION: Intra-articular injections of HA combined with PDRN can also be considered in the treatment of knee osteoarthritis. However, further large-scale and multi-center studies are required to demonstrate the potential of the proposed combination.

Polydeoxyribonucleotides Improve Diabetic Wound Healing in Mouse Animal Model for Experimental Validation

BACKGROUND: Wound healing mechanisms is believed to have effects similar to wound healing disorders in diabetic patients, including abnormal inflammatory cells, angiogenesis disorders, and reduced collagen synthesis. Therefore, reestablishment of structural and promoted angiogenesis could be beneficial to promote wound healing process. OBJECTIVE: Therefore, we investigated whether the polydeoxyribonucleotide (PDRN) that was self-production in Korea, could be useful as an intradermal injection for promoting wound healing. Also, we validate for wound healing effect of PDRN using healing-impaired (db/db) mice. METHODS: In this study, we confirmed the effects of PDRN by creating wound models in in vitro and in vivo model. Using an in vitro wound healing assay, we observed that PDRN stimulated closure of wounded monolayers of human fibroblast cells. PDRN (8.25 mg/ml) or phosphate-buffered saline (0.9% NaCl) was injected once daily into the dermis adjacent to the wound for 12 days after skin injury. RESULTS: Time course observations revealed that mice treated with PDRN showed accelerated wound closure and epidermal and dermal regeneration, enhanced angiogenesis. The wound area and depth decreased at 3, 6, 9, and 12 days after skin injury. Histological evaluation showed an increase of vascular endothelial growth factor, CD31, and collagen fibers in the PDRN group compared with the control group, indicating that PDRN was effective in the treatment of delayed wound healing caused by diabetes. CONCLUSION: This study suggests that our PDRN has a wound healing effect in transgenic animal models with cells and diabetes through angiogenesis.

The Effect of Polydeoxyribonucleotide on Chronic Non-healing Wound of an Amputee: A Case Report

Polydeoxyribonucleotide (PDRN) is safe and effective in wound healing, cellular growth, synthesis of extracellular matrix protein, and inflammation reduction via activation of adenosine A2 receptors. We report a 28-year-old male patient treated with PDRN injections for chronic non-healing wound refractory to negative pressure wound therapy, skin graft, or growth factors. Three injections of PDRN were administered at the wound site into the anterior and medial sides of the left stump on the 1st, 4th, and 9th days of hospitalization. The PDRN ameliorated wound healing by enhancing cell growth, tissue repair, and angiogenesis. PDRN application represents a potential treatment for non-healing wounds obviating the need for additional therapies, and hospitalization, as well as improve patient’s activities of daily living.

Efecto aditivo de las células madre mesenquimales y del defibrótido en un modelo de trombosis arterial en ratas / Additive effect of mesenchymal stem cells and defibrotide in an arterial rat thrombosis model

Introducción. El objetivo de este estudio consistió en investigar el efecto aditivo de las células madre mesenquimales (MSC, por sus siglas en inglés) y del defibrótido (DFT) en un modelo de trombosis arterial femoral en ratas. Métodos. Se incluyeron 30 ratas Sprague Dawley. Se desarrolló un modelo de trombosis arterial mediante cloruro de hierro (FeCl3) en la arteria femoral izquierda. Las ratas se asignaron equitativamente en cinco grupos: grupo 1, intervención quirúrgica simulada (sin lesión arterial); grupo 2, inyección de solución salina tamponada con fosfato (PBS); grupo 3, MSC; grupo 4, DFT; grupo 5, MSC + DFT. Todas las ratas recibieron dos inyecciones intraperitoneales de 0,5 ml: la primera se administró 4 horas después del procedimiento y la segunda 48 horas después de la primera. Se sacrificó a las ratas siete días después de la segunda inyección. Resultados. Aunque el uso por separado de MSC derivadas de médula ósea humana (hBM-MSC) o de DFT permitió una resolución parcial del trombo, la combinación de ambos tuvo como resultado la resolución casi completa. La neovascularización fue doblemente mejor en las ratas tratadas con hBM-MSC + DFT (11,6 ± 2,4 canales) en comparación con los grupos asignados por separado a hBM-MSC (3,8 ± 2,7 canales) y DFT (5,5 ± 1,8 canales) (P < 0,0001 y P= 0,002, respectivamente). Conclusión. El uso combinado de hBM-MSC y DFT en un modelo de trombosis arterial en ratas mostró que el efecto aditivo tuvo como resultado la resolución casi completa del trombo.

Background/aim. In this study, we aimed to investigate the additive effect of mesenchymal stem cells (MSC) and defibrotide (DFT) in a rat model of femoral arterial thrombosis. Methods. Thirty Sprague Dawley rats were included. An arterial thrombosis model by ferric chloride (FeCl3) was developed in the left femoral artery. The rats were equally assigned to 5 groups: Group 1-Sham-operated (without arterial injury); Group 2-Phosphate buffered saline (PBS) injected; Group 3-MSC; Group 4-DFT; Group 5-MSC + DFT. All had two intraperitoneal injections of 0.5 ml: the 1st injection was 4 h after the procedure and the 2nd one 48 h after the 1st injection. The rats were sacrificed 7 days after the 2nd injection. Results. Although the use of human bone marrow-derived (hBM) hBM-MSC or DFT alone enabled partial resolution of the thrombus, combining them resulted in near-complete resolution. Neovascularization was two-fold better in hBM-MSC + DFT treated rats (11.6 ± 2.4 channels) compared with the hBM-MSC (3.8 ± 2.7 channels) and DFT groups (5.5 ± 1.8 channels) (P < 0.0001 and P= 0.002, respectively). Conclusion. The combined use of hBM-MSC and DFT in a rat model of arterial thrombosis showed additive effect resulting in near-complete resolution of the thrombus.

Polydeoxyribonucleotide Improves Peripheral Tissue Oxygenation and Accelerates Angiogenesis in Diabetic Foot Ulcers

BACKGROUND: Polydeoxyribonucleotide (PDRN) is known to have anti-inflammatory and angiogenic effects and to accelerate wound healing. The aim of this study was to investigate whether PDRN could improve peripheral tissue oxygenation and angiogenesis in diabetic foot ulcers. METHODS: This was a prospective randomized controlled clinical trial. Twenty patients with a non-healing diabetic foot ulcer were randomly distributed into a control group (n=10) and a PDRN group (n=10). Initial surgical debridement and secondary surgical procedures such as a split-thickness skin graft, primary closure, or local flap were performed. Between the initial surgical debridement and secondary surgical procedures, 0.9% normal saline (3 mL) or PDRN was injected for 2 weeks by the intramuscular (1 ampule, 3 mL, 5.625 mg, 5 days per week) and perilesional routes (1 ampule, 3 mL, 5.625 mg, 2 days per week). Transcutaneous oxygen tension (TcPO2) was evaluated using the Periflux System 5000 with TcPO2/CO2 unit 5040 before the injections and on days 1, 3, 7, 14, and 28 after the start of the injections. A pathologic review (hematoxylin and eosin stain) of the debrided specimens was conducted by a pathologist, and vessel density (average number of vessels per visual field) was calculated. RESULTS: Compared with the control group, the PDRN-treated group showed improvements in peripheral tissue oxygenation on day 7 (P < 0.01), day 14 (P < 0.001), and day 28 (P < 0.001). The pathologic review of the specimens from the PDRN group showed increased angiogenesis and improved inflammation compared with the control group. No statistically significant difference was found between the control group and the PDRN group in terms of vessel density (P=0.094). Complete healing was achieved in every patient. CONCLUSIONS: In this study, PDRN improved peripheral tissue oxygenation. Moreover, PDRN is thought to be effective in improving inflammation and angiogenesis in diabetic foot ulcers.

Polydeoxyribonucleotide Improves Peripheral Tissue Oxygenation and Accelerates Angiogenesis in Diabetic Foot Ulcers

BACKGROUND: Polydeoxyribonucleotide (PDRN) is known to have anti-inflammatory and angiogenic effects and to accelerate wound healing. The aim of this study was to investigate whether PDRN could improve peripheral tissue oxygenation and angiogenesis in diabetic foot ulcers. METHODS: This was a prospective randomized controlled clinical trial. Twenty patients with a non-healing diabetic foot ulcer were randomly distributed into a control group (n=10) and a PDRN group (n=10). Initial surgical debridement and secondary surgical procedures such as a split-thickness skin graft, primary closure, or local flap were performed. Between the initial surgical debridement and secondary surgical procedures, 0.9% normal saline (3 mL) or PDRN was injected for 2 weeks by the intramuscular (1 ampule, 3 mL, 5.625 mg, 5 days per week) and perilesional routes (1 ampule, 3 mL, 5.625 mg, 2 days per week). Transcutaneous oxygen tension (TcPO2) was evaluated using the Periflux System 5000 with TcPO2/CO2 unit 5040 before the injections and on days 1, 3, 7, 14, and 28 after the start of the injections. A pathologic review (hematoxylin and eosin stain) of the debrided specimens was conducted by a pathologist, and vessel density (average number of vessels per visual field) was calculated. RESULTS: Compared with the control group, the PDRN-treated group showed improvements in peripheral tissue oxygenation on day 7 (P < 0.01), day 14 (P < 0.001), and day 28 (P < 0.001). The pathologic review of the specimens from the PDRN group showed increased angiogenesis and improved inflammation compared with the control group. No statistically significant difference was found between the control group and the PDRN group in terms of vessel density (P=0.094). Complete healing was achieved in every patient. CONCLUSIONS: In this study, PDRN improved peripheral tissue oxygenation. Moreover, PDRN is thought to be effective in improving inflammation and angiogenesis in diabetic foot ulcers.

Delayed Hepatic Veno-Occlusive Disease after Haploidentical Hematopoietic Stem Cell Transplantation:A Report of Six Cases / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the morbidity, risk factors, clinical characterisitics, treatments and prognosis of delayed hepatic veno-occlusive disease(HVOD) after haploidentical hematopoietic stem cell transplantation (hi-HSCT).</p><p><b>METHODS</b>The clinical data of 208 patients undergoing hi-HSCT were retrospectively analyzed.</p><p><b>RESULTS</b>Six patients were diagnosed with delayed VOD, among them 4 patients were moderate VOD and 2 patients were severe VOD. The incidence of VOD after hi-HSCT was 2.88%, the median onset time was 44.5(30-57) days after transplant, 2 patients died of multiple organ failure (MOF) due to rapid progress of disease. With intravenous administration of defibrotide, 4 patients displayed encouraging response, but 2 patients died of hepatic acute graft-versus-host disease (aGVHD), 1 had bone marrow relapse and the other one was cured.</p><p><b>CONCLUSION</b>Norethindrone is one of the high risk factors, while sex, age and disease status are irrelevant to the occurrence of VOD. Unfractionated heparin (UH) can effectively decrease the morbidity. Pretransplant hepatic function reserve, high dose preconditioning regimens and pharmacotherapy may result in delayed VOD onset. The delayed VOD has the same clinical features and treatment-response as early VOD, but a poorer prognosis is usually observed. A larger amount of samples (patients) is needed to research the relationship of the delayed VOD with hi-HSCT. Defibrotide can effectively increase the survival rate of VOD patients.</p>

Treatment for Acute Stage Complex Regional Pain Syndrome Type II with Polydeoxyribonucleotide Injection

Complex regional pain syndrome (CRPS) type II is a syndrome that develops after nerve injury. Symptoms may be severe, and vary depending on the degree of sympathetic nerve involvement. As yet, there is no satisfactory treatment. We report the case of a female patient who had an L5 left transverse process fracture and an S2 body fracture, who developed symptoms of CRPS type II in her left lower leg that were aggravated during ambulation in spite of absolute bed rest for one month after the trauma. Several treatments, including bed rest, medication, and numerous nerve blocks were attempted, but the pain persisted. We finally tried injection of polydeoxyribonucleotide (PDRN) solution at the left L5 transverse process fracture site because we knew of the anti-inflammatory effect of PDRN. One day after this treatment, her symptoms had almost disappeared and three days later, she was discharged. We will also further discuss the possibility of using PDRN solution for the treatment of CRPS.

Static body postural misalignment in individuals with temporomandibular disorders: a systematic review

BACKGROUND: The association between body postural changes and temporomandibular disorders (TMD) has been widely discussed in the literature, however, there is little evidence to support this association. OBJECTIVES: The aim of the present study was to conduct a systematic review to assess the evidence concerning the association between static body postural misalignment and TMD. METHOD: A search was conducted in the PubMed/Medline, Embase, Lilacs, Scielo, Cochrane, and Scopus databases including studies published in English between 1950 and March 2012. Cross-sectional, cohort, case control, and survey studies that assessed body posture in TMD patients were selected. Two reviewers performed each step independently. A methodological checklist was used to evaluate the quality of the selected articles. RESULTS: Twenty studies were analyzed for their methodological quality. Only one study was classified as a moderate quality study and two were classified as strong quality studies. Among all studies considered, only 12 included craniocervical postural assessment, 2 included assessment of craniocervical and shoulder postures,, and 6 included global assessment of body posture. CONCLUSION: There is strong evidence of craniocervical postural changes in myogenous TMD, moderate evidence of cervical postural misalignment in arthrogenous TMD, and no evidence of absence of craniocervical postural misalignment in mixed TMD patients or of global body postural misalignment in patients with TMD. It is important to note the poor methodological quality of the studies, particularly those regarding global body postural misalignment in TMD patients. .

Effects of Polydeoxyribonucleotide in the Treatment of Pressure Ulcers

This study aimed to examine the positive effects of polydeoxyribonucleotide (PDRN) on the wound-healing process in pressure ulcers. In this randomized controlled trial, the effects of PDRN were compared over time between an experimental group (n=11) and a control group (n=12). The former was administered the same dose of PDRN intramuscularly (1 ampule, 3 mL, 5.625 mg, for 5 days) for 2 weeks and perilesionally (1 ampule, 3 mL, 5.625 mg, twice a week) for 4 weeks. The primary endpoint for determining efficacy was wound healing in the pressure ulcers, which was reflected by the wound surface area determined using VISITRAK Digital (Smith & Nephew, Largo, FL). The secondary endpoint was the pressure ulcer scale for healing score, determined using pressure ulcer scale for healing (PUSH Tool 3.0 developed by the National Pressure Ulcer Advisory Panel). After the 4-week treatment period, PDRN therapy was found to significantly reduce the wound size and PUSH score, without adverse effect during the treatment. The findings indicate that PDRN can positively modify the wound healing process in pressure ulcers, and its use could improve the clinical outcomes of patients and lower the need for additional therapies or hospital stay.

The Effects of Polydeoxyribonucleotide on the Survival of Random Pattern Skin Flaps in Rats

BACKGROUND: Partial or complete necrosis of a skin flap is a common problem. Polydeoxyribonucleotide (PDRN) can be extracted from trout sperm and used as a tissue repair agent. The aim of this study was to investigate whether PDRN could improve the survival of random pattern skin flaps in rats. METHODS: Twenty-two male Sprague-Dawley rats were randomly divided into two groups: the PDRN treatment group (n=11) and the control group (n=11). Caudally pedicled random pattern skin flaps were elevated on their dorsal skin and resutured. The treatment group received daily intraperitoneal administration of PDRN (8 mg/kg/day), and the control group received fluid vehicle (NaCl 0.9%, 8 mg/kg/day) from day 0 to day 6. On day 7, the flap survival was evaluated and the harvested tissue surrounding the demarcation line of the necrotic area was stained with H&E, anti-rat vascular endothelial cell growth factor (VEGF) antibody, and PECAM-1/CD31 antibody. RESULTS: The average necrotic area of the flap in the PDRN group was significantly smaller when compared with that of the control group. Histologic and immunohistochemical evaluation showed that granulation thickness score and VEGF-positive staining cells were marked higher in the PDRN group than in the control group. PECAM-1/CD31-positive microvascular densities were significantly higher in the PDRN group when compared with the control group. CONCLUSIONS: This study confirms that PDRN improves the survival of random pattern skin flaps in rats. These results may represent a new therapeutic approach to enhancing flap viability and achieving faster wound repair.

6-Thioguanine-Induced Hepatic Sinusoidal Obstruction Syndrome in a Leukemic Child with TPMT Heterozygote / 임상소아혈액종양

Hepatic sinusoidal obstruction syndrome (hSOS) can be developed as a common complication after hematopoietic stem cell transplantation (HSCT), and rarely after 6-thioguanine-based chemotherapy without HSCT. A four-year-old boy with heterozygotic polymorphism for thiopurine methyltransferase (TPMT) developed hSOS after he received chemotherapy with cytarabine, cyclophosphamide, intrathecal methotrexate and 6-thioguanine (6-TG) as reconsolidation chemotherapy of acute lymphoblastic leukemia (ALL). He was treated with defibrotide, N-acetylcysteine, urusodeoxycholic acid, glutathione, and supportive care. He recovered completely in nine days without long-term complication, and completed chemotherapy with 6-mercaptopurine without severe complication. We report a case of hSOS developed in an ALL patient with TPMT heterozygote after 6-TG based reconsolidation chemotherapy.

6-Thioguanine-Induced Hepatic Sinusoidal Obstruction Syndrome in a Leukemic Child with TPMT Heterozygote / 임상소아혈액종양

Hepatic sinusoidal obstruction syndrome (hSOS) can be developed as a common complication after hematopoietic stem cell transplantation (HSCT), and rarely after 6-thioguanine-based chemotherapy without HSCT. A four-year-old boy with heterozygotic polymorphism for thiopurine methyltransferase (TPMT) developed hSOS after he received chemotherapy with cytarabine, cyclophosphamide, intrathecal methotrexate and 6-thioguanine (6-TG) as reconsolidation chemotherapy of acute lymphoblastic leukemia (ALL). He was treated with defibrotide, N-acetylcysteine, urusodeoxycholic acid, glutathione, and supportive care. He recovered completely in nine days without long-term complication, and completed chemotherapy with 6-mercaptopurine without severe complication. We report a case of hSOS developed in an ALL patient with TPMT heterozygote after 6-TG based reconsolidation chemotherapy.

Protective Effect of Defibrotide on Splanchnic Injury following Ischemia and Reperfusion in Rats

A splanchic artery occlusion for 90 min followed by reperfusion of the mesenteric circulation resulted in a severe form of circulatory shock, characterized by endothelial dysfunction, severe hypotension, marked intestinal tissue injury, and a high mortality rate. The effect of defibrotide, a complex of single-stranded polydeoxyribonucleotides having antithrombotic effect, was investigated in a model of splanchnic artery occlusion (SAO) shock in urethane anesthetized rats. Occlusion of the superior mesenteric artery for 90 min produced a severe shock state, resulting in a fatal outcome within 120 min of reperfusion in many rats. Defibrotide (10 mg/kg body weight) 10 min prior to reperfusion significantly improved mean arterial blood pressure in comparison to vehicle treated rats (p<0.05). Defibrotide treatment also significantly attenuated in the increase of plasma amino nitrogen concentration, intestinal myeloperoxidase activity, intestinal lipid peroxidation, infiltration of neutrophils in intestine and thrombin induced adherence of neutrophils to superior mesentric artery segments. Superoxide anion and hydrogen peroxide production in 1 micrometer formylmethionylleucylphenylalanine (fMLP)-activated PMNs was inhibited by defibrotide in a dose-dependent fashion. Defibrotide effectively scavenged hydrogen peroxide, but not hydroxyl radical. Treatment of SAO rats with defibrotide inhibited tumor necrosis factor-alpha, and interleukin-1beta productions in blood in comparison with untreated rats. These results suggest that defibrotide partly provides beneficial effects by preserving endothelial function, attenuating neutrophil accumulation, and antioxidant in the ischemic reperfused splanchnic circulation.

Sensitivity of non-cooperative binding parameters of ligand-DNA complex by computer analysis.

A simple user-friendly computer programme has been developed to operate on an IBM-PC compatible machine to aid in the process of curve fitting using the McGhee and von Hippel equation [J. Mol Biol, 86 (1974), 469-489] for the analysis of ligand-DNA interactions and the experimental data on berberine-calf thymus DNA and berberine-poly(dI-dC). poly(dI-dC) for non-cooperative binding. A sensitivity analysis on binding constant (K0) and the number of binding sites (N0) show that a small variation in the latter remarkably alters the fitting curve. At a level of 1% change in N0 from the best fit value, the standard deviation grows by almost 4%, while, on the other hand, a 1% change in K0 affects the same value only by less than 0.4%.

Conformational transitions and specific heat anomaly in poly(dG-dC) and its methylated analogue.

The Zimm and Bragg theory of the helix<-->coil transition has been modified to explain order<-->order transitions in polynucleotides, in particular B<-->Z<-->psi(-)<-->coil transitions in poly(dG-dC) and poly(dG-me5dC). Lambda anomalies in specific heat measurements around the transition point have also been explained by a further modification of the same theory. Theoretical results are found to be in good agreement with the experimental data. The nucleation parameter is consistent with the stabilization/destabilization of the ordered states (Z helix) under various environmental conditions, e.g. methylation of cytosine residue at C5 position or change in the cationic concentration of the solvent.

Interaction of berberine chloride with deoxyribonucleic acids: evidence for base and sequence specificity.

The interaction of berberine chloride with natural and synthetic DNAs of differing base composition and sequences was followed by various spectroscopic and viscometric studies. The binding of berberine chloride was characterized by hypochromism and bathochromism in the absorption bands, enhancement of fluorescence intensity, stabilization against thermal denaturation, perturbations in the circular dichroic spectrum, increase in the contour length of sonicated rod-like DNA and induction of unwinding-rewinding process of covalently closed superhelical DNA, depending on the base composition and sequences of base pairs. Binding parameters determined from absorbance and fluorescence titration by Scatchard analysis, according to an excluded-site model, indicated a very high specificity of berberine to AT-rich DNAs and alternate AT polymer. Fluorescence quantum yield was maximum for the complexes with AT-rich DNAs and alternate AT polymer. Taken together, these results suggest that berberine chloride exhibits considerable specificity towards alternating AT polymer and binds to AT-rich DNAs by a mechanism of classical intercalation.

Interaction of antitumour agent mitoxantrone with poly[d(G-C)]--a circular dichroic study.

The binding of antitumour drug mitoxantrone [1,4-dihydroxy-5, 8-bis [2-(2-hydroxy ethyl)amino)ethyl)amino)-9, 10-anthracenedione] to the synthetic polynucleotide poly[d(G-C)] was studied by circular dichroic titrations. The interaction induced intense chiroptical properties in the visible (688 nm) as well as the ultraviolet (260, 320 nm) region in an otherwise optically inactive drug. The interaction occurs in two stages, one below a drug/nucleotide ratio of 0.11 and other above this value. The second mode of interaction causes an almost cooperative enhancement of the visible induced circular dichroism (ICD).