Injection Of Phosphate Research Articles

Immune challenge-induced oxidative damage may be mitigated by biliverdin.

An effective immune response results in the elimination of pathogens, but this immunological benefit may be accompanied by increased levels of oxidative damage. However, organisms have evolved mechanisms to mitigate the extent of such oxidative damage, including the production and mobilization of antioxidants. One potential mechanism of mitigating immune challenge-induced changes in oxidative physiology is increasing biliverdin production. Biliverdin is chemically an antioxidant, but within-tissue correlations between biliverdin concentration and oxidative damage have never been directly examined. To test how biliverdin tissue concentrations are associated with physiological responses to an immune challenge, we exposed northern bobwhite quail (Colinus virginianus) to one of four treatments: injection of a non-pathogenic antigen - either lipopolysaccharide or phytohemagglutinin, control injection of phosphate-buffered saline or a sham procedure with no injection. Twenty-four hours later, we quantified oxidative damage and triglyceride concentration in the plasma, and biliverdin concentration in the plasma, liver and spleen. We found that both types of immune challenge increased oxidative damage relative to both non-injected and vehicle-injected controls, but treatment had no effects on any other metric. However, across all birds, oxidative damage and biliverdin concentration in the plasma were negatively correlated, which is consistent with a localized antioxidant function of biliverdin. Additionally, we uncovered multiple links between biliverdin concentration, change in mass during the immune challenges and triglyceride levels, suggesting that pathways associated with biliverdin production may also be associated with aspects of nutrient mobilization. Future experiments that manipulate biliverdin levels or oxidative damage directly could establish a systemic antioxidant function or elucidate important physiological impacts on body mass maintenance and triglyceride storage, mobilization or transport.

Increased placental T cell trafficking results in adverse neurobehavioral outcomes in offspring exposed to sub-chronic maternal inflammation

Interleukin-1 beta (IL-1β) is a cytokine mediator of perinatal brain injury. The effect of sub-chronic systemic IL-1β exposure in perinatal and offspring outcomes is unclear. The aim of this study was to examine the effects of maternal IL-1β exposure on pregnancy and offspring outcomes. At E15, CD1 dams were allocated to receive intraperitoneal injection of phosphate buffered saline or mouse recombinant IL-1β (1 mcg) for four consecutive days. We analyzed pup survivaland neurobehavioral status. At E18, placental H&E staining and fetal brain Nissl staining was performed. Placental gene expression was analyzed by qPCR and T cell infiltration was analyzed by flow cytometry. Effects of inflammation on feto-placental blood flow were analyzed by Doppler ultrasonography. IL-1β decreased pup survival (P < .0001) and adversely affected offspring performance on neurodevelopmental tests (P < .05). Placentas of exposed dams exhibited significant thinning of maternal and fetal sides, and fetal brain exhibited cortical thinning. Placental qPCR analysis revealed significant upregulation of NFκB2 (P = .0021) and CXCL11 (P = .0401). While maternal IL-1β exposure did not affect feto-placental blood flow, placental flow cytometry showed an increase in placental infiltration of CD4+ T cells at 24 h post-injection (hpi, P < .0001) and CD8+ T cells at 72 hpi (P = .0217). Maternal sub-chronic, systemic inflammation with IL-1β decreased pup survival and played a key role in perinatal brain injury. The mechanisms behind these outcomes may involve immune system activation and alterations in placental T cell trafficking.

Hitting the Mark: Optimizing the Use of Calcium Phosphate Injections for the Treatment of Bone Marrow Lesions of the Proximal Tibia and Distal Femur

Increased contact pressures of the osteoarthritic joint can lead to underlying osseous injury, with resultant marrow edema changes of the subchondral bone. These osteoarthritis-related bone marrow lesions can subsequently lead to persistent pain and further disability. Limited joint preservation treatment options exist to alleviate symptoms or potentially alter the natural history of the affected joint; however, recent success with injectable calcium phosphate has provided early pain relief and may provide a scaffold for endogenous repair mechanisms. In this Technical Note, a comprehensive surgical approach using injectable calcium phosphate to target bone marrow lesions of the proximal tibia and distal femur is presented. Critical technique considerations include the use of magnetic resonance and fluoroscopic imaging to target the area of the subchondral bone while refraining from overfilling and/or forced pressurization during delivery and the use of postinjection arthroscopy to prevent potential injurious sequelae.

AB084. intracavernous administration of embryonic stem cell-derived exosome restores erectile function through penile neurovascular regeneration in the streptozotocin-induced diabetic mouse

BackgroundExosome contains a variety of protein, mRNA, and miRNA and is known to play an important role in intercellular communication as a bio-nanoparticle with a diameter of 40 to 100 nm. Recent studies have demonstrated the therapeutic potential of exosome in a variety of animal models for cardiovascular diseases and neuropathies. The aim of this study was to investigate effectiveness of embryonic stem cell (ESC)-derived exosome in restoring erectile function in diabetic mice.MethodsDiabetes was induced by intraperitoneal injection of streptozotocin into 8-week-old C57BL/6 male mice. At 8 weeks after the induction of diabetes, the animals were distributed into 3 groups: control nondiabetic mice and diabetic mice receiving two successive intracavernous injections of phosphate buffered saline (PBS) (days −3 and 0; 20 µL) or ESC-derived exosome (days −3 and 0; 1 µg in 20 µL of PBS). Two weeks after treatment, we measured erectile function by electrical stimulation of the cavernous nerve. The penis was harvested and stained with antibodies to PECAM-1, smooth muscle α-actin, NG2, and βIII tubulin. We also determined angiogenic potential of ESC-derived exosome in an ex vivo aortic ring assay and in primary cultured mouse cavernous endothelial cell (MCEC) and pericyte (MCP) mono-culture or co-culture system in vitro.ResultsIntracavernous injections of ESC-derived exosome significantly improved erectile function in diabetic mice, which reached up to 90% of control values. ESC-derived exosome induced significant restoration of cavernous contents of endothelial cells, smooth muscle cells, pericytes, and neuronal cells in diabetic condition. Moreover, ESC-derived exosome promoted microvascular sprouting from aortic ring and accelerated tube formation in primary cultured MCEC and MCP mono-culture or co-culture system in vitro.ConclusionsESC-derived exosome successfully restored erectile function through enhanced cavernous angiogenesis and neural regeneration in diabetic mice. Further studies are needed to clarify mechanism by which ESC-derived exosome induces neurovascular repair.

Radioprotective effect of epimedium on neurogenesis and cognition after acute radiation exposure

The radioprotective effect of herb epimedium (or yin yang huo) extract (5 g/kg, oral administration daily for 4 weeks) on neurogenesis and cognition after acute radiation exposure with 5.5 Gy was evaluated in Balb/c mice by behavioral tests and immunohistochemical study. The results indicated that epimedium extract could improve animal weight loss, locomotor activity and spatial learning and memory which are similar to pre-irradiation intraperitoneal injection (100 mg/kg) of amifostine phosphate, a well- known radioprotective drug. Immunohistochemical study showed that epimedium extract prevented the loss of proliferation cells, newly generated neurons, and interneurons in the hilus, in particular, the subgranular zone of the dentate gyrus. It suggests that herb epimedium may be a promising radio-neuro-protective drug to prevent radiation-induced neuropsychological disorders.

Antitumor activity of RT2 peptide derived from crocodile leukocyte peptide on human colon cancer xenografts in nude mice.

RT2, derived from the leukocyte peptide of Crocodylus siamensis, can kill human cervical cancer cells via apoptosis induction, but no evidence has shown in vivo. In this study, we investigated the antitumor effect of RT2 on human colon cancer xenografts in nude mice. Twenty-four mice were injected subcutaneously with human colon cancer HCT 116 cells. Eleven days after cancer cell implantation, the mice were treated with intratumoral injections of phosphate buffered saline (PBS) or RT2 (0.01, 0.1, and 1 mg/mouse) once every 2 days for a total of 5 times. The effect of a 10-day intratumoral injection of RT2 on body weight, biochemical, and hematological parameters in BALB/c mice showed no significant difference between the groups. Tumor volume showed a significant decrease only in the treatment group with RT2 (1 mg/mouse) at day 6 (P < .05), day 8 (P < .01), and day 10 (P < .01) after the first treatment. The protein expression levels of cleaved poly (ADP-ribose) polymerase (PARP), apoptosis-inducing factor (AIF), and the p53 tumor suppressor protein (p53) in xenograft tumors increased after treatment with RT2 (1 mg/mouse) compared to those in the PBS-injected group. Moreover, RT2 increased the expression of Endo G and Bcl-2 family proteins. Therefore, the peptide RT2 can inhibit tumor growth via the induction of apoptosis in an in vivo xenograft model.

A randomized controlled study of the efficacy and safety of dexamethasone in the treatment of pain in patients with emergency trauma

Objective To study the efficacy and safety of dexamethasone in the treatment of pain in patients with emergency trauma. Methods From December 2015 to December 2016, two hundred and sixty-four patients were enrolled in the study and randomly divided into the experimental group and the control group, each with 132 cases.In the experimental group, dexamethasone sodium phosphate injection was used as a test drug, and the control group was treated with glucose injection as a placebo.VAS score, overall evaluation, IL-6/ blood glucose and painkillers intake were recorded by physicians at the time of admission (0 h), 6, 12, 24, 48 h after injury.At the same time, wound healing, hospitalization time, fracture healing time and adverse events were recorded. Results The 24 h and 48 h VAS scores in the experimental group were (2.31±0.37) and (1.21±0.13) points, which were significantly lower than those in the control group ((4.12±0.54) points, (3.01±0.51) points). The difference was significant (F (time)=2 917.519, F(Group)=137.507, F (time *group)=83.927, P 0.05). Conclusion The emergency use of dexamethasone can safely and effectively alleviate preoperative pain and expand the indications of dexamethasone.It is beneficial to relieve postoperative analgesia, to reduce the use of morphine and NSAID drugs, and to provide a new way for multimodal analgesia. Key words: Dexamethasone; Pain; Emergency trauma

276 Adipose-derived stem cell-derived exosomes ameliorate erectile dysfunction in a rat model of type 2 diabetes

The efficacy of adipose-derived stem cells (ADSCs) in alleviating erectile dysfunction (ED) of diabetic rats has been demonstrated mainly through a paracrine effect. This study aimed to investigate the effect of ADSC-derived Exosomes (EXOs) on erectile function in a type 2 diabetic ED rat model. EXOs were isolated from the supernatants of cultured ADSCs by ultracentrifugation. In total, 24 type 2 diabetic rats were randomly assigned to three groups and were treated with an intracavernous injection of ADSC-derived EXOs, ADSCs, or phosphate buffered saline. Another eight age-matched rats underwent sham operation and composed the normal control group. Intracavernous pressure and mean arterial pressure testing and histologic and western blot analyses were performed 4 weeks after the intracavernous injection. ADSC-derived EXOs and ADSCs administered led to an increase in the ratio of intracavernous pressure to mean arterial pressure compared with that for phosphate buffered saline treatment. Histologic and western blot analyses demonstrated an increased ratio of smooth muscle to collagen, increased expression of an endothelial marker (CD31), a smooth muscle marker (a-smooth muscle actin), and antiapoptotic protein Bcl-2 and decreased the expression of the apoptotic protein cleaved caspase-3 and apoptosis of endothelial and smooth muscle cells in the corpus cavernosum tissue after EXO or ADSC injection compared with values for the phosphate buffered saline treatment.

Local combined with systemic glucocorticoid treatment of young and middle-aged patients with severe and profound hearing loss

Objective:To investigate the clinical effect of local combined with systemic glucocorticoid injection on young and middle-aged patients with severe and profound hearing loss. Method:Ninety-three cases of young and middle-aged patients with severe and profound hearing loss admitted to our hospital from May 2013 to May 2017 were retrospectively analyzed. Patients were divided into two groups according to the different treatment. Forty-six cases in control group were treated with systemic injection of glucocorticoids.Forty-seven cases in observation group were treated with local administration combined with systemic glucocorticoid therapy. The efficacy, changes of average hearing threshold and the incidence of adverse reactions were evaluated.Result:The total effective rate in the observation group was 68.09%, which was significantly higher than that in the control group (45.65%,P<0.05).After treatment, the average hearing thresholds in the observation groups of 0.5,1.0,2.0 and 4.0kHz were(35.77±13.28)dB HL,(39.71±14.09)dB HL, (32.99±10.73)dB HL and (30.17±9.29)dB HL respectively,which were significantly higher than that in the control group(26.30±14.03)dB HL,(25.14±13.28)dB HL,(20.54±9.27)dB HL,and (20.98±8.71)dB HL(P<0.05).Among all the patients received systemic injection of dexamethasone sodium phosphate injection,there was no other complications were found except for one patient whose blood pressure instability was relieved after treatment stopping. In the observation group, intratympanic methylprednisolone sodium succinate did not cause serious complications such as tympanic cavity infection and perforation of the tympanic membrane on the basis of systemic injection.While in the observation group,one case complained of earache and one case complained of a slight burning sensation in the ear, and both of them eased after rest.Conclusion:Local combined with systemic glucocorticoid treatment of young and middle-aged patients with severe and profound hearing loss can significantly improve the efficiency and increase the average hearing threshold changes at 0.5,1.0,2.0 and 4.0 kHz,and there was no serious adverse reactions.

Daily therapy with a slow-releasing H2S donor GYY4137 enables early functional recovery and ameliorates renal injury associated with urinary obstruction

ObjectivesTo assess the effects of slow-releasing H2S donor GYY4137 on post-obstructive renal function and injury following unilateral ureteral obstruction (UUO) by using the UUO and reimplantation (UUO-R) model in rats and to elucidate potential mechanisms by using an in vitro model of epithelial-mesenchymal transition (EMT). MethodsMale Lewis rats underwent UUO at the left ureterovesical junction. From post-operative day (POD) 1–13, rats received daily intraperitoneal (IP) injection of phosphate buffered saline (PBS, 1 mL) or GYY4137 (200 μmol/kg/day in 1 mL PBS, IP). On POD 14, the ureter was reimplanted back into the bladder, followed by a right nephrectomy. Urine and serum samples were collected to monitor renal function. On POD 30, the left kidney was removed and tissue sections were stained with H&E, TUNEL, CD68, CD206, myeloperoxidase, and Masson's trichrome to determine cortical thickness, apoptosis, inflammation, and fibrosis. In our in vitro model of EMT, NRK52E cells were treated with 10 ng/mL TGF-β1, 10 μM GYY4137 and/or 50 μM GYY4137. Western blot analysis was performed to determine the expression of E-cadherin, vimentin, Smad7 and TGF-β1 receptor II (TβRII). ResultsGYY4137 led to a moderate decrease in post-obstructive serum creatinine, cystatin C and FENa. We also observed a trend towards a decrease in post-obstructive proteinuria following GYY4137 treatment. Histologically, we observed a significant decrease in apoptosis, inflammation, and fibrosis. Furthermore, our in vitro studies demonstrate that in the presence of TGF-β1, GYY4137 significantly decreases vimentin and TβRII and significantly increases E-cadherin and Smad7. ConclusionsH2S may help to accelerate the recovery of renal function post-obstruction and attenuates renal injury associated with UUO. It is possible that H2S mitigates fibrosis by regulating the TGF-β1-mediated EMT pathway. Taken together, our data suggest that H2S may be a potential novel therapy for improving renal function and limiting renal injury associated with obstructive uropathy.

Monitoring of macrophage recruitment enhanced by Toll-like receptor 4 activation with MR imaging in nerve injury.

Macrophage recruitment is critical for nerve regeneration after an injury. The aim of this study was to investigate whether ultrasmall superparamagnetic iron oxide (USPIO) nanoparticle-based MRI could be used to monitor the enhanced macrophage recruitment by Toll-like receptor 4 (TLR4) activation in nerve injury. Rats received intraperitoneal injections of either lipopolysaccharide (LPS) or phosphate buffered saline (PBS) or no injection (controls) after a sciatic nerve crush injury. After intravenous injection of the USPIOs (LPS and PBS groups) or PBS (control group), MRI was performed and correlated with histological findings. LPS group showed more remarkable hypointense signals on T2*-weighted imaging and lower T2 values in the crushed nerves than PBS group. The hypointense signal areas were associated with an enhanced recruitment of iron-loaded macrophages to the injured nerves. USPIO-enhanced MRI can be used to monitor the enhanced macrophage recruitment by means of TLR4 signal pathway activation in nerve injury. Muscle Nerve, 2018.

Efficacy of supratarsal injection of triamcinolone acetonide and dexamethasone sodium phosphate in treating paediatric vernal keratoconjunctivitis in suburban Malaysia

Efficacy of supratarsal injection of triamcinolone acetonide and dexamethasone sodium phosphate in treating paediatric vernal keratoconjunctivitis in suburban Malaysia

Comparative Clinical Study of Local Submucosal Corticosteriod Versus Diclofenac Sodium Injections Before Odontectomy of Mandibular Impacted Third Molar

The aim of this comparative clinical study was to investigate the effects of preoperative injections of Dexamethasone Sodium Phosphate versus Diclofenac Sodium. The study includes 60 patients (age range 16-35) who were randomly distributed into three equal groups. Edema and trismus was measured preoperatively, immediate, 24 h, 2 week, 1 month postoperatively. Pain was assessed by visual analogue scale. Noteworthy reduction in postoperatively complication associated with Dexamethasone group was noticed. Edema & trismus in control group was greater followed by Diclofenac groups then group I. Pain was greater score in control group followed by Diclofenac group then group I.

Supratarsal injections versus topical steroids in resistant vernal keratoconjunctivitis

Purpose The aim of this study was to compare the safety and efficacy of supratarsal triamcinolone acetonide injection versus supratarsal dexamethasone sodium phosphate injection versus topical prednisolone acetate 1% eye drops in the treatment of resistant cases of vernal keratoconjunctivitis (VKC).Patients and methods This is a comparative, prospective randomized study that was done on 120 eyes (60 patients) having resistant VKC. The eyes included in the study were divided into three groups. Group 1 included 40 eyes (20 patients) that received supratarsal injection of 20 mg of triamcinolone acetonide; group 2 included 40 eyes (20 patients) that received supratarsal injection of 2 mg of dexamethasone sodium phosphate; and group 3 included 40 eyes (20 patients) that received topical prednisolone acetate 1% eye drops.Results Supratarsal steroid injections provided more effective resolution of symptoms and signs in comparison to topical steroid eye drops. No statistically significant difference was found between triamcinolone acetonide and dexamethasone sodium phosphate supratarsal injections (P>0.05). However, the supratarsal injections showed a statistically significant higher resolution of symptoms and signs than those of topical steroids. Eyes in groups 1 and 2 (supratarsal injections) had a lower recurrence rate and a delayed recurrence in comparison to group 3 (topical steroid eye drops) (P<0.05). No complications related to the drugs or to the technique of injection were reported in all groups.Conclusion Supratarsal injection of steroids is an effective, easy, well-tolerated, and safe technique in the treatment of resistant VKC, which provides an effective and longer relief of symptoms and signs with a delayed and lower rate of recurrence.

Hydrogen sulfide provides intestinal protection during a murine model of experimental necrotizing enterocolitis

BackgroundNecrotizing enterocolitis (NEC) continues to be a morbid surgical condition among preterm infants. Novel therapies for this condition are desperately needed. Hydrogen sulfide (H2S) is an endogenous gasotransmitter that has been found to have beneficial properties. We therefore hypothesized that intraperitoneal injection of various H2S donors would improve clinical outcomes, increase intestinal perfusion, and reduce intestinal injury in an experimental mouse model of necrotizing enterocolitis. MethodsNEC was induced in five-day-old mouse C57BL/6 mouse pups through maternal separation, formula feeding, and intermittent hypoxic and hypothermic stress. The control group (n=10) remained with their mother and breastfed ad lib. Experimental groups (n=10/group) received intraperitoneal injections of phosphate buffered saline (PBS) vehicle or one of the following H2S donors: (1) GYY4137, 50mg/kg daily; (2) Sodium sulfide (Na2S), 20mg/kg three times daily; (3) AP39, 0.16mg/kg daily. Pups were monitored for weight gain, clinical status, and intestinal perfusion via transcutaneous Laser Doppler Imaging (LDI). After sacrifice on day nine, intestinal appearance and histology were scored and cytokines were measured in tissue homogenates of intestine, liver, and lung. Data were compared with Mann–Whitney and p<0.05 was considered significant. ResultsClinical score and weight gain were significantly improved in all three H2S-treated groups as compared to vehicle (p<0.05 for all groups). Intestinal perfusion of the vehicle group was 22% of baseline while the GYY4137 group was 38.7% (p=0.0103), Na2S was 47.0% (p=0.0040), and AP39 was 43.0% (p=0.0018). The vehicle group had a median histology score of 2.5, while the GYY4137 group's was 1 (p=0.0013), Na2S was 0.5 (p=0.0004), and AP39 was 0.5 (p=0.0001). Cytokine analysis of the intestine of the H2S-treated groups revealed levels closer to breastfed pups as compared to vehicle (p<0.05 for all groups). ConclusionIntraperitoneal administration of H2S protects against development of NEC by improving mesenteric perfusion, and by limiting mucosal injury and altering the tissue inflammatory response. Further experimentation is necessary to elucidate downstream mechanisms prior to clinical implementation.

Control of Citrus Huanglongbing via Trunk Injection of Plant Defense Activators and Antibiotics.

Citrus huanglongbing (HLB) or greening is a devastating disease of citrus worldwide and no effective control measure is currently available. Plant defense activators environmentally friendly compounds capable of inducing resistance against many plant pathogens. Earlier studies showed that foliar spray of plant defense inducers could slow down HLB disease progress. In this study, eight plant defense activators and three antibiotics were evaluated in three field trials for their effect to control HLB by trunk injection of young and mature sweet orange trees. Results showed that four trunk injections of several activators, including salicylic acid, oxalic acid, acibenzolar-S-methyl, and potassium phosphate, provided significant control of HLB by suppressing 'Candidatus Liberibacter asiaticus' titer and disease progress. Trunk injection of penicillin, streptomycin, and oxytetracycline hydrochloride resulted in excellent control of HLB. In general, antibiotics were more effective in reduction of 'Ca. L. asiaticus' titer and HLB symptom expressions than plant defense activators. These treatments also resulted in increased yield and better fruit quality. Injection of both salicylic acid and acibenzolar-S-methyl led to significant induction of pathogenesis-related (PR) genes PR-1 and PR-2 genes. Meanwhile, injection of either potassium phosphate or oxalic acid resulted in significant induction of PR-2 or PR-15 gene expression, respectively. These results suggested that HLB diseased trees remained inducible for systemic acquired resistance under field conditions. In summary, this study presents information regarding controlling HLB via trunk injection of plant defense activators and antibiotics, which helps citrus growers in decision making regarding developing an effective HLB management program.

Construction and evaluation in vitro and in vivo of tedizolid phosphate loaded cationic liposomes

First, the SA-TDZA-Lips were prepared by reverse-phase evaporation method. Then, the drug release behaviour was evaluated by dynamic membrane dialysis in vitro and the preliminary safety was evaluated by haemolysis method. Finally, with tedizolid phosphate injection (TDZA-Inj) and tedizolid phosphate loaded liposomes (TDZA-Lips) as the control groups, the pharmacokinetic characteristic and tissues distribution of SA-TDZA-Lips were evaluated after intravenous injection. As a result, the stearylamine modified tedizolid phosphate liposomal delivery system was constructed successfully and the particle size was 194.9 ± 2.93 nm. The encapsulation efficiency (EE) was 53.52 ± 2.18%. The in vitro release of SA-TDZA-Lips was in accordance with Weibull equation. And there was no haemolysis happened, which indicated good preliminary safety for injection. The results of pharmacokinetics showed that the t1/2β increased by 0.74 times and 0.51 times higher than that of TDZA-Inj group and TDZA-Lips group, respectively. The MRT of SA-TDZA-Lips was 1.30 and 1.09 times higher than that of TDZA-Inj group and TDZA-Lips group, respectively. The AUC was 2.40 times and 0.23 times higher than that of TDZA-Inj group and TDZA-Lips group, respectively. The tissue distribution results showed that the relative uptake rate (Re) of TDZA in the lung was 1.527, which indicated the targeting. In conclusion, the SA-TDZA-Lips prepared in this study had several advantages like positive charge, strong cell affinity, prolonged circulation time in vivo, sustained release effect, and increased drug concentration in lungs. All advantages above provided significant clinical value of application for the treatment of bacterial pneumonia with tedizolid phosphate.

A Time Limit for Initiating Anti-Inflammatory Treatment for Improved Olfactory Function after Head Injury.

We previously reported that treatment with an anti-inflammatory drug, specifically a steroid, is effective in improving recovery during the acute phase of head injury. Clinically, however, patients with head injury usually become aware of their olfactory loss several weeks or months after the injury, which may be a critical factor in poor recovery from olfactory dysfunction. This raises an important question: When should steroid administration begin in order to achieve optimum improvement of olfactory dysfunction? The present study was designed to reveal the time limit for starting anti-inflammatory treatment for better improvement of post-traumatic olfactory dysfunction. Olfactory nerve transection (NTx) was performed in olfactory marker protein (OMP)-tau-lacZ mice and subcutaneous injections of dexamethasone sodium phosphate for 5 consecutive days was started at 7, 14, 28, and 42 days after the NTx (7-, 14-, 28-, and 42-day time-points). Histological assessment of olfactory nerve recovery in the olfactory bulb was made at 5, 14, and 42 days after the start of drug treatment. Olfactory function assessments using both an olfactory avoidance behavioral test and evoked potential testing also were performed. Animals treated at 7 days post-injury had less injury-associated tissue with fewer astrocytes and macrophages and better histological and functional nerve recovery, compared with control mice. However, those treated at 14, 28, or 42 days post-injury did not show significant histological or functional differences between saline control and treatment groups. These findings suggest that an anti-inflammatory treatment using steroids for traumatic olfactory dysfunction may be effective if started at least by 7 days, but may be ineffective at 14 days or later after head injury.

Subchondroplasty for the Treatment of Post-Traumatic Bone Marrow Lesions of the Medial Femoral Condyle in a Pre-Clinical Canine Model

The objective of this study was to assess the safety and efficacy of subchondroplasty for the treatment of bone marrow lesions of the femoral condyle in a canine model of post-traumatic osteoarthritis. Impaction injury to the femoral condyles is often a pre-cursor to post-traumatic osteoarthritis (PTOA). Appropriate treatment of the pain and dysfunction associated with bone marrow lesions (BML) in this setting is unknown. Subchondroplasty (SCP) involves the percutaneous injection of calcium phosphate into the region of the BML. The objective of this study was to assess the safety and efficacy of SCP for the treatment of BMLs of the femoral condyle in a canine PTOA model. With ACUC approval, purpose-bred research hounds (n=12) underwent an arthroscopic-assisted impact injury (40N) to both medial femoral condyles (MFC) to result in focal articular cartilage damage with associated BML. Twelve weeks after impact injury, dogs were treated by injection (3 ml) of calcium phosphate SCP (AccuFill BSM, Zimmer) into one MFC and sham injection in the contralateral MFC using fluoroscopic and arthroscopic guidance. Dogs were evaluated over the following year for clinical function and kinetics, radiographic and MRI scoring and at 3, 6 and 12-month endpoints (n=4/time) by arthroscopic, gross, microCT, and histologic assessments. Data were compared for statistically significant (p<0.05) differences to pre-operative measures and between groups. Twelve weeks after impact and prior to treatment, all dogs had MFC BMLs and significantly (p<0.01) more lameness/less limb loading, greater pain and joint effusion, and decreased knee range of motion compared to pre-operative values, with no significant difference between limbs. Transient pain and lameness requiring extended oral analgesic treatment were noted for up to 3 months after SCP injection. At all time points after treatment, all dogs had radiographic evidence of mild osteoarthritis and MRI-evident BMLs of the MFC with associated clinical symptoms, with no significant differences between groups until 1 year after treatment. At the 1-year endpoint, SCP was associated with significantly (p<0.05) less pain, better knee range of motion, and better arthroscopic scores. BMLs were present in both groups based on one-year MRI, microCT, and histology with SCP-treated MFCs showing subjectively better trabecular bone architecture. SCP injectate was retained in all MFCs at one year and was well integrated with trabecular bone with no evidence of associated pathology. Both groups showed MRI, gross, and histologic evidence for medial compartment gonarthrosis at all time points with no significant differences noted between groups for these assessments. In this pre-clinical canine model of PTOA, the treatment of BMLs of the medial femoral condyle with SCP was associated with superior clinical results and better trabecular bone architecture at the one year time point when compared with sham controls. Despite improvements in clinical outcomes and bony histology, there were no difference detected in the progression of medial compartment post-traumatic gonarthrosis between groups. Clinical trials are warranted to further validate these results and to determine the efficacy of SCP in human subjects.

Treatment of pulmonary fibrosis with siRNA against a collagen-specific chaperone HSP47 in vitamin A-coupled liposomes

ABSTRACTBackground: Pulmonary fibrosis is a life-threatening pathological state of progressive interstitial lung diseases, such as idiopathic pulmonary fibrosis. Myofibroblasts are known to play a critical role in the pathogenesis of pulmonary fibrosis. This study aimed to evaluate the inhibitory effect of a small interfering RNA (siRNA) on a collagen-specific chaperone heat shock protein 47 (HSP47). The siRNA was preferentially delivered to myofibroblasts in a bleomycin (BLM)-induced pulmonary fibrosis rat model using siRNA against HSP47, encapsulated in a vitamin A-coupled liposome (VA-lip-siRNA HSP47). Methods and Results: Male Sprague–Dawley rats were treated with an intratracheal injection of BLM or phosphate buffered saline followed by an intravenous injection of VA-lip-siRNA HSP47 three times per week under preventive administration schedules from day 1 to day 21 and therapeutic administration schedules from day 15 to day 35. The expression of HSP47 after the treatment was assessed by immunoblotting. The specific delivery of VA-lip-siRNA HSP47 conjugated with 6′-carboxyfluoresce into myofibroblasts was examined by immunofluorescence staining. The effect of VA-lip-siRNA HSP47 on fibrosis was analyzed by morphological and biochemical methods. Preferential delivery of VA-lip-siRNA HSP47 to myofibroblasts in fibrotic areas in BLM-treated rats was verified by immunofluorescence staining. Treatment of VA-lip-siRNA HSP47 clearly suppressed HSP47 expression and induced apoptosis of myofibroblasts in the lung of BLM-treated rats. Hydroxyproline levels and inflammatory cytokines in the lungs, and the number of inflammatory cells in the bronchial alveolar lavage of BLM-treated rats were significantly suppressed by the treatment. Morphological assessment showed that VA-lip-siRNA HSP47 also significantly improved the morphological pulmonary fibrosis of BLM-treated rats in both preventive and therapeutic schedules. Conclusions: These results suggest that VA-lip-siRNA HSP47 improves pulmonary fibrosis in not only preventive, but also therapeutic schedules, and thus, this drug delivery system should provide a novel therapy for refractory pulmonary fibrosis.