Benefit of Local Anesthesia in Reducing Pain during Collagenase Injection for Dupuytren's Contracture.

Collagenase injection for Dupuytren's contracture is commonly administered without anesthesia. The authors studied the benefit of injecting local anesthesia before collagenase in reducing treatment-related pain. This prospective cohort study included 187 patients (mean age, 69 years; 80 percent men) at two orthopedic departments in Sweden. At one center, 161 consecutive patients scheduled for collagenase injection were assigned to two groups by alternating outpatient clinics; 78 received collagenase without local anesthesia using a modified method (injecting 0.80 mg in multiple spots in the cord) and 83 received local anesthesia injected in the proximal palm before collagenase. At the other center, 26 consecutive patients received collagenase using the standard method (0.58 mg injected in one spot) without local anesthesia. Immediately after the first injection (collagenase or local anesthesia), the patients rated the severity of injection-related pain on a visual analogue scale from 0 (no pain) to 10 (worst pain). Before finger manipulation 1 or 2 days after injection, the patients rated the pain experienced since injection. Mean score ± SD for pain experienced during modified collagenase injection was 4.3 ± 2.5 without local anesthesia and 2.3 ± 1.7 during injection of local anesthesia (before collagenase) (age- and sex-adjusted mean difference, 2.1; 95 percent CI, 1.5 to 2.7; p < 0.001). Mean pain score ± SD during standard collagenase injection without local anesthesia was 4.8 ± 1.8. Mean pain score ± SD during the injection-manipulation interval was 2.9 ± 1.9 in the group without local anesthesia and 2.9 ± 2.3 in the local anesthesia group (p = 0.79). This study shows that local anesthesia significantly reduces the patient's overall pain experience during collagenase treatment for Dupuytren's contracture. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, II.

α7 Nicotinic Acetylcholine Receptor Stimulation Attenuates Neuroinflammation through JAK2-STAT3 Activation in Murine Models of Intracerebral Hemorrhage.

Accounting for high mortality and morbidity rates, intracerebral hemorrhage (ICH) remains one of the most detrimental stroke subtypes lacking a specific therapy. Neuroinflammation contributes to ICH-induced brain injury and is associated with unfavorable outcomes. This study aimed to evaluate whether α7 nicotinic acetylcholine receptor (α7nAChR) stimulation ameliorates neuroinflammation after ICH. Male CD-1 mice and Sprague-Dawley were subjected to intracerebral injection of autologous blood or bacterial collagenase. ICH animals received either α7nAChR agonist PHA-543613 alone or combined with α7nAChR antagonist methyllycaconitine (MLA) or Janus kinase 2 (JAK2) antagonist AG490. Neurobehavioral deficits were evaluated at 24 hours, 72 hours, and 10 weeks after ICH induction. Perihematomal expressions of JAK2, signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor-α (TNF-α), and myeloperoxidase (MPO) were quantified via Western blot. Histologic volumetric analysis of brain tissues was conducted after 10 weeks following ICH induction. PHA-543613 improved short-term neurobehavioral (sensorimotor) deficits and increased activated perihematomal JAK2 and STAT3 expressions while decreasing TNF-α and MPO expressions after ICH. MLA reversed these treatment effects. PHA-543613 also improved long-term neurobehavioral (sensorimotor, learning, and memory) deficits and ameliorated brain atrophy after ICH. These treatment effects were reduced by AG490. α7nAChR stimulation reduced neuroinflammation via activation of the JAK2-STAT3 pathway, thereby ameliorating the short- and long-term sequelae after ICH.

Functional characterization of a mouse model for central post-stroke pain.

BACKGROUND: Stroke patients often suffer from a central neuropathic pain syndrome called central post-stroke pain. This syndrome is characterized by evoked pain hypersensitivity as well as spontaneous, on-going pain in the body area affected by the stroke. Clinical evidence strongly suggests a dysfunction in central pain pathways as an important pathophysiological factor in the development of central post-stroke pain, but the exact underlying mechanisms remain poorly understood. To elucidate the underlying pathophysiology of central post-stroke pain, we generated a mouse model that is based on a unilateral stereotactic lesion of the thalamic ventral posterolateral nucleus, which typically causes central post-stroke pain in humans. RESULTS: Behavioral analysis showed that the sensory changes in our model are comparable to the sensory abnormalities observed in patients suffering from central post-stroke pain. Surprisingly, pharmacological inhibition of spinal and peripheral key components of the pain system had no effect on the induction or maintenance of the evoked hypersensitivity observed in our model. In contrast, microinjection of lidocaine into the thalamic lesion completely reversed injury-induced hypersensitivity. CONCLUSIONS: These results suggest that the evoked hypersensitivity observed in central post-stroke pain is causally linked to on-going neuronal activity in the lateral thalamus.

Leukocyte invasion of the brain after experimental intracerebral hemorrhage in mice.

BACKGROUND AND PURPOSE: Neuroinflammatory processes contribute to secondary neuronal damage after intracerebral hemorrhage. We aimed to characterize the time course of brain immigration of different leukocyte subsets after striatal injection of either autologous blood or collagenase in mice. METHODS: Intracerebral hemorrhage was induced by injection of either autologous blood (20 µL) or collagenase (0.03 U) in C57Bl/6J mice. Hematoma volumetry was performed on cryosections. Blood volume was measured by hemoglobin spectrophotometry. Leukocytes were isolated from hemorrhagic hemisphere 1, 3, 5, and 14 days after intracerebral hemorrhage, stained for leukocyte markers, and measured by flow cytometry. Heterologous blood injection from CD45.1 mice was used to investigate the origin of brain-invading leukocytes. RESULTS: Collagenase injection induced a larger hematoma volume but a similar blood content compared with blood injection. Cerebral leukocyte infiltration in the hemorrhagic hemisphere was similar in both models. The majority of leukocytes isolated from the brain originated from the circulation. CD4+ T lymphocytes were the predominant brain leukocyte population in both models. However, cerebral granulocyte counts were higher after collagenase compared with blood injection. CONCLUSIONS: Brain infiltration of systemic immune cells is similar in both murine intracerebral hemorrhage models. The pathophysiological impact of invading leukocytes and, in particular, of T cells requires further investigation.

Collagenase injections for treatment of Dupuytren disease.

Palmodigital fasciectomy remains the gold standard. The initial outcome is, in my experience, far more predictable than either NA or enzyme fasciotomy (EF). It is also a more durable treatment. NA and EF can be conceptualized as similar procedures--one uses a needle and the other an enzyme to weaken a cord sufficient to be able to rupture it and thus straighten a contracted joint. Both are less invasive and the hand is quick to recover. Both procedures are equally initially effective. CHH seems to offer greater durability. Today's patients are often better educated and seek a specific type of treatment, in particular, effective nonoperative treatment. Pharmaceutical companies now market directly and effectively to patients, and this strategy and Internet use have already resulted in an increase in the number of patients searching for practitioners willing to administer and capable of administering collagenase treatment.

Electrical homogenization of ventricular scar by application of collagenase: a novel strategy for arrhythmia therapy.

BACKGROUND: Radiofrequency ablation for ventricular tachycardia is an established therapy. Use of chemical agents for scar homogenization represents an alternative approach. The purpose of this study was to characterize the efficacy of collagenase (CLG) application on epicardial ventricular scar homogenization. METHODS AND RESULTS: Myocardial infarcts were created in Yorkshire pigs (n=6) by intracoronary microsphere injection. After 46.6±4.3 days, CLG type 2, type 4, and purified CLG were applied in vitro (n=1) to myocardial tissue blocks containing normal myocardium, border zone, and dense scar. Histopathologic studies were performed to identify the optimal CLG subtype. In vivo high-density electroanatomic mapping of the epicardium was also performed, and border zone and dense scar surface area and late potentials were quantified before and after CLG-4 application (n=5). Of the CLG subtypes tested in vitro, CLG-4 provided the best scar modification and least damage to normal myocardium. During in vivo testing, CLG-4 application decreased border zone area (21.3±14.3 to 17.1±11.1 mm(2), P=0.043) and increased dense scar area (9.1±10.3 to 22.0±20.6 mm(2), P=0.043). The total scar area before and after CLG application was 30.4±23.4 and 39.2±29.5 mm(2), respectively (P=0.08). Late potentials were reduced by CLG-4 application (28.8±21.8 to 13.8±13.1, P=0.043). During CLG-4 application (50.0±15.5 minutes), systolic blood pressure and heart rate were not significantly changed (68.0±7.7 versus 61.8±5.3 mmHg, P=0.08; 77.4±7.3 versus 78.8±6.0 beats per minute, P=0.50, respectively). CONCLUSIONS: Ventricular epicardial scar homogenization by CLG-4 application is feasible and effective. This represents the first report on bioenzymatic ablation of arrhythmogenic tissue as an alternative strategy for lesion formation.

Low-level laser therapy in experimental model of collagenase-induced tendinitis in rats: effects in acute and chronic inflammatory phases.

A variety of treatments for tendinopathies is currently used or has been trialed. However, in fact, there is a remarkably little evidence that any conventional therapies are effective. In the last years, low-level laser therapy (LLLT) has been showing interesting results in inflammatory modulation in different musculoskeletal disorders, but the optimal parameters and mechanisms behind these effects are not fully understood. The aim of this study is to investigate if the LLLT modulates the acute and chronic phase of collagenase-induced tendinitis in rat by interfering in mRNA expression for matrix metalloproteinases (MMP13 and MMP1), vascular endothelial growth factor (VEGF), and anti-inflammatory mediator (interleukin (IL)-10). For such, tendinitis was induced by collagenase injection in male Wistar rats. Animals were treated with LLLT (780 nm, potency of 22 mW, 107 mW/cm(2), energy density of 7.5 J/cm(2), and energy delivered of 1.54 J) with different number of treatments in accordance with the inflammatory phase analyzed. LLLT was able to modulate mRNA gene expression of IL-10, VGEF, MMP1, and MMP13 both in acute than in chronic inflammatory phase (p<0.05). Our results suggest that LLLT with parameters employed in the present study was able to modulate IL-10, VEGF, MMP1, and MMP13 mRNA gene expression both in acute than in chronic tendon inflammation. However, further studies are needed to establish optimal parameters for LLLT.

Review of non-surgical treatment options for Peyronie's disease.

Non-surgical treatment of Peyronie's disease (PD) has come a long way since it was first described in 1743. A myriad of treatment options are currently available, including oral, intralesional and external energy therapies. The purpose of this article is to review the contemporary literature on non-surgical therapies for PD, and where possible, focus on randomized, placebo-controlled trials, as well as review the latest guidelines for the management of PD from the International Committee on Sexual Medicine, which conveyed its findings in July 2009. At this time, it appears that a combination of oral agents and/or intralesional injection with traction therapy may provide a synergy between the chemical effects of the drugs and the mechanical effects of traction. Until a reliable treatment emerges, it does appear that some of the non-surgical treatments discussed can be used to stabilize the scarring process and may result in some reduction of deformity with improved sexual function.

Low-level laser therapy (LLLT; 780 nm) acts differently on mRNA expression of anti- and pro-inflammatory mediators in an experimental model of collagenase-induced tendinitis in rat.

Low-level laser therapy (LLLT) has been found to produce anti-inflammatory effects in a variety of disorders. Tendinopathies are directly related to unbalance in expression of pro- and anti-inflammatory cytokines which are responsible by degeneration process of tendinocytes. In the current study, we decided to investigate if LLLT could reduce mRNA expression for TNF-α, IL-1ß, IL-6, TGF-ß cytokines, and COX-2 enzyme. Forty-two male Wistar rats were divided randomly in seven groups, and tendinitis was induced with a collagenase intratendinea injection. The mRNA expression was evaluated by real-time PCR in 7th and 14th days after tendinitis. LLLT irradiation with wavelength of 780 nm required for 75 s with a dose of 7.7 J/cm(2) was administered in distinct moments: 12 h and 7 days post tendinitis. At the 12 h after tendinitis, the animals were irradiated once in intercalate days until the 7th or 14th day in and them the animals were killed, respectively. In other series, 7 days after tendinitis, the animals were irradiated once in intercalated days until the 14th day and then the animals were killed. LLLT in both acute and chronic phases decreased IL-6, COX-2, and TGF-ß expression after tendinitis, respectively, when compared to tendinitis groups: IL-6, COX-2, and TGF-ß. The LLLT not altered IL-1ß expression in any time, but reduced the TNF-α expression; however, only at chronic phase. We conclude that LLLT administered with this protocol reduces one of features of tendinopathies that is mRNA expression for pro-inflammatory mediators.

Dynamics of hydration of cicatricial tissue during local therapy with collagenase preparations.

The components of complex dielectric permeability of cicatricial tissue at the frequency of 55 GHz were studied during local therapy of cicatrices with Fermenkol (a complex of collagenolytic proteases). Electrical characteristics of tissues in the microwave range were interpreted in terms of hydration parameters (total content of water in tissues and the ratio of structured and free water). The rightfulness of interpretations is discussed on the basis of the results of measurements of cell suspension models (native blood and its fractions brought to a standard hematocrit). The results attest to a relationship between hydration of the cicatricial tissue and its morphology and function.

Therapeutic effects of PG201, an ethanol extract from herbs, through cartilage protection on collagenase-induced arthritis in rabbits.

In order to assess the therapeutic effects of PG201 (an ethanol extract from herbs) on osteoarthritis, we investigated whether PG201 could suppress the disease progression of collagenase-induced arthritis (CNIA) in rabbits. The right knees of rabbits were injected intra-articularly with collagenase, and the rabbits were orally treated with distilled water (DW), PG201 (200 mg/kg) or diclofenac (DCF, 10 mg/kg) once a day for 8 weeks. Oral administration of PG201 significantly suppressed the stiffness and bone space narrowing. Cartilage erosion and GAG release (p<0.01) were considerably reduced in the knee joints. As well, the mRNA expression of matrix degradation enzymes including MMP-1, -3, and -13 was decreased. On the contrary, the concentrations of TIMP-2 in the synovial fluids were considerably amplified in the PG201 treated group (p<0.01), but not in the DCF treated group. The pathologic inflammatory molecules involved in cartilage destruction such as IL-1beta, PGE2, and NO were also diminished by PG201. Taken together, these results indicate that PG201 has therapeutic effects on CNIA through the prominent protection of cartilage. PG201 indeed has great potential as a form of treatment for osteoarthritis.

Effect of hyperbaric oxygen therapy on patellar tendinopathy in a rabbit model.

BACKGROUND: Hyperbaric oxygen therapy is a method for augmenting oxygen availability to tissues. This study investigated the effect of hyperbaric oxygen therapy on the collagenase-induced tendinopathy in the rabbit patellar tendon. METHODS: In this study, 13 rabbits were treated by ultrasound-guided injection of 0.025 mL collagenase into the patellar tendon at the right knee, with the left knee serving as a control condition. The rabbits were randomly divided into two groups. After tendinopathy had been confirmed by histologic examination 3 weeks after treatment, hyperbaric oxygen therapy was initiated for group 1. The hyperbaric oxygen therapy involved 30 daily sessions of 2.5 ATA for 120 minutes starting 6 weeks after treatment. The rabbits in group 2 were put in normobaric room air. Both groups were killed 10 weeks after treatment. Histologic examinations as well as mechanical and biochemical tests were performed after the animals were killed. RESULTS: The ultimate tensile load in the tendon that had hyperbaric oxygen therapy was 34.8% greater than that in the control tendon 10 weeks after treatment (p < 0.05). Hydroxyproline concentrations increased 82.2% simultaneously in the tendons that had hyperbaric oxygen therapy, as compared with the concentrations in the control tendons (p < 0.05). However, no statistical difference was found between the two groups in terms of pyridinoline concentration at the 10th week (p > 0.05). The histologic examination demonstrated an increase in blastlike tenocytes in group 1, with more mature phenotype, more organized collagen matrix, absence of myxoid degeneration, and increased vascularity at the 10th week, as compared with the control knee. CONCLUSIONS: The results validate the effectiveness of hyperbaric oxygen therapy in the treatment of tendinopathy. Hyperbaric oxygen therapy may increase collagen synthesis and collagen cross-link formation during the early healing process.

Effects of SKI 306X, a new herbal agent, on proteoglycan degradation in cartilage explant culture and collagenase-induced rabbit osteoarthritis model.

OBJECTIVE: Protective effects of SKI 306X, a natural herbal product extracted from three herbs Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris, on articular cartilage was examined and compared with other osteoarthritis (OA) drugs using in vitro and in vivo models. METHODS: In vitro culture of rabbit articular cartilage explants was used as a model to measure the effects of drugs on the matrix degradation. The recombinant human interleukin-1alpha (rhIL-1alpha, 5 ng/ml) was added to induce proteoglycan (PG) degradation and the degree of PG degradation was assessed by measuring the amount of glycosaminoglycan (GAG) released into the culture medium. In in vivo experiment, collagenase was intraarticularly injected twice into the right knee joint of rabbits to induce OA-like change, and test agents were orally administered once a day for 28 days. The degrees of OA-like changes were evaluated through a histological examination. RESULTS: In vitro study revealed SKI 306X inhibited the degradation of PG in a concentration-dependent manner. Trichosanthes kirilowii, which is one of the major components of SKI 306X, also significantly inhibited the GAG release in cartilage explant culture at 0.3 and 0.1 mg/ml. Dexamethasone and NSAIDs, such as diclofenac and rofecoxib, had no significant effects on the suppression of PG degradation. In in vivo studies, OA-like degeneration of the articular cartilage and synovial tissue was induced by injecting collagenase into the right knee joint of mature rabbits. At a dose of 200 mg/kg, SKI 306X reduced the OA-like histological changes, whereas diclofenac had no effect at 10 mg/kg. CONCLUSION: These results indicate that SKI 306X inhibited PG degradation in cartilage explant culture, and its prophylactic administration significantly protected the knee joint of rabbit from OA-like change in collagenase-induced experimental OA model. This strongly suggests that SKI 306X can be a good OA agent with some cartilage protection activity.

Enhanced adjuvantic property of polymerized liposome as compared to a phospholipid liposome.

Liposome, although intensively researched as vaccine or drug delivery vehicle, has been of limited use due to the low and unpredictable long-term stability. In order to overcome such problems, polymerized liposome (PL) that could initiate polymerization under very mild reaction condition was examined and compared to a conventional liposome. The polymerizable lipid, 1,2-bis[12-(lipoyloxy)dodecanoyl]-sn-glycero-3-phosphorylcholine (DLL), was synthesized according to the literature, and 1,2-distearoyl-sn-glycero-3-phosphorylcholine (DSPC) was used as the conventional lipid counterpart. Polymerization of liposome was as easy and convenient as just shaking in pH 7.4 buffer. The protein encapsulation efficiency of DLL was higher than that of DSPC, and its protein release rate was lower. Immunoglobulin G (IgG) activity examined after intraperitoneal injection of antigen encapsulated by either DLL or DSPC showed that ca. 2 times as much antibody was formed by DLL-encapsulated lysozyme compared with DSPC-encapsulated form. The reasons for the superior adjuvantic properties of DLL and its future application as a drug delivery system are briefly discussed.

In vitro dentin pretreatment: surface roughness and adhesive shear bond strength.

The aim was to define the morphology and roughness of dentin from different tooth areas after various pretreatments to identify the effect of hybrid layer, resin tags, and mineralised dentin surface on shear bond strength. Thirty-eight extracted molars were used, each providing two sections of cervical (c) and lateral (l) dentin. Five pretreatments were performed: A) 0.2% EDTA; B) abrasion with Al2O3 particles, 0.2% EDTA; C) 10% H3PO4; D) 10% H3PO4 and immersion in a collagenase solution; E) control: no treatment. Z100 composite resin cylinders were bonded to the specimens with All Bond 2 bonding resin and tested for shear bond strength. Twelve other specimens from each group were analysed with an optical profilometer and an atomic force microscope, and four were further examined by scanning electron microscopy (SEM). Mean shear strength values in MPa were: Ac: 8.36 +/- 4.23; Al: 8.77 +/- 3.68; Bc: 6.05 +/- 3.62; Bl: 8.39 +/- 4.60; Cc: 6.87 +/- 3.45; Cl: 9.00 +/- 5.62; Dc: 13.30 +/- 5.45; Dl: 8.44 +/- 4.47; Ec: 4.10 +/- 1.54; El: 6.09 +/- 4.34. No statistically significant difference for cervical versus lateral dentin was found within treatments except for group D. Treatments performed on lateral dentin did not differ significantly. In cervical dentin, A differed from E; C from E; and D from A, B, C and E. An increased surface roughness was found in group D. Shear bond strength to dentin did not seem to depend on a hybrid layer formation, but on the direct contact of the adhesive with the mineralised dentinal surface and partly on the orientation of the dentinal tubules.

Effects of chondroitin sulfate-C on bradykinin-induced proteoglycan depletion in rats.

Depletion of the proteoglycan content of articular cartilage was induced by injecting bradykinin (30-300 mumol/l, 50 microliters/knee) into the left knee articular cavities of rats 3 times a day for 2 days. The degree of the reduction in the intensity of histopathological safranin O staining was used as an index of proteoglycan depletion. Bradykinin reduced the cartilage proteoglycan contents of the knee joints of non-injected limbs in a dose-dependent manner and at 300 mumol/l markedly reduced these contents, but evoked no inflammatory changes. The extent of the reduction of the cartilage proteoglycan contents induced by bradykinin injection depended on the dose and injection frequency. Chondroitin sulfate-C (CAS 25322-46-7, Chondroitin ZS Tab) (30-1,000 mg/kg/day) administered orally to rats for 14 days inhibited the bradykinin-induced proteoglycan depletion of the articular cartilage in a dose-dependent manner. These results suggest that a reduction of the proteoglycan content of cartilage, like that associated with osteoarthritis, was induced by injecting bradykinin into the knee articular cavities of rats and chondroitin sulfate-C protected against this effect.

[Experimental study of therapeutical properties and safety of king crab collagenase-containing ointment]. / Eksperimental'noe izuchenie lechebnykh svoistv mazi, soderzhashchei kollagenazu kamchatskogo kraba.

The effects of ointment containing king crab (Paralithodes camtschatica) collagenase on intact skin, thermal, and pyonecrotic wounds were studied in rats by using hematological, biochemical, immunological, and morphological methods. The ointment for the skin and viscera was shown to be safe. It is highly effective in debriding the infected wounds. Different concentrations of collagenase were tested. The concentration of collagenase was recommended to be 0.2 mg/g ointment for use.

Rat tendon morphologic and functional changes resulting from soft tissue mobilization.

Augmented Soft Tissue Mobilization (ASTM) is a new non-invasive soft tissue mobilization technique which has been used successfully to treat a variety of musculoskeletal disorders. The purpose of this study was to determine the effects of ASTM therapy on the morphological and functional characteristics of enzyme induced injured rat Achilles tendons. Four groups of five rats were allocated as follows: (A) control, (B) tendinitis, (C) tendinitis plus ASTM, and (D) ASTM alone. Collagenase injury was induced, and the surgical site was allowed to heal for 3 wk. ASTM was performed on the Achilles tendon of groups C and D for 3 min on postoperative days 21, 25, 29, and 33 for a total of four treatments. Gait data were gathered prior to each treatment. The Achilles tendons of each group were harvested 1 wk after the last treatment. Specimens were prepared for light and electron microscopy, and immunostaining for type I and type III collagen and fibronectin was performed. Light microscopy showed increased fibroblast proliferation in the tendinitis plus ASTM treatment group. Although healing in rats may not translate directly to healing in humans, the findings of this study suggest that ASTM may promote healing via increased fibroblast recruitment.