Selective deletion of SHIP-1 in hematopoietic cells in mice leads to severe lung inflammation involving ILC2 cells.

Src homology 2 domain-containing inositol 5-phosphatase 1 (SHIP-1) regulates the intracellular levels of phosphotidylinositol-3, 4, 5-trisphosphate, a phosphoinositide 3-kinase (PI3K) product. Emerging evidence suggests that the PI3K pathway is involved in allergic inflammation in the lung. Germline or induced whole-body deletion of SHIP-1 in mice led to spontaneous type 2-dominated pulmonary inflammation, demonstrating that SHIP-1 is essential for lung homeostasis. However, the mechanisms by which SHIP-1 regulates lung inflammation and the responsible cell types are still unclear. Deletion of SHIP-1 selectively in B cells, T cells, dendritic cells (DC) or macrophages did not lead to spontaneous allergic inflammation in mice, suggesting that innate immune cells, particularly group 2 innate lymphoid cells (ILC2 cells) may play an important role in this process. We tested this idea using mice with deletion of SHIP-1 in the hematopoietic cell lineage and examined the changes in ILC2 cells. Conditional deletion of SHIP-1 in hematopoietic cells in Tek-Cre/SHIP-1 mice resulted in spontaneous pulmonary inflammation with features of type 2 immune responses and airway remodeling like those seen in mice with global deletion of SHIP-1. Furthermore, when compared to wild-type control mice, Tek-Cre/SHIP-1 mice displayed a significant increase in the number of IL-5/IL-13 producing ILC2 cells in the lung at baseline and after stimulation by allergen Papain. These findings provide some hints that PI3K signaling may play a role in ILC2 cell development at baseline and in response to allergen stimulation. SHIP-1 is required for maintaining lung homeostasis potentially by restraining ILC2 cells and type 2 inflammation.

Quantitative proteomics reveal the protective effects of EDS against osteoarthritis via attenuating inflammation and modulating immune response.

ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium brevicornu Maxim, Dioscorea nipponica Makino, and Salvia miltiorrhiza Bunge formula (EDS) are three traditional Chinese medicines commonly combined and used to treat osteoarthritis (OA). However, the mechanism of its therapeutic effect on OA is still unclear. AIM OF THE STUDY: The aim of this study was to investigate the potential anti osteoarthritis mechanism of EDS in the treatment of OA rats' model by quantitative proteomics. MATERIALS AND METHODS: A papain-induced rat OA model was established, and then EDS was intragastrically administered for 28 days. A label-free quantification proteomics was performed to evaluate the holistic efficacy of EDS against OA and identify the possible protein profiles mechanisms. The expression levels of critical changed proteins were validated by RT-qPCR and Western blotting. The effects of EDS were then assessed by evaluating pathologic changes in the affected knee joint and measuring pressure pain threshold, acoustic reflex threshold, angle of joint curvature. RESULTS: Proteomics analysis showed that 62 proteins were significantly upregulated and 208 proteins were downregulated in OA group compared to control group. The changed proteins were involved in activation of humoral immunity response, complement cascade activation, leukocyte mediated immunity, acute inflammatory response, endocytosis regulation, and proteolysis regulation. The EDS treatment partially restored the protein profile changes. The protective effects of EDS on pathologic changes in OA rats' knee joint and pain threshold assessment were consisted with the proteomics results. CONCLUSIONS: The results suggest that EDS exerted synergistic therapeutic efficacies to against OA through suppressing inflammation, modulating the immune system, relieving joint pain, and attenuating cartilage degradation.

Maintenance of Type 2 Response by CXCR6-Deficient ILC2 in Papain-Induced Lung Inflammation.

Innate lymphoid cells (ILC) are important players of early immune defenses in situations like lymphoid organogenesis or in case of immune response to inflammation, infection and cancer. Th1 and Th2 antagonism is crucial for the regulation of immune responses, however mechanisms are still unclear for ILC functions. ILC2 and NK cells were reported to be both involved in allergic airway diseases and were shown to be able to interplay in the regulation of the immune response. CXCR6 is a common chemokine receptor expressed by all ILC, and its deficiency affects ILC2 and ILC1/NK cell numbers and functions in lungs in both steady-state and inflammatory conditions. We determined that the absence of a specific ILC2 KLRG1+ST2- subset in CXCR6-deficient mice is probably dependent on CXCR6 for its recruitment to the lung under inflammation. We show that despite their decreased numbers, lung CXCR6-deficient ILC2 are even more activated cells producing large amount of type 2 cytokines that could drive eosinophilia. This is strongly associated to the decrease of the lung Th1 response in CXCR6-deficient mice.

CBCT analysis of rabbit temporomandibular joints with osteoarthrosis induced by monoiodoacetate and papain / Análisis por TCCB de la articulación temporomandibular de conejos con osteoartrosis inducida por monoiodoacetato y papaina

Osteoarthrosis (OA) is a degenerative disease characterized by loss of joint cartilage, remodelling of the subchondral bone, narrowing of joint spaces and the formation of osteophytes. Animal models are used to study the mechanisms of OA, as well as to test the effects of anti-osteoarthrosis drugs. The objective of the present study was to determine the changes identifiable by imaging techniques occurring in rabbit temporomandibular joints (TMJ) at 15, 25 and 45 days after OA inducement by monoiodoacetate (MIA) and papain. The imaging technology used was cone-beam computerised tomography (CBCT). The model animals were 22 young adult male New Zealand rabbits, divided randomly into three study groups: Four rabbits in the control group, nine in the papain experimental group and nine in the monoiodoacetate (MIA) experimental group. OA was induced by arthrocentesis in the lower compartment of both TMJs. The rabbits were examined by CBCT at 15, 25 and 45 days after the injection of MIA and papain. The mandibular condyles presented loss of their rounded shape, deformation of the condyle or mandibular fossa, cortical irregularity, cortical wear and changes in the dimensions of the condyle. OA induction by MIA and papain generates changes observable by CBCT in the dimensions of the mandibular condyle in rabbits. Both inducers promote signs compatible with OA on the joint surfaces of the TMJ; MIA promotes more expressive changes.

La osteoartrosis (OA) es una enfermedad degenerativa caracterizada por la pérdida de cartílago articular, remodelación ósea subcondral, estrechamiento del espacio articular y formación de osteofitos. El modelo animal es utilizado para estudiar los mecanismos de la OA, así como testar el efecto de drogas anti-osteoartrosis. El objetivo de este estudio fue determinar los cambios imagenológicos, mediante tomografía computarizada cone-beam (TCCB), que se generan en 15, 25 y 45 días, luego de la inducción de OA por medio de Monoiodoacetato (MIA) y Papaína sobre la ATM de conejos. Fueron utilizados 22 conejos machos, adultos jóvenes, de raza New Zealand divididos aleatoriamente en 3 grupos de estudio: 4 conejos para un grupo control, 9 conejos para el grupo experimental con Papaína y 9 conejos para el grupo experimental con monoiodoacetato (MIA). Se realizó la inducción de OA por la técnica de artrocentesis en el compartimiento inferior de ambas ATMs. Se les realizó examen de TCCB en los días 15, 25 y 45 tras la inyección de MIA y de papaina. Los cóndilos mandibulares presentaron pérdida de forma redondeada de cóndilo, deformidad de cóndilo o fosa mandibular, irregularidad de corticales, desgaste de corticales, cambio de dimensiones de cóndilo. La inducción de OA por medio de MIA y papaína genera cambios en la dimensión del cóndilo mandibular de conejos observables a través de TCCB. Además, ambos inductores promueven signos compatibles con OA en las superficies articulares de la ATM, siendo que la MIA promueve cambios más expresivos.

Characterization of a novel, papain-inducible murine model of eosinophilic rhinosinusitis.

BACKGROUND: Eosinophilic chronic rhinosinusitis (ECRS) is a disease characterized by eosinophilic inflammatory infiltrate and a local type 2 cytokine milieu. Current animal models fail to recapitulate many of the innate and adaptive immunologic hallmarks of the disease, thus hindering the development of effective therapeutics. In the present study, mice were exposed intranasally to the cysteine protease papain, which shares functional similarities with parasitic proteases and aeroallergens, to generate a rapidly inducible murine model of eosinophilic rhinosinusitis. METHODS: C57BL/6 mice were intranasally instilled with 20 µg papain or heat-inactivated papain (HP) on days 0-2 and days 7-10, and then euthanized on day 11. Nasal lavage fluid (NALF) was analyzed to quantify eosinophils and inflammatory cytokine secretion. Sinonasal tissue was sectioned and stained for goblet cells or homogenized to analyze cytokine levels. Serum samples were assayed for immunoglobulin E (IgE) by enzyme-linked immunoassay. Sinonasal mucosal tissue was dissociated and analyzed by flow cytometry. RESULTS: Compared with HP treatment, papain induced significant eosinophilia in NALF, goblet cell hyperplasia, innate and adaptive immune cell infiltration, type 2 cytokine production, and IgE responses. Flow cytometric analysis of sinonasal tissues revealed significant inflammatory cell infiltration and interleukin-13-producing cell populations. CONCLUSION: In this study, we demonstrated that the cysteine protease papain induces allergic sinonasal eosinophilic rhinosinusitis and resembles T-helper 2 cell inflammation and innate immune characteristics of ECRS. This model permits further study into the molecular mechanisms underlying ECRS pathology and provides a model system for the evaluation of potential pharmacologic interventions.

A promising novel formulation for articular cartilage regeneration: Preclinical evaluation of a treatment that produces SOX9 overexpression in human synovial fluid cells.

Osteoarthritis (OA) is a chronic disorder of synovial joints, in which there is progressive softening and disintegration of the articular cartilage. OA is the most common form of arthritis, and is the primary cause of disability and impaired quality of life in the elderly. Despite considerable medical necessity, no treatment has yet been proven to act as a disease­modifying agent that may halt or reverse the structural progression of OA. The replacement of the joint with a prosthesis appears to be the best option in the advanced stages of the disease. A formulation (BIOF2) for cartilage regeneration has been recently developed. The present study evaluated the effects of BIOF2 on gene expression in human cell cultures, followed by efficacy trials in three OA animal models. Human synovial fluid cells that were exposed to the formulation exhibited increased transcription factor SOX­9 (SOX9; chondrogenic factor) expression, and decreased mimecan (mineralization inducer) and macrophage­stimulating protein receptor (osteoclastogenic factor) expression. The intra­articular application of BIOF2 in the animal models significantly increased cartilage thickness from 12 to 31% at 28 days, compared with articular cartilage treated with saline solution. The articular area and number of chondrocytes additionally increased significantly, maintaining an unaltered chondrocyte/mm2 proportion. Evaluation of the histological architecture additionally displayed a decrease in the grade of articular damage in the groups treated with BIOF2. In conclusion, BIOF2 has proven to be effective for treating OA in animal models, most likely due to SOX9 overexpression in articular cells.

R-Ras deficiency does not affect papain-induced IgE production in mice.

INTRODUCTION: R-Ras GTPase has recently been implicated in the regulation of immune functions, particularly in dendritic cell (DC) maturation, immune synapse formation, and subsequent T cell responses. METHODS: Here, we investigated the role of R-Ras in allergen-induced immune response (type 2 immune response) in Rras deficient (R-Ras KO) and wild type (WT) mice. RESULTS: Initially, we found that the number of conventional DC's in the lymph nodes (LNs) was reduced in R-Ras KO mice. The expression of co-stimulatory CD80 and CD86 molecules on these cells was also reduced on DC's from the R-Ras KO mice. However, there was no difference in papain-induced immune response between the R-Ras WT and KO as measured by serum IgE levels after the immunization. Interestingly, neither the DC number nor co-stimulatory molecule expression was different between WT and R-Ras KO animals after the immunization. CONCLUSIONS: Taken together, despite having reduced number of conventional DC's in the R-Ras KO mice and low expression of CD80 on DC's, the R-Ras KO mice are capable of mounting papain-induced IgE responses comparable to that of the WT mice. To our knowledge, this is the first report addressing potential differences in in vivo allergen responses regulated by the R-Ras GTPase.

Itk is required for Th9 differentiation via TCR-mediated induction of IL-2 and IRF4.

Th9 cells produce interleukin (IL)-9, a cytokine implicated in allergic asthma and autoimmunity. Here we show that Itk, a mediator of T cell receptor signalling required for Th2 immune responses and the development of asthma, is a positive regulator of Th9 differentiation. In a model of allergic lung disease, Itk-deficient mice show reduced pulmonary inflammation and IL-9 production by T cells and innate lymphoid type 2 cells (ILC2), despite normal early induction of ILC2s. In vitro, Itk(-/-) CD4(+) T cells do not produce IL-9 and have reduced levels of IRF4 (Interferon Regulator Factor 4), a critical transcription factor for effector T cell function. Both IL-9 and IRF4 expression are rescued by either IL-2 or constitutively active STAT5, but not NFATc1. STAT5 binds the Irf4 promoter, demonstrating one mechanism by which IL-2 rescues weakly activated T cells. Itk inhibition also reduces IL-9 expression by human T cells, implicating ITK as a key regulator of Th9 induction.

A papain-induced disc degeneration model for the assessment of thermo-reversible hydrogel-cells therapeutic approach.

Nucleus pulposus (NP) regeneration by the application of injectable cell-embedded hydrogels is an appealing approach for tissue engineering. We investigated a thermo-reversible hydrogel (TR-HG), based on a modified polysaccharide with a thermo-reversible polyamide [poly(N-isopropylacrylamide), pNIPAM], which is made to behave as a liquid at room temperature and hardens at > 32 °C. In order to test the hydrogel, a papain-induced bovine caudal disc degeneration model (PDDM), creating a cavity in the NP, was employed. Human mesenchymal stem cells (hMSCs) or autologous bovine NP cells (bNPCs) were seeded in TR-HG; hMSCs were additionally preconditioned with rhGDF-5 for 7 days. Then, TR-HG was reversed to a fluid and the cell suspension injected into the PDDM and kept under static loading for 7 days. Experimental design was: (D1) fresh disc control + PBS injection; (D2) PDDM + PBS injection; (D3) PDDM + TR-HG (material control); (D4) PDDM + TR-HG + bNPCs; (D5) PDDM + TR-HG + hMSCs. Magnetic resonance imaging performed before and after loading, on days 9 and 16, allowed imaging of the hydrogel-filled PDDM and assessment of disc height and volume changes. In gel-injected discs the NP region showed a major drop in volume and disc height during culture under static load. The RT-PCR results of injected hMSCs showed significant upregulation of ACAN, COL2A1, VCAN and SOX9 during culture in the disc cavity, whereas the gene expression profile of NP cells remained unchanged. The cell viability of injected cells (NPCs or hMSCs) was maintained at over 86% in 3D culture and dropped to ~72% after organ culture. Our results underline the need for load-bearing hydrogels that are also cyto-compatible.

Increased physical activity severely induces osteoarthritic changes in knee joints with papain induced sulfate-glycosaminoglycan depleted cartilage.

INTRODUCTION: Articular cartilage needs sulfated-glycosaminoglycans (sGAGs) to withstand high pressures while mechanically loaded. Chondrocyte sGAG synthesis is regulated by exposure to compressive forces. Moderate physical exercise is known to improve cartilage sGAG content and might protect against osteoarthritis (OA). This study investigated whether rat knee joints with sGAG depleted articular cartilage through papain injections might benefit from moderate exercise, or whether this increases the susceptibility for cartilage degeneration. METHODS: sGAGs were depleted from cartilage through intraarticular papain injections in the left knee joints of 40 Wistar rats; their contralateral joints served as healthy controls. Of the 40 rats included in the study, 20 rats remained sedentary, and the other 20 were subjected to a moderately intense running protocol. Animals were longitudinally monitored for 12 weeks with in vivo micro-computed tomography (µCT) to measure subchondral bone changes and single-photon emission computed tomography (SPECT)/CT to determine synovial macrophage activation. Articular cartilage was analyzed at 6 and 12 weeks with ex vivo contrast-enhanced µCT and histology to measure sGAG content and cartilage thickness. RESULTS: All outcome measures were unaffected by moderate exercise in healthy control joints of running animals compared with healthy control joints of sedentary animals. Papain injections in sedentary animals resulted in severe sGAG-depleted cartilage, slight loss of subchondral cortical bone, increased macrophage activation, and osteophyte formation. In running animals, papain-induced sGAG-depleted cartilage showed increased cartilage matrix degradation, sclerotic bone formation, increased macrophage activation, and more osteophyte formation. CONCLUSIONS: Moderate exercise enhanced OA progression in papain-injected joints and did not protect against development of the disease. This was not restricted to more-extensive cartilage damage, but also resulted in pronounced subchondral sclerosis, synovial macrophage activation, and osteophyte formation.

Papain-induced asthma: a man with dyspnea from dawn till dust.

A 52-year-old man, a current smoker (40 pack years) with unremarkable medical history, was referred to the outpatient pneumology clinic because of recent complaints of shortness of breath and wheezing, which were relieved by inhaled bronchodilators. Serial peak expiratory flow (PEF) measurements showed a clear rise in PEF during the weekend and a fall on the evening after the first day of the week. It also showed that evening values were always lower than morning values. During a holiday, a slow but persistent rise in PEF was observed. Such a pattern is highly suggestive for occupational asthma. A detailed description of his job revealed papain exposure. After a positive specific IgE and skin prick test for papain the diagnosis of papain induced asthma was made. When an allergy and serious lung function impairment is proven against products encountered in a work related situation, not improving after maximal preventive measures, the patient is advised to change job.

[Comparison study on knee osteoarthritis in rabbits induced by different concentrations of papain].

OBJECTIVE: To compare the knee osteoarthritis (OA) models in rabbits by different concentrations of papain and provide data for exploring pathogenesis and treatments of this disease. METHODS: Sixty New Zealand white rabbits were randomly divided into four groups of 15 each and given injections into the right knee on days 1, 3 and 5 including intra-articular injections of 2%, 5% or 10% (w/v) papain and 0.03 mol/L L-cysteine at the dose of 0.1 ml/kg (experimental groups). The 0.9% NaCl (w/v) with a dose of 0.1 ml/kg were injected intra-articularly into the right knees of rabbits in the control group. The rabbits were sacrificed at 2, 4, 6 weeks respectively after the initiation of papain injection and these OA models were evaluated through recording the width of knee joint, performing the morphological observation and histological evaluation of articular cartilage and synovium. RESULTS: The degenerative changes were demonstrated in knee joints of rabbit in all experimental groups, such as thinner articular cartilage, fibrillation and destroyed cartilage matrix, and inflammation, proliferation, and degeneration of the synovial tissue. All these changes were much worse with increased concentration and prolonged observation time. CONCLUSION: Different severity of OA are established through intra-articular injections of 2%, 5% or 10% papain and 0.03 mol/L L-cysteine at the dose of 0.1 ml/kg. These models are of the characters of short period and a good reproducibility.

Toxicity reduction and MMP-2 stimulation of papain and bromelain loaded in elastic niosomes.

The elastic niosomes (Tween 61/cholesterol/sodium cholate at 1:1:0.1 molar ratio) loaded with the protease enzymes (papain and bromelain) gave the vesicular sizes of 109.5 to 143.9 nm with the negative zeta potential of -14.7 to -30.1 mv. The elastic niosomes loaded with the standard papain (PS), extracted papain (PE), standard bromelain (BS) and extracted bromelain (BE) showed deformability index (DI values) of 1.35, 1.81, 1.22 and 1.61 times higher than their corresponding non-elastic niosomes, respectively. The elastic niosomes did not only improve the entrapment efficiency of the enzymes over the non-elastic niosomes of about 1.35 times, but also reduced the toxicity on skin human fibroblasts by SRB assay of the PS, PE, BS and BE at 1.68, 2.10, 1.56 and 1.52 times, respectively. The relative MMP-2 stimulation of PS, PE, BS and BE loaded in elastic niosomes were 1.26 +/- 0.14, 1.34 +/- 0.15, 1.09 +/- 0.09 and 1.20 +/- 0.04 for the pro MMP-2 and 1.26 +/- 0.12, 1.41 +/- 0.23, 1.01 +/- 0.08 and 1.03 +/- 0.12 for the active MMP-2, respectively in comparing to the control which were similar activity to their free enzymes. The PE loaded in elastic niosomes gave superior characteristics (low cytotoxicity and high MMP-2 stimulation) to other enzymes. The elastic niosomes can enhance the chemical stability of PE, which exhibited higher remaining contents than the free PE of 1.36 times when kept at 27 +/- 2 degrees C after 8 weeks. Therefore, the extracted papain loaded in elastic niosomes appeared to have potential to be developed as a topical product for scar treatment.

Biocompatibility analysis of chemomechanical caries removal material Papacárie on cultured fibroblasts and subcutaneous tissue.

Chemomechanical caries removal allies an atraumatic technique with antimicrobiotic characteristics, minimizing painful stimuli and maximally preserving healthy dental structures. The purpose of this study was to compare the cytotoxic effects of papain-based gel (Papacarie) and another caries-removing substance, Carisolv, to a nontreatment control on cultured fibroblasts in vitro and the biocompatibility in subcutaneous tissue in vivo. The cytotoxicity analysis was performed on fibroblast cultures (NIH-3T3) after 0-, 4-, 8-, and 12-hour exposure (cell viability assay) and after 1-, 3-, 5-, and 7-day exposure (survival assay). In the in vivo study, the 2 compounds were introduced into polyethylene tubes that were implanted into subcutaneous tissues of rats. After 1, 7, 14, 30, and 60 days, tissue samples were examined histologically. Cell viability did not differ between the 2 experimental groups. The control group, however, showed significantly higher percentage viability. There were no differences in cell survival between the control and experimental groups. The histological analysis revealed a moderate inflammatory response at 2 and 7 days and a mild response at 15 days, becoming almost imperceptible by 30 and 60 days in both experimental groups. The 2 tested substances exhibited acceptable biocompatibilities and demonstrated similar responses in the in vitro cytotoxicity and in vivo implantation assay.

[Study of therapeutic effect and mechanism of Sihuang powder treating acute synovitis in experimental rabbit induced by papain injection].

OBJECTIVE: To prove the therapeutic effects of Sihuang powder (composed by four traditional Chinese herbs: root of baikal skullcap, bark of amur corktree, root of sorrel rhubarb, fruit of cape jasmine, which were mixed with wild Chrysanthemum flower solution)in treating acute synovitis in experimental rabbit knee osteoarthritic models induced by papain injection and to explore its mechanism. METHODS: Thirty-two New-Zealand white rabbits were divided into 6 groups: blank group, model group, Sihuang powder with high dosage group (2 g/kg), Sihuang powder with low dosage group (1 g/kg), Yingtaiqing group and wild Chrysanthemum flower group. The latter four groups were treated respectively with low and high dose Sihuang powder synovium and cartilage were tested concentrations of nitrogen monoxide (NO) and IL-1 level and then were prepared for pathologic and histologic observation 10 days later. Cartilage pathologic changes were record and synovium pathologic changes were valued by means of Mankin's value system. RESULTS: The NO concentration of synovium in Sihuang powder with high dosage group was lower than that of model group, and there was significantly differences between the two groups (P < 0.01). The IL-1 level of synovium was failed after treated with Sihuang powder with high dosage (P < 0.05). Sihuang powder with low dosage and Yingtaiqing also could restrain IL-1's release (P < 0.05). In Mankin's value system, Sihuang powder with high dosage almost eliminated inflammatory cells infiltrating in synovium, which was seldom found in other groups. The value of Sihuang powder with high dosage group was the lowest in treatment groups (P < 0.005). Sihuang powder with low dosage group and wild Chrysanthemum flower group also decreased the degree of inflammatory in synovium (P < 0.05). CONCLUSION: Sihuang powder can reduce the concentration of NO and IL-1 and improve inflammatory cell infiltrate in lining cells of synovium. Moreover, it can alleviate swelling and pain of joint, improve joint movement and postpone degeneration of the cartilage.

Mesenchymal stem cells transplantation protects against rat pulmonary emphysema.

Pulmonary emphysema is characterized by loss of alveolar structure. Bone marrow mesenchymal stem cells (MSCs) have been shown to differentiate into alveolar epithelial cells. However, the effect of MSCs transplantation on pulmonary emphysema is unknown. To address this question, cultured bone marrow MSCs from male donor rats were infused into female recipients treated with irradiation and instillation of papain. We found that the emphysematous changes in rats received MSCs transplantation were ameliorated when compared with the rats without MSCs transplantation. Y chromosome fluorescent in situ hybridization (FISH) and immunohistochemical staining for SP-C, confirmed that MSCs engrafted in recipient lungs and differentiated into type II alveolar epithelial cells. Additionally, MSCs transplantation reduced the extent of irradiation and papain-induced alveolar cell apoptosis, likely due to the up-regulation of the expression of Bcl-2 and Bax gene. We conclude that MSCs transplantation protects against the irradiation and papain-induced pulmonary emphysema. The mechanisms of protection may involve the engraftment of MSCs in the lungs, differentiation of MSCs into type II alveolar epithelial cells and suppression of alveolar cell apoptosis.

Resposta de fibroblastos pulpares humanos em cultura ao gel de papacárie / Response of cultured pulpal fibroblasts to papacárie gel

Introdução: O gel de Papacárie® é um produto nacional que contém na sua formulação a papaína, uma proteinase da família das cisteínas, que permite a remoção químico-mecânica da cárie de forma prática, indolor e seletiva, agindo apenas sobre as fibrilas colágenas desestruturadas do tecido cariado. Apesar disso, pouco se sabe sobre a citotoxicidade do gel sobre células da polpa dentária, principalmente em casos de cáries muito profundas, onde poderia ocorrer de forma involuntária o contato indireto ou direto. Métodos: Dessa forma, decidiu-se analisar in vitro a citotoxicidade do gel de Papacárie® utilizando-se células da linhagem celular FP5, provenientes de uma polpa dental humana. Simulando o contato direto, o gel foi depositado em lamínulas de vidro e aplicado sobre culturas confluentes. Para simular o contato indireto, essas células cultivadas foram submetidas à ação do meio de cultura DME previamente condicionado por esse gel. Todos os experimentos foram realizados em triplicata, e os resultados comparados a um grupocontrole, onde as células não entraram em contato com a droga. A contagem e a análise de viabilidade celular foram realizadas 50 segundos e 24 horas após o contato das células com o produto. Resultados: O gel de Papacárie® mostrou ser citotóxico quando em contato direto por 50 segundos, apresentando porcentagem de viabilidade celular menor que a do grupo-controle. Conclusão: Baseando-se nas condições experimentais deste estudo, concluiu-se que o gel de Papacárie é um produto biocompatível.

Introduction: Papain, the main component of Papacárie®, is a proteinase from the cysteine family that allows the chemomechanical removal of carious lesions without pain and in a more selective way than conventional means of demineralized dentinal removal, based on high speed burs. Little is known about the cytotoxic effects of Papacárie on pulp cells, mainly in deep cavities where an involuntary and direct or indirect contact with these cells could occur. Methods: Thus, we decided to analyze in vitro the cytotoxity of Papacárie using human pulp fibroblasts (FP5 cell line). Simulating the direct contact, the gel was applied on round glass coverslips and left in contact with confluent cultures. To simulate the indirect contact, cells were submitted to culture medium previously conditioned by the gel. All the experiments were conducted in triplicate, and the results compared with the controls groups using ANOVA the test. The cell viability percentages were obtained 50 seconds and 24 hours after the cell contact with Papacárie. Results: The results showed that direct contact with the gel for 50 seconds presenting lower cell viability percentages than those of control cells. Conclusion: Based on the experimental conditions of this study, it was concluded that the Papacárie gel is a biocompatible product.

[Modification of red cell membranes with perftoran in papaine emphysema in rats].

Papaine emphysema model on 75 mongrel mature white male rats (10 intact rats were control) was used to study the size, form, surface architechtonics, deformability and state of membrane-receptor erythrocyte complex before and after perftoran intraperitoneal administration. Perftoran emulsion produced a membrane-modulating effect with recovery of hormonal reception sensitivity, PHA-, cAMP-receptor systems as well as restoration of erythrocytic normocytosis and diskocytosis.

Proteolytic enzyme conjugated to SC-glucan as an enzymatic transdermal drug penetration enhancer.

The objective of this study was to investigate the effect of papain, a proteolytic enzyme, on the percutaneous absorption of drugs. To guarantee the enzyme stability during the skin penetration, papain was modified by the conjugation to SC-glucan. The enhancing activity of drug penetration was evaluated using antipyrine and indomethacin as hydrophilic and hydrophobic model drugs, respectively. The SC-glucan-papain conjugate was found to be very effective for facilitating the percutaneous absorption of antipyrine. Microscopic observations showed that the thickness of stratum corneum and viable epidermis was increased by the treatment of the SC-glucan-papain conjugate. Moreover, it induced phase separation, lacuna formation, and lamellar disruption within the stratum corneum interstices. These structural changes by the SC-glucan-papain conjugate are likely to be induced from hydrolysis of extensive crosslinking of corneocyte envelopes and intracellular proteins. However, the SC-glucan-papain conjugate showed no skin irritation according to the Draize test, which may be due to the difficulty of the SC-glucan-papain conjugate in penetrating into the skin.

Efeito da atividade física sobre a evolução do enfisema pulmonar: um estudo experimental em ratos Wistar / Effects of physical activity on the development pulmonar emphysema in rats: experimental study in rats Wistar

O propósito da presente investigação foi avaliar o papel da atividade física no desenvolvimento de enfisema induzido por papaina em ratos. Para tanto, ratos Wistar foram radomicamente divididos em quatro grupos (n = 10 para cada grupo) que receberam, respectivamente, infusão intra-traqueal de papaína (6 mg em 1 ml de NaCI 0,9 por cento) ou veículo e foram submetidos ou não ao protocolo de exercício em uma esteira ergométrica. Os ratos exercitaram-se a 13,3 m/min, 6 dias por semana, durante 9 semanas (o tempo de exercício foi aumentado gradualmente, de 10 a 35 min) / The purpose of the present study was to evaluate the role of exercise trainning on the development of papain-induced emphysema in rats. Wistar rats were randomly assigned to four groups (n = 10 for each group) that receiveid, respectively, intratracheal infusion of papain (6 mg in 1 ml NaCI 0,9 per cent) or vehicle and were submitted or not to a protocol of exercise on a treadmill...