RESUMO
We have previously shown that a single application of the growth factors ciliary neurotrophic factor (CNTF) or fibroblast growth factor 2 (FGF-2) to the crushed optic nerve of the frog, Rana pipiens, increases the numbers and elongation rate of regenerating retinal ganglion cell axons. Here we investigate the effects of these factors on the numbers and types of macrophages that invade the regeneration zone. In control PBS-treated nerves, many macrophages are present 100 µm distal to the crush site at 1 week after injury; their numbers halve by 2 weeks. A single application of CNTF at the time of injury triples the numbers of macrophages at 1 week, with this increase compared to control being maintained at 2 weeks. Application of FGF-2 is equally effective at 1 week, but the macrophage numbers have fallen to control levels at 2 weeks. Immunostaining with a pan-macrophage marker, ED1, and a marker for M2-like macrophages, Arg-1, showed that the proportion of the putative M2 phenotype remained at approximately 80% with all treatments. Electron microscopy of the macrophages at 1 week shows strong phagocytic activity with all treatments, with many vacuoles containing axon fragments and membrane debris. At 2 weeks with PBS or FGF-2 treatment the remaining macrophages are less phagocytically active, containing mainly lipid inclusions. With CNTF treatment, at 2 weeks many of the more numerous macrophages are still phagocytosing axonal debris, although they also contain lipid inclusions. We conclude that the increase in macrophage influx seen after growth factor application is beneficial for the regenerating axons, probably due to more extensive removal of degenerating distal axons, but also perhaps to secretion of growth-promoting substances.
Assuntos
Fator Neurotrófico Ciliar/farmacologia , Fator Neurotrófico Ciliar/uso terapêutico , Fator 2 de Crescimento de Fibroblastos/farmacologia , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Traumatismos do Nervo Óptico/tratamento farmacológico , Traumatismos do Nervo Óptico/metabolismo , Animais , Axônios/efeitos dos fármacos , Axônios/metabolismo , Axônios/ultraestrutura , Imuno-Histoquímica , Microscopia Eletrônica , Rana pipiens , Vacúolos/efeitos dos fármacos , Vacúolos/metabolismo , Vacúolos/ultraestruturaRESUMO
AIM: The goal of this study was to evaluate if a biological hydrogel of recombinant human Fibroblast Growth Factor type 2 (rhFGF-2) in a hyaluronic acid (HA) carrier applied in periodontal intrabony defects would enhance the clinical parameters of regeneration of the periodontal attachment apparatus MATERIALS AND METHODS: Thirty adult patients were evaluated. Two intrabony defects present in contra-lateral quadrants in each patient were randomly allocated for each of the treatment methods employed. Control group (n = 30) were treated by open debridement with the papilla preservation flaps, while the test group (n = 30) also received a topical application of rhFGF-2/HA in the intrabony defect. The parameters evaluated, at baseline and after one year, were, were probing depth (PD), gingival recession (REC), probing attachment level (PAL) and probing bone level (PBL). The primary outcome measures was PAL gain RESULTS: Test sites exhibited significantly more PD reduction (5.5 versus 2.9 mm), PAL gains (4.8 versus 2.2 mm) and shallower residual PD (4.2 versus 6.6 mm) than controls. Moreover, residual PD smaller than 5 mm (100 versus 0%) and PAL gain > 4 mm (60 versus 20%) was significantly more frequent in the test group CONCLUSION: Application of rhFGF-2/HA significantly improved clinical parameters of periodontal wound healing one year after treatment.
Assuntos
Perda do Osso Alveolar/tratamento farmacológico , Regeneração Óssea/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Ácido Hialurônico/química , Adulto , Idoso , Processo Alveolar/efeitos dos fármacos , Periodontite Crônica/tratamento farmacológico , Desbridamento/métodos , Portadores de Fármacos , Feminino , Seguimentos , Retração Gengival/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Perda da Inserção Periodontal/tratamento farmacológico , Bolsa Periodontal/tratamento farmacológico , Estudos Prospectivos , Radiografia Dentária Digital/métodos , Proteínas Recombinantes , Retalhos Cirúrgicos/cirurgia , Resultado do TratamentoRESUMO
PURPOSE: To construct a new biomaterial-small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor, and to evaluate the new biomaterials for the reconstruction of abdominal wall defects. METHODS: Thirty six Sprague-Dawley rats were used in the animal experiments and randomly divided into three groups. The new biomaterial was constructed by combining small intestinal submucosa with gelatin hydrogel for basic fibroblast growth factor release. Abdominal wall defects were created in rats, and repaired using the new biomaterials (group B), compared with small intestinal submucosa (group S) and ULTRAPROTM mesh (group P). Six rats in each group were sacrificed at three and eight weeks postoperatively to examine the gross effects, inflammatory responses, collagen deposition and neovascularization. RESULTS: After implantation, mild adhesion was caused in groups B and S. Group B promoted more neovascularization than group S at three weeks after implantation, and induced significantly more amount of collagen deposition and better collagen organization than groups S and P at eight weeks after implantation. CONCLUSION: Small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor could promote better regeneration and remodeling of host tissues for the reconstruction of abdominal wall defects.
Assuntos
Parede Abdominal/cirurgia , Materiais Biocompatíveis/uso terapêutico , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Gelatina/uso terapêutico , Hidrogel de Polietilenoglicol-Dimetacrilato/uso terapêutico , Mucosa Intestinal/transplante , Parede Abdominal/patologia , Animais , Colágeno/análise , Imuno-Histoquímica , Intestino Delgado , Teste de Materiais , Distribuição Aleatória , Ratos Sprague-Dawley , Regeneração , Reprodutibilidade dos Testes , Fatores de Tempo , Aderências Teciduais , Resultado do TratamentoRESUMO
Chronic kidney disease (CKD) is characterized by loss of renal function. The pathological processes involved in the progression of this condition are already known, but the molecular mechanisms have not been completely explained. Recent reports have shown the intrinsic capacity of the kidney to undergo repair after acute injury through the reexpression of repairing proteins (Villanueva S, Cespedes C, Vio CP. Am J Physiol Regul Integr Comp Physiol 290: R861-R870, 2006). Stimulation with basic fibroblast growth factor (bFGF) could accelerate this process. However, it is not known whether bFGF can induce this phenomenon in kidney cells affected by CKD. Our aim was to study the evolution of renal damage in animals with CKD treated with bFGF and to relate the amount of repairing proteins with renal damage progression. Male Sprague-Dawley rats were subjected to 5/6 nephrectomy (NPX) and treated with bFGF (30 µg/kg, NPX+bFGF); a control NPX group was treated with saline (NPX+S). Animals were euthanized 35 days after bFGF administration. Functional effects were assessed based on serum creatinine levels; morphological damage was assessed by the presence of macrophages (ED-1), interstitial α-smooth muscle actin (α-SMA), and interstitial collagen through Sirius red staining. The angiogenic factors VEGF and Tie-2 and the epithelial/tubular factors Ncam, bFGF, Pax-2, bone morphogenic protein-7, Noggin, Lim-1, Wnt-4, and Smads were analyzed. Renal stem cells were evaluated by Oct-4. We observed a significant reduction in serum creatinine levels, ED-1, α-SMA, and Sirius red as well as an important induction of Oct-4, angiogenic factors, and repairing proteins in NPX+bFGF animals compared with NPX+S animals. These results open new perspectives toward reducing damage progression in CKD.
Assuntos
Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Rim/efeitos dos fármacos , Insuficiência Renal Crônica/tratamento farmacológico , Animais , Proteínas Morfogenéticas Ósseas/metabolismo , Creatinina/sangue , Fator 2 de Crescimento de Fibroblastos/farmacologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Rim/metabolismo , Rim/patologia , Masculino , Nefrectomia , Ratos , Ratos Sprague-Dawley , Receptor TIE-2/metabolismo , Insuficiência Renal Crônica/metabolismo , Insuficiência Renal Crônica/patologia , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
PURPOSE: Failure of severed adult central nervous system (CNS) axons to regenerate could be attributed with a reduced intrinsic growing capacity. Severe spinal cord injury is frequently associated with a permanent loss of function because the surviving neurons are impaired to regrow their fibers and to reestablish functional contacts. Peripheral nerves are known as good substrate for bridging CNS trauma with neurotrophic factor addition. We evaluated whether fibroblastic growth factor 2 (FGF-2) placed in a gap promoted by complete transection of the spinal cord may increase the ability of sciatic nerve graft to enhance motor recovery and fibers regrow. METHODS: We used a complete spinal cord transection model. Rats received a 4 mm-long gap at low thoracic level and were repaired with saline (control) or fragment of the sciatic nerve (Nerve) or FGF-2 was added to nerve fragment (Nerve+FGF-2) to the grafts immediately after complete transection. The hind limbs performance was evaluated weekly for 8 weeks by using motor behavior score (BBB) and sensorimotor tests-linked to the combined behavior score (CBS), which indicate the degree of the motor improvement and the percentage of functional deficit, respectively. Neuronal plasticity were evaluated at the epicenter of the injury using MAP-2 and GAP-43 expression. RESULTS: Spinal cord treatment with sciatic nerve and sciatic nerve plus FGF-2 allowed recovery of hind limb movements compared to control, manifested by significantly higher behavioral scores. Higher amounts of MAP-2 and GAP-43 immunoreactive fibers were found in the epicenter of the graft when FGF-2 was added. CONCLUSIONS: FGF-2 added to the nerve graft favored the motor recovery and fiber regrowth. Thus, these results encourage us to explore autologous transplantation as a novel and promising cell therapy for treatment of spinal cord lesion.
Assuntos
Fator 2 de Crescimento de Fibroblastos/fisiologia , Regeneração Nervosa/fisiologia , Nervo Isquiático/transplante , Traumatismos da Medula Espinal/tratamento farmacológico , Traumatismos da Medula Espinal/cirurgia , Transplante de Tecidos/métodos , Animais , Comportamento Animal/fisiologia , Modelos Animais de Doenças , Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Masculino , Ratos , Ratos Wistar , Recuperação de Função Fisiológica/fisiologia , Nervo Isquiático/citologia , Nervo Isquiático/fisiologia , Traumatismos da Medula Espinal/fisiopatologiaRESUMO
Introducción: Debido a los cambios recientes en torno al tratamiento de las fracturas, en los últimos años se ha priorizado una combinación de métodos no sólo mecánicos sino también biológicos. En este estudio se evalúan los resultados de un tratamiento combinado mediante osteosíntesis y aplicación de injerto enriquecido con agregado plaquetario, con resultados alentadores. Materiales y métodos: Nuestra serie comprendió 29 pacientes tratados entre 1999 y 2006, laboralmente activos, con una edad promedio de 42 años (rango, 26 a 62 años). En todos los casos se efectuó osteosíntesis con el agregado plaquetario rico en factores de crecimiento plaquetario. Los resultados se analizaron en función de la formación de callo fracturario a los 6 meses. La obtención de injerto esponjoso fue dificultosa en los pacientes reintervenidos. Resultados: La obtención y preparación del agregado plaquetario no presentó inconvenientes. La consolidación clínica y radiológica se alcanzó en los 29 casos al término de 4 meses (2-6 meses); en 2 casos fue necesario repetir el procedimiento de aporte sin recambio del implante a los 2 meses de la primera intervención. Conclusiones: El injerto autólogo enriquecido con plasma rico en factores de crecimiento pudo haber contribuido de manera favorable a la consolidación de estos casos complejos, con gran ausencia biológica, en los que habían fracasado otros métodos(AU)
Assuntos
Humanos , Adulto , Pessoa de Meia-Idade , Substâncias de Crescimento/uso terapêutico , Fraturas Ósseas/terapia , Fraturas não Consolidadas/terapia , Transplante Autólogo , Regeneração Óssea , Consolidação da Fratura , Terapia Combinada , Fator de Crescimento Derivado de Plaquetas/uso terapêutico , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Fator de Crescimento Transformador beta/uso terapêutico , Resultado do TratamentoRESUMO
Introducción: Debido a los cambios recientes en torno al tratamiento de las fracturas, en los últimos años se ha priorizado una combinación de métodos no sólo mecánicos sino también biológicos. En este estudio se evalúan los resultados de un tratamiento combinado mediante osteosíntesis y aplicación de injerto enriquecido con agregado plaquetario, con resultados alentadores. Materiales y métodos: Nuestra serie comprendió 29 pacientes tratados entre 1999 y 2006, laboralmente activos, con una edad promedio de 42 años (rango, 26 a 62 años). En todos los casos se efectuó osteosíntesis con el agregado plaquetario rico en factores de crecimiento plaquetario. Los resultados se analizaron en función de la formación de callo fracturario a los 6 meses. La obtención de injerto esponjoso fue dificultosa en los pacientes reintervenidos. Resultados: La obtención y preparación del agregado plaquetario no presentó inconvenientes. La consolidación clínica y radiológica se alcanzó en los 29 casos al término de 4 meses (2-6 meses); en 2 casos fue necesario repetir el procedimiento de aporte sin recambio del implante a los 2 meses de la primera intervención. Conclusiones: El injerto autólogo enriquecido con plasma rico en factores de crecimiento pudo haber contribuido de manera favorable a la consolidación de estos casos complejos, con gran ausencia biológica, en los que habían fracasado otros métodos
Assuntos
Humanos , Adulto , Pessoa de Meia-Idade , Substâncias de Crescimento/uso terapêutico , Fraturas Ósseas/terapia , Fraturas não Consolidadas/terapia , Transplante Autólogo , Regeneração Óssea , Consolidação da Fratura , Terapia Combinada , Fator de Crescimento Derivado de Plaquetas/uso terapêutico , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Fator de Crescimento Transformador beta/uso terapêutico , Resultado do TratamentoRESUMO
PURPOSE: The infection is one of the main factors that affect the physiological evolution of the surgical wounds. The aim of this work is to evaluate the effects of fibroblast growth factor (FGFâ) and anti-FGFâ in the healing, synthesis and maturation of collagen when topically used on infected skin wounds of rats. METHODS: An experimental study was performed in 60 male Wistar rats. All animals were divided in two groups (A and B). Each group was divided in three subgroups A1, B1; A2, B2 and A3, B3. After anesthesia with pentobarbital, two open squared wounds (1cm(2)), 4 cm distant to each other, were done in the dorsal skin of all the rats. In group A (n=30) the wounds were contaminated with multibacterial standard solution, and in group B(n=30) the wounds were maintained sterile. These wounds were named F1 (for inflammation analysis) and F2 (for collagen study). The open wounds of A1 and B1 rats were topically treated with saline solution, A2 and B2 were treated with FGFâ and subgroups A3 and B3 were treated with FGFâ and anti-FGFâ. The rats were observed until complete epitelization of F2 wounds for determination of healing time and the expression of types I and III collagen, using Picro Sirius Red staining. Inflammatory reaction in F1 wounds was studied using hematoxilineosin staining. The three variable was measured by the Image Pro-Plus Média Cybernetics software. The statistical analysis was performed by ANOVA and Tukey test, considering p<0.05 as significant. RESULTS: It was observed that infection retarded significantly (p<0.05) the time of wound scarring and the topical application of FCFb reverted the inhibition of healing caused by bacteria. The inflammatory reaction was greater in the subgroup B2 than in B1 and A3, and the difference was significant (p<0.05). It was observed greater expression of type I collagen in all the subgroups treated with FCFb, when compared with the untreated subgroups. Type III collagen was significantly decreased in wounds of B3 rats, comparing to the other subgroups. CONCLUSIONS: The FCFb accelerated the healing of open infected wounds and contributed with maturation of collagen, enhancing the type I collagen density. The anti-FCFb antibody was able to attenuate the production of both type I and III collagen.
Assuntos
Colágeno/metabolismo , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Fibroblastos/fisiologia , Infecção da Ferida Cirúrgica/tratamento farmacológico , Cicatrização/efeitos dos fármacos , Análise de Variância , Animais , Colágeno/farmacologia , Inflamação/fisiopatologia , Masculino , Ratos , Ratos Wistar , Pele/patologia , Cloreto de Sódio , Resistência à Tração , Fatores de TempoRESUMO
PURPOSE: The infection is one of the main factors that affect the physiological evolution of the surgical wounds. The aim of this work is to evaluate the effects of fibroblast growth factor (FGFâ) and anti-FGFâ in the healing, synthesis and maturation of collagen when topically used on infected skin wounds of rats. METHODS: An experimental study was perfomed in 60 male Wistar rats. All animals were divided in two groups (A and B). Each group was divided in three subgroups A1, B1; A2, B2 and A3, B3. After anesthesia with pentobarbital, two open squared wounds (1cm²), 4cm distant to each other, were done in the dorsal skin of all the rats. In group A (n=30) the wounds were contaminated with multibacterial standard solution, and in group B(n=30) the wounds were maintained sterile. These wounds were named F1 (for inflammation analysis) and F2 (for collagen study). The open wounds of A1 and B1 rats were topically treated with saline solution, A2 and B2 were treated with FGFâ and subgroups A3 and B3 were treated with FGFâ and anti-FGFâ. The rats were observed until complete epitelization of F2 wounds for determination of healing time and the expression of types I and III collagen, using Picro Sirius Red staining. Inflammatory reaction in F1 wounds was studied using hematoxilineosin staining. The three variable was measured by the Image Pro-Plus Média Cybernetics software. The statistical analysis was performed by ANOVA and Tukey test, considering p<0.05 as significant. RESULTS: It was observed that infection retarded significantly (p<0.05) the time of wound scarring and the topical application of FCFb reverted the inhibition of healing caused by bacteria. The inflammatory reaction was greater in the subgroup B2 than in B1 and A3, and the difference was significant (p<0.05). It was observed greater expression of type I collagen in all the subgroups treated with FCFb, when compared with the untreated subgroups. Type III collagen was significantly...(AU)
OBJETIVO: Avaliar os efeitos do fator de crescimento de fibroblastos básico (FCFâ) e do anti-FCFâ na cicatrização e maturação do colágeno em feridas infectadas na pele de ratos. MÉTODOS: Um estudo experimental foi realizado em 60 ratos Wistar, divididos em dois grupos (A e B). Cada grupo foi divididos em 03 subgrupos A1,B1; A2,B2 e A3,B3. Após anestesia com pentobarbital sódico intraperitoneal, foram feitas duas feridas abertas de 1cm² na pele no dorso distando 4cm uma da outra. Essas feridas foram denominadas feridas F1 (para análise inflamatória) e F2 (para estudo do colágeno). No grupo A(n=30), as feridas foram contaminadas com solução multibateriana e no grupo B (n=30) as feridas não foram contaminadas. As feridas receberam tratamento tópico com aplicação única. Nos subgrupos A1 e B1 foram tratadas com solução salina tópica, as dos subgrupos A2 e B2 foram tratadas com o FCFâ e nos subgrupos A3 e B3 foram tratadas com FCFâ e com o anti-FCFâ. Os dados formam analisados pelos testes ANOVA de Tukey, considerando p<0,05 como significante. RESULTADOS: A infecção retardou de modo significante o tempo de cicatrização e a aplicação do FCFâ foi capaz de reverter a inibição da cicatrização provocada pela infecção(p<0.05). A resposta inflamatória foi maior nos grupos tratados com o FCFâ, e a aplicação do anti-FCFâ inibiu a reação inflamatória(p<0.05). Houve aumento significante dos colágenos tipo I e III em todos os subgrupos tratados com FCFâ, comparando com os não tratados, sendo a expressão do tipo I mais intensa do que do tipo III (p<0.05). A aplicação do anti-FCFâ inibiu a expressão das moléculas do colágeno. CONCLUSÕES: O FCFâ foi capaz de acelerar a cicatrização de feridas abertas infectadas e contribui para a maturação do colágeno, ao aumentar a expressão do colágeno tipo I, fenômeno que foi atenuado pela ação do anti-FCFâ.(AU)
Assuntos
Animais , Masculino , Ratos , Colágeno/metabolismo , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Fibroblastos/fisiologia , Infecção da Ferida Cirúrgica/tratamento farmacológico , Cicatrização , Análise de Variância , Colágeno/farmacologia , Inflamação/fisiopatologia , Ratos Wistar , Pele/patologia , Cloreto de Sódio , Resistência à Tração , Fatores de TempoRESUMO
BACKGROUND: The poor predictability of periodontal regenerative treatment of Class III furcation defects stimulates the study of alternatives to improve its results, such as the use of polypeptide growth factors. The objective of this study was to evaluate, both histologically and histometrically, the effects of topical application of basic fibroblast growth factor (b-FGF) associated with guided tissue regeneration (GTR) in the treatment of Class III defects surgically induced in dogs. METHODS: All second and fourth premolars of 5 mongrel dogs were used and randomly assigned to one of three treatment groups: group 1 (control), treated with scaling and root planing, tetracycline hydrochloride (125 mg/ml) conditioning, and GTR with a collagen membrane; group 2, same treatment as group 1 plus 0.5 mg of b-FGF; group 3, same treatment as group 1 plus 1.0 mg of b-FGF. After a 90-day healing period, routine histologic processing and staining with hematoxylin and eosin and Masson trichrome were performed. RESULTS: The descriptive analysis indicated better regenerative results in both groups treated with b-FGF while the histometric data, analyzed by means of analysis of variance (ANOVA), showed greater filling of the defects in group 2 in comparison to the defects in groups 3 and 1, respectively, which was represented by a smaller area of plaque-occupied space (P = 0.004) as well as a greater amount of newly formed cementum (P = 0.002). CONCLUSIONS: These results indicate that b-FGF, especially in smaller doses, may enhance the regenerative results in Class III furcation lesions, leading to greater filling of these defects with both mineralized and non-mineralized tissues.