Valproic acid and bladder healing: an experimental study in rats.

PURPOSE: to recognize the effects of valproic acid (VPA), an epigenetic drug, on the bladder healing process, in rats. METHOD: twenty male Wistar rats were divided in two groups: experimental (A), treated with VPA (150mg/Kg/day), and control (B) with 0.9% sodium chloridrate. Healing was analyzed on the third and seventh days, evaluating the inflammatory reaction, collagen synthesis and angiogenesis. RESULTS: inflammatory reaction on the third day was minimal and acute in both groups. On the seventh day, it was subacute in both groups, moderate intensity in group A and minimal in group B (p=0.0476). Collagen III intensity, marked by immunohistochemistry, was similar in both groups. Collagen I intensity on the third day was similar in both groups, but on the seventh day it was higher in experimental than control (p=0.0476). Collagen evaluation by picrosiriusred allowed to verify that the presence of collagen III was similar in both groups (p=0.3312) on the third day, and it was higher in control on the seventh day (p=0.0015). Collagen I showed similarity on the third day (p=0.3100), and it was higher in control on the seventh day (p=0.0015). Vessel marked with anti-SMA counting showed fewer vessels on the third (p=0.0034) and seventh day (p=0.0087) in experimental group. The lower intensity of angiogenesis was confirmed with anti-CD34, on the third day (p=0,0006) and on the seventh day (p=0,0072). CONCLUSION: VPA determined alterations in the bladder healing process, in rats, with lower collagen density and less angiogenic activity, but without compromising the integrity of the organ.

The effects of valproic acid on skin healing: experimental study in rats.

PURPOSE: To recognize the effects of valproic acid (VPA), an epigenetic drug, on the skin healing process. METHODS: Sixty male Wistar rats were divided into two groups: the experiment treated with VPA (100 mg/kg/day); and the control, with 0.9% sodium chloride by gavage. Skin healing was studied in three moments (the third, the seventh, and the 14th day), evaluating the parameters: inflammatory reaction and its intensity (anti-LCA), angiogenesis (anti-CD34), collagen I and III (anti-collagen I, anti-collagen III and Picrosirius-red F3BA) and myofibroblasts (anti-alpha-AMS). RESULTS: The inflammatory reaction was acute or sub-acute in both groups on the third day. On the seventh and the 14th day, chronic predominated in the control (p=0.006), and sub-acute in the experiment (p=0.020). There was a greater number of leukocytes in the group treated only on the third day (p=0.036). The number of vessels was lower in the treated group at the three times (p3=0.002, p7<0.001, and p14=0.027). Myofibroblasts were rare in the third day and moderate quantity in the remaining periods. Collagen I density was higher in the control at the three times (p<0.001) and collagen III in the treated group (p<0.001). CONCLUSIONS: VPA led to a more intense inflammatory reaction, decreased angiogenesis and collagen deposition, especially type I collagen.

Biological compatibility of oxidized cellulose vs. porcine gelatin to control bleeding in liver lesions in rats.

PURPOSE: To compare biological compatibility, hemostasis, and adhesion formation between oxidized regenerated cellulose and lyophilized hydrolyzed porcine collagen in liver trauma. METHODS: Forty male Wistar rats constituted two groups: group A (oxidized cellulose) and group B (lyophilized hydrolyzed collagen). Standardized liver trauma was made, and the hemostatic agent was applied. Animals in subgroups A7 and B7 were submitted to euthanasia and relaparotomy after seven days, and in subgroups A14 and B14 after 14 days. Macroscopic and microscopic results were evaluated. RESULTS: There was no fluid in the cavity in any of the animals, and adhesions were present in all of them. In the analysis after seven days, the adhesions were grades 3 or 4 and consisted of omentum, small intestine, and abdominal wall (p<0.05). In both groups, the mesh was surrounded by a capsule, which was not observed after 14 days. In the evaluation after 14 days, adhesions were grades 2 or 3 (p>0.05). The microscopic examination showed subacute and chronic reactions, in both groups and in both timepoints, with similar frequency. The intensity of fibrosis always presented positive scores. Microabscesses and xanthomatous macrophages were observed in both groups. CONCLUSIONS: There was no superiority of one agent over the other.

Valproic acid and bladder healing: an experimental study in rats / Ácido valpróico e cicatrização em bexiga: estudo experimental em ratos

ABSTRACT Purpose: to recognize the effects of valproic acid (VPA), an epigenetic drug, on the bladder healing process, in rats. Method: twenty male Wistar rats were divided in two groups: experimental (A), treated with VPA (150mg/Kg/day), and control (B) with 0.9% sodium chloridrate. Healing was analyzed on the third and seventh days, evaluating the inflammatory reaction, collagen synthesis and angiogenesis. Results: inflammatory reaction on the third day was minimal and acute in both groups. On the seventh day, it was subacute in both groups, moderate intensity in group A and minimal in group B (p=0.0476). Collagen III intensity, marked by immunohistochemistry, was similar in both groups. Collagen I intensity on the third day was similar in both groups, but on the seventh day it was higher in experimental than control (p=0.0476). Collagen evaluation by picrosiriusred allowed to verify that the presence of collagen III was similar in both groups (p=0.3312) on the third day, and it was higher in control on the seventh day (p=0.0015). Collagen I showed similarity on the third day (p=0.3100), and it was higher in control on the seventh day (p=0.0015). Vessel marked with anti-SMA counting showed fewer vessels on the third (p=0.0034) and seventh day (p=0.0087) in experimental group. The lower intensity of angiogenesis was confirmed with anti-CD34, on the third day (p=0,0006) and on the seventh day (p=0,0072). Conclusion: VPA determined alterations in the bladder healing process, in rats, with lower collagen density and less angiogenic activity, but without compromising the integrity of the organ.

RESUMO Objetivo: reconhecer os efeitos do ácido valpróico (VPA), uma droga epigenética, no processo de cicatrização da bexiga, em ratos. Método: vinte ratos Wistar machos foram divididos em dois grupos: experimental (A), utilizando VPA (150mg/Kg/dia), e controle (B), tratados com cloreto de sódio 0,9% por gavagem. A cicatrização da bexiga foi analisada no terceiro e sétimo dia, estudando-se a reação inflamatória, síntese de colágeno, reepitelização e angiogênese. Resultados: a reação inflamatória no terceiro dia foi mínima e aguda em ambos os grupos. No sétimo dia, foi subaguda em ambos os grupos com intensidade moderada no grupo A e mínima no grupo B (p=0,0476). A intensidade do colágeno III, marcada pela imuno-histoquímica, foi semelhante nos dois grupos, nos dois tempos estudados. A intensidade de colágeno I no terceiro dia foi semelhante nos dois grupos, e maior no sétimo dia no grupo experimental (p=0,0476). A avaliação do colágeno pelo picrosiriusred mostrou que a presença de colágeno III foi semelhante em ambos os grupos (p=0,3312) no terceiro dia, e maior no controle no sétimo dia (p=0,0015). O colágeno I foi semelhante no terceiro dia (p=0,3100), e maior no controle no sétimo dia (p=0,0015). A contagem de vasos marcados pelo anti-SMA mostrou menos vasos no terceiro (p=0,0034) e sétimo dia (p=0,0087) no grupo experimental, confirmado pelo anti-CD34, no terceiro (p=00006) e no sétimo dia (p=0,0072). Conclusão: o VPA determinou alterações no processo de cicatrização da bexiga, em ratos, com menor densidade de colágeno e menor atividade angiogênica, mas sem comprometer a integridade do órgão.

The effects of valproic acid on skin healing: experimental study in rats

Purpose: To recognize the effects of valproic acid (VPA), an epigenetic drug, on the skin healing process. Methods: Sixty male Wistar rats were divided into two groups: the experiment treated with VPA (100 mg/kg/day); and the control, with 0.9% sodium chloride by gavage. Skin healing was studied in three moments (the third, the seventh, and the 14th day), evaluating the parameters: inflammatory reaction and its intensity (anti-LCA), angiogenesis (anti-CD34), collagen I and III (anti-collagen I, anti-collagen III and Picrosirius-red F3BA) and myofibroblasts (anti-alpha-AMS). Results: The inflammatory reaction was acute or sub-acute in both groups on the third day. On the seventh and the 14th day, chronic predominated in the control (p=0.006), and sub-acute in the experiment (p=0.020). There was a greater number of leukocytes in the group treated only on the third day (p=0.036). The number of vessels was lower in the treated group at the three times (p3=0.002, p7<0.001, and p14=0.027). Myofibroblasts were rare in the third day and moderate quantity in the remaining periods. Collagen I density was higher in the control at the three times (p<0.001) and collagen III in the treated group (p<0.001). Conclusions: VPA led to a more intense inflammatory reaction, decreased angiogenesis and collagen deposition, especially type I collagen.

Acetylsalicylic acid (Aspirin®) and liver regeneration: experimental study in rats.

OBJECTIVE: to evaluate the influence of acetylsalicylic acid (ASA) on cell proliferation after partial hepatectomy in rats. METHODS: 40 male Wistar rats were separated into four groups of ten rats each. Groups 1 and 2 (controls): undergoing 30% partial hepatectomy and, after one day (group 1) and seven days (group 2), to euthanasia; daily administration of 0.9% saline solution (1mL per 200g of body weight). Groups 3 and 4 (experimental): undergoing 30% partial hepatectomy and, after one day (group 3) and seven days (group 4), to euthanasia; daily administration of ASA (40mg/mL, 1mL per 200g of body weight). The absolute number of cells stained with PCNA was counted in photomicrographs, in five fields, and it was calculated the mean of positive cells per animal and per group. RESULTS: the final mean of PCNA+ cells per group was: in group 1, 17.57 ± 6.77; in group 2, 19.31 ± 5.30; in group 3, 27.46 ± 11.55; and, in group 4, 12.40 ± 5.23. There was no significant difference at the two evaluation times in the control group (p=0.491), but there was in the experimental group (p=0.020), with a lower number of PCNA+ cells on the seventh day. The comparison between the two groups, on the first day, showed more PCNA+ cells in the livers of the animals that received ASA (p=0.047), and on the seventh day the number was lower in the experimental group (p=0.007). CONCLUSION: ASA induced greater hepatocyte proliferation.

Histological evaluation of the effect of low-frequency electric stimulation on healing Achilles tendons in rats.

PURPOSE: Histologically evaluate the effects of low frequency electrical stimulation in the treatment of Achilles tendon injuries in rats. METHODS: Thirty-four rats underwent Achilles tendon tenotomy and tenorrhaphy. They were randomly allocated in two groups. Half of the sample constituted the experiment group, whose lesions were stimulated with 2 Hz, nonpolarized current and 1 mA, for 14 days. The other animals formed the control group. They were evaluated at 2, 4 and 6 weeks. The histological study was carried out, the collagen density and the wound maturity index were measured. RESULTS: The healing score was higher in the group stimulated at the 6th week (p = 0.018). The density collagen 1 was higher in the group treated at the three times (p = 0.004) and that collagen 3 was higher in the group treated at 6 weeks (p = 0.004). Together, collagen 1 and 3 were higher in the group stimulated at 4 and 6 weeks (p = 0.009, p = 0.004). The maturity index was higher in this group at the three moments (p = 0.017 p = 0.004 and p = 0.009). CONCLUSION: Low frequency electric stimulation improved healing and increased the quantity of collagen.

Histological evaluation of the effect of low-frequency electric stimulation on healing Achilles tendons in rats

Purpose: Histologically evaluate the effects of low frequency electrical stimulation in the treatment of Achilles tendon injuries in rats. Methods: Thirty-four rats underwent Achilles tendon tenotomy and tenorrhaphy. They were randomly allocated in two groups. Half of the sample constituted the experiment group, whose lesions were stimulated with 2 Hz, nonpolarized current and 1 mA, for 14 days. The other animals formed the control group. They were evaluated at 2, 4 and 6 weeks. The histological study was carried out, the collagen density and the wound maturity index were measured. Results: The healing score was higher in the group stimulated at the 6th week (p = 0.018). The density collagen 1 was higher in the group treated at the three times (p = 0.004) and that collagen 3 was higher in the group treated at 6 weeks (p = 0.004). Together, collagen 1 and 3 were higher in the group stimulated at 4 and 6 weeks (p = 0.009, p = 0.004). The maturity index was higher in this group at the three moments (p = 0.017 p = 0.004 and p = 0.009). Conclusion: Low frequency electric stimulation improved healing and increased the quantity of collagen.(AU)

Acetylsalicylic acid (Aspirin®) and liver regeneration: experimental study in rats / Ácido acetilsalicílico (Aspirina®) e regeneração hepática: estudo experimental em ratos

ABSTRACT Objective: to evaluate the influence of acetylsalicylic acid (ASA) on cell proliferation after partial hepatectomy in rats. Methods: 40 male Wistar rats were separated into four groups of ten rats each. Groups 1 and 2 (controls): undergoing 30% partial hepatectomy and, after one day (group 1) and seven days (group 2), to euthanasia; daily administration of 0.9% saline solution (1mL per 200g of body weight). Groups 3 and 4 (experimental): undergoing 30% partial hepatectomy and, after one day (group 3) and seven days (group 4), to euthanasia; daily administration of ASA (40mg/mL, 1mL per 200g of body weight). The absolute number of cells stained with PCNA was counted in photomicrographs, in five fields, and it was calculated the mean of positive cells per animal and per group. Results: the final mean of PCNA+ cells per group was: in group 1, 17.57 ± 6.77; in group 2, 19.31 ± 5.30; in group 3, 27.46 ± 11.55; and, in group 4, 12.40 ± 5.23. There was no significant difference at the two evaluation times in the control group (p=0.491), but there was in the experimental group (p=0.020), with a lower number of PCNA+ cells on the seventh day. The comparison between the two groups, on the first day, showed more PCNA+ cells in the livers of the animals that received ASA (p=0.047), and on the seventh day the number was lower in the experimental group (p=0.007). Conclusion: ASA induced greater hepatocyte proliferation.

RESUMO Objetivo: avaliar a influência do ácido acetilsalicílico (AAS) na proliferação celular após hepatectomia parcial em ratos. Métodos: 40 ratos Wistar machos foram separados em quatro grupos com dez ratos cada. Grupos 1 e 2 (controles): submetidos à hepatectomia parcial de 30% e, após um (grupo 1) e sete dias (grupo 2), à eutanásia; administração diária de solução fisiológica 0,9% (1mL por 200g de peso). Grupos 3 e 4 (experimentos): submetidos à hepatectomia parcial de 30% e, após um (grupo 3) e sete dias (grupo 4), à eutanásia; administração diária de AAS (40mg/mL, 1mL por 200g de peso). Realizou-se a contagem do número absoluto de células coradas com PCNA em fotomicrografias, em cinco campos e cálculo da média de células positivas por animal e por grupo. Resultados: A média final de células PCNA+ por grupo foi: no grupo 1, de 17,57 ± 6,77; no grupo 2 de 19,31 ± 5,30; no grupo 3, de 27,46 ± 11,55; e, no grupo 4, de 12,40 ± 5,23. Não houve diferença significante nos dois tempos de avaliação no grupo controle (p=0,491), mas houve no grupo experimento (p=0,020), observando-se menor número de células PCNA+ no sétimo dia. A comparação entre os dois grupos, no primeiro dia, mostrou mais células PCNA+ nos fígados dos animais que receberam AAS (p=0,047), e no sétimo dia o número foi menor no grupo experimento (p=0,007). Conclusão: O AAS induziu maior proliferação hepatocitária.

Biological compatibility of oxidized cellulose vs. porcine gelatin to control bleeding in liver lesions in rats

ABSTRACT Purpose: To compare biological compatibility, hemostasis, and adhesion formation between oxidized regenerated cellulose and lyophilized hydrolyzed porcine collagen in liver trauma. Methods: Forty male Wistar rats constituted two groups: group A (oxidized cellulose) and group B (lyophilized hydrolyzed collagen). Standardized liver trauma was made, and the hemostatic agent was applied. Animals in subgroups A7 and B7 were submitted to euthanasia and relaparotomy after seven days, and in subgroups A14 and B14 after 14 days. Macroscopic and microscopic results were evaluated. Results: There was no fluid in the cavity in any of the animals, and adhesions were present in all of them. In the analysis after seven days, the adhesions were grades 3 or 4 and consisted of omentum, small intestine, and abdominal wall (p<0.05). In both groups, the mesh was surrounded by a capsule, which was not observed after 14 days. In the evaluation after 14 days, adhesions were grades 2 or 3 (p>0.05). The microscopic examination showed subacute and chronic reactions, in both groups and in both timepoints, with similar frequency. The intensity of fibrosis always presented positive scores. Microabscesses and xanthomatous macrophages were observed in both groups. Conclusions: There was no superiority of one agent over the other.

Tensiometric evaluation of the effect of lowfrequency electric stimulation on healing Achilles tendons in rats.

PURPOSE: To evaluate the effects of low-frequency electric stimulation on biomechanics following surgical treatment of the Achilles tendon in rats. METHODS: Forty-two rats were divided into two groups. One was given electric stimulation and the other was not. All were submitted to Achilles tenotomy and tenorrhaphy performed with a modified Kessler stitch. The experiment group underwent electric stimulation with 2 Hz, a nonpolarized current of 1 mA intensity for 14 days. The animals were euthanized at 2, 4 and 6 weeks for the biomechanical study. RESULTS: The work performed, that is, the tendon's capacity to absorb energy until rupture, was greater in the electrically stimulated group in the 2nd (p = 0.032) and in the 6th week (p = 0.010). The maximum tension, which is the capacity to support a load, was higher in the treated group in the 2nd (p = 0.030) and the 6th week (p = 024). These results indicate greater resistance of the electrically stimulated tendons. An analysis of the elastic module showed no differences. CONCLUSION: Low-frequency electric stimulation increased the resistance of the tendons at 2 and 6 weeks of evolution in rats.

Tensiometric evaluation of the effect of low frequency electric stimulation on healing Achilles tendons in rats

Purpose: To evaluate the effects of low-frequency electric stimulation on biomechanics following surgical treatment of the Achilles tendon in rats. Methods: Forty-two rats were divided into two groups. One was given electric stimulation and the other was not. All were submitted to Achilles tenotomy and tenorrhaphy performed with a modified Kessler stitch. The experiment group underwent electric stimulation with 2 Hz, a nonpolarized current of 1 mA intensity for 14 days. The animals were euthanized at 2, 4 and 6 weeks for the biomechanical study. Results: The work performed, that is, the tendons capacity to absorb energy until rupture, was greater in the electrically stimulated group in the 2nd (p = 0.032) and in the 6th week (p = 0.010). The maximum tension, which is the capacity to support a load, was higher in the treated group in the 2nd (p = 0.030) and the 6th week (p = 024). These results indicate greater resistance of the electrically stimulated tendons. An analysis of the elastic module showed no differences. Conclusion: Low-frequency electric stimulation increased the resistance of the tendons at 2 and 6 weeks of evolution in rats.(AU)

Comparative analysis of the effects of honey, copaiba oil-resin and a commercial product (fibrinolysin, deoxyribonuclease and chloramphenicol) on second intention healing, in rats. / Análise comparativa dos efeitos do mel, do óleo-serina de copaíba e de um produto comercial (fibrinolisina, desoxirribonuclease e cloranfenicol) na cicatrização por segunda intenção, em ratos.

OBJECTIVE: to compare the healing by second intention under the effects of topical application of honey, copaíba oil-resin and a commercial product (fibrinolysin, deoxyribonuclease and chloramphenicol) with a control group in rats. METHODS: we carried out a skin resection, 1cm in diameter, on the back of 40 rats allocated to four groups of ten animals. All wounds were cleaned daily with 2ml of 0.9% NaCl solution. The first group (control - GC) was restricted to such procedure. In the wounds of the second (GM), third (GO) and fourth groups (GF), after cleaning, we respectively applied 1ml of honey, 1ml of copaíba oil-resin and 1ml of cream containing fibrinolysin, deoxyribonuclease and chloramphenicol. The wounds were occluded with sterile gauze. Immediately after the incision and on days three, seven and 14 of the experiment, the wounds were copied and contraction was analyzed using planimetry. After euthanasia, we histologically evaluated the inflammatory reaction and collagen in the scars. RESULTS: the reduction of the wound area of GM (p=0.003), GO (p=0.011) and GF (p=0.002) were higher than the GC. The amount of type-I collagen present in GM and GO was higher than in GC and GF groups (p<0.05). There was a predominance of chronic inflammatory stage in GM (p=0.004), GO (p<0.001) and GF (p=0.003) when compared with GC. CONCLUSION: the topical use of honey and copaíba oil-resin increases wound contraction, the presence of type-I collagen and accelerates healing.

OBJETIVO: comparar a cicatrização, por segunda intenção, sob os efeitos da aplicação tópica de mel, óleo-resina de copaíba e um produto comercial (fibrinolisina, desoxirribonuclease e cloranfenicol) a um grupo controle, em ratos. MÉTODOS: ressecção de pele, com 1cm de diâmetro, foi realizada no dorso de 40 ratos alocados em quatro grupos de dez animais. Todas as feridas foram limpas, diariamente, com 2ml de solução de NaCl 0,9%. O primeiro grupo (controle - GC) ficou restrito a tal procedimento. Nas feridas do segundo (GM), terceiro (GO) e quarto grupos (GF), após limpeza, aplicou-se, respectivamente, 1ml de mel, 1ml de óleo-resina de copaíba e 1ml de creme contendo fibrinolisina, desoxirribonuclease e cloranfenicol. Ocluíram-se as feridas com gaze estéril. Imediatamente após a incisão e nos dias três, sete e 14 do experimento, as feridas foram copiadas e, usando planimetria, analisou-se a contração. Após a eutanásia, a histologia foi utilizada para avaliação da reação inflamatória e do colágeno nas cicatrizes. RESULTADOS: a redução da área da ferida do GM (p=0,003), GO (p=0,011) e GF (p=0,002) foram superiores ao do GC. A quantidade de colágeno tipo I presente no GM e no GO foi superior aos grupos GC e GF (p<0,05). Houve predominância do estágio inflamatório crônico no GM (p=0,004), GO (p<0,001) e GF (p=0,003) quando comparados ao GC. CONCLUSÃO: o uso tópico do mel e do óleo-resina de copaíba aumenta a contração da ferida, a presença de colágeno tipo I e acelera a cicatrização.

Differentiation of myofibroblasts in wounds after topical use of metronidazole: an experimental study. / Diferenciação de miofibroblastos em feridas após uso tópico do metronidazol: estudo experimental.

OBJECTIVE: to assess the effects of topical administration of metronidazole on fibroblast differentiation and on wound contraction during experimental secondary intention wound healing in rats. METHODS: we submitted 108 rats to a circular wound on the back, 2cm in diameter, and divided them into six groups: control group, with application of saline solution on the wound and five experimental groups, divided according to the concentration of metronidazole solution used (4%, 6%, 8%, 10% and 12%). We changed the dressings daily throughout the trial period, which comprised three stages of analysis: three, seven and 14 days. We evaluated wound contraction by digital planimetry, and identified myofibroblasts and protomyofibroblasts using CD34 and α-SMA immunohistochemistry techniques. RESULTS: wound contraction was not different between the experimental and the control groups. Protomyofibroblasts were significantly more numerous at seven days (p=0.022) in the 4%, 6% and 8% metronidazole groups. After 14 days, in the same groups, myofibroblasts predominated significantly (p=0.01). CONCLUSION: the topical administration of metronidazole solution in skin wounds healing by secondary intention was able to improve the differentiation of fibroblasts. The contraction phase of wound healing remained unchanged, without significant reduction of the contraction evaluated by digital planimetry. These results can be used in favor of the wound healing process.

OBJETIVO: avaliar os efeitos da administração tópica do metronidazol na diferenciação de fibroblastos e na contração da ferida durante cicatrização experimental por segunda intenção em ratos. MÉTODOS: cento e oito animais foram submetidos a uma ferida circular no dorso, com 2cm de diâmetro e divididos em seis grupos: grupo controle, com aplicação de solução salina sobre a ferida e cinco grupos experimentais divididos de acordo com a concentração da solução do metronidazol utilizada (4%, 6%, 8%,10% e 12%). Curativos foram realizados diariamente durante todo o período do experimento, que foi subdividido em três momentos de análise: três, sete e 14 dias. A contração da ferida foi avaliada por planimetria digital e os miofibroblastos e protomiofibroblastos foram identificados usando técnicas de imuno-histoquímica CD34 e a-SMA. RESULTADOS: a contração da ferida não apresentou diferença entre os grupos e o controle. Os protomiofibroblastos foram significativamente mais numerosos aos sete dias (p=0,022) nos grupos metronidazol de 4%, 6% e 8%. Após 14 dias, nos mesmos grupos, os miofibroblastos predominaram significativamente (p=0,01). CONCLUSÃO: a administração tópica de solução de metronidazol em feridas de pele com cicatrização por segunda intenção foi capaz de melhorar a diferenciação de fibroblastos. A fase de contração da cicatrização de feridas permaneceu inalterada, sem redução significativa da contração avaliada pela planimetria digital. Estes resultados podem ser utilizados em favor do processo de cicatrização de feridas.

Metronidazole concentration in the bloodstream following its topical application, at different concentration levels, on experimental skin wounds during healing by secondary intention1.

PURPOSE: To characterize qualitatively and quantitatively the absorption of metronidazole solution, in greater concentrations and for longer periods, when applied topically to an experimental open skin wound model. METHODS: An open skin wound, 2 cm in diameter and total skin thickness was prepared, under anesthetic, in the dorsal region of 108 Wistar rats weighing between 300 and 350 grams. The animals were allocated to groups of 18 animals in accordance with the concentration of metronidazole in the solution to be applied daily to the wound. In the control group (CG), 0.9% sodium chloride solution was used for application, and in the experimental groups (GI, GII, GIII, GIV and GV) metronidazole solution at 4%, 6%, 8%, 10% and 12%, respectively, was applied. After 3, 7 and 14 days of treatment. Blood samples collected through cardiac puncture were examined for the existence or non-existence of metronidazole, using high performance liquid chromatography (HPLC). Detected metronidazole values were compared statistically within each group (temporal analysis 3 days X 7 days X 14 days) and between the groups that used topical metronidazole (4% X 6% X 8% X 10% and 12%) using the Kruskal-Wallis test, considering a statistical significance of 95% (p<0.05). RESULTS: Metronidazole was detected in all the samples at all times in all the groups in which topical metronidazole was applied to the wounds. Characteristically, there was no significant difference between the doses obtained within each group over time (3 days X 7 days X 14 days) GI=0.461; GII=0.154; GIII=0.888; GIV= 0.264 and GV=0.152. In the evaluation between groups, a similar degree of absorption was found after 3 days (p=0.829) and 14 days (p=0.751). CONCLUSION: The serum concentration of metronidazole that was achieved was not influenced by the concentration of the solution applied to the skin wound, with similar extend, or by the duration of the application.

Metronidazole concentration in the bloodstream following its topical application, at different concentration levels, on experimental skin wounds during healing by secondary intention

Purpose:To characterize qualitatively and quantitatively the absorption of metronidazole solution, in greater concentrations and for longer periods, when applied topically to an experimental open skin wound model.Methods:An open skin wound, 2 cm in diameter and total skin thickness was prepared, under anesthetic, in the dorsal region of 108 Wistar rats weighing between 300 and 350 grams. The animals were allocated to groups of 18 animals in accordance with the concentration of metronidazole in the solution to be applied daily to the wound. In the control group (CG), 0.9% sodium chloride solution was used for application, and in the experimental groups (GI, GII, GIII, GIV and GV) metronidazole solution at 4%, 6%, 8%, 10% and 12%, respectively, was applied. After 3, 7 and 14 days of treatment. Blood samples collected through cardiac puncture were examined for the existence or non-existence of metronidazole, using high performance liquid chromatography (HPLC). Detected metronidazole values were compared statistically within each group (temporal analysis 3 days X 7 days X 14 days) and between the groups that used topical metronidazole (4% X 6% X 8% X 10% and 12%) using the Kruskal-Wallis test, considering a statistical significance of 95% (p<0.05).Results:Metronidazole was detected in all the samples at all times in all the groups in which topical metronidazole was applied to the wounds. Characteristically, there was no significant difference between the doses obtained within each group over time (3 days X 7 days X 14 days) GI=0.461; GII=0.154; GIII=0.888; GIV= 0.264 and GV=0.152. In the evaluation between groups, a similar degree of absorption was found after 3 days (p=0.829) and 14 days (p=0.751).Conclusion:The serum concentration of metronidazole that was achieved was not influenced by the concentration of the solution applied to the skin wound, with similar extend, or by the duration of the application.(AU)

Diferenciação de miofibroblastos em feridas após uso tópico do metronidazol: estudo experimental / Differentiation of myofibroblasts in wounds after topical use of metronidazole: an experimental study

RESUMO Objetivo: avaliar os efeitos da administração tópica do metronidazol na diferenciação de fibroblastos e na contração da ferida durante cicatrização experimental por segunda intenção em ratos. Métodos: cento e oito animais foram submetidos a uma ferida circular no dorso, com 2cm de diâmetro e divididos em seis grupos: grupo controle, com aplicação de solução salina sobre a ferida e cinco grupos experimentais divididos de acordo com a concentração da solução do metronidazol utilizada (4%, 6%, 8%,10% e 12%). Curativos foram realizados diariamente durante todo o período do experimento, que foi subdividido em três momentos de análise: três, sete e 14 dias. A contração da ferida foi avaliada por planimetria digital e os miofibroblastos e protomiofibroblastos foram identificados usando técnicas de imuno-histoquímica CD34 e a-SMA. Resultados: a contração da ferida não apresentou diferença entre os grupos e o controle. Os protomiofibroblastos foram significativamente mais numerosos aos sete dias (p=0,022) nos grupos metronidazol de 4%, 6% e 8%. Após 14 dias, nos mesmos grupos, os miofibroblastos predominaram significativamente (p=0,01). Conclusão: a administração tópica de solução de metronidazol em feridas de pele com cicatrização por segunda intenção foi capaz de melhorar a diferenciação de fibroblastos. A fase de contração da cicatrização de feridas permaneceu inalterada, sem redução significativa da contração avaliada pela planimetria digital. Estes resultados podem ser utilizados em favor do processo de cicatrização de feridas.

ABSTRACT Objective: to assess the effects of topical administration of metronidazole on fibroblast differentiation and on wound contraction during experimental secondary intention wound healing in rats. Methods: we submitted 108 rats to a circular wound on the back, 2cm in diameter, and divided them into six groups: control group, with application of saline solution on the wound and five experimental groups, divided according to the concentration of metronidazole solution used (4%, 6%, 8%, 10% and 12%). We changed the dressings daily throughout the trial period, which comprised three stages of analysis: three, seven and 14 days. We evaluated wound contraction by digital planimetry, and identified myofibroblasts and protomyofibroblasts using CD34 and α-SMA immunohistochemistry techniques. Results: wound contraction was not different between the experimental and the control groups. Protomyofibroblasts were significantly more numerous at seven days (p=0.022) in the 4%, 6% and 8% metronidazole groups. After 14 days, in the same groups, myofibroblasts predominated significantly (p=0.01). Conclusion: the topical administration of metronidazole solution in skin wounds healing by secondary intention was able to improve the differentiation of fibroblasts. The contraction phase of wound healing remained unchanged, without significant reduction of the contraction evaluated by digital planimetry. These results can be used in favor of the wound healing process.

Análise comparativa dos efeitos do mel, do óleo-serina de copaíba e de um produto comercial (fibrinolisina, desoxirribonuclease e cloranfenicol) na cicatrização por segunda intenção, em ratos / Comparative analysis of the effects of honey, copaiba oil-resin and a commercial product (fibrinolysin, deoxyribonuclease and chloramphenicol) on second intention healing, in rats

RESUMO Objetivo: comparar a cicatrização, por segunda intenção, sob os efeitos da aplicação tópica de mel, óleo-resina de copaíba e um produto comercial (fibrinolisina, desoxirribonuclease e cloranfenicol) a um grupo controle, em ratos. Métodos: ressecção de pele, com 1cm de diâmetro, foi realizada no dorso de 40 ratos alocados em quatro grupos de dez animais. Todas as feridas foram limpas, diariamente, com 2ml de solução de NaCl 0,9%. O primeiro grupo (controle - GC) ficou restrito a tal procedimento. Nas feridas do segundo (GM), terceiro (GO) e quarto grupos (GF), após limpeza, aplicou-se, respectivamente, 1ml de mel, 1ml de óleo-resina de copaíba e 1ml de creme contendo fibrinolisina, desoxirribonuclease e cloranfenicol. Ocluíram-se as feridas com gaze estéril. Imediatamente após a incisão e nos dias três, sete e 14 do experimento, as feridas foram copiadas e, usando planimetria, analisou-se a contração. Após a eutanásia, a histologia foi utilizada para avaliação da reação inflamatória e do colágeno nas cicatrizes. Resultados: a redução da área da ferida do GM (p=0,003), GO (p=0,011) e GF (p=0,002) foram superiores ao do GC. A quantidade de colágeno tipo I presente no GM e no GO foi superior aos grupos GC e GF (p<0,05). Houve predominância do estágio inflamatório crônico no GM (p=0,004), GO (p<0,001) e GF (p=0,003) quando comparados ao GC. Conclusão: o uso tópico do mel e do óleo-resina de copaíba aumenta a contração da ferida, a presença de colágeno tipo I e acelera a cicatrização.

ABSTRACT Objective: to compare the healing by second intention under the effects of topical application of honey, copaíba oil-resin and a commercial product (fibrinolysin, deoxyribonuclease and chloramphenicol) with a control group in rats. Methods: we carried out a skin resection, 1cm in diameter, on the back of 40 rats allocated to four groups of ten animals. All wounds were cleaned daily with 2ml of 0.9% NaCl solution. The first group (control - GC) was restricted to such procedure. In the wounds of the second (GM), third (GO) and fourth groups (GF), after cleaning, we respectively applied 1ml of honey, 1ml of copaíba oil-resin and 1ml of cream containing fibrinolysin, deoxyribonuclease and chloramphenicol. The wounds were occluded with sterile gauze. Immediately after the incision and on days three, seven and 14 of the experiment, the wounds were copied and contraction was analyzed using planimetry. After euthanasia, we histologically evaluated the inflammatory reaction and collagen in the scars. Results: the reduction of the wound area of GM (p=0.003), GO (p=0.011) and GF (p=0.002) were higher than the GC. The amount of type-I collagen present in GM and GO was higher than in GC and GF groups (p<0.05). There was a predominance of chronic inflammatory stage in GM (p=0.004), GO (p<0.001) and GF (p=0.003) when compared with GC. Conclusion: the topical use of honey and copaíba oil-resin increases wound contraction, the presence of type-I collagen and accelerates healing.

Metronidazole concentration in the bloodstream following its topical application, at different concentration levels, on experimental skin wounds during healing by secondary intention

Abstract Purpose: To characterize qualitatively and quantitatively the absorption of metronidazole solution, in greater concentrations and for longer periods, when applied topically to an experimental open skin wound model. Methods: An open skin wound, 2 cm in diameter and total skin thickness was prepared, under anesthetic, in the dorsal region of 108 Wistar rats weighing between 300 and 350 grams. The animals were allocated to groups of 18 animals in accordance with the concentration of metronidazole in the solution to be applied daily to the wound. In the control group (CG), 0.9% sodium chloride solution was used for application, and in the experimental groups (GI, GII, GIII, GIV and GV) metronidazole solution at 4%, 6%, 8%, 10% and 12%, respectively, was applied. After 3, 7 and 14 days of treatment. Blood samples collected through cardiac puncture were examined for the existence or non-existence of metronidazole, using high performance liquid chromatography (HPLC). Detected metronidazole values were compared statistically within each group (temporal analysis 3 days X 7 days X 14 days) and between the groups that used topical metronidazole (4% X 6% X 8% X 10% and 12%) using the Kruskal-Wallis test, considering a statistical significance of 95% (p<0.05). Results: Metronidazole was detected in all the samples at all times in all the groups in which topical metronidazole was applied to the wounds. Characteristically, there was no significant difference between the doses obtained within each group over time (3 days X 7 days X 14 days) GI=0.461; GII=0.154; GIII=0.888; GIV= 0.264 and GV=0.152. In the evaluation between groups, a similar degree of absorption was found after 3 days (p=0.829) and 14 days (p=0.751). Conclusion: The serum concentration of metronidazole that was achieved was not influenced by the concentration of the solution applied to the skin wound, with similar extend, or by the duration of the application.

Adhesions on polypropylene versus Sepramesh® meshes: an experimental study in rats. / Aderências em telas de polipropileno versus telas Sepramesh®: estudo experimental em ratos.

OBJECTIVE: to compare the formation of induced intraperitoneal adhesions in rats when using polypropylene and Sepramesh® meshes. METHODS: we used 20 male Wistar rats, randomly grouped in two groups of ten animals each. We arranged two 10x20mm meshes intraperitoneally into each animal, one being the polypropylene (PP), and the other, Sepramesh®. In Group 1, the polypropylene mesh was positioned to the right, and the Sepramesh®, to the left. In Group 2, the meshes' layout was reversed. After 14 days of the procedure, we euthanized the animals and analyzed the incorporation and percentages of adhesions macroscopically in each mesh. We submitted the collected data to statistical analysis with a significance level of 5% (p<0.05). RESULTS: all meshes showed adhesions. In the Sepramesh® ones, the percentage of surface covered by adhesions ranged from 2% to 86%, with a mean of 18.6±18.6%, while in the polypropylene meshes, it varied between 6% and 86%, with an average of 57.4%±34.9% (p<0.05). The preferred adhesion sites on both meshes were the edges. CONCLUSION: although no mesh was able to completely inhibit the development of adhesions, the Sepramesh® mesh presented less adhesions to the polypropylene mesh. The most common sites of adhesion formation were the edges of the prosthesis, which evidences the importance of the adequate fixation of the meshes.

OBJETIVO: comparar a formação de aderências intraperitoneais, induzidas em ratos, quando utilizadas as telas de polipropileno e Sepramesh®. MÉTODOS: foram utilizados 20 ratos Wistar, machos, agrupados randomicamente em dois grupos de dez animais cada. Duas telas de dimensão 10x20mm foram dispostas intraperitonealmente em cada animal, uma de polipropileno (PP) e a outra Sepramesh®. No Grupo 1, a tela de polipropileno foi posicionada à direita e a tela Sepramesh® à esquerda. No Grupo 2, a disposição das telas foi invertida. Após 14 dias do procedimento, os animais foram eutanasiados e a incorporação e a porcentagem de aderências, em cada tela, analisadas macroscopicamente. Os dados coletados foram submetidos à análise estatística com nível de significância adotado de p<0,05. RESULTADOS: todas as telas apresentaram aderências. Nas telas Sepramesh®, a porcentagem de superfície coberta por aderências variou entre 2% e 86%, com média de 18,6±18,6%, enquanto que, nas telas de polipropileno, variou entre 6% e 86%, com média de 57,4%±34,9% (p<0,05). Os sítios preferenciais de formação de aderências, em ambas as telas, foram as bordas. CONCLUSÃO: embora nenhuma tela tenha sido capaz de inibir completamente o desenvolvimento de aderências, a tela Sepramesh® apresentou menos aderências em relação à tela de polipropileno. A preferência da formação de aderências nas bordas das próteses evidencia a importância da fixação adequada das telas.