Conjunctival changes induced by prostaglandin analogues and timolol maleate: a histomorphometric study

PURPOSE: To compare histological changes induced by antiglaucoma medications in the rabbit conjunctiva. METHODS: Fifty New Zealand rabbits were divided in 5 groups of 10 animals. The left eyes were treated daily with one drop of bimatoprost 0.03 percent, travoprost 0.004 percent, latanoprost 0.005 percent, timolol maleate 0.5 percent or artificial tears containing benzalkonium chloride (BAK) for 30 days. The right eyes served as controls. Superior limbic conjunctival biopsies were performed at the 8th and 30th day in 5 rabbits of each group. The conjunctiva was fixed with 10 percent formaldehyde, followed by HE and PAS staining. Morphohistometric quantitative analyses were performed to evaluate the following parameters: inflammatory infiltrate, epithelial thickness, number of goblet cells, diameter and number of blood vessels. RESULTS: At the 8th and 30th posttreatment days, all groups, except one that received artificial tears, exhibited a diffuse inflammatory infiltrate, composed by lymphocytes and neutrophils, which was denser in the timolol group than in the prostaglandin (PG) analogues groups. At the 30th day, the timolol group also showed an increased subepithelial collagen density and a significant increase in epithelial thickness (p=0.0035). The goblet cell density was significantly increased at the 8th day in the group treated with travoprost (p=0.0006), and at the 30th day in those treated with bimatoprost (p=0.0021) and latanoprost (p=0.009). CONCLUSIONS: Although a moderate, diffuse inflammatory infiltrate was observed in PG-treated eyes, no changes in conjunctival epithelial thickness or subconjunctival collagen density were observed with these medications, suggesting that these drugs induce fewer changes than timolol maleate in the rabbit conjunctiva.

OBJETIVOS: Comparar alterações histológicas induzidas por medicação anti-glaucomatosa na conjuntiva de coelhos. MÉTODOS: Cinqüenta coelhos da raça Nova Zelândia foram divididos em 5 grupos de 10 animais. Os olhos esquerdos foram tratados com uma gota diária de bimatoprosta 0,03 por cento, travoprosta 0,004 por cento, latanoprosta 0,005 por cento, maleato de timolol 0,5 por cento ou lágrimas artificiais contendo cloreto de benzalcônio (BAK) por 30 dias. Os olhos direitos serviram como controles. Foram realizadas biópsias conjuntivais límbicas superiores no 8º e 30º dias em 5 coelhos de cada grupo. A conjuntiva foi fixada com formaldeído 10 por cento, seguido por coloração de HE e PAS. Foi realizada análise quantitativa morfohistométrica para avaliar os seguintes parâmetros: infiltrado inflamatório, espessura epitelial, número de células caliciformes, diâmetro e número de vasos sanguíneos. RESULTADOS: No 8º e 30º dias de tratamento, todos os grupos, exceto aquele que recebeu lágrimas artificiais, exibiram infiltrado inflamatório difuso, composto por linfócitos e neutrófilos, sendo mais denso no grupo timolol do que nos grupos dos análogos de prostaglandinas. No 30º dia, o grupo timolol apresentou um aumento na densidade de colágeno subepitelial e um aumento significativo da espessura epitelial (p=0,0035). A densidade de células caliciformes aumentou significativamente no 8º dia no grupo tratado com travoprosta (p=0,0006), e no 30º dia nos grupos tratados com bimatoprosta (p=0,0021) e latanoprosta (p=0,009). CONCLUSÕES: Embora tenha sido observado um infiltrado inflamatório difuso e moderado nos olhos tratados com análogos de prostaglandinas, não houve alterações na espessura epitelial conjuntival ou densidade colágena subepitelial com essas medicações, sugerindo que essas drogas induzem menores alterações que o maleato de timolol na conjuntiva de coelhos.

Conjunctival changes induced by prostaglandin analogues and timolol maleate: a histomorphometric study.

PURPOSE: To compare histological changes induced by antiglaucoma medications in the rabbit conjunctiva. METHODS: Fifty New Zealand rabbits were divided in 5 groups of 10 animals. The left eyes were treated daily with one drop of bimatoprost 0.03%, travoprost 0.004%, latanoprost 0.005%, timolol maleate 0.5% or artificial tears containing benzalkonium chloride (BAK) for 30 days. The right eyes served as controls. Superior limbic conjunctival biopsies were performed at the 8th and 30th day in 5 rabbits of each group. The conjunctiva was fixed with 10% formaldehyde, followed by HE and PAS staining. Morphohistometric quantitative analyses were performed to evaluate the following parameters: inflammatory infiltrate, epithelial thickness, number of goblet cells, diameter and number of blood vessels. RESULTS: At the 8th and 30th posttreatment days, all groups, except one that received artificial tears, exhibited a diffuse inflammatory infiltrate, composed by lymphocytes and neutrophils, which was denser in the timolol group than in the prostaglandin (PG) analogues groups. At the 30th day, the timolol group also showed an increased subepithelial collagen density and a significant increase in epithelial thickness (p=0.0035). The goblet cell density was significantly increased at the 8th day in the group treated with travoprost (p=0.0006), and at the 30th day in those treated with bimatoprost (p=0.0021) and latanoprost (p=0.009). CONCLUSIONS: Although a moderate, diffuse inflammatory infiltrate was observed in PG-treated eyes, no changes in conjunctival epithelial thickness or subconjunctival collagen density were observed with these medications, suggesting that these drugs induce fewer changes than timolol maleate in the rabbit conjunctiva.

[Evaluation of the aroreira (Schinus terebinthifolius Raddi) in the healing process of surgical incision in the bladder of rats]. / Avaliação da ação da Aroeira (Schinus terebinthifolius Raddi) na cicatrização de feridas cirúrgicas em bexiga de ratos.

PURPOSE: To evaluate the hydroalcoholic aroeira extract in the healing process of surgical incisions in the bladder of rats in an experimental comparative study. METHODS: Forty male Wistar rats were used. All of them underwent a median abdominal incision, with an 1cm cystotomy, followed by one plain suture with separated stitches of poliglactine 910 5-0. The animals were then divided into two groups of 20 rats each, and named the aroeira group (GA) and the control group (GC). In the first one, 80% hydro-alcoholic plant extract was injected in the peritoneal cavity, in a one dose of 100mg/kg. The second group, a 0.9% saline solution was injected in place of aroeira extract. Each group was divided into two subgroups (SGA and SGC) of 10 animals. According to their scheduled death, they were named subgroups SGA3 and SGC3 when killed in the 3th day after surgery and subgroups SGA7 and SGC7 in the 7th day. The abdominal cavity and the bladder suture were macroscopically evaluated. RESULTS: The microscopic analysis of the surgical incision in the bladder was performed by means of the hematoxilin-eosine stain and the trichrome of Masson. The macroscopic analysis showed an infection in the surgical incision in three rats of the SGC group and in one of the SGA, and peritoneal adhesion in the 26 rats belonging nine to SGC and 17 in the SGA. The microscopic evaluation revealed a more severe acute inflammation process in the SGC on the 3th (p=0.045) and in the 7th (0=0.002) days. In the SGA, a more significant collagenization (p=0.001) could be seen, as far as the evidence of vascular neoformation (p=0.002) on the 3rd day. Chronic inflammatory reaction (p=0.006) and a more intense vascular neoformation (p=0.001) were observed in the 7th day. CONCLUSION: The hydroalcoholic aroeira extract had a favorable effect in the healing process of cystotomies done in rats.

[Comparative study of the healing process of gastrographies with and without the use of Jatropha gossypiifolia L. (bellyache bush) extract in rats]. / Estudo comparativo da cicatrização de gastrorrafias com e sem o uso do extrato de Jatropha gossypiifolia L. (pião roxo) em ratos.

INTRODUCTION: The healing process of the gastrointestinal tract is of huge importance in the surgical field history, as well as the search for appropriate types of threads and suture techniques and the many factors that influence it. The use of phytotherapic drugs as accelerators of the healing process has been done for many years and is one of the current objectives of scientific researches trying to prove its therapeutic properties. PURPOSE: To evaluate the use of Jatropha gossypiifolia L. extract on the healing process of gastrographies in rats. METHODS: Forty wistar male rats were divided in 2 groups of 20 rats, named control and Jatropha groups. Ten animals of each group were killed in the third day post-surgery and were named control and Jatropha groups of the 3rd day. The remaining 10 animals of each group were killed in the seventh day and were named accordingly for the 7th day. In each animal, gastrostomy and gastrography were performed in a single plane using polypropylene thread 6-0 (Prolene). The animals from the Jatropha group were given a single dose of 200 mg/kg of the Jatropha gossypiifolia L. extract intraperitoneally on the same day of the procedure and the ones from the control group were given the same quantity in milliliters (ml), but of saline solution (sodium chloride 0.9%). The following parameters were evaluated: 1) macroscopic alterations; 2) the suture's resistance to atmospheric air insufflation (pressure of rupture); 3) histologic characteristics. RESULTS: No animal died during the clinical follow-up and optimal healing of the abdominal wall was seen without any signs of infection, dehiscence, abscesses or peritonitis. Healing of the serous surface was considered good in all animals, without occurrence of fistulas; however, intraperitoneal adhesions occurred in 7 rats of the sub-group control and 9 of the sub-group Jatropha on the 3rd day post-operative and in 9 of the sub-group control and 8 of the sub-group Jatropha on the 7th day, but the differences were not statistically significant between the groups. Healing of the mucous surface was classified as good in all the animals. The resistance of the gastrographies to the atmospheric air insufflation showed statistically significant increase of the rupture pressure in the Jatropha group during the 3rd day of observation. Histologic evaluation showed differences that were statistically significant, considered the criteria, as well as reduced acute inflammatory reaction and better coaptation of the edges in the sub-group Jatropha of 7th post-surgery, when compared to the sub-group control of the same period. CONCLUSION: The raw extract of Jatropha gossypiifolia L. aids the healing on the 3rd day post-surgery, concerning the enhanced resistance of the gastrographies to pressure of rupture, and on the 7th day, presenting better coaptation of the edges and reducing acute inflammatory reaction by microscopic analyses.

Estudo comparativo da cicatrização de gastrorrafias com e sem o uso do extrato de Jatropha gossypiifolia L. (pião roxo) em ratos / Comparative study of the healing process of gastrorrhaphies with and without the use of Jatropha gossypiifolia L. (bellyache bush) extract in rats

INTRODUÇÃO:A cicatrização do trato gastrintestinal é de grande importância na história da cirurgia, buscando-se tipos de fios e pontos apropriados e os diversos fatores que a influenciam. O uso de fitoterápicos como aceleradores da cicatrização tem sido feito por muitos anos sendo objetivo atual de pesquisas para a comprovação científica dessas propriedades terapêuticas. OBJETIVO: Avaliar o uso do extrato bruto de Jatropha gossypiifolia L. na cicatrização de gastrorrafias em ratos. MÉTODOS: Foram utilizados 40 ratos Wistar, machos, divididos em dois grupos de 20, denominados de grupos controle e Jatropha. Dez animais de cada grupo foram mortos no 3º dia pós-operatório e denominados subgrupos controle e Jatropha do 3º dia e os 10 restantes de cada grupo foram mortos no 7º dia com a mesma denominação do 7º dia. Em cada animal foi realizado gastrotomia e gastrorrafia em plano único com fio polipropileno 6-0 (Prolene®, Ethicon). Os animais do grupo Jatropha receberam dose única de 200mg/Kg do extrato bruto de Jatropha gossypiifolia L via intraperitoneal no dia do procedimento e os do grupo controle a mesma quantidade em mililitros de solução salina (cloreto de sódio à 0,9%). Foram avaliados os seguintes parâmetros: 1) alterações macroscópicas; 2) a resistência à insuflação de ar atmosférico (pressão de ruptura) da sutura; 3) características histológicas. RESULTADOS: Não houve morte dos animais na evolução clínica, ocorrendo boa cicatrização da parede abdominal, com ausência de sinais de infecção, deiscência, abscessos ou peritonites. A cicatrização da superfície serosa foi considerada boa em todos os animais, não ocorrendo fístulas, porém, as aderências intra-peritoneais ocorreram em sete ratos do subgrupo controle e nove do subgrupo Jatropha do 3º dia pós-operatório e em nove do subgrupo controle e oito do subgrupo Jatropha do 7º dia, não havendo diferenças estatisticamente significantes entre os grupos. A cicatrização da superfície mucosa foi classificada como boa em todos os animais. A resistência das gastrorrafias à insuflação de ar atmosférico demonstrou aumento estatisticamente significante da pressão de ruptura no grupo Jatropha do 3º dia de observação. A avaliação histológica demonstrou diferenças estatisticamente significantes, quanto aos critérios, reação inflamatória aguda menor e coaptação das bordas maior no subgrupo Jatropha do 7º dia pós-operatório em relação ao grupo controle do mesmo período. CONCLUSÃO: O extrato bruto de Jatropha gossypiifolia L. favorece a cicatrização no 3º dia pós-operatório em relação à maior resistência das gastrorrafias à pressão de ruptura, e no 7º dia com melhor coaptação das bordas e reduzindo reação inflamatória aguda à microscopia.

INTRODUCTION: The healing process of the gastrointestinal tract is of huge importance in the surgical field history, as well as the search for appropriate types of threads and suture techniques and the many factors that influence it. The use of phytotherapic drugs as accelerators of the healing process has been done for many years and is one of the current objectives of scientific researches trying to prove its therapeutic properties. PURPOSE: To evaluate the use of Jatropha gossypiifolia L. extract on the healing process of gastrorraphies in rats. METHODS: Forty wistar male rats were divided in 2 groups of 20 rats, named control and Jatropha groups. Ten animals of each group were killed in the third day post-surgery and were named control and Jatropha groups of the 3rd day. The remaining 10 animals of each group were killed in the seventh day and were named accordingly for the 7th day. In each animal, gastrostomy and gastrorraphy were performed in a single plane using polypropylene thread 6-0 (Prolene). The animals from the Jatropha group were given a single dose of 200 mg/kg of the Jatropha gossypiifolia L. extract intraperitoneally on the same day of the procedure and the ones from the control group were given the same quantity in milliliters (ml), but of saline solution (sodium chloride 0.9%). The following parameters were evaluated: 1) macroscopic alterations; 2) the suture's resistance to atmospheric air insufflation (pressure of rupture); 3) histologic characteristics. RESULTS: No animal died during the clinical follow-up and optimal healing of the abdominal wall was seen without any signs of infection, dehiscence, abscesses or peritonitis. Healing of the serous surface was considered good in all animals, without occurrence of fistulas; however, intraperitoneal adhesions occurred in 7 rats of the sub-group control and 9 of the sub-group Jatropha on the 3rd day post-operative and in 9 of the sub-group control and 8 of the sub-group Jatropha on the 7th day, but the differences were not statistically significant between the groups. Healing of the mucous surface was classified as good in all the animals. The resistance of the gastrorraphies to the atmospheric air insufflation showed statistically significant increase of the rupture pressure in the Jatropha group during the 3rd day of observation. Histologic evaluation showed differences that were statistically significant, considered the criteria, as well as reduced acute inflammatory reaction and better coaptation of the edges in the sub-group Jatropha of 7th post-surgery, when comparet to the sub-group control of the same period. CONCLUSION: The raw extract of Jatropha gossypiifolia L. aids the healing on the 3rd day post-surgery, concerning the enhanced resistance of the gastrorraphies to pressure of rupture, and on the 7th day, presenting better coaptation of the edges and reducing acute inflammatory reaction by microscopic analyses.

Avaliação da ação da Aroeira (Schinus terebinthifolius Raddi) na cicatrização de feridas cirúrgicas em bexiga de ratos / Evaluation of the aroreira (Schinus terebinthifolius Raddi) in the healing process of surgical incision in the bladder of rats

OBJETIVO: Avaliar a ação cicatrizante do extrato hidroalcoólico da aroeira em bexigas de ratos. MÉTODOS: Foram utilizados 40 ratos machos da linhagem Wistar, divididos em dois grupos de 20, denominados grupo aroeira (GA) e grupo controle (GC). Todos foram submetidos à incisão abdominal mediana com cistotomia de 1 cm, seguida de cistorrafia em plano único com pontos separados de poliglactina 910 5-0 (Vicryl®). Após este procedimento, nos animais GA injetou-se 100mg por quilo de peso de extrato hidroalcoólico da aroeira na cavidade peritoneal e nos GC injetou-se 1 ml por quilo de peso de solução salina a 0,9 por cento . Cada grupo foi dividido em dois sub-grupos de 10 animais de acordo com a data da morte: sub-grupo A3 e C3, sacrificados no 3° dia pós-operatório e sub-grupo A7 e C7, sacrificados no 7° dia. A parede, a cavidade abdominal e a sutura da bexiga foram avaliadas macroscopicamente. Amostras de tecido da bexiga foram retiradas e analisadas histologicamente, utilizando a coloração de Hematoxilina-Eosina (HE) e tricrômio de Masson. RESULTADOS: Na análise macroscópica observou-se infecção na incisão cirúrgica em três ratos do GC e em um do GA, e aderências peritoneais em 29 ratos do GC controle e 17 no GA. A avaliação microscópica mostrou processo inflamatório agudo mais severo no 3° (p=0,045) e no 7° dia (p=0,002). Evidenciou-se ainda diferença estatística nos parâmetros utilizados para a avaliação histológica da cicatrização da bexiga nas variáveis colagenização (p = 0.001), reepitelização (p = 0,046) e neoformação (p = 0) nos subgrupos GC e GA no 3° dia e na variável neoformação vascular (p=0,001) no subgrupo do 7° dia. CONCLUSÃO: O uso de extrato hidroalcoólico de aroeira mostrou efeito cicatrizante favorável nas cistotomias em ratos.

PURPOSE: To evaluate the hydroalcoholic aroeira extract in the healing process of surgical incisions in the bladder of rats in an experimental comparative study. METHODS: Forty male Wistar rats were used. All of them underwent a median abdominal incision, with an 1cm cistotomy, followed by one plan suture with separated stitches of poliglactine 910 5-0. The animals were then divided into two groups of 20 rats each, and named the aroeira group (GA) and the control group (GC). In the first one, 80 percent hydro-alcoholic plant extract was injected in the peritoneal cavity, in a one dose of 100mg/kg. The second group, a 0.9 percent saline solution was injected in place of aroeira extract. Each group was divided into two subgroups (SGA and SGC) of 10 animals. According to their scheduled death, they were named subgroups SGA3 and SGC3 when killed in the 3th day after surgery and subgroups SGA7 and SGC7 in the 7th day. The abdominal cavity and the bladder suture were macroscopically evaluated. RESULTS: The microscopic analysis of the surgical incision in the bladder was performed by means of the hematoxilin-eosine stain and the trichrome of Masson. The macroscopic analysis showed an infection in the surgical incision in three rats of the SGC group and in one of the SGA, and peritoneal adhesion in the 26 rats belonging nine to SGC and 17 in the SGA. The microscopic evaluation revealed a more severe acute inflammation process in the SGC on the 3th (p=0.045) and in the 7th (0=0.002) days. In the SGA, a more significant colagenization (p=0.001) could be seen, as far as the evidence of vascular neoformation (p=0.002) on the 3th day. Chronic inflammatory reaction (p=0.006) and a more intense vascular neoformation (p=0.001) were observed in the 7th day. CONCLUSION: The hydroalcoholic aroeira extract have a favorable effect in the healing process of cystotomies done in rats.

Citofotometria do ADN de células do colo do útero infectadas e näo infectadas pelo papiloma vírus humano / Cytophotometric of the cervix cells DNA infected and non infected by the papiloma human virus

Este estudo tem como objetivo determinar características citofotométricas dos núcleos das células do colo uterino infectadas ou näo pelo HPV, considerando o número de cópias virais. Incluiu-se 69 pacientes entre 16 e 48 anos, que realizavam avaliaçäo ginecológica de rotina, citologia oncótica e colcoscopia. As suspeitas de lesäo por HPV à colcoscopia, foram consideradas grupo de estudo e submetidas à biópsia (n=40); as que näo apresentaram lesäo foram consideradas grupo controle (CT, n=29). Confirmou-se a infecçäo ou näo de HPV pela captura híbrida, quantificando ainda em baixa cópias virais (BC ,100) e altas cópias(AC.100). Realizou-se citofotometria pela coloraçäo de FEULGEN e uso do sistema SAMBA 4000. Os resultados demonstram: área nuclear menor em AC que CT e BC (p<0,05), com alta sensibilidade na diferenciaçäo de CT x AC e BC x AC; densidade óptica integrada maior em BC que CT e AC (p<0,05); BC e AC apresentaram maior índice de pequenos agrupamentos de cromatina e de contraste que CT; BC e AC apresentaram menor índice de grandes agrupamentos de cromatina e de energia que CT; a taxa de ploidia foi semelante entre os tres; BC apresentou mais núcleos nas fases S, G2 e M que CT. Concluiu-se que a citofotometria do ADN do núcleo das células do colo uterino foi capaz de diferenciar entre CT, BC e AC, em diferentes níveis de sensitividade e especificidade, segundo os diversos parâmetros analisados, dos núcleos infectados pelo HPV em comparaçäo aos näo infectados