Fernando Hintz Greca

Correlação entre o fluxo sangüíneo intestinal e a cicatrização de anastomoses colônicas: estudo experimental em cães

Laser Doppler Flowmetry in the medical field provides a progress to quantify blood flow. Colonic anastomosis deishence is sometimes associated with hipovolemic shock. The aim of the present study was to correlate the bowel blood flow alterations due to hypovolemic shock and colonic anastomosis healing. Sixteen dogs were submited to colonic anastomosis. In eight of them shock was induced. The blood flow was measured during all surgical procedure except when anastomosis was being performed. Also cardiac frequency and medium arterial pressure were measured during all the experiment and hematocrit was obtained from three different samples. On the 7th post operative day, dogs were reoperated to evaluate the anastomosis. Fifteen centimeters of colon with the anastomosis were resected to access anastomosis rupture pressure, colagen densitometry and histology features. Infection complication were twice higher in the shock group compared to the non-shock group. There was no statistic difference between groups relating to rupture pressure. However the total colagen concentration was higher in the non-shock group and histologic study showed better healing parameters in the non-shock group. We concluded that low intestinal flow provides worse anastomosis healing, a high incidence of clinical complications, low colagen concentration and a low standard histology parameters.

Colite do cólon excluso: modelo experimental em ratos

In 1981 Glotzer et al. described a kind of proctocolitis, limited to a segment of colon, excluded from the fecal stream, with pathological features similar to the ulcerative colitis. The purpose of the present study was to establish a experimental model in order to evaluate the pathologic alterations of the colonic mucosa, excluded from the intestinal transit. Thirty five PUCPR Wistar male rats, with a mean age of 120 days were allocated in 4 groups. Under ether anesthesia, the animals were submitted to a 0,5cm resection of the left colon. A proximal terminal colostomy was performed and the distal rectal stump was closed. The excluded segment of the bowel was evaluated macro and microscopically a week (A group), 2 weeks (D group), 4 weeks (B group) and 8 weeks (C group) after the operation. The inflammatory reaction, collagen concentration and changes in the goblets cells were analyzed .Mo was considered the features of the excluded colon seen at the beginning of the experiment and Mf was these features seen at the end of experiment. When we compared these two different phases of the experiment, we verified that a mild acute and cronic inflammatory reaction was present in all of the groups. Ulcers were seen in 11 colons (p=0,0010) and they were not related to the exclusion phase. A significant reduction in the thickness of the colonic wall was observed in all the specimens. In Mo, the mature collagen (type I) was the predominant one (p=0.008) whereas in Mf, the immature collagen ( type III) was the predominant one (p=0.008). Reduction of total collagen, loss of collagen type I and the increase of collagen type III were constant. There was no significant change in the percentage of area occupied by the goblets cells. It was concluded that in rats, the exclusion of the distal colon by colostomy of up to 8 weeks, determines: colon atrophy and the occurrence of superficial ulcerated lesions with low-level inflammatory reaction.

Influência da peritonite sobre a síntese de colágeno em anastomoses do cólon distal: estudo experimental em ratos

The purpose of the present study was to evaluate the burst strength and the collagen concentration in a left colonic anastomosis when peritonitis was present. Forty Wistar male rats were used, with a mean age of 120 days and a mean weight of 302 g. The animals were allocated in two groups of 20 animals each: a control group and an experimental group. Under ether anesthesia, the rats of the experimental group were submitted to a laparotomy and the abdominal sepsis was induced according to the Wichterman method, while those in the control group were submitted to laparotomy and manipulation of the intestinal segments involved in the experiment. Twenty four hours later, the animals underwent to a new laparotomy and a left colonic anastomosis, with a single layer of polypropilene interrupted suture, 2,0 cm above the peritoneal reflection. On the 3rd and 7th post-operative day, the colonic segment with the anastomosis was resected in order to evaluate the bursting strength . The Red Sirius dye was used to evaluate collagen concentration in the histopathological study. Results showed in both groups an increase in the bursting strength , relating to post-operative day. Collagen concentration was poorer in the peritonitis group on the 3rd and 7th post-operative days (p = 0.000168) (p = 0.0020) respectively. The percentages of collagen I and III were similar on the 3rd post-operative day. On the 7th day, however, collagen type III (p =0.000079) predominated in the peritonitis group. The results above showed that the concentration and maturation of collagen are reduced in a colonic anastomosis performed when peritonitis is present. These alterations does not interfere in the bursting strength.

Retalho de submucosa de intestino delgado autólogo para aumento da capacidade da bexiga: estudo experimental em cães

BACKGROUND: Many materials, including organic and synthetic prosthesis, have been described as substitutes of the genitourinary tract. The most commonly used is the ileum, which has many advantages, as easy mobilization and regular vascular pattern. However, the ideal substitute for urinary tract has not been found yet. Ileocystoplasty is not a harmless procedure, and so, investigations are constantly being made to find new substitutes. Small intestinal submucosa (SIS) is an extracelular matrix that can be used as a reabsorbable scaffold for tissue engineering. It allows blood vessels ingrowth, epithelial and connective cells proliferation, induces growing factors production and acquires macro and microscopic characteristics similar to the native tissue. In the urinary tract, previous experimental studies have demonstrated that SIS promotes bladder regeneration. The aim of the present study was to evaluate the biocompatibility of an autologous SIS implant used for bladder augmentation. METHOD: Eight mongrel dogs, weighing between 8 and 15 kg, were prepared for surgery under general anesthesia. A 3cm full-thickness midline cystotomy was performed, followed by immediate augmentation with autologous small intestine submucosa patch graft. On the 30th post-operative day, the animals were re-operated and a fragment of the urinary bladder wall including its patch was removed for histopathogical study. RESULTS: All implants were well incorporated. Macroscopically, SIS could not be distinguished from the native bladder tissue. Mild adherences occurred on the anterior parietal peritoneum. No other macroscopic complications were found. Acute inflammatory response was absent, and chronic inflammatory cells were seen in mild degree. Foreign body reaction was not present, implying satisfactory biological acceptation of the SIS patch graft. Epithelialization was complete in 7/8 implants. Fibroblastic proliferation was moderate. The densitometry of collagen revealed a greater amount of collagen III. Submucosa patch absorption was moderate in most cases. CONCLUSION: We concluded that autologous small intestine submucosa promotes bladder tissue regeneration. Small intestinal submucosa can represent a new option in urinary tract reconstruction.

Influência do buflomedil em retalhos cutâneos isquêmicos: estudo experimental em ratos

Vasoactive drugs have been employed in an attempt to increase the viability of ischemic skin flaps. The aim of the present study was to evaluate the effects of the vasoactive drug, Buflomedil, in the prevention of ischemic skin flaps of rats. Twenty male rats were divided into na experimental group B and a control group A. In the animals of group B, 3mg/kg of Buflomedil was given intraperitoneally every 12 hours for 2 days before surgery and for 7 days after surgery. An abdominal skin flap was used as a model, with a cephalic base measuring 9 x 4cm. The viability of the flaps was evaluated with a laser fluxometer in predetermined points in the early pre and postoperative period and on the 7th post operative day Planimetric analysis of the total area of the flap, of the vascularized area, and of the necrotic area were done on the 7th day after the operation. Inferior necrosis of the flap was observed in all of the animals in both groups. The statistical analysis of the results obtained did not show a significant difference in the planimetry between the control and the experimental groups. Fluxometry did not show a significant difference in relation to the median points between the two groups. It was concluded that Buflomedil, in the manner in which it was used in this experiment, does not reduce necrosis in ischemic skin flaps of rats.

A influência do azul de metileno na prevenção da lesão pulmonar após isquemia-reperfusão intestinal

OBJETIVO: Estudar a acao do azul de metileno como supressor da producao de radicais livres de oxigenio, atuando como receptor alternativo de eletrons na enzima xantina oxidase. METODOS: Foram utilizados 32 ratos Wistar (Rattus norvegicus albinus, Rodentia mammalia) divididos em 2 grupos de 16 animais, os quais foram denominados grupos: experimento e controle. Ambos os grupos foram submetidos a laparotomia mediana e oclusao da arteria mesenterica cranial por 60 minutos. A reperfusao foi confirmada por meio da verificacao do reaparecimento da pulsacao na arcada mesenterica. Foi entao administrado no grupo experimento 2 ml de azul de metileno 1 % esteril intraperitonealmente, enquanto que no grupo controle foi administrado solucao salina isotonica esteril em mesmo volume e pela mesma via de administracao. Apos 4 horas de reperfusao, os animais foram sacrificados. Amostras dos pulmoes foram obtidas para: analise histopatologica, avaliacao do edema e para determinacao da atividade da xantina oxidase. RESULTADOS: O dano pulmonar encontrado no grupo controle foi superior ao encontrado no grupo experimento. Observou-se uma maior formacao de edema nos pulmoes do grupo controle. A atividade da xantina oxidase foi semelhante em ambos os grupos. CONCLUSAO: O azul de metileno diminui a lesao pulmonar apos isquemia-reperfusao intestinal.

Cicatrização de anastomoses do cólon esquerdo com doença inflamatória: estudo experimental em ratos

Complications related to anastomosis failure are frequently described in the surgery of inflammatory bowel disease. The aim of the present study was to evaluated colonic wound healing in an inflamed bowel. Forty Wistar rats were divided in 2 groups: the control and experimental groups. In the experimental group, colitis was induced by the infusion of 10% acetic acid per rectum to understand how inflammation interferes with the healing process of intestinal. On the7th post-operative day an end to end colonic anastomosis was performed with interrupted suture. On the 3rd and 7th post-operative days, the anastomosis were evaluated. We observed that mortality and the number of complications were greater in the group of animals with inflammatory bowel disease. Dehiscence with peritonitis was the most common complication(p=0.0222). The bursting strength in the colons without leakage was lesser than those in the control group, however this difference was not statistically significant (p=0,0836). It was verified that the anastomosis performed in colons with inflammatory bowel disease showed greater total collagen concentration, with a predominance of immature collagen (type III) (p=0.0000). The mature collagen (type I) was the predominant collagen in the wounds of normal colon. (p=0.0102). On the 3th post-operative day It was also observed that the organization of collagen was poorer on the third day in the colonic wounds with inflammatory bowel disease. Nevertheless, the analysis of the inflammatory reaction at the level of the anastomosis was similar in both groups. These results suggest that inflammatory bowel disease increases the risks of anastomosis dehiscence, probably because of the delay in the maturation and the organisation of collagen.

A ação dos ácidos graxos de cadeia curta na cicatrização de anastomoses colônicas: estudo experimental em ratos

Short-chain fatty acids (SCFA) derive from the fermentation of fibbers ingested in the diet and act as a cell substrate of the colonic mucosa. The object aim of the present study was to evaluate the action of SCFA in colonic wound healing .Sixteen female rats were used, divided into 2 groups of 8 animals each. In the control group (HC) the animals underwent Hartmann procedure and post-operative infusion of isotonic saline solution per rectum for 7 days. In the experimental group (HA) saline solution was replaced by isomolar solution with SCFA. In the animals of the control group, the average bursting pressure of the rectal stump was 128.37 mm Hg, and 137.25 mm Hg in the experimental group (p=0.5693). Histologic evaluation with hematoxicilin and eosin was similar in both groups studied. The collagen densitometry analysis showed a significantly greater concentration of the mature collagen (type I) in the experimental group when compared with the control group (p=0.0094). Greater concentration of total collagen was found in the experimental group when compared to the control group (p=0.0371). It was concluded that rats who underwent intraluminal infusion of SCFA have greater concentration of mature and total collagen in the suture line of the rectal stump, when compared with to the control group.

O papel do tamanho dos poros na biocompatibilidade de duas próteses de polipropileno usadas na retopexia pré-sacra estudo experimental em cães

Rectal prolapse is a socially disabling condition afflicting both the young and old individuals. Among the most used surgical procedures are the rectopexies with polypropylene meshes. To compare the biological behavior of two polypropylene, macroporous and monofilament meshes, of different pore size and manufactured with different structural configurations, when fixed on the pre-sacral fascia. Prolene® mesh, a woven mesh with pore size of 164 x 96µ; and T mesh, a non-woven experimental test mesh, with pore size of 4 x 3 mm and pore area 762 times larger than that of Prolene®. Eighteen mongrel dogs, weighing between 8 and 15 kg, were divided in two groups: Prolene® mesh (n=9) and T mesh (n=9). A 4,5 x 2 cm patch of mesh was used to perform the pre-sacral rectopexy by placing nonabsorbable sutures. On the 30th post-operative day, the animals were re-operated and a fragment of the rectal wall including its mesh patch was removed. The macroscopic study evaluated the presence of seroma, hematoma, abscess, fistula, stricture, adhesions and the degree of incorporation of the mesh. The microscopic analysis estimated the inflammatory reaction, foreign body reaction, fibroblastic penetration and the collagen densitometry. One dog of the Prolene® mesh group developed seroma and the mesh was not incorporated. Adhesions were present in all dogs. The other macroscopic parameters were absent in both groups. The microscopic analysis showed lesser inflammatory reaction and higher migration of fibroblasts, attesting the favorable incorporation of the prosthesis. Low foreign body reaction was present, implying satisfactory biological acceptation of the prosthetic material. The densitometry of collagen revealed that the T mesh, with pore area 762 x larger than that of the Prolene® mesh, incorporated a greater amount of total collagen. Whereas the amount of type III or immature collagen was similar in both meshes, the type I or mature collagen was greater in T mesh group. There was no difference between the two groups in macroscopic and histologic studies. Both meshes presented successful incorporation and good biological acceptation. There was a significantly greater incorporation of total collagen and type I (mature) collagen in the T mesh.

A antibioticoterapia prévia sobre o infiltrado inflamatório pulmonar após isquemia e reperfusão intestinal: estudo experimental em ratos

Ischemia following intestinal reperfusion is a common place event that favors bacterial translocation in remote organs, such as the lungs. Previous antibioticotherapy may be an efficient procedure to lessen the hazards of this process. The purpose of this study was to analyse the concentration of polimorphonuclears (PMN) in the lung alveolar septum of rats, after induced ischemia followed by intestinal reperfusion, with and without previous antibiotictoherapy. Forty-eight rats were used, randomly divided into 6 groups of 8 animals each (A, B, C, D, E, and F). Groups A, C and E did not receive antibiotic and groups B, D and F received 500mg/Kg of erythromycin orally. Groups A and B were the control groups. In the groups C and D intestinal ischemia was induced for 30 min; and in groups E and F reperfusion for 30 min followed the ischemia. A thoracothomy was performed and the lungs were removed and sent for histopathologic and histometric analysis. The number of PMN in the alveolar septum was observed. The control group (A) showed a mean number of 4.2 PMN per field in the septum; in the ichemia group (C) , the mean number of PMN was 13.6 and in the group where the reperfusion followed ischemia (E), the number of PMN was 17,5. The comparison between the mean number of PMN in groups A and C and A and E showed a statistically significant difference (p<0.0001). However, the difference between the groups C and E was not significant (p=0.095). The mean number of PMN per field was of 8.6 in group B; 9.9 in group D and 12.9 in group F. The comparison between the groups B and D demonstrated a p=0.007. The comparison between the groups that received antibiotics or not demonstrated that the number of PMN in group B was twice as much as in group A (p=0.0049) . Group C showed a greater concentration of PMN than did group D (p=0.019). In group E there was a greater concentration of PMN than in group F (p=0.014). It may be concluded that: (1) intestinal ischemia leads to a greater PMN concentration in the alveolar septum; (2) previous antibioticotherapy increased lung inflammatory infiltrate in the alveolus of rats with normal intestines; (3) PMN density was greater in alveolar septum of animals who underwent I/R than those who were not reperfused, and (4) previous antibioticotherapy reduced significantly the PMN concentration after I/R.