Joaquim Coutinho Netto

Evaluation of the biocompatibility of a new biomembrane

Biocompatibility has been considered one of the most important items to validate a biomaterial for its application in human organisms. The present work evaluates the biocompatibility of a new biomembrane using in vivo assay in different animal species. The experiments to evaluate the cellular reaction were carried out through the implantation of the material into the subcutaneous tissue of animals and the results showed a good reaction of the host tissue without any signal of fibrosis or rejection. The cell adhesion experiments were done by means of the measure of the DNA content on the material surface after its implantation into the subcutaneous tissue of animals and the results showed a growing number of DNA that was proportional to the time of implantation. The healing process was evaluated using a dermal ulcer model and the results showed a good tissue repair resembling a physiologic process. The overall results presented here lead to the conclusion that this new biomembrane is a biocompatible material but more research must be done, as it is a new material desired for medical use.

Miringoplastia com a utilização de um novo material biossintético

Myringosplaty is a surgery that is performed to control infections in middle ear, to reconstruct the sound conducting mechanism for the oval window and protection of the round window. Some materials are used to reconstruct the tympanic membrane like autologous temporalis fascia, cartilage-perichondium graft taken from the tragus, cartilage only, dura mater human placental graft. It is important the vascular support to the graft with well-vascularized flap in wide perforations. AIM: The main goal of this work is shown the use of a new biomaterial, the natural latex membrane with polylysin. STUDY DESIGN: Longitudinal Cohort. MATERIAL AND METHOD: This material was developed from Department of Biochemistry from Faculdade de Medicina de Ribeirao Preto Universidade de Sao Paulo. The natural latex membrane is used stimulating neovessels and organized tecidual growing in different parts and tissues of the human body. This material promotes any allergic reaction and is innocuous to the human tissue. The 238 ears with tympanic membrane perforation resulted from chronic middle ear infection and thek was going through myringoplasty with autologous temporalis fascia and natural latex membrane. The ages were 7 to 76 years. RESULTS: The first results show 181 (90,5%) healing tympanic membrane perforation, 96 healing of wide perforation 73 medium perforation and 12 small one. Neovessels could be seen in all grafts and remaining tympanic membrane. This is not a habitual funding when the natural latex membrane is not used. CONCLUSION: According to these results we can conclude that the natural latex membrane can be used as a temporary implant in myringoplasty, improving the well vascularized of remaining tympanic membrane.

Substituição parcial do pericárdio de cães por membrana de látex natural

BACKGROUND: There are numerous advantages in closing the pericardial cavity after heart surgery and intrapericardial pneumonectomies. OBJECTIVE: To study the behavior of a natural latex membrane as a partial pericardium substitute. MATERIAL AND METHODS: Sixteen dogs divided into 3 groups were submitted to resection of an elliptical flap of the left anterolateral portion of the pericardium (7 x 5 cm): Group A (n=4) - the removed flap was immediately reimplanted; Group B (n=8) - the flap was replaced with a 0.3 mm thick natural latex membrane of equivalent area; Group C (n=4) - the latex flap was 0.7 mm thick. In all animals the flap was fixed with continuous 5-0 or 6-0 polypropylene sutures. In group C, 4 additional U-shaped stitches anchored with dacron pledgets were applied. All animals were submitted to ECG and to white cell counts during the preoperative period as well as to macro- and microscopic post mortem study. RESULTS: Group A - the self-graft was macro and microscopically intact, strongly adhering to the lung and loosely adhering to the epicardium; Group B - total suture dehiscence occurred in 1 animal and partial dehiscence in 2. The latex membrane did not adhere to the lung or to the epicardium. In 3 animals (37.5%) there was full regeneration of the pericardium underlying the natural latex, microscopically identical to the native pericardium. Group C - the sutures were intact and total pericardium regeneration was observed in 3 animals (75%). In the remaining animals of Groups B and C, the epicardium underlying the latex was slightly thickened, permitting easy visualization of the coronary vessels. Some areas with dense lymphocytes and plasma cells infiltrates, fibroblast and vascular proliferation were microscopically identified in its thickness; in some other areas the fibroblasts surrounded slits with mesothelial lining indicating irregular pericardium regeneration. No local infection or changes in white cell counts were observed and the ECG only showed T wave inversion in all groups. CONCLUSION: The natural latex membrane proved to be adequate for partial pericardium replacement in dogs kept under observation for up to 345 days, favoring regeneration of the native pericardium.

Evaluation of rhBMP-2 and Natural Latex as Potential Osteogenic Proteins in Critical Size Defects by Histomorphometric Methods

This in vivo study evaluated the osteogenic potential of two proteins, recombinant human bone morphogenetic protein‐2 (rhBMP‐2) and a protein extracted from natural latex (Hevea brasiliensis, P‐1), and compared their effects on bone defects when combined with a carrier or a collagen gelatin. Eighty‐four (84) Wistar rats were divided into two groups, with and without the use of collagen gelatin, and each of these were divided into six treatment groups of seven animals each. The treatment groups were: (1) 5 μg of pure rhBMP‐2; (2) 5 μg of rhBMP‐2/monoolein gel; (3) pure monoolein gel; (4) 5 μg of pure P‐1; (5) 5 μg of P‐1/monoolein gel; (6) critical bone defect control. The animals were anesthetized and a 6 mm diameter critical bone defect was made in the left posterior region of the parietal bone. Animals were submitted to intracardiac perfusion after 4 weeks and the calvaria tissue was removed for histomorphometric analysis. In this experimental study, it was concluded that rhBMP‐2 allowed greater new bone formation than P‐1 protein and this process was more effective when the bone defect was covered with collagen gelatin (P < 0.05). Anat Rec, 2010.

The vegetal biomembrane in the healing of chronic venous ulcers

BACKGROUND: The vegetal biomembrane has been used to treat cutaneous ulcers. OBJECTIVES: To assess the role of the vegetal biomembrane on the chronic venous ulcers treatment compared to treatment with collagenase cream. METHODS: Fourteen patients were selected to be treated with vegetal biomembrane and 7 with Fibrase®(CONTROL), followed clinically and photographically by the Wound Healing Index by ImageJ during 120 days and biopsied on the 1stand 30thdays for histological examination. RESULTS: The vegetal biomembrane was better in promoting healing of the ulcers, especially on the inflammatory phase, confirmed by abundant exudation and wound debridement than the CONTROL group, on the 30thday. There was a greater tendency to angiogenesis followed by re-epithelialization with highest wound healing index on the 90thand 120thdays. CONCLUSION: A combined analysis of clinical and histopathological findings suggests that the vegetal biomembrane acted as a factor inducing wound healing, especially on the inflammatory phase, confirmed by abundant exudation of the lesions promoting the transformation of the microenvironment of the chronic venous ulcers, and also stimulating angiogenesis and subsequent re-epithelialization.

The role of polar phytocomplexes on anticonvulsant effects of leaf extracts of Lippia alba (Mill.) N.E. Brown chemotypes

Objectives The purpose of the present work was to characterize the pharmacological profile of different L. alba chemotypes and to correlate the obtained data to the presence of chemical constituents detected by phytochemical analysis.

Prótese vascular derivada do látex

BACKGROUND: The development of vascular grafts has been crucial for advances and achievements in reconstructive vascular surgery over the past 5 decades. OBJECTIVES: To develop a new model of microperforated vascular graft using fabric covered with a natural latex-derived polymer taken from Hevea brasiliensis and assess its patency, thrombogenicity, biocompatibility and healing process, besides some mechanical properties (adaptability, elasticity, impermeability and possibility of suture), using expanded polytetrafluoroethylene graft as control. METHODS: Fifteen dogs were divided into three groups of five animals. The microperforated latex graft was implanted in all dogs and the expanded polytetrafluoroethylene graft was implanted in the contralateral pelvic limb. Postoperative follow-up was 4, 8 and 12 weeks. Analysis of results was performed according to clinical evaluation of pulses, complications (fluid collection, dehiscence, granuloma and infection), arteriography, macroscopic analysis and scanning electron micrography. RESULTS: Statistical tests revealed no significant differences (p > 0.05) concerning post-operative complications and graft patency. Both grafts were properly integrated to surrounding tissues, with connective tissue formed by collagen fibers. A neointimal layer covering all extension of the luminal surface was observed in the microperforated latex graft. Conversely, the endothelial development over the neointimal surface was limited to regions adjacent to the anastomoses in the expanded polytetrafluoroethylene graft. CONCLUSIONS: The microperforated latex graft showed satisfactory structural qualities (adaptability, elasticity, impermeability and possibility of suture) as a vascular substitute. It stimulated endothelial growth beyond contact regions with the artery in anastomoses and was biocompatible in the dogs arterial system, presenting adequate tissue integration.

Avaliação da isquemia cerebral focal induzida pela oclusão da artéria cerebral média e a ação neuroprotetora do cetoprofeno em ratos

Cerebral ischaemia is eventualy observed during neurosurgical procedures and in several clinical entities that may cause severe neurological deficits and even death. Because it is a severe and complex problem, several studies have been done aiming to elucidate the mechanisms of the ischemic phenomenon and aiming to abolish or to diminish its effects, using drugs that protect the neurons from ischaemia-induced damage. Several neurotransmitters play a role in cerebral ischaemia with emphasis to glutamate by its high concentration in the central nervous system. The purpose of this study was to evaluate the effect of focal cerebral ischaemia in the rat through the dosage of the glutamate and morphological findings, and to evaluate a possible protective effect of the ketoprofen to ischemic neurons. Thirty-six rats Wistar were divided into four groups. The first was a control group, the second a sham group and the animals of the third and fourth groups were submitted to induced cerebral ischaemia through selective obstruction of the midlle cerebral artery during 15, 30 and 45 minutes. Animals of the fourth group were previously treated with ketoprofen 15 minutes before the ischaemia. The ischaemia was evaluated through the histopathological examination and through dosage of the extracellular glutamate in vitro. The histopathological examination showed that there was no difference between the animals of the control and of the sham groups. In the animals submitted to ischemia histopathological alterations appeared at 30 minutes and become more intense at 45 minutes of ischaemia. The main findings were interstitial edema, chromatinic disorganization, vacuolization and nuclear desintegration. The animals treated with ketoprofen showed similar alterations, but they were less intense. Decrease in the dosage of glutamate in the parietal cortex of the animals submitted to ischaemia started at 30 minutes and became more intense at 45 minutes of ischaemia and was similar for animals previously treated or not with ketoprofen, indicating that this drug seems not to interfere with the metabolism of the glutamate at the synapses. The morphological findings in the parietal cortex of the animals submitted to ischaemia, previously treated or not with ketoprofen, suggest that this drug has a neuroprotective effect.

Bone Repair Investigation Using rhBMP-2 and Angiogenic Protein Extracted From Latex

Background: The aim of this work was to study the new bone tissue formation after bone morphogenetic protein type 2 (rhBMP‐2) and P‐1 application, using 5 and 10 μg of each, combined to a material carrier, in critical bone defects. Methods: It was used 70 Wistar rats (male, ∼250 g) that were divided in 10 groups with seven animals on each. Groups are the following: critical bone defect only, pure monoolein gel, 5 μg of pure P‐1, 5 μg of pure rhBMP‐2, 5 μg of P‐1/monoolein gel, 5 μg of rhBMP‐2/monoolein gel, 10 μg of pure P‐1, 10 μg of pure rhBMP‐2, 10 μg of P‐1/monoolein gel, 10 μg of rhBMP‐2/monoolein gel. Animals were sacrificed after 4 weeks of the surgical procedure and the bone samples were submitted to histological, histomorphometrical, and immunohistochemical evaluations. Results: Animals treated with pure P‐1 protein, in both situations with 5 μg and 10 μg, had no significant difference (P > 0.05) for new bone formation; other groups treated with 10 μg were statistically significant (P < 0.05) among themselves and when compared with groups in which it was inserted the monoolein gel or critical bone defect only (P < 0.05). In the group involving the 10 μg rhBMP‐2/monoolein gel association, it was observed an extensive bone formation, even when compared with the same treatment without the gel carrier. Conclusion: Using this experimental animal model, more new bone tissue was found when it was inserted the rhBMP‐2, especially when this protein was combined to the vehicle, and this process seems to be dose dependent. Microsc. Res. Tech., 2011.© 2011 Wiley Periodicals, Inc.

Curativo de biomembrana vegetal e hipersensibilidade

BACKGROUND: The natural biomembrane of latex extracted from Hevea brasiliensis has been used as a dressing for skin ulcers. OBJECTIVES: To evaluate how safe the natural biomembrane is in relation to hypersensitivity to latex when used as a dressing. METHODS: We selected patients with skin ulcers, forming the following groups: control - low occupational exposure to latex (n = 17); latex-exposed control - high occupational exposure (n = 14); ulcerated, using the natural biomembrane (n = 13); ulcerated control, not using the natural biomembrane (n = 14); and new cases (n = 9), assessed before and after 3 months of using the natural biomembrane. All patients underwent clinical and epidemiological evaluation for latex hypersensitivity and specific IgE (UniCap®), and the control and latex-exposed control groups underwent the patch test. RESULTS: Hypersensitivity was positive in 64.7% of the patients in the control group, 71.4% of the patients in the latex-exposed control group, 61.5% of the ulcerated using the natural biomembrane, 35.7% of the ulcerated control, and only 22 , 2% of the new cases. In the patch test of the control and latex-exposed control groups, only one individual in the control group (low contact) showed erythema in the first reading, which became negative in the second. The mean contact with latex in the latex-exposed control group was 3.42 hours / day. In the fluoroimmunoenzymatic assay, most of the sera was classified as zero (range 0-6). No serum was rated above 2, which is not considered significant for hypersensitivity (classification > 4). CONCLUSION: The natural biomembrane proved to be safe as a dressing, for it did not induce hypersensitivity reactions among the volunteers who underwent the patch test or among users of the natural biomembrane, as it was clinically and immunologically demonstrated by IgE levels.