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A device to apply user-specified strains to biomaterials in culture
Mitchell, S. B., J. E. Sanders, et al. (2001), IEEE Trans Biomed Eng 48(2): 268-73.
Abstract: An apparatus was developed to apply user-specified displacements to biomaterial samples in culture. The device allowed cyclic waveforms of bandwidth 0 Hz to 20 Hz to be applied under physiologic thermal (37.5 degrees C) and [CO2] (5%) conditions. For a 0 Hz to 20 Hz bandwidth signal similar in shape to a ventricular pressure waveform, the mean displacement error was 0.26% of the full-scale output. The maximum overshoot was 0.700%. Environmental system evaluation tests demonstrated a specimen cartridge temperature of 37.20 +/- 0.15 degrees C during cyclic loading and 37.23 +/- 0.21 degrees C during static conditions. [CO2] was 5.29 +/- 0.54% during cyclic loading and 5.25 +/- 0.61% during static conditions. Laminar flow applied at the loading rod entrances to the specimen cartridge ensured the sample remained sterile during testing. As a preliminary evaluation, polyurethane samples were seeded with fetal foreskin fibroblasts and subject to intermittent cyclic displacements. Results demonstrated enhanced cell proliferation and increased [PGE2] for samples subjected to 10% strain compared with unstrained controls. A next step will be to evaluate cell response sensitivity to strain magnitude, duration, direction, and frequency. The long-term intent is to establish mechanical loading configurations that induce acceptable or adaptation-inducing responses for use in implant design and tissue engineering applications.

A dual constant-composition titration system as an in vitro resorption model for comparing dissolution rates of calcium phosphate biomaterials
Chow, L. C., M. Markovic, et al. (2003), J Biomed Mater Res B Appl Biomater 65(2): 245-51.
Abstract: It has been postulated that the in vivo resorption rates of calcium-phosphate bone-graft materials are closely related to their dissolution rates in demineralizing solutions having ionic compositions mimicking the acidic environment produced by osteoclasts. Thus, it should be possible to use an in vitro model to produce dissolution-rate data of calcium-phosphate materials as a starting point for predicting in vivo resorption properties. Direct pH measurements of the extracellular fluid from bone-resorbing cells showed that the pH was as low as 3. In the present study, a dual constant-composition dissolution system was used as an in vitro resorption model to compare dissolution rates of different calcium-phosphate materials. NIST standard reference hydroxyapatite (HA), dicalcium-phosphate dihydrate (DCPD), and calcium-phosphate cement (CPC) discs of known dimensions (6-mm d x 3-mm h) were allowed to dissolve at 37 degrees C in a solution that had an inorganic composition similar to that of serum ([Ca] = 1.15 mmol/l; [P] = 1.2 mmol/l; [KCl] = 133 mmol/l) and a pH of 3.0. A Ca ion-specific electrode and a pH electrode were used to control the addition of titrants to compensate for the increases in calcium and phosphate concentrations, respectively, in the demineralizing solution. The rate and stoichiometry (Ca/P molar ratio) of dissolution were obtained from the titration data. Each solid dissolved at an approximately constant rate during the dissolution process. The dissolution rates, expressed in mg cm(-2) min(-1), (mean +/- standard deviation, n = 5) were for HA: 6.58 +/- 1.22; DCPD: 21.0 +/- 2.6; and CPC: 8.21 +/- 0.73. DCPD dissolved three times faster than HA (p < 0.05). CPC dissolved 1.2 times faster than HA but the difference was not statistically significant (p > 0.05). This model can be used to study the rate and stoichiometry of dissolution of calcium-phosphate bone-graft materials and coatings under a wide range of mineral saturation conditions.

A follow-up study of patients with subjective symptoms related to dental materials
Lygre, G. B., N. R. Gjerdet, et al. (2005), Community Dent Oral Epidemiol 33(3): 227-34.
Abstract: OBJECTIVES: The extent to which substances released from dental materials cause adverse health effects and whether removal of dental materials results in improvement of health is a matter of dispute. The aim of the study was to investigate changes in the intensity of subjective symptoms after replacement of dental materials in patients referred for adverse reactions related to dental materials, and to compare the profiles of symptoms with those found in the general population. METHODS: Information was obtained from 142 patients referred to the Dental Biomaterials Adverse Reaction Unit in Bergen, Norway. At the time of examination, all patients completed a questionnaire regarding a range of subjective symptoms. A follow-up questionnaire was sent to all patients 1 to 2 years later. Similar questionnaires were sent to a reference group of 800 persons drawn from the general population. RESULTS: The follow-up questionnaires were completed and returned by 84 patients, and 441 persons in the reference group. The patient group presented higher symptom indices than the reference group (P<0.001). Generally, there was some decrease in the intensity of different symptoms in patients who had replaced dental materials. The decrease was evident regarding intraoral symptoms (P=0.022) and total symptom index (P=0.041). The group of patients who had replaced materials still had significantly higher symptom indices than those of the reference group. Patients who had not replaced dental materials did not present any reduction in symptom indices. CONCLUSION: The pattern of symptoms was similar for the groups investigated. At the group level, the intensity of local and some general subjective symptoms was reduced after replacement of the materials, but not to the level found in the general population.

A general classification scheme for "hydrophilic" and "hydrophobic" biomaterial surfaces
Hoffman, A. S. (1986), J Biomed Mater Res 20(9): ix-xi.

A HER-2/neu peptide admixed with PLA microspheres induces a Th1-biased immune response in mice
Nikou, K. N., N. Stivaktakis, et al. (2005), Biochim Biophys Acta 1725(2): 182-9.
Abstract: The elimination of cancer cells requires strong cellular immune responses, and these responses are induced by the activation of Th1 lymphocytes. In this work, the possibility of inducing a Th1 type of immune response in vivo by mixing a HER-2/neu synthetic CTL (cytotoxic T lymphocyte) peptide [HER-2/neu (789-797)], with poly-lactide (PLA) microspheres was investigated. Various formulations of the peptide were administered to HLA-A2.1 transgenic (HHD) mice. Cellular experiments, assessing proliferation and cytokine determination in splenocyte culture supernatants, were carried out in order to evaluate the type of immune response to the antigen. The in vivo administration of the peptide antigen admixed with the PLA microspheres induced a potent immune response which was comparable to that induced by the combination of the antigen in complete Freund's adjuvant (CFA). Furthermore, the cytokine profile produced by the T lymphocytes of the immunized animals indicated that the combination of the peptide antigen with the PLA microspheres induced a strong Th1 biased immune response to the antigen. The time of peptide incubation with the microspheres prior to administration did not affect the immune response, which further simplifies the preparation of this type of vaccine. The results justify further investigation of the possibility of inducing effective cellular immune responses against cancer cells overexpressing HER-2/neu molecules by simply mixing appropriate HER-2/neu peptide antigens with PLA microspheres.

A hip wear simulator for the evaluation of biomaterials in hip arthroplasty components
Mejia, L. C. and T. J. Brierley (1994), Biomed Mater Eng 4(4): 259-71.
Abstract: A multistation (8-station) hip simulator has been designed and tested that provides a practical imitation of the motions and loads seen by the hip joint during a typical walking cycle. A biaxial rocking motion of +/- 23 degrees is synchronized with the respective resultant forces of the extension-flexion (heel-strike to toe-off) movements of the leg. This particular simulator provides a practical engineering in vitro implementation of the walking cycle. It also provides a realistic and practical compromise between general wear screening devices (such as pin-on-disk systems) and the intensive research accomplished through full scale simulation (full 6 degree-of-freedom systems) and modeling. Evaluation of system performance shows that control of rpm (revolutions-per-minute) for the desired axial rotation of 1 Hz was kept to 60 cpm +/- 1 cpm for axial loads (per actuator) as high as 4500 Newtons. Although loading error was 2% in the peak load areas of interest (3000 Newton), station-to-station load control variability was less than.6%. Baseline wear studies with this simulator using ultra-high-molecular-weight polyethylene and Cobalt-Chromium (UHMWPE/CoCr) hip systems indicate an average specimen-to-specimen wear variability of less than 7% range after 5 million test cycles. Testing was performed in a calf serum environment at an equilibrium temperature of 33 degrees C.

A histological evaluation for guided bone regeneration induced by a collagenous membrane
Taguchi, Y., N. Amizuka, et al. (2005), Biomaterials 26(31): 6158-66.
Abstract: This study was designed to evaluate the histological changes during ossification and cellular events including osteogenic differentiation responding to collagenous bioresorbable membranes utilized for GBR. Standardized artificial bony defects were prepared at rat maxillae, and covered with a collagenous bioresorbable membrane. These animals were sacrificed at 1, 2, 3 and 4 weeks after the GBR-operation. The paraffin sections were subject to tartrate resistant acid phosphatase (TRAP) enzyme histochemistry and immunohistochemistry for alkaline phosphatase (ALP), osteopontin (OP) and osteocalcin (OC). In the first week of the experimental group, woven bone with ALP-positive osteoblasts occupied the lower half of the cavity. The collagenous membrane included numerous ALP-negative cells and OP-immunoreactive extracellular matrices. At 2 weeks, the ALP-, OP- and OC-immunoreactivity came to be recognizable in the region of collagenous membrane. Since ALP-negative soft tissue separated the collagenous membrane and the new bone originating from the cavity bottom, the collagenous membrane appeared to induce osteogenesis in situ. At 3 weeks, numerous collagen fibers of the membrane were embedded in the adjacent bone matrix. At 4 weeks, the membrane-associated and the cavity-derived bones had completely integrated, showing the same height of the periosteal ridge as the surrounding alveolar bones. The collagen fibers of a GBR-membrane appear to participate in osteogenic differentiation.

A histomorphometric evaluation of factors influencing the healing of bony defects surrounding implants
Pretorius, J. A., B. Melsen, et al. (2005), Int J Oral Maxillofac Implants 20(3): 387-98.
Abstract: PURPOSE: The authors' aim was to perform a histomorphometric study of the healing of bone defects created adjacent to titanium and hydroxyapatite (HA) -coated implants and covered with either a resorbable or a nonresorbable membrane in combination with different filler materials and to evaluate to what degree coating, membrane, and/or filler influenced the healing of the defects. MATERIALS AND METHODS: Posterior teeth were extracted from the mandibles of 10 baboons, and 12 implants were placed in each animal in the edentulous areas. The implants were either titanium or HA-coated, the membranes were either Vicryl, Gore-Tex, or Resolut, and the filler was either demineralized freeze-dried bone (DFDB), autogenous bone, or Biocoral. The implants were observed for either 3, 6, 9, 12, or 18 months. The volume of newly generated tissue and the relative contribution of bone, marrow, and filler were evaluated, as was relative extension of resorption, formation, and quiescent surface. RESULTS: The results indicated that autogenous bone is still the gold standard, but both the DFDB and Biocoral compared favorably to it. Both filler materials were being gradually replaced by bone; this process was not yet finished at 18 months postsurgery. DISCUSSION: Since even the sterilization of DFDB cannot exclude the possibility of a disease transmission, it is important to find an appropriate substitute. Both filler and membranes contributed to the re-establishment of the original volume; better results were achieved with the Vicryl and Gore-Tex membranes than with the Resolut. Biocoral can be considered an effective material. CONCLUSION: A bony defect is not necessarily a contraindication for the placement of an implant. (More than 50 references.)

A hybrid coating of biomimetic apatite and osteocalcin
Krout, A., H. B. Wen, et al. (2005), J Biomed Mater Res A 73(4): 377-87.
Abstract: A novel hybrid coating of biomimetic apatite(BAp) and osteocalcin (OC) was prepared by incubating BAp-coated Ti6A14V coupons in an osteocalcin-containing medium. A significant amount (up to 1.0 wt %) of OC was adsorbed by the BAp coating within 3 h of incubation as demonstrated by high-performance liquid chromatography.Characterizations of the hybrid coating with environmental scanning microscopy and X-ray diffraction indicated that protein adsorption does not alter the microstructure of the coating. The presence of OC in the hybrid coating was visualized with fluorescence microscopy using an immuno-labeling procedure. The affinity of OC to the BAp coating was examined using a 20-h elution test in phosphate-buffered saline and only a minimal amount (<10%) of the loaded OC was eluted out. When the coating was fully dissolved in hydrochloric acid solution after elution, about 78% of the loaded OC could be recovered. Enzyme-linked immunosorbent assay and peptide sodium dodecylsulfate-polyacrylamide gel electrophoresis confirmed the integrity and activity of OC molecules throughout the tests. The preliminary cell culture tests showed a significant effect of OC on the attachment and proliferation of osteoblasts. The quick loading profile and high affinity of OC to the BAp coating make it an ideal candidate for the hybrid coating preparation in clinical environment.

A hydrogel prepared by in situ cross-linking of a thiol-containing poly(ethylene glycol)-based copolymer: a new biomaterial for protein drug delivery
Qiu, B., S. Stefanos, et al. (2003), Biomaterials 24(1): 11-8.
Abstract: A new poly(ethylene glycol)-based copolymer containing multiple thiol (-SH) groups was cross-linked in situ to form a polymer hydrogel under mild conditions. No organic solvent, elevated temperature, or harsh pH is required in the formulation or patient administration processes, making it particularly useful for delivery of fragile therapeutics, such as proteins. The in vitro release of fluorescein-labeled bovine serum albumin and the in vivo release of the model proteins, erythropoietin, RANTES and three PEG-conjugated RANTES derivatives showed sustained release for 2-4 weeks and demonstrated prolonged biological activity of the released proteins in animals.

A hydrophilic plasma polymerized film composite with potential application as an interface for biomaterials
Marchant, R. E., S. D. Johnson, et al. (1990), J Biomed Mater Res 24(11): 1521-37.
Abstract: A hydrophilic polymer composite film (approx. 420 nm thick), with potential application as an interface for biomaterials has been prepared on nonorganic substrates, which include glass, silicon, and aluminum foil, using a glow discharge plasma polymerization technique. A thin film (110 nm thick) polymerized from hexane provided an adherent protective coating for the substrate material, and covalent bonding sites for the outer layer polymerized from N-vinyl-2-pyrrolidone. This outer layer provided the hydrophilic surface or interface. The two layers were copolymerized for a short period during transition between monomers to provide an intimate covalently bonded diffuse interphase. Preliminary in vitro and in vivo biocompatibility studies indicate that the hydrophilic film is non-cytotoxic, and does not increase the inflammatory response when compared with negative controls.

A hydroxyapatite-coated Insall-Burstein II total knee replacement: 11-year results
Oliver, M. C., O. D. Keast-Butler, et al. (2005), J Bone Joint Surg Br 87(4): 478-82.
Abstract: We report the clinical and radiographic outcome of a consecutive series of 138 hydroxyapatite-coated total knee replacements with a mean follow-up of 11 years (10 to 13). The patients were entered into a prospective study and all living patients (76 knees) were evaluated. The Hospital for Special Surgery knee score was obtained for comparison with the pre-operative situation. No patient was lost to follow-up. Radiographic assessment revealed no loosening. Seven prostheses have been revised, giving a cumulative survival rate of 93% at 13 years. We believe this to be the longest follow-up report available for an hydroxyapatite-coated knee replacement and the first for this design of Insall-Burstein II knee.

A hydroxyapatite-coated total knee replacement: prospective analysis of 1000 patients
Cross, M. J. and E. N. Parish (2005), J Bone Joint Surg Br 87(8): 1073-6.
Abstract: We prospectively reviewed 1000 consecutive patients who underwent a cementless, hydroxyapatite-coated, stemless, total knee replacement over a period of nine years. Regular post-operative clinical follow-up was performed using the Knee Society score. The mean pre-operative score was 96, improving to 182 and 180 at five and ten years, respectively. To date, there have been seven (0.5%) cases which required revision, primarily for septic loosening (four cases), with low rates of other post-operative complications. The cumulative survival at ten years with revision as the end-point, was 99.14% (95% confidence interval 92.5 to 99.8). These results support the use of hydroxyapatite in a cementless total knee replacement since it can give reliable fixation with an excellent clinical and functional outcome.

A kinetic approach to osteoblast adhesion on biomaterial surface
Bigerelle, M. and K. Anselme (2005), J Biomed Mater Res A 75(3): 530-40.
Abstract: An incompletely understood question in the field of biomaterials is how eucaryotic cells adhere on material surfaces. The adhesion of cells on materials is generally studied after some hours. Because this evaluation after some hours cannot let us presume about the future of the cells on the material, we have developed a culture model that does allow study in the long term of an elaborate cell/material interface closer to the in vivo situation. For that, we used a progressive trypsin-based detachment method. Here we report on the mathematical modeling of long-term human primary osteoblastic cell adhesion on metallic substrates, which allows us to quantify the real adhesion simultaneously by taking into account the effect of cell proliferation. A time-dependent adhesion index t(d) is proposed, which varies with culture time t according to the power law: t(d)(t) = at(b), a being independent of b. The exponent b is equal to 0.5 +/- 0.03 and is independent of the substrate's characteristics, meaning that the long-term adhesion increases proportionally to the square root of culture time. On the contrary, the parameter a significantly depends on the material's nature, the surface's topography, and the surface chemistry of the substrate and is sufficient to characterize cell adhesion. From this relationship, we suggest that a diffusion-based process related to the kinetic of formation of extracellular matrix should be involved in long-term adhesion on materials. (c) 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005.

A laboratory investigation of the effectiveness of various skin and surface decontaminants for aliphatic polyisocyanates
Bello, D., S. R. Woskie, et al. (2005), J Environ Monit 7(7): 716-21.
Abstract: Isocyanates may cause contact dermatitis and respiratory sensitization leading to asthma. Dermal exposure to aliphatic isocyanates in auto body shops is very common. However, little is known about the effectiveness of available commercial products used for decontaminating aliphatic polyisocyanates. This experimental study evaluated the decontamination effectiveness of aliphatic polyisocyanates for several skin and surface decontaminants available for use in the auto body industry. The efficiency of two major decontamination mechanisms, namely (i) consumption of free isocyanate groups via chemical reactions with active hydrogen components of the decontaminant and (ii) physical removal processes such as dissolution were studied separately for each decontaminant. Considerable differences were observed among surface decontaminants in their rate of isocyanate consumption, of which those containing free amine groups performed the best. Overall, Pine-Sol(R) MEA containing monoethanolamine was the most efficient surface decontaminant, operating primarily via chemical reaction with the isocyanate group. Polypropylene glycol (PPG) had the highest physical removal efficiency and the lowest reaction rate with isocyanates. All tested skin decontaminants performed similarly, accomplishing decontamination primarily via physical processes and removing 70-80% of isocyanates in one wiping. Limitations of these skin decontaminants are discussed and alternatives presented. In vitro testing using animal skins and in vivo testing with field workers are being conducted to further assess the efficiency and identify related determinants.

A laboratory model biofilm fermenter: design and initial trial on a single species biofilm
Wirthlin, M. R., P. K. Chen, et al. (2005), J Periodontol 76(9): 1443-9.
Abstract: BACKGROUND: The minimum inhibitory concentration (MIC) does not provide information on the efficacy of antimicrobial agents against infections involving biofilms, which are many times more resistant than planktonic forms of bacteria. This report is on the design and initial trial of a device for growing standard biofilms and testing antimicrobial agents. METHODS: We constructed a durable, autoclaveable laboratory model biofilm fermenter (LMBF) that holds hydroxyapatite discs 300 microm below a surface onto which an artificial saliva medium drips at a rate comparable to human salivary flow. Inoculated with Streptococcus sanguinis, the device formed biofilms that were swept with a Teflon wiper under aerobic conditions. Five-day-old biofilm-coated discs were aseptically removed and placed in 3 ml of sterile saline, 0.12% chlorhexidine gluconate, or 0.1% phosphate-buffered chlorine dioxide mouthwash for 1 minute. The discs and test agent were immediately diluted with saline to 10 ml, vortexed for 30 seconds, serially diluted, plated on blood agar, and incubated anaerobically 2 days. Bacterial counts were done, and the MIC of each mouthwash was determined. RESULTS: In tests with sterile water and sterile medium, the device maintained a closed system. After inoculation with S. sanguinis, a steady state was reached at day 5. Chlorhexidine at stock concentration achieved about a 2 log10 reduction (P = 0.002), but never achieved complete killing. Chlorine dioxide had no significant effect. The MIC against planktonic S. sanguinis was 112.8 microg/ml for chlorhexidine and 9.0 microg/ml for chlorine dioxide. CONCLUSIONS: The LMBF generates and maintains a single-species oral model biofilm to a steady state and enables in vitro tests of disinfectant mouthwashes in simulated clinical use. It should be usable for more advanced tests of multiple species biofilms.

A long-term in vitro biocompatibility study of a biodegradable polyurethane and its degradation products
van Minnen, B., B. Stegenga, et al. (2006), J Biomed Mater Res A 76(2): 377-85.
Abstract: The biological safety of degradation products from degradable biomaterials is very important. In this study a new method is proposed to test the cytotoxicity of these degradation products with the aim to save time, laboratory animals, and research funds. A biodegradable polyurethane (PU) foam was subjected to this test method. The PU had soft segments of DL-lactide/epsilon-caprolactone and hard segments synthesized from butanediol and 1,4-butanediiosocyanate. Copolymer foams without urethane segments, consisting of DL-lactide/epsilon-caprolactone, were tested as well. Accumulated degradation products were collected by degrading the foams in distilled water at 60 degrees C up to 52 weeks. Cell-culture medium was prepared from powder medium with this water. In different tests the cytotoxicity of this medium was established. The first signs of cytotoxicity were observed after 3-5 weeks of degradation. This accounts for both materials and reestablishes the good short-term biocompatibility of these materials. The PU showed more toxicity toward the end stages of degradation in comparison with the copolymer. This is probably related to the accumulation of degradation products of the urethane segments. In the in vivo situation the degradation of the PU and the metabolism and excretion of degradation products may differ. Therefore, long-term in vivo studies will have to establish whether these in vitro results are representative for the in vivo behavior of the degrading PU. (c) 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2006.

A low-flow adaptation phase improves shear-stress resistance of artificially seeded endothelial cells
Gulbins, H., A. Pritisanac, et al. (2005), Thorac Cardiovasc Surg 53(2): 96-102.
Abstract: INTRODUCTION: The purpose of this study was to evaluate the effect of different adaptation phases on the shear-stress resistance of endothelial cells seeded artificially onto vascular prostheses and biological heart valves. MATERIAL AND METHODS: Human endothelial cells (EC), fibroblasts (FB), and smooth muscle cells (SMC) were isolated from vena saphena magna pieces and expanded in culture. Group A: 15 polyurethane vascular grafts (20 mm diameter) were seeded with FB and SMC (53 +/- 1.2 million cells), followed by EC seeding (39 +/- 0.9 million cells). Group B: eight stentless porcine valves (Freestyle, Medtronic, USA) were seeded with FB (68 +/- 1.5 million cells) and EC (42 +/- 1.1 million cells). Shear-stress testing was done under pulsatile flow (pulse rate: 80 pulses/min.). Adaptation phase: flow was set to 0.9 +/- 0.3 l/min (systolic pressure: 40 - 50 mm Hg). High flow was 3.2 +/- 0.6 l/min. (systolic pressure: 140 - 160 mm Hg) and lasted over four hours in all groups. The vascular grafts were divided into three groups (n = 5 each): group 1 (high flow immediately), group 2 (adaptation phase of 15 minutes), and group 3 (adaptation phase of 30 minutes). The valves either were given high flow immediately (n = 4) or had an adaptation phase of 30 minutes (n = 4). Specimens were obtained after cell seeding, before, and after perfusion. RESULTS: A confluent EC layer was achieved on all grafts. After perfusion without adaptation, large defects within the cell layer were found. No FB and SMC were seen at the bottom of these defects. In group B, the defects were largest on the ventricular surface of the leaflets. After an adaptation phase of 15 minutes in group A, only a few defects within the EC layer were detected with a still confluent FB and SMC. After a 30-minute adaptation phase defects within the EC layer were very rare and no interruption of the underlying FB and SMC layer was seen. Immunohistochemical staining for factor VIII and CD31 proved the EC to be viable and staining for collagen IV and laminin revealed the formation of a basement membrane. After perfusion, the specimen also stained positive for eNOS. CONCLUSION: An adaptation phase of 30 minutes proved to be sufficient to allow artificially seeded endothelial cells to adapt to shear stress. The formation of a basement membrane was of great importance for the maintenance of a confluent EC layer.

A method for evaluation of initial tissue response to biomaterials
Wennberg, A., G. Hasselgren, et al. (1978), Acta Odontol Scand 36(2): 67-73.
Abstract: In the present paper an implantation technique is described whereby the effect of the surgical operation is eliminated and initial tissue reactions to materials may be studied. A teflon body was implanted intramuscularly in rabbits. After six weeks the overlaying tissue was excised and the implant removed. An intact, nonepithelialized tissue surface was exposed, which due to the shape of the implant showed three indentations. Materials were placed in the indentations for 15 minutes and the tissue reaction was registered by enzyme histochemical methods. Silicate cement, zinc phosphate cement and a 4% phenol solution caused an inhibition in the dehydrogenase enzyme activity in the tissue subjacent to the indentations. The severity of the tissue reaction, indicated by the width of the inhibition zone, varied among the test materials. Silicate cement caused the widest inhibition zone and the phenol solution the narrowest one. These results correlate well with previous tissue compatibility studies and indicate that the method is applicable for in vivo screening of initial tissue response to biomaterials.

A method for histomorphometric characterization of a peri-implantar new formed tissue to biomaterials
Cannas, M., M. Bosetti, et al. (1995), Ital J Anat Embryol 100 Suppl 1: 605-12.
Abstract: The spatial distribution and concentration of distinct cellular elements are of interest in the evaluation of the biological reaction to implant materials, the so called biomaterials. We present a method, based on a computer assisted cell-counting program, developed to quantify the soft tissue reaction to implanted material. Five pseudomembranes formed around silicone skin expanders were studied; they were subjected to histological immunohistochemical and histomorphometrical investigation. This quantitative approach overcomes weakness due to the subjectivity of the observers and offers new possibilities when assaying the local tissue compatibility; moreover, it shows a good applicability to a wide variety of implant materials.

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