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Collagen nerve conduits--assessment of biocompatibility and axonal regeneration.

Bridging of nerve gaps is still a major problem in peripheral nerve surgery. Alternatively to autologous nerve grafts tissue engineering of peripheral nerves focuses on biocompatible conduits to reconstruct nerves. Such non-neural conduits fail to support regeneration over larger gaps due to lacking viable Schwann cells that promote regeneration by producing growth factors and cell guiding molecules. This problem may be overcome by implantation of cultivated Schwann cells into suitable scaffolds. In the present experiments we tested a collagen type I/III tube as a potential nerve guiding matrix. Revascularization, tolerance and Schwann cell settlement were evaluated by light, fluorescence and scanning electron microscopy after different implantation times. The conduits were completely revascularized between day 5 and 7 post-operatively and well integrated into the host tissue. Implanted Schwann cells adhered, survived and proliferated on the inner surface of the conduits. Nevertheless, bridging a 2 cm gap of the sciatic nerve of adult Wistar rats with these collagen/Schwann cell conduits led to a disappointing regeneration compared to controls with autologous grafts. From these results, we conclude that a sufficient biocompatibility of bioartificial nerve conduits is a necessary prerequisite, however, it remains only one of several parameters important for peripheral nerve regeneration.

Animals↗

Selective laser sintering of biocompatible polymers for applications in tissue engineering.

The ability to use biological substitutes to repair or replace damaged tissues lead to the development of Tissue Engineering (TE), a field that is growing in scope and importance within biomedical engineering. Anchorage dependent cell types often rely on the use of temporary three-dimensional scaffolds to guide cell proliferation. Computer-controlled fabrication techniques such as Rapid Prototyping (RP) processes have been recognised to have an edge over conventional manual-based scaffold fabrication techniques due to their ability to create structures with complex macro- and micro-architectures. Despite the immense capabilities of RP fabrication for scaffold production, commercial available RP modelling materials are not biocompatible and are not suitable for direct use in the fabrication of scaffolds. Work is carried out with several biocompatible polymers such as Polyetheretherketone (PEEK), Poly(vinyl alcohol) (PVA), Polycaprolactone (PCL) and Poly(L-lactic acid) (PLLA) and a bioceramic namely, Hydroxyapatite (HA). The parameters of the selective laser sintering (SLS) process are optimised to cater to the processing of these materials. SLS-fabricated scaffold specimens are examined using a Scanning Electron Microscope (SEM). Results observed from the micrographs indicate the viability of them being used for building TE scaffolds and ascertain the capabilities of the SLS process for creating highly porous scaffolds for Tissue Engineering applications.

Biocompatible Materials↗

[In vitro biocompatibility of novel absorbable hydroxyapatite and AO artificial bone beta-tricalcium phosphate with rhesus bone marrow stromal cells].

OBJECTIVE: To study the in vitro biocompatibility of novel hydroxyapatite (HA) and AO artificial bone beta-tricalcium phosphate (beta-TCP) with rhesus bone marrow stromal cells (rBMSCs) . METHODS: The third passage of rBMSCs were cultured with HA and beta-TCP respectively, with the cells cultured without the materials as the control. The morphology and proliferation of cells were observed by inverted phase-contrast microscope and scanning electron microscope (SEM). MTT assay was used to semiquantitatively evaluate the cell proliferation. RESULTS: The rBMSC cocultured with HA exhibited good growth as observed under inverted phase-contrast microscope, without significant difference from the cells in the control group. Some small particles were seen pealing off from beta-TCP, and some of the cells died. Under SEM, rBMSCs showed good adhesion to HA with obvious proliferation, but the ratio of adhesive cells was not as high as that in beta-TCP group. MTT assay showed no significant difference in the cell number between HA and the control groups, but the cell number in beta-TCP group was notably less than that of control group. CONCLUSION: Novel HA has good biocompatibility with rBMSCs for bone tissue engineering, and AO artificial bone still needs improvement to serve as scaffold material for BMSCs.

Absorbable Implants↗

Biocompatibility and recellularization potential of an acellular porcine heart valve matrix.

BACKGROUND AND AIM OF THE STUDY: Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. The study aim was to investigate the biocompatibility and recellularization potential of an acellular porcine valve matrix. METHODS: Acellular porcine valve matrix contact and extract cytotoxicity was tested against porcine fibroblasts and smooth muscle cells (SMC). Porcine cells were incubated with decellularized aortic valve leaflets and aortic wall, and then assessed for changes in morphology and contact inhibition of growth. Soluble tissue extracts were prepared from decellularized leaflets and aortic wall, and assessed for their effect on the viability of cultured porcine cells. Acellular leaflets were seeded with either fibroblasts or SMC at 1 x 10(3) to 1 x 10(6) cells/cm2 for 24 h, or 5 x 10(4) cells/cm2 for 1-4 weeks. Cell attachment onto, and migration into, the acellular matrix was assessed by scanning electron microscopy and histology. RESULTS: No contact inhibition of growth, or changes in fibroblast or SMC morphology, were observed following contact with the acellular valve matrix. No soluble extract cytotoxicity was found. Intermediate cell-seeding densities (2.5 x 10(4) to 7.5 x 10(4) cells/cm2) of both cell types produced confluent cell attachment; at the lowest concentration (1 x 10(3) cells/cm2) cell attachment was sparse, and at the highest (1 x 10(6) cells/cm2) it was multilayered. The SMC migrated throughout the leaflet matrix over four weeks, but there was no fibroblast migration into the matrix. CONCLUSION: The absence of contact and extract cytotoxicity indicated that the acellular valve matrix was biocompatible in vitro. The failure of porcine fibroblasts to grow on, or infiltrate into, the matrix suggested that the SMC may be the preferred cell type for future leaflet recellularization studies in the development of a tissue-engineered heart valve replacement.

Animals↗

[Biocompatibility study on tendon mixed extraction of bovine collagen for periodontal tissue engineering].

OBJECTIVE: To study the biocompatibility of tendon mixed extraction of bovine collagen (tMEBC) and to explore the feasibility of using the three-dimensional framework as periodontal tissue engineering scaffold. METHODS: After being prepared, the tMEBC were cultured with the P4-P6 of human periodontal ligament fibroblasts (HPDLFs) in vitro. Three-dimensional framework was prepared from bovine tendon. The P4-P6 of HPDLFs (with an initial density of 5 x 10(6) cells/ml) were cultured in vitro. Cell attachment and proliferation were measured by cell counting 1 day, 3, 5, and 10 days after cell seeding. Histological examination was performed with light microscope and scanning electron microscope 5 and 10 days after cell seeding. RESULTS: Porous structure, which supported the proliferation and attachment of HPDLFs, was found in tMEBC. The density of cell increased from 0.556 x 10(4) cells/ml 24 hours after cell seeding to 3.944 X 10(4) cells/mI 10 days after seeding. Light and scanning electron microscope examination indicated that HPDLFs were attached and extended on the three-dimensional scaffolds and were well embedded in the newly formed tissue matrix. CONCLUSION: tMEBC has good biocompatibility with the HPDLFs, and can be used as scaffold for cell transplantation in periodontal tissue engineering.

Animals↗

[Biocompatibility of a special hydrogel orbital implant (PHEMA and MMA) in rabbits].

OBJECTIVE: To investigate the biocompatibility of a hydrogel orbital implant and its rate of vascularization in an experimental study. METHODS: The implant was made of copolymerization of 2-hydroxyethyl methacrylate (HEMA) and methyl methacrylate (MMA). Thirty-two hydrogel orbital implants were implanted into the right orbit of 32 New Zealand albino rabbits. The rate of vascularization was examined by SPECT, immunohistochemistry and electron microscopy. RESULTS: Single Positron Emission Computerized Tomography (SPECT) examination showed that a radiotracer could be found locally concentrated on hydrogel orbital implants in vivo along with time. On light microscopy, implants fibro vascular tissue invade the pores of the hydrogel orbital implant after 2 weeks and gradually invaded the implant deeper and deeper from 4 to 8 weeks, almost all implants were fully vascularized after 12 weeks with a relatively spare inflammatory reaction. Only one case needed additional surgery. CONCLUSIONS: The hydrogel orbital implant has many advantages, such as well biocompatible, fast vascularization, simple operation and low complication of surgery. It is safe and applicable for clinical use as a new type of orbital implant material.

Animals↗

[Biocompatibility of polyester net in the cornea of rabbits].

OBJECTIVE: To explore biocompatibility and biohealing characteristics of a polyester net with cornea after the net implanted into the cornea lamina. METHODS: Twenty New Zealand rabbits weighing 2.0 - 2.5 kg were randomly divided into experimental and control group. Surgery was performed in right eye of all these animals and the left one as control. A square of implant of 5 - 6 mm in diameter was cut from polyester net (0.2 mm in thickness and 1 mm pore diameter) and implanted into the cornea lamina of experimental group; only a sack was made without implantation in control group. Cornea edema and growth status of new blood vessels were observed at scheduled times after operation; animals were sacrificed 2 weeks, 1, 3, 5 and 8 months after operation and their cornea was removed and examined under light microscopy. RESULTS: Different degrees of cornea edema and new vessels were revealed after surgery. The edema and neovascularization were all automatically disappeared in the period of post-operation or after suture removed. The implant was stable inside the cornea and lasted as longest as 8 months. The number of fibroblasts was markedly increased at the upper and bottom surface of the cornea lamina close to the implant and in the area of net pore particularly. And finally, a close connection of collagen fibers was formed. No serious complications such as necrosis, ulcer, as well as implant dislocation from cornea were seen. CONCLUSION: The polyester net possesses a well biocompatibility with the cornea tissue.

Animals↗

[Synthesis of a bio-active bone-matrix material and study of the cellular biocompatibility].

To prepare poly(lactic acid/glycolic acid/ asparagic acid-co- polyethylene glycol) (PLGA-[ASP-PEG]) and examine the cellular biocompatibility. PLGA-[ASP-PEG] was obtained by bulk ring-opening copolymerization method, examined by infrared spectrometry (IR) and 1H nuclear magnetic resonance spectroscopy (1H NMR). Bone marrow stromal cells(BMSCs) were cultured with PLGA-[ASP-PEG] (experiment gruop) and PLGA (control group) in vitro respectively, and were observed by phase-contrast microscopy and scanning electron microscopy. The resuls showed that PLGA-[ASP-PEG] was obtained and proved by IR and 1H NMR. The BMSCs of the experiment group could well attach to and extend on the surface of the PLGA-[ASP-PEG], and could proliferate and secrete better extracellular matrix, compared with control. The PLGA-[ASP-PEG] has good cellular a biocompatibility. It can be used as a biomaterial for bone tissue engineering.

Aspartic Acid↗

Biocompatibility of a bicarbonate/lactate-buffered PD fluid tested with a double-chamber cell culture system.

OBJECTIVE: In peritoneal dialysis (PD), neutrally buffered PD fluids with lower concentrations of glucose degradation products (GDP) have tested superior to conventional fluids in terms of biocompatibility. However, conventional in vitro studies provoke debate because, due to the lack of subsequent equilibration with the blood, they do not resemble the true intraperitoneal situation of PD. METHODS: We established a double-chamber cell culture system with peritoneal mesothelial cells seeded on top of a permeable membrane, with a physiological buffer below. Thus adequately reflecting the in vivo equilibration pattern, we compared a conventional fluid with a neutral bicarbonate/lactate-buffered PD solution. Using an exchange pattern adapted from an 8-hour continuous ambulatory PD regimen, cell viability was assessed with an MTT assay, and cell function via constitutive and stimulated interleukin (IL)-6 release. As an indicator of potential induction of fibrosis and as a parameter of mesothelial cell integrity, respectively, transforming growth factor-beta 1 (TGF-beta1) generation and cancer antigen 125 (CA125) release were measured. RESULTS: The conventional solution significantly compromised mesothelial cell viability and function in terms of mitochondrial activity (p < 0.05) and stimulated IL-6 release (p < 0.05). The bicarbonate/lactate fluid had no effect on cell viability or IL-6 release and turned out to be equivalent to the properties of the growth medium. Whereas lactate-incubated cells did not respond to IL-1beta stimulation, bicarbonate/lactate-treated cells adequately increased IL-6 release after stimulation (p < 0.0005). Release of TGF-beta1 and CA125 did not differ between the different fluids and the control. CONCLUSIONS: Due to the sustained equilibration process, the double-chamber cell culture model allows a more realistic insight into mesothelial cell viability and function in terms of PD. As in classic in vitro studies, an adverse effect of conventional PD solutions on mesothelial cells was overt in the present cell culture system. The neutral bicarbonate/lactate-buffered fluid with low GDP content, however, did not interfere with mesothelial cell vitality or function, indicating superior biocompatibility.

Bicarbonates↗

[Study of biocompatibility of polybutylcyanoacrylate nanoparticles].

OBJECTIVE: To evaluate the biocompatibility of polybutylcyanoacrylate nanoparticles (PBCA-NP) manufactured by emulsion polymerization technique. METHODS: MTT assay was performed for evaluation of the cytotoxicity of PBCA-NP (e.g. cell proliferation and hemolytic test) experiments of long-term subcutaneous implantation and implantation in the muscular tissues of PBCA-NP was conducted in rabbits to observe the inflammatory reactions due to the implantation. RESULTS: No obvious cytotoxicity or hemolytic reactions were observed (with the hemolytic rate at different concentrations of PBCA-NP(5%). Three months after PBCA-NP implantation, obvious lymphocyte infiltration was not observed in the tissues around the implants which underwent gradual degradation. CONCLUSION: PBCA-NP possesses good biocompatibility.

Animals↗

Evaluation of biocompatible osteoconductive polymer as an orthopedic implant in dogs.

The biocompatibility and osteoconductive properties of biocompatible osteoconductive polymer (BOP), a synthetic implant, were evaluated. Bilateral oval cortical defects (1 x 2 cm) were made in the lateral subtrochanteric area of the proximal portion of the femur in 16 dogs that later were treated with BOP fiber (n = 16) or autogenous cancellous bone (n = 11), or were not treated (n = 5). The BOP block was attached extraperiosteally to the proximal portion of the humerus in 6 dogs. Radiographic assessment of surgery sites was performed at 4-week intervals, and histologic evaluation was performed at 4, 8, 16, and 24 weeks after surgery. Radiographic signs of bone healing were not observed in defects treated with BOP fiber. Defects treated with cancellous bone or not treated had radiographic signs of progressive bone ingrowth. Radiographic evidence of periosteal new bone formation near control and BOP-treated defects was observed 4 weeks after surgery; increased periosteal reaction was associated with BOP fiber. This new bone had resorbed by week 24, except bone adjacent to BOP fiber, where continued periosteal reaction was apparent. Histologic evidence of bone formation was observed extending to, but not incorporating, BOP fibers. The BOP fibers became surrounded by a fibrous capsule, and fibrovascular connective tissue infiltrated between and into BOP fibers, but minimal bone formation incorporated the BOP material during the follow-up period. During that time, active periosteal new bone formation was evident adjacent to the BOP fibers. Defects treated with cancellous bone or not treated healed by ingrowth of cancellous bone during the first 12 weeks after surgery and by reformation of the lateral cortical wall by week 24. The BOP blocks became surrounded by a fibrous capsule, but connective tissue or bone ingrowth into BOP blocks was not observed. Results indicate that BOP is not osteoconductive within a 6-month time frame when used in subtrochanteric femoral defects or when placed extraperiosteally on the proximal portion of the humerus of clinically normal dogs.

Animals↗

[The study on biocompatibility of diamond-like carbon coated nickel-titanium shape memory alloy with osteoblasts cultured in vitro].

OBJECTIVE: To investigate the biocompatibility of diamond-like carbon (DLC) coated Nickel-Titanium shape memory alloy with osteoblasts cultured in vitro. METHODS: Rabbit's osteoblasts were incubated with DLC-coated Nickel-Titanium shape memory alloy disks and uncoated ones of equal size for 5 days. The control group (without shape memory alloy in culture media) was performed simultaneously. The cultured cells were counted and graphed. The samples from culture media were collected and the concentrations of alkaline phosphatase (ALP) and nickel (Ni2+) were measured from the 1st to 5th day respectively. RESULTS: The proliferation of osteoblasts and the concentration of ALP in both DLC-coated group and control group was higher than uncoated group. The proliferation of osteoblasts on the 3rd, 4th, and 5th day in both DLC-coated group and control group was significantly higher than that in the uncoated group (P < 0.05). The concentration of ALP in DLC-coated group on the 2nd, 3rd, and 5th day and in the control group on the 3rd, 4th, and 5th day was significantly higher than that in the uncoated group (P < 0.05). The concentration of Ni2+ on the 3rd, 4th, and 5th day was significantly lower than that in the uncoated group (P < 0.05). CONCLUSION: DLC- coated Nickel-Titanium shape memory alloys appears to have better biocompatibility with osteoblast cultured in vitro compared to uncoated ones.

Alloys↗

[A comparative study on biocompatibility of acellular corneal stroma materials prepared by serial digestion methods].

OBJECTIVE: To observe the biocompatibility of the acellular corneal stroma materials prepared by three different methods. METHODS: Three different serial digestion methods were used to produce the acellular corneal stroma materials. The biocompatibility of the materials was investigated by the cell seeding and the materials were implanted into the rabbit corneal stroma layer. RESULTS: The cells in the materials 1 and 2 were not decellularized completely. The rabbit corneal fibroblasts died on the materials 1 and 2 after the cell seeding for 3-4 days. An obvious rejection could be observed after the implantation. The cells in material 3 were decellularized completely and the collagen fibers or elastic fibers were reserved integrally, showing a typical three-dimensional network. The rabbit corneal fibroblasts could expand on the materials in vitro. No obvious rejection could be observed and the materials were gradually absorbed. CONCLUSION: The acellular porcine cornea stroma materials prepared by trypsin-Dnase-Rnase are suitable for reconstruction of the tissue engineered cornea.

Animals↗

[Biocompatibility of acellular urinary bladder submucosa].

OBJECTIVE: To investigate the biocompatibility of acellular urinary bladder submucosa (AUBS). METHODS: The acellular collagen matrix of human urinary bladder submucosa was developed using freeze-thawed enzymatic treatment and freeze-drying technique. Human oral keratinocytes were cultured and seeded on AUBS at a density of 2 x 10(6)/ml in vitro. The proliferation of the cells were observed. Pockets were created in the abdominal muscle wall of 18 SD rats. AUBS in size 1 cm x 1 cm was implanted into the pocket. The grafts were observed by light microscope 3, 6, 10, 14, 21 and 28 days after operation. RESULTS: AUBS mainly consisted of collagen fibers with a three-dimensional network structure. After the oral keratinocytes were seeded, continuous oral epithelium layer was formed on the surface of AUBS after 10 days in vitro. Histological observation of the grafted AUBS showed progressive cell infiltration at 6 days. New capillaries formed at 14 days. The collagen fibers arranged regularly at 28 days after implantation. CONCLUSION: Freeze-dried AUBS may be used as a suitable scaffold for tissue regeneration, which can induce cell proliferation both in vivo and in vitro and has good biocompatibility.

Animals↗

[Mechanical properties and biocompatibility of nanometer calcium carbonate/poly (L-lactide) composites].

The purpose of this study is to synthesize nanometer calcium carbonate/poly(L-lactide) composites and evaluate its biocompatibility, and mechanical properties. The nanometer calcium carbonate/poly(L-lactide) composites was prepared by the method of solution mixing and cell culture. Its properties were analyzed using, mechanical tests, MTT assay, transmission electron microscope and scanning electron microscope. It was found that the new kind copolymer had good biocompatibility, nanometer calcium carbonate could enhance the strength of the pure poly (L-lactide) matrix. The nanometer calcium carbonate/poly (L-lactide) composites is a promising bioabsorbable material. It is worth further medical research.

Animals↗

[Activation of nuclear factor kappaB (NF-kappaB), induction of proinflammatory cytokines in vitro and evaluation of biocompatibility of the carbonate ceramic in vivo].

UNLABELLED: Ceramic materials based on calcium carbonate have been prepared in response for the demand for resorbable materials for use in bone surgery. Calcium carbonate in the form of crystalline aragonite or calcite with various amount of lithium fluoride was the raw material. Material CC-1FA from crystalline aragonite (99% CaCO3 and 1% LiF), CC-5FA - from crystalline aragonite (95% CaCO3 and 5% LiF) and CC-1FK -material from crystalline calcite (99% CaCO3 and 1% LiF) was studied. To evaluate their biocompatibility and inflammatory effect we investigated the activity of nuclear factor kappaB (NF-kappaB) and proinflammatory cytokine concentrations: tumor necrosis factor TNF-alpha, interleukins 11-6 and IL-8 in peripheral human leukocytes (PBL) after stimulation in vitro with tested calcite materials. Evaluation of local soft tissue reaction after implantation was also the aim of our study. Proinflammatory cytokines take part in inflammatory reaction caused by biomaterials. Expression of these proteins is controlled by proinflammatory regulatory transcription factors including the commonly appearing NF-kappaB (Nuclear Factor kappaB). In a quiescent cell NF-kappaB resides in the cytosol in an inactive form which its activated under the influence of kinases. The activated NF-kappaB protein translocates from cytosol to nucleus of cell and binding to specific DNA sequence it initiates transcriptions. Thanks to the quick regulation of immunological response it ensures the survival of cells in unfavorable reactions of environmental factors. It regulates the expression of many genes mainly connected with the course of the inflammatory process (of some cytokines, proteins of acute phase, collagenasis, stromilozine other enzymes decomposing the elements of matrix) and with proliferation and differentiating of cells. However its excessive activity can lead to unfavorable reactions, for example uncontrollable division of cells, appearance of giant cells of foreign body type. In our study protein expression of NF-kappaB in PBL were assessed using anti-c-Rel-antibody (PBL expressing c-Rel in the nucleus = labelled NF-kappaB (+) cells). The NF-kappaB activation in PBL was expressed as: the percentage of NF-kappaB (+) cells. The level of cytokines: IL-6, IL-8 and TNF-alpha level in the supernatants from leukocytes culture with tested materials was determined by an enzyme-linked immunoabsorbent assay (ELISA) after 24 and 72 hours incubation. The local tissue reaction in vivo was evaluated 1 and 3 months after implantation calcite devices into dorsal muscles of rats. The achieved results showed that the tested calcite material incubated 24 and 72 hours with PBL culture didn't synthesized higher amounts of the cytokines IL-6 and IL-8 in comparison with the untreated cells. All tested materials stimulated after 24 and 72 hours PBL culture to produce a significant level of TNF-alpha (p < 0.05) (higher level after CC-5FA and CC-1FA materials and lower after CC-1FK). Activation of transcription factor nuclear factor NF-kappaB in leukocytes after 24 hours incubation with CC-5FA, CC-1FA was higher than CC-1FK but significantly was only after CC-5FA stimulating. The expression of NF-kappaB after 72 hours of incubation decreased and was on the level comparable with control group. Histological observation 1 month after implantation showed that the carbonate ceramic devices were surrounded with connective tissue surrounding implants capsule with presence of macrophages. Additionally after 3 month in the dense connective tissue cartilaginous and calcification focus was visible. Soft tissue reaction showed chronic inflammatory reaction connected with resorbtion of biomaterials, the most increased after implantation of CC-5FA, than CC-1FA and CC-1FK. CONCLUSION: Transcription nuclear factor NF-kappaB was activated after in vitro stimulation in short time by the tested calcite material CC-5FA. This activity was correlated with induction in vitro of TNF-alpha in short and long time. The most increased soft tissue reaction 1 and 3 month after implantation of CC-5FA material was found. Our study showed that calcite materials can activate NF-kappaB and demonstrated differences in biocompatibility of tested materials.

Animals↗

[Study on all-round evaluation of biocompatibility of biomaterial].

Biocompatibility has always been the focal point in the study of biomaterials applied to medical apparatus. But at present, the standard system hasn't formed completely to evaluate the biomaterials. In recent years scientists tend to use general standard of evaluation. In this article, based on the recalcification time, the adhesion of platelets as well as the total quantity of plasma protein and some other evidences, combining with the Analytic hierarchy process (AHP), the method of general evaluation on the biocompatibility of anticoagulant biomaterials was discussed.

Anticoagulants↗

Biocompatibility and membrane development.

Development of new biocompatible membranes for clinical application needs the expertise of various specialists, chemists, chemical engineers, and clinicians. As the biocompatibility of a membrane cannot be considered in terms of a single biochemical pathway, due to the interrelation between the complement, coagulation, and inflammatory systems, it is difficult to achieve optimal results with one single polymer. A compromise between different approaches has therefore to be found. Membrane development also needs sophisticated test systems, which simulate the clinical situation as closely as possible. Adequate results are achieved with the 'ex vivo' model, which represents open-loop haemodialysis. The ex vivo model gives good results which are close to those from clinical experiments.

Biocompatible Materials↗