Evaluation of the tissue reaction to a percutaneous access device using titanium fibre mesh anchorage in goats

The tissue reaction to a percutaneous access device, applicable as a carrier for an implantable glucose sensor, was evaluated in goats. Titanium fibre mesh structures were used for anchorage of the device in superficial as well as deeper soft-tissue locations. The percutaneous part was subcutaneously anchored with a fibre mesh sheet. The distal part was placed intraperitoneally and anchored in deeper soft-tissue layers using a fibre mesh cuff. All implants showed good healing with the surrounding tissue. Histological evaluation showed that the subcutaneous fibre mesh sheets and peritoneal fibre mesh cuffs were filled with immature connective tissue, generally free of inflammation. Problems concerning disconnection of the silicone catheter from the titanium holding element and filling of part of the peritoneal fibre mesh cuff with silicone glue have to be overcome by more appropriate preclinical testing and improved implant design. Our results demonstrate that titanium fibre mesh structures can be used effectively for soft-tissue anchorage of percutaneous access devices. A sufficient ingrowth of connective tissue was obtained in superficial as well as in deeper soft-tissue layers. The access device could have application as a carrier for an implantable glucose sensor for glucose monitoring in different tissue compartments.     

Effect of thin carbonate-containing apatite (CA) coating of titanium fiber mesh on trabecular bone response

The influence of thin carbonate-containing apatite (CA) coating on trabecular bone response to cylindrical titanium fiber mesh (porosity of 85%, pore size of 200–300 μm, 2.8 mm diameter × 6 mm length) implants was investigated. Thin CA coatings were deposited by the so-called molecular precursor method. Molecular precursor solution was obtained by adding dibutylammonium diphosphate salt to Ca–EDTA/amine ethanol solution by adjusting Ca/P = 1.67. Sintered cylindrical titanium fiber mesh was immersed into molecular precursor solution and then tempered at 600 °C for 2 h. The immersion and tempering process was repeated three times. An adherent thin CA film could be deposited on the inside of titanium fiber mesh. After the immersion of a CA-coated titanium fiber mesh in simulated body fluid, apatite crystals precipitated on the titanium fiber mesh. Uncoated and CA-coated titanium fiber mesh was inserted into the trabecular bone of the left and right femoral condyles of rabbits. Histological and histomorphometrical evaluation revealed a significantly greater amount of bone formation inside the porous area of the CA-coated titanium fiber mesh after 12 weeks of implantation. The present results suggested that a thin CA-coated titanium mesh has better osteoconductivity and will be useful for a three-dimensional scaffold.     

A percutaneous device to study glucose kinetics in subcutaneous tissue fluid

In the current study subcutaneous glucose kinetics were investigated in tissue fluid collected with a percutaneous device (PD). PDs containing a subcutaneous tissue chamber were implanted subcutaneously in New Zealand white rabbits. Sintered titanium fiber mesh sheets were used for subcutaneous anchorage of the PD. The bottom of the subcutaneous tissue chamber was either covered with a titanium fiber mesh sheet, a cellulose acetate membrane, or left uncovered. Subcutaneous glucose kinetics were determined after injection of octreotide and glucagon. The tissue reaction to the implants was evaluated histologically. No dynamic relationship was observed between glycaemia and subcutaneous tissue fluid glucose for all membrane covered devices. Histological evaluation showed that the presence of a seroma cavity in combination with obstruction of the membrane prevented adjustment of the subcutaneous glucose concentration in response to changes in glycaemia. In the uncovered devices, on the other hand, changes in glycaemia were reflected in subcutaneous tissue fluid. Our results prove that it is possible to measure changes in the glucose concentration in subcutaneous tissue fluid collected with a percutaneous device. Therefore, we conclude that a percutaneous device has an application as model to study the in vivo performance of implantable glucose sensors. The use of porous membranes in such devices has to be avoided.     

Surface properties of hydroxyapatite ceramic as new percutaneous material in skin tissue

Hydroxyapatite ceramic (HAC) was studied for utilization as a percutaneous device to prevent exit-site and tunnel infection along peritoneal dialysis catheters. As a result, it was found that HAC had good compatibility with skin tissue compared with silicone rubber and glassy carbon. In the present study, the surface properties of HAC after long-term implantation in skin of dogs was evaluated by scanning electron microscopic observation, solubility and bending tests. At 12 weeks after implantation, the surface of HAC was eroded severely by body fluid or cells, and the grain boundaries were clearly relieved. Ingrowth and adhesion of collagen into the etched ditch of HAC were observed. At 12 months, the eroded surface had been tightly covered and adhered to by a collagenous layer in spite of ripping off the surrounding fibrous capsule. In solubility tests, HAC dissolved at the rate of 0.23 mg cm^–2 HAC surface area per year. On the other hand, the bending strength decreased by about 10% at 4 weeks and scarcely decreased after that time. From these results, it was confirmed that the solubility of HAC produced tight contact and strong adhesion with skin tissue to prevent bacterial infection, and HAC was practical for a permanent percutaneous device.     

A quantitative comparison of the cell response to commercially pure titanium and Ti-6Al-4V implants in the abdominal wall of rats

Commercially pure (c.p.) titanium and Ti-6Al-4V implants were inserted in the abdominal wall of rats. The surrounding fluid space, inflammatory cells and fibrous capsule were evaluated after 1, 6 and 12 weeks. Light-microscopic morphometry demonstrated a fluid space around both implant materials which gradually decreased with time. Macrophages were preferentially distributed close to the implant surface in the innermost zone (0–25 µm from the surface). In contrast, fibroblasts and endothelial cells were located mainly in the outer three zones (25–100 µm from the surface). At all time periods studied and around both materials, lymphocytes were detected throughout the surrounding tissue. The outer border of the fibrous capsule, which consisted of macrophages, fibroblasts, endothelial cells and collagen, was difficult to define, in particular during the early phase of healing. At later time stages, 6 and 12 weeks, no difference in width (60–90 µm) was observed between the two materials. No major quantitative differences with respect to the number of different cells, fluid space width and fibrous capsule thickness were noted between the two materials studied. The observed mild inflammatory reaction and the absence of statistically significant differences between c.p. titanium and Ti-6Al-4V in soft tissue indicate that both materials could be suitable for use in soft tissues. In the context of previous comparative studies it may be concluded that the animal species as well as the different implantation locals play an important role in the determination of biocompatibility.     

A percutaneous device as model to study the in vivo performance of implantable amperometric glucose sensors

Glucose kinetics were investigated in subcutaneous tissue of rabbits, in which a percutaneous device was implanted. The device was used for collection of tissue fluid and as carrier of an amperometric glucose sensor. Changes in glycaemia were reflected in subcutaneous tissue fluid. However, a limited number of responses of the implanted sensors were observed. Histologic evaluation showed thin fibrous capsules surrounding the implants. Accumulations of inflammatory cells were observed inside the subcutaneous chamber. The experiments again showed that changes in blood glucose concentration can be measured in subcutaneous tissue fluid collected with a percutaneous device. Nevertheless, implanted glucose sensors could not reliably monitor these changes. Supported by our histological observations and sufficient in vitro performance, we suppose that the cellular reaction to the sensor plays an important role in this poor in vivo performance. In combination with adsorption of tissue fluid proteins, this results in a reversible deactivation of implanted sensors. The exact mechanisms involved in this process are currently unknown and need further investigation.     

On the formation of fibrous capsule and fluid space around machined and porous blood plasma clot coated titanium

Machined and machined submicron porous titanium, with and without a thin blood plasma coating (100 nm), were implanted for 7 or 28 days in subcutaneous pockets on the back of the rat. After explantation the specimens were analyzed by light microscopy with respect to thickness of the fibrous capsule, the fluid space width between implants and fibrous capsule, and formation of blood vessels. The results at 7 days indicate a thinnest fluid space for the plasma clot coated porous titanium surface, and the spaces vanished at the light microscopic level after 28 days outside all the analyzed surfaces. The thickness of the fibrous capsule increased outside the different surfaces at 7-28 days, and in this respect no significant differences were observed between the different surfaces at any time. Analysis of neovascularization showed that the number of vessels and proportion of vessels in the fibrous capsule increased with time at all surfaces, except machined Ti where the number instead decreased from 7 to 28 days. The average distance between the blood vessels and the fluid space increased with time for all types of surfaces. The results in the present study indicate that the healing process around titanium can be modulated by porosity and thin pre-prepared plasma coatings.     

Tissue reactions to bioabsorbable ciprofloxacin-releasing polylactide-polyglycolide 80/20 screws in rabbits’ cranial bone

The aim of this study was to assess tissue reactions to bioabsorbable self-reinforced ciprofloxacin-releasing polylactide/polyglycolide (SR-PLGA) 80/20 screws in rabbits' cranial bone. Two screws were implanted in each rabbit, one screw on either side of the sagittal suture (n = 28 rabbits). Animals were sacrificed after 2, 4, 8, 16, 24, 54 and 72 weeks, four animals per group. On histological examination the number of macrophages, giant cells, active osteoblasts and fibrous tissue layers were assessed and degradation of the screws was evaluated. At 2 weeks, the highest number of macrophages and giant cells were seen near the heads of the screws. After 4 and 8 weeks, the number of giant cells decreased but that of macrophages decreased from 16 weeks and on. Screws were surrounded by fibrous tissue capsule that progressively was growing in thickness by time. Active osteoblasts were seen around the shaft of the screws with the highest number seen at 4 weeks postoperatively. At 16 weeks, compact fragmentation of the screw heads was seen with macrophages seen inside the screw matrices. After 24 weeks, no polarization of the screws was seen. After one year, PLGA screws had been replaced by adipose tissue, fibrous tissue and "foamy macrophages" which had PLGA particles inside them. After 1(1/2) years, the amount of biomaterial remaining had decreased remarkably. The particles of biomaterial were inside "foamy macrophages." Ciprofloxacin-releasing SR-PLGA 80/20 screws elicited a mild inflammatory reaction but did not interfere with osteoblast activity. No complications were seen when implanted in cranial bone of rabbit.     

Morphology and mechanical behavior of TTCP-derived calcium phosphate cement subcutaneously implanted in rats

A pre-hardened, TTCP-derived CPC was immersed in Hanks’ solution as well as subcutaneously implanted into abdomen of rats. The implant-soft tissue interfacial morphology was examined and properties of the CPC were evaluated and compared under in vitro and in vivo conditions. The results indicate that the surface of immersed samples appeared rougher and more porous than that of implanted samples and was covered with a layer of fine apatite crystals. The CPC samples implanted for 4 weeks or longer were surrounded by a layer of fibrous tissue, which was further surrounded by a soft tissue capsule comprising numerous fat cells. The soft tissue capsule had a non-uniform distribution in thickness, which increased most significantly between 4 weeks and 12 weeks after implantation. None of polymorphic cells, osteoblast cells or bone cells adjacent to the implant were observed. The majority of original TTCP powder was transformed into apatite after 1 day of either immersion in Hanks’ solution or implantation. The average porosity values of samples immersed in Hanks’ solution for 4 weeks or longer were significantly larger than those immersed for 1 day or 1 week. The porosity values of samples implanted for different times were not significantly different. The DTS values of Hanks’ solution-immersed samples largely decreased after a few weeks of immersion. The implanted samples maintained their strengths throughout the study.     

Maintenance of blood flow rate on dialysis with self‐centering CentrosFLO catheter: A multicenter prospective study

Introduction Chronic central venous catheters (CVC) for dialysis lose patency and deliver lower blood flow over time, often due to fibrous sheathing that covers the lumen tips. The CentrosFLO central venous catheter has a shape that directs the arterial and venous tips away from the walls of the vena cava and right atrium, making sheathing of the tips less likely. Methods A prospective, multicenter, single arm, non‐controlled, observational study was conducted at eight sites in the United States. All consenting dialysis patients receiving CentrosFLO catheters through the right or left internal jugular veins were accepted in the study, as long as the catheter was expected to be used for 45 days and was not an over‐the‐wire replacement for a previous CVC. Data were automatically collected on initial and average dialysis blood flow rate and initial arterial and venous pressures, for up to 26 weeks of dialysis therapy. Findings 75 patients were enrolled. Kaplan‐Meier analysis indicated that 87% of patients maintained blood flow rate over 300 mL/min throughout 26 weeks of follow‐up. There was no decline in average dialysis blood flow rate and no significant change in hydraulic resistance of the arterial or venous lumens of the catheters during the study. Discussion The CentrosFLO catheter demonstrates long term patency with good flow rates on dialysis, which, by comparison with previous studies, shows a clinically significant improvement in blood flow rate vs. other catheters. Stable hydraulic resistance of the catheter lumens showed no evidence of tip encroachment by fibrous sheaths.     

Long-term in vivo response to citric acid-based nanocomposites for orthopaedic tissue engineering

The disadvantages of current bone grafts have triggered the development of a variety of natural and synthetic bone substitutes. Previously, we have described the fabrication, characterization, and short-term tissue response of poly(1,8-octanediol-co-citrate) (POC) with 60 weight % hydroxyapatite nanocrystals (POC-HA) at 6 weeks. In order to better understand the clinical potential, longer term effects, and the biodegradation, biocompatibility, and bone regenerative properties of these novel nanocomposites, POC-HA, POC, and poly-L-lactide (PLL) were implanted in osteochondral defects in a rabbit model and assessed at 26 weeks. Explants were stained with Masson Goldner Trichrome and the fibrous capsule and tissue ingrowth measured. In addition, the bone-implant and bone-cartilage response of POC-HA, POC, and PLL were assessed through histomorphometry and histological scoring. Upon histological evaluation, both POC-HA and POC implants were biocompatible, but PLL implants were surrounded by a layer of leukocytes at 26 weeks. In addition, due to the degradation properties of POC-HA, tissue grew into the implant and had the highest area of tissue ingrowth although not statistically significant. Histomorphometric analyses supported a similar osteoid, osteoblast, and trabecular bone surface area among all implants although the fibrous capsule thickness was the largest for POC. Moreover, histological scoring demonstrated comparable scores among all three groups of the articular cartilage and subchondral bone. This study provides the long-term bone and cartilage response of novel, citric acid-based nanocomposites and their equivalence to FDA-approved biomaterials. Furthermore, we provide new insights and further discussion of these nanocomposites for orthopaedic applications.     

The influence of bone formation on anchoring percutaneous devices with titanium fibre mesh flanges

For man-made percutaneous devices (PD), it is known that anchoring will improve the clinical success. Previously, our Department has designed PDs that use a sheet of titanium (Ti) fibre mesh for anchoring. In nature, a very successful natural PD occurs, namely the tooth. Teeth are anchored in the alveolar bone. In the current study, we evaluated whether a sheet of (ectopic) bone can be made, and be used to anchor a skin-penetrating device. Using available tissue engineering techniques, sheets of Ti fibre mesh were loaded with osteoblast-like cells. These sheets, and non-loaded controls, were placed subcutaneously in 20 syngeneic rats. After four weeks 10 rats were sacrificed, and tissues were prepared for histology. On the other 10 rats, a percutaneous Ti bar was screwed. These rats were evaluated clinically up to eight weeks. Finally, also their tissues were prepared for histology. The results showed, that bone formation was only established in one cell-loaded implant, of the four-week group. Clinical evaluation, and the histomorphometrical data, showed no differences between cell-loaded and unloaded samples. We concluded that the combination of Ti mesh with rat bone marrow cells was not able to generate bone formation after subcutaneous implantation predictably. Thus, our original aim could not be met. On the other hand, our results did confirm the biocompatible behaviour of a PD equipped with a Ti fibre mesh anchoring flange.     

Cast titanium as implant material

The tissue response in rats to implants made of machined and cast titanium was evaluated after 1 and 12 weeks. The implants consisted of a circular plate portion, located in the abdominal wall, and a cylindrical rod portion protruding into the peritoneal cavity. The chemical and topographical surface properties of the two types of implants differed considerably. The implants with surrounding tissue were processed en bloc for light and electron microscopy. The bulk metal was removed by an electrochemical procedure which permitted the sectioning and evaluation of the intact implant-tissue interface. The general distribution of macrophages and fibroblasts was the same around the plate portion of both types of implants. Macrophages constituted the predominating cell type with the highest concentration in the innermost cell zone closest to the implant. The number of macrophages per section area was significantly higher around machined implants. Multinuclear giant cells, always located at the implant surface, were more frequent around cast implants. The majority of the intraperitoneal rod portions were partially (1 week) or completely (12 weeks) covered by tissue; partial or complete overgrowth of tissue was rare for machined rod portions. Imaging electron energy loss spectroscopy demonstrated the presence of titanium in macrophages in the peripheral part of the tissue capsule around cast, but not machined implants. We conclude that the tissue responses to the two types of titanium implants differed considerably in the two biological environments (soft tissue in abdominal wall; peritoneal cavity) examined and that the response in one environment does not predict the response in the other. We also believe that improvements have to be made in the casting procedure in order to reduce surface roughness and contamination before cast implants can be used in clinical applications.     

Carbon Nanotubes: Biocompatibility of Immobilized Aligned Carbon Nanotubes (Small 8/2011)

In vivo host responses to an electrode‐like array of aligned carbon nanotubes (ACNTs) embedded within a biopolymer sheet are reported. This biocompatibility study assesses the suitability of immobilized carbon nanotubes for bionic devices. Inflammatory responses and foreign‐body histiocytic reactions are not substantially elevated when compared to negative controls following 12 weeks implantation. A fibrous capsule isolates the implanted ACNTs from the surrounding muscle tissue. Filamentous nanotube fragments are engulfed by macrophages, and globular debris is incorporated into the fibrous capsule with no further reaction. Scattered leukocytes are observed, adherent to the ACNT surface. These data indicate that there is a minimal local foreign‐body response to immobilized ACNTs, that detached fragments are phagocytosed into an inert material, and that ACNTs do not attract high levels of surface fouling. Collectively, these results suggest that immobilized nanotube structures should be considered for further investigation as bionic components.     

Biocompatibility of immobilized aligned carbon nanotubes

In vivo host responses to an electrode-like array of aligned carbon nanotubes (ACNTs) embedded within a biopolymer sheet are reported. This biocompatibility study assesses the suitability of immobilized carbon nanotubes for bionic devices. Inflammatory responses and foreign-body histiocytic reactions are not substantially elevated when compared to negative controls following 12 weeks implantation. A fibrous capsule isolates the implanted ACNTs from the surrounding muscle tissue. Filamentous nanotube fragments are engulfed by macrophages, and globular debris is incorporated into the fibrous capsule with no further reaction. Scattered leukocytes are observed, adherent to the ACNT surface. These data indicate that there is a minimal local foreign-body response to immobilized ACNTs, that detached fragments are phagocytosed into an inert material, and that ACNTs do not attract high levels of surface fouling. Collectively, these results suggest that immobilized nanotube structures should be considered for further investigation as bionic components.     

Evaluation of PHBHHx and PHBV/PLA fibers used as medical sutures

Two types of fibers were prepared by using bio-based materials: a mono-filament made from poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) and a multi-filament made from poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and polylactic acid (PLA) blend. The two fibers were evaluated for mechanical properties, biocompatibility and degradability for the potential application as medical sutures. The PHBHHx fiber showed remarkable biocompatibility by H.E. Stainning, with very little impact to the surrounding tissues. The degradation of the fiber was observed by SEM after implantation for 36 weeks, and the major degradation product was detected after 96 weeks. Consistently, the PHBHHx fiber maintained more than half of the mechanical properties after 96 weeks. The other fiber was prepared by twisting PHBV/PLA blend strands to a bunch, and showed high biocompatibility and relatively high degradability. The bunched structure loosed after 36 weeks of implantation. These low-cost and easily prepared fibers have great potential in medical applications, since they could avoid the formation of fibrous capsule, reduce the size of scar, and degrade into non-toxic and even beneficial products.     

不同表面特征的多孔钛与皮下软组织间生物力学性质的研究

为解决经皮器械长期生物密封问题, 选择具有三维支架结构和合适的表面微孔多孔钛作为其皮下固定材料, 表面预沉积一层类骨磷灰石, 空白样品作为对照, 皮下植入兔子的背部. 术后40天和70天对样品与周围软组织间的结合强度进行测定, 并对断口处组织与材料间的结合形貌和区域成分进行分析. 结果表明, 随着时间的延长, 不同表面特征的材料与组织的结合强度都不断增大; 表面具有类骨磷灰石层的样品无论是结合强度还是结合形态都优于空白样品, 涂层样品40天时与组织的结合强度几乎与对照样品70天时与组织的结合强度相当. 说明类骨磷灰石层在体内有利于较早形成活跃的细胞材料表面, 有助于皮下结缔组织的长入以及与材料表面的牢固结合.     

An easy way to remove a stuck hemodialysis catheter

The replacement of tunneled hemodialysis catheters (CVCs) is a common procedure. In some cases, the CVC cannot be removed from the central vein because of tight adhesions to the surrounding fibrin sheath. A tight fibrin sheath leads to firm adherence between the catheter and the central veins or right atrial wall. Such stuck catheters cannot be removed from the central vein using standard methods. We present here a case demonstrating the successful removal of such a stuck CVC using the reverse Seldinger method.     

Pre-treatment of titanium implants with fluoride improves their retention in bone

Fluoride pre-treatment of titanium improved the bone response to this material in the present study. Fluoride pre-treated titanium implants had a four times increased retention in rabbits ulna after four and eight weeks healing periods as measured by a push out technique. Scanning electron microscopic evaluation of the implants revealed that the F-treated implants were partly covered with bone after the push out procedure indicating that an internal fracture had occurred in the bone rather than between the bone and the implant. This was not observed in the titanium control group. It is suggested that the presence of a fluoride coat on the surface of titanium implants stimulates the bone response leading to a connection between titanium and phosphate from tissue fluids. Free fluoride ions will catalyse this reaction and induce the formation of fluoridated hydroxyapatite and fluorapatite in the surrounding bone.     

Nano-structured titanium coating for improving biological performance

Nano-structured titanium coating was obtained by alkali treating the vacuum plasma sprayed samples following hot water immersing for 24 h. The influences of the surface microstructure on the biological performance were studied. A canine model was applied for in vivo evaluation of the bone bonding ability of the coatings. The histological examination results demonstrate that new bone was formed more rapidly on the nano-structured coating implants and grew into the porosity than the as-sprayed one. After 4 weeks implantation, the nano-structured implants were found to appose directly to the surrounding bone while large lacunae could still be observed at the interface between the as-sprayed samples and bone. All these results indicate that a nano-structured surface on the porous titanium coating is favorable for bone bonding.