Extracellular Fluid Flow Research Articles

Molecular mechanisms for signal-transmission of mechanical stress into bone-forming cells

One function of bone in organism is to mechanically support the body. The bone is always exposed to mechanical stress such as gravity and locomotion, and the shape of bone is adapted to the mechanical loading. Mechanical loading on bone generates extracellular deformation and fluid flow, and the mechanical stimuli are translated to mechanical signals such as mechanical strain and fluid shear stress. Bone-forming cells such as osteocytes and osteoblasts are mechanosensors. When these cells receive the mechanical stress, they stimulate the production of local regulators for bone metabolism such as prostaglandins, and various growth factors and cytokines. By the actions of these factors on bone-forming cells and bone-resorbing cells in bone microenvironment, the bone metabolism is turn over in conformity with the mechanical stress.

A case for strain-induced fluid flow as a regulator of BMU-coupling and osteonal alignment.

Throughout life, human bone is renewed continuously in a tightly controlled sequence of resorption and formation. This process of bone remodeling is remarkable because it involves cells from different lineages, collaborating in so-called basic multicellular units (BMUs) within small spatial and temporal boundaries. Moreover, the newly formed (secondary) osteons are aligned to the dominant load direction and have a density related to its magnitude, thus creating a globally optimized mechanical structure. Although the existence of BMUs is amply described, the cellular mechanisms driving bone remodeling-particularly the alignment process-are poorly understood. In this study we present a theory that explains bone remodelling as a self-organizing process of mechanical adaptation. Osteocytes thereby act as sensors of strain-induced fluid flow. Physiological loading produces stasis of extracellular fluid in front of the cutting cone of a tunneling osteon, which will lead to osteocytic disuse and (continued) attraction of osteoclasts. However, around the resting zone and the closing cone, enhanced extracellular fluid flow occurs, which will activate osteocytes to recruit osteoblasts. Thus, cellular activity at a bone remodeling site is well related to local fluid flow patterns, which may explain the coordinated progression of a BMU.

The pathophysiology of syringomyelia - historical overview and current concept.

Various ideas and hypotheses have been brought forward to explain the development of syringomyelia in the past two centuries. None of them offers a sufficient basis to serve as a concept for the treatment of all affected patients. Apart from a discussion of the different hypotheses this paper proposes a new pathophysiological concept based on clinical, experimental and literature studies. Syringomyelia is understood as a state of chronic interstitial edema of the spinal cord due to accumulation of extracellular fluid (ECF). This accumulation is caused by a cascade of events starting with obstruction of cerebrospinal fluid (CSF) flow and/or spinal cord tethering which ultimately alter ECF flow and increase ECF volume. Treatment should be targeted against the pathological process which causes CSF flow obstruction and cord tethering to inhibit this pathophysiological process at a decisive point.

Estimation of the poroelastic parameters of cortical bone.

Cortical bone has two systems of interconnected channels. The largest of these is the vascular porosity consisting of Haversian and Volkmann's canals, with a diameter of about 50 microm, which contains a.o. blood vessels and nerves. The smaller is the system consisting of the canaliculi and lacunae: the canaliculi are at the submicron level and house the protrusions of the osteocytes. When bone is differentially loaded, fluids within the solid matrix sustain a pressure gradient that drives a flow. It is generally assumed that the flow of extracellular fluid around osteocytes plays an important role not only in the nutrition of these cells, but also in the bone's mechanosensory system. The interaction between the deformation of the bone matrix and the flow of fluid can be modelled using Biot's theory of poroelasticity. However, due to the inhomogeneity of the bone matrix and the scale of the porosities, it is not possible to experimentally determine all the parameters that are needed for numerical implementation. The purpose of this paper is to derive these parameters using composite modelling and experimental data from literature. A full set of constants is estimated for a linear isotropic description of cortical bone as a two-level porous medium. Bone, however, has a wide variety of mechanical and structural properties; with the theoretical relationships described in this note, poroelastic parameters can be derived for other bone types using their specific experimental data sets.

Does an equivalent of the ”ventral node" exist in chick embryos? A scanning electron microscopic study

The internal organs of vertebrates show species-specific left-right (L-R) asymmetries. Questions on the embryonic origin of these asymmetries have been fascinating embryologists since the 19th century. During the past years, remarkable progress has been made in answering these questions. Evolutionary highly conserved molecular signaling cascades have been identified that start from Hensen's node and transfer side-specific identity to the embryonic left and right halves. However, the question of what initiates these signaling cascades has remained unanswered. Studies on mouse embryos have shown that the ventral surface of Hensen's node consists of a ciliated epithelium called the ventral node. Recent findings suggest that the monocilia of ventral nodel cells generate a leftward flow of extracellular fluid possibly leading to the accumulation of an unknown morphogen at the left of the node, which then might start the signaling cascades. This hypothesis might explain the fact that gene defects causing ciliary dyskinesia are frequently associated with situs anomalies. Studies on chick embryos led to the discovery of the L-R signaling cascades. However, whether an equivalent of the ventral node exists in avian embryos remained unknown. Therefore, I examined the endoderm and epiblast of early chick embryos for the presence of monociliated cells. In the endoderm, a population of monociliated cells indeed was present. These cells, however, were neither confined to the area of Hensen's node nor did they form the predominant cell population at this location. In the epiblast, monociliated cells formed the predominant cell population at the periphery of the blastodisc but only a relatively small subpopulation of epiblast cells at Hensen's node. These findings suggest that, in the early chick embryo, an equivalent of the ventral node of mouse embryos neither exists on the ventral nor the dorsal surface of Hensen's node. It is unlikely that nodal cilia are required for initiating the L-R patterning in chick embryos.

Disturbances of cerebrospinal fluid flow attributable to arachnoid scarring cause interstitial edema of the cat spinal cord.

Spinal arachnoid scarring may be caused by trauma, inflammation, surgery, spinal instability, degenerative diseases, or malformations and may lead to progressive neurological deficits and syringomyelia. We wanted to investigate the effects of focal arachnoid scarring in the cervical spinal canal of cats on pressures in the subarachnoid space and spinal cord tissue, as well as on spinal cord histological features. Twenty-nine adult cats were used for this study. Nine animals served as control animals, whereas 20 animals received a focal arachnoid scar at C1-C2, which was produced by placement of a kaolin-soaked fibrin sponge on the posterior surface of the spinal cord. After 4 months, pressure recordings above and below the scar, in the subarachnoid space and spinal cord, were performed. Elasticity measurements were performed with small bolus injections. Morphometric analyses of brain and ventricle volumes, sizes of the central canal, and sizes of the perivascular spaces in gray and white matter were also performed. No animal developed clinical or neurophysiological evidence of neurological symptoms at any time. In the kaolin-treated group, pressure recordings revealed a significant increase in the subarachnoid pressure at C1, because of the cerebrospinal fluid flow obstruction. Pressure gradients tended to increase at all measuring points. A significant difference was detected between the spinal cord and subarachnoid space at C2, where the intramedullary pressure exceeded the subarachnoid pressure. Elasticity was significantly increased in the spinal cord at C2. Intracranially, no evidence of hydrocephalus was observed. In the spinal cord, perivascular spaces were significantly enlarged in the posterior white matter above the arachnoid scar and in the central gray matter below the area of scarring in the cervical cord. Arachnoid scarring at C1-C2 produces an interstitial type of edema in the central gray matter below the area of scarring in the cat cervical cord, because of altered cerebrospinal fluid and extracellular fluid flow dynamics. These changes may be interpreted as the initial stage in the development of syringomyelic cavities.

Mean pressures and flows in the human intracranial system as determined by mathematical simulations of a steady-state infusion test

In order to understand the fluid dynamics within the human intracranial system, the relatively small flow of extracellular fluid into, and out of, the interstitial brain tissue must be determined. Due to the magnitude of these flows, it is difficult to measure them clinically. Through a steady-state infusion simulation run on a mathematical model, values for these small flows may be calculated based on clinical data regarding the conductance of cerebrospinal fluid outflow. In this way, the mathematical model allows information to be obtained regarding these small mean flows, as well as the remaining mean flows and mean pressures throughout the Intracranial space, with minimal reliance on data from intrusive procedures. [Neurol Res 2000; 22: 809-814]

LaminOss immediate-load implants: I. Introducing osteocompression in dentistry.

Osteocompression is a physiologic principle that has been clinically practiced in orthopedics since the early 1900s. In dentistry, controlled functional osteocompression is the compaction created by the tapping procedure and bone lamination achieved by a sinusoidal screw implant design providing physiologic stimulation due to streaming potentials. Functionally, there is always an applied force acting on bone modified by an implant design, and there is always a resisting force acting on the implant through the viscoelastic properties of trabecular structure. Through biomechanical events in bone, osseous tissue can be stimulated within physiologic limitations by implant design to develop along the lines of compressive forces dependent on the implant load-bearing area to sustain equilibrium. At the cellular level, these biomechanical events act on the cells through a phenomenon known as streaming potentials. This is an electrochemical potential created by the flow of extracellular fluid past a positively charged cell surface. Streaming potentials have a stimulating effect on osteoblasts and osteocytes. This stimulation under acceptable physiologic limits translates to an ordered deposition of osseous tissue that aids in the support of these compressive forces. As a sinusoidal thread design, the LaminOss osteocompressive immediate-load implant (Impladent Ltd, Holliswood, NY) has shown in animal histologic observation 2.5 times greater bone lamination achieved by the function of osteocompression due to the benefits of streaming potentials created by the LaminOss implant design. No evidence of bone necrosis was observed by any of the eight implants.

A uniform strain criterion for trabecular bone adaptation: Do continuum-level strain gradients drive adaptation?

In this paper, it is postulated that the apparent density of trabecular bone adapts so that continuum-level strains within the bone are uniform and, as a consequence, spatial strain gradients within the bone/marrow continuum are minimized. The feasibility of a uniform strain criterion was tested using computational finiteelement analysis of the proximal femur. We demonstrated that (1) this criterion produced a realistic apparent density distribution in the proximal femur, (2) the solutions for apparent density were convergent and unique, (3) predicted apparent densities compared well to experimental measurements, and (4) strain gradients within the bone/marrow continuum were reduced substantially. Thus, a possible goal of trabecular bone adaptation may be the reduction of strain gradients within the bone/marrow continuum. Osteocytes within the bone tissue and bone cells on the surface of a trabeculum are mechanosensitive and play a role in bone adaptation. In addition, the bone marrow is rich in osteoprogenitor cells near the bone surface that are mechanosensitive. Strain gradients within bone/marrow continuum cause pressure gradients in the marrow, causing extracellular fluid flow which could stimulate osteoprogenitor cells and contribute to bone adaptation.

Electrical properties of extracted rat liver tissue.

We attempted to investigate the process of ischemia-induced disturbances in the rat liver, employing the electrical bio-impedance technique. The electrical bio-impedance was measured continuously over 6 h by the 4-electrode method, at various incubation temperatures, in six liver samples extracted from male Wistar rats. The electrical properties of biological tissues can be expressed in terms of three parameters: extracellular resistance (Re), intracellular resistance (Ri) and cell membrane capacitance (Cm). These three parameters were calculated from the measured values of the electrical impedance by the curve-fitting technique, using a computer program. The Re value increased rapidly after the rat livers were extracted, and then decreased slowly. The Re value reached a peak after about 13 min at 36 degrees C, and then decreased slowly, becoming constant after 3 h. There was a negative correlation between the Tmax of Re (the time when Re reached a maximum) and the incubation temperature (R = -0.973, P < 0.001). The Ri value decreased once in the early stage after extraction, followed by almost no change and then an increase after 4 h at 36 degrees C. The Cm showed a similar pattern of change to the Re value, and a negative correlation was also found between the Tmax of Cm and the incubation temperature (R = -0.969, P < 0.001). The increase in the Re and Cm values, and the decrease in the Ri value for quite long periods after the blood flow has stopped, suggest an increase in the resistance of extracellular fluid due to a decrease in its volume, an increase in cell membrane capacitance due to cell swelling, and a decrease in cellular fluid resistance due to an increase in its volume. The time when the Cm value decreases rapidly after an initial gradual decrease after the peak corresponds well with the time when the Ri value begins to increase, from which it is estimated that cell lysis proceeds and that the flow of extracellular fluid into the cell begins at this time. The findings of this study suggest the possibility of estimating the changes in liver tissue or the tissue structure due to ischemia.

Mechanotransduction and the functional response of bone to mechanical strain.

Mechanotransduction plays a crucial role in the physiology of many tissues including bone. Mechanical loading can inhibit bone resorption and increase bone formation in vivo. In bone, the process of mechanotransduction can be divided into four distinct steps: (1) mechanocoupling, (2) biochemical coupling, (3) transmission of signal, and (4) effector cell response. In mechanocoupling, mechanical loads in vivo cause deformations in bone that stretch bone cells within and lining the bone matrix and create fluid movement within the canaliculae of bone. Dynamic loading, which is associated with extracellular fluid flow and the creation of streaming potentials within bone, is most effective for stimulating new bone formation in vivo. Bone cells in vitro are stimulated to produce second messengers when exposed to fluid flow or mechanical stretch. In biochemical coupling, the possible mechanisms for the coupling of cell-level mechanical signals into intracellular biochemical signals include force transduction through the integrin-cytoskeleton-nuclear matrix structure, stretch-activated cation channels within the cell membrane, G protein-dependent pathways, and linkage between the cytoskeleton and the phospholipase C or phospholipase A pathways. The tight interaction of each of these pathways would suggest that the entire cell is a mechanosensor and there are many different pathways available for the transduction of a mechanical signal. In the transmission of signal, osteoblasts, osteocytes, and bone lining cells may act as sensors of mechanical signals and may communicate the signal through cell processes connected by gap junctions. These cells also produce paracrine factors that may signal osteoprogenitors to differentiate into osteoblasts and attach to the bone surface. Insulin-like growth factors and prostaglandins are possible candidates for intermediaries in signal transduction. In the effector cell response, the effects of mechanical loading are dependent upon the magnitude, duration, and rate of the applied load. Longer duration, lower amplitude loading has the same effect on bone formation as loads with short duration and high amplitude. Loading must be cyclic to stimulate new bone formation. Aging greatly reduces the osteogenic effects of mechanical loading in vivo. Also, some hormones may interact with local mechanical signals to change the sensitivity of the sensor or effector cells to mechanical load.

Surgical results of 100 intramedullary tumors in relation to accompanying syringomyelia.

During the period from 1977 to August 1992, 100 intramedullary tumors in 94 patients were operated on in the Department of Neurosurgery at the Nordstadt Hospital in Hannover, Germany. Of these, 45% presented with associated syringes. A syrinx was more likely to be found above (49%) than below (11%) the tumor level. In 40%, a syrinx could be identified above and below the tumor level. Ependymomas and hemangioblastomas were the most common tumor types to be associated with syringes. Astrocytomas tended to demonstrate syringes less often. Regardless of histology, the higher the spinal level, the more likely a syrinx was encountered. In general, the presence of an associated syrinx favored the resectability of the tumor, because it indicated a displacing rather than an infiltrating tumor. Patients with syringomyelia tended to recover from surgery sooner. However, surgical results and long-term prognosis were not influenced significantly by an associated syrinx. The most important factor determining long-term outcome was the preoperative level of neurological function. We propose that factors independent of the tumor, disturbances of cerebrospinal fluid and extracellular fluid flow in particular, have major roles in the pathogenesis of syrinx formation associated with intramedullary tumors.

Numerical analysis of extracellular fluid flow and chemical species transport around and within porous bioactive glass.

Modeling of the physical phenomena present at the biomaterial-tissue interface provides a valuable tool for examining the underlying mechanisms which influence the overall behavior of the implant-host system. Based on histological data from a previous implantation study (E. Schepers, M. De Clercq, P. Ducheyne, and R. Kempeneers, "Bioactive glass particulate materials as a filler for bone lesions," J. Oral Rehab.; 18, 439-452, 1991, Ref. 1) which documented the differentiation of mesenchymal cells to cells expressing the osteoblastic phenotype in porous bioactive glass, a finite element momentum and mass transport model was constructed. In this analysis, the extracellular compositional variations and fluid flow conditions around and within porous bioactive glass granules were determined. Numerical simulations demonstrated that the interstitial fluid flow around these granules (300-360 microns) is viscosity dominated (low Reynolds number flow) and that the fluid inside the granules remains stagnant. This velocity field results in shear stresses proportional to the velocity gradient at the granule-fluid interface outside the particles and no shear stresses inside the particles. A parametric study on the effect of interstitial fluid flow on chemical species (Na+, Ca+2, HPO(4)-2) transport outside the granules revealed three domains. At low velocities (0-0.1 micron/s), the transport of species is diffusion controlled. At intermediate velocities (1.0-10 microns/s), diffusion and convection contribute to the species transport. The concentration of chemical species is nearly uniform at high velocities (100-800 microns/s). For all three cases, the transport of chemical species within the granules is diffusion controlled. The differences in transport mechanisms and interstitial fluid flow conditions lead to variations in concentrations, reaction rates, and shear stresses between the inside and the outside of the glass granules. These differences may influence cellular migration, attachment, differentiation, and the overall response to these bioactive materials.

Biomechanics of Brain Tissue : Simulation of Cerebrospinal Fluid Flow

The brain consists of brain tissue, blood and extracellular fluid. We can thus consider that brain tissue has multi-phasic properties. There is a very important interrelation between the tissue and the fluid. Thus, we consider that the respective factors on brain neurosurgery, the distribution of brain tissue pressure and extracellular fluid flow are very important, and we cannot neglect those factors closely related to metabolism of the brain tissue. This study constructs a two-dimensional consolidation model of the brain using the finite element method (FEM) and simulates the flow and distribution of cerebrospinal fluid (CSF). The two dominant equations on the consolidation theory are approximated numerically in time by the finite difference method and in space by the FEM. The results obtained by computer simulation regarding brain edema using the FEM are compared with the pathological observation with regrad to the flux distribution and flow direction of CSF, and we conclude that they are much the same as those from the view point of the pathological and clinical surgery. We furthermore propose that we should take the boundary conditions of pressure of the subarchanoid space and ventricle into consideration in the simulation of brain edema.

Diffusion-weighted imaging differentiates ischemic tissue from traumatized tissue.

Diffusion-weighted magnetic resonance imaging (MRI) has been shown to be particularly effective in detecting early (0 to 4 hours) pathophysiological changes in localized brain regions after cerebral ischemia. The present study sought to establish whether diffusion-weighted MRI would be similarly effective in predicting outcome after traumatic brain injury. Diffusion-weighted MRI images and T2-weighted MRI images were obtained over 4 hours after either moderate fluid percussion-induced traumatic brain injury or unilateral carotid ligation in rats. Diffusion-weighted MRI images of traumatic brain injury demonstrated focal regions of image hypointensity as early as 1 hour after trauma. The relative diffusion coefficient in these hypointense regions was significantly increased (P < .005) by 4 hours after trauma compared with the noninjured hemisphere, but only in the transverse plane in the x direction. In contrast, induction of diffuse, nonfocal ischemia by unilateral carotid ligation resulted in scattered regions of hyperintensity with a significant (P < .001) decrease in relative diffusion coefficient as early as 1 hour after ligation compared with the noninjured hemisphere. This decrease exhibited no directionality. We conclude that traumatic brain injury results in an increased water diffusion distance with the directionality indicative of bulk flow of extracellular fluid toward the lateral ventricles (vasogenic edema). In contrast, the decreased water diffusion distance with no apparent directionality observed in ischemia is most likely indicative of cytotoxic edema. Diffusion-weighted MRI therefore has the potential to differentiate cases of traumatic brain injury with no focal ischemia from those instances of traumatic brain injury in which focal ischemia is a complication.

Biomechanics of Brain Tissue. Simulation of Cerebrospinal Fluid Flow.

The brain consists of brain tissue, blood and extracellular fluid. We can thus consider that brain tissue has multi-phasic properties. There is a very important interrelation between the tissue and the fluid. Thus, we consider that the respective factors on brain neurosurgery, the distribution of brain tissue pressure and extracellular fluid flow are very important, and we cannot neglect those factors closely related to metabolism of the brain tissue. This study constructs a two-dimensional consolidation model of the brain using the finite element method (FEM) and simulates the flow and distribution of cerebrospinal fluid (CSF). The two dominant equations on the consolidation theory are approximated numerically in time by the finite difference method and in space by the FEM. The results obtained by computer simulation regarding brain edema using the FEM are compared with the pathological observation with regrad to the flux distribution and flow direction of CSF, and we conclude that they are much the same as those from the view point of the pathological and clinical surgery. We furthermore propose that we should take the boundary conditions of pressure of the subarchanoid space and ventricle into consideration in the simulation of brain edema.

Chapter 1: A role for centrally-released vasopressin in brain ion and volume regulation: a hypothesis

Vasopressin is synthesized and released within the central nervous system (CNS). It may then act locally on adjacent target cells or on more distant intracerebral sites following distribution through brain extracellular fluid (ECF) and/or cerebrospinal fluid (CSF). Systemically released vasopressin increases body water and electrolyte content. Vasopressin functions in an analogous way in the CNS, to increase brain water and electrolyte content. This chapter presents results of two series of experiments consistent with this hypothesis; one uses an in vivo rat model developed in this laboratory; the other employs cultured glial cells. Brain tissue and plasma are normally in osmotic equilibrium, and brain volume depends on the number of osmotically active solutes in the nervous tissue. In vivo model allows evaluating mechanisms of brain volume regulation in terms of a four-compartment model of fluid and electrolyte exchange among blood plasma, brain ECF, brain intracellular fluid (ICF) and CSF. The model includes two features unique to the regulation of volume in the CNS. These are: (1) secretion of ECF from plasma, based on the activity of Na + - pumps located at the blood–brain barrier; and (2) bulk flow of ECF between brain and CSF.

The possible role of poroelasticity in the apparent viscoelastic behavior of passive cardiac muscle

This paper investigates the contribution of extracellular fluid flow to the apparent viscoelastic behavior of passive cardiac muscle. The muscle is modeled as an incompressible, isotropic, poroelastic solid saturated by an incompressible viscous fluid. Based on Biot's linear and nonlinear consolidation theories, solutions are presented for general time-dependent uniaxial loading of unconfined cylindrical muscle specimens. The nonlinear analysis includes the effects of large strain, material nonlinearity, and strain-dependent permeability. The computed results show that, for axial stretch ratios greater than 1.1, the changing permeability and the loading rate strongly affect the total stress relaxation and the short-time relaxation rate. Comparisons of theoretical and published experimental results show that extracellular fluid flow can account for several observed biomechanical features of passive myocardium, including the insensitivity of stress-strain curves to loading rate and of stress-relaxation curves to the amount of stretch. Theoretical hysteresis loops, however, are too small. Thus, both poroelastic and tissue viscoelastic effects must be considered in studies of passive cardiac muscle.

Ca2+-activated K+ conductance causes membrane hyperpolarizations in a monkey kidney cell line (JTC-12).

We have previously reported hyperpolarizing membrane potential changes in a monkey kidney cell line (JTC-12) which has characteristics resembling proximal tubular cells. These hyperpolarizations could be observed spontaneously or evoked by mechanically touching adjacent cells. In this report, we have shown further evidence that these hyperpolarizations are elicited by an increase in membrane conductance to K+ which is caused by an increase in cytosolic Ca2+ concentration. In addition, we have found another type of hyperpolarization which is evoked by applying flow of extracellular fluid to the cell. Intracellular injection of Ca2+ and Sr2+ evoked hyperpolarizations, while intracellular injection of Mn2+ and Ba2+ did not. Intracellular injection of EGTA suppressed both spontaneous and mechanically evoked hyperpolarizations. In Ca2+-free medium, both spontaneous and flow-evoked hyperpolarizations were not observed, while mechanical stimuli consistently evoked hyperpolarization. In Na+-free medium, the incidence of cells showing the spontaneous or flow-evoked hyperpolarization increased, and the amplitude and the duration of the mechanically evoked hyperpolarization became greater. Quinidine inhibited all types of hyperpolarization. These data suggest that hyperpolarizations in JTC-12 cells are due to an increase in Ca2+-activated K+ conductance.

13] Opsonins and dysopsonins: An overview

Publisher Summary This chapter provides an overview of opsonins and dysopsonins. “Phagocytosis” refers to the process by which a particle is recognized by the phagocyte as being foreign and is ingested by the cell and subsequently sequestered in an intracellular vacuole. This process is initiated by the migration (chemotaxis) of the phagocyte to the site of microbial infiltration. Once contact between the particle and granulocyte has been achieved, a cohesive force prevents their separation despite extracellular fluid flow. This critical step in the phagocytic process is known as the “adhesive phase.” The prerequisite for ingestion and subsequent digestion is the formation of the critical adhesive bond between the particle to be ingested and the phagocytic cell. Several biological entities are known to either promote (opsonins) or retard (dysopsonins) this event. Opsonins (and dysopsonins) primarily affect the critical first step in phagocytosis—that is, the attachment phase between the phagocyte and the particle. The heat-stable opsonins that appear in the serum of immunized animals are specific for the immunizing agent and are adsorbed from the serum by the strain used for immunization but not by other organisms.